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RESEARCH ARTICLE
Genetic Guidelines for Captive Breeding and
Reintroductions of the Endangered Black-Fronted
Piping Guan, Aburria jacutinga (Galliformes,
Cracidae), an Atlantic Forest Endemic
Paulo R.R. Oliveira-Jr.,
1
Mariellen C. Costa,
2
Luis F. Silveira,
3
and Mercival R. Francisco
4
*
1
Programa de P
os-GraduaSc~
ao em Diversidade Biol
ogica e ConservaSc~
ao, Universidade Federal de S~
ao Carlos,
Campus de Sorocaba, Sorocaba, S~
ao Paulo, Brazil
2
Programa de P
os-GraduaSc~
ao em Ecologia e Recursos Naturais, Universidade Federal de S~
ao Carlos, S~
ao
Carlos, S~
ao Paulo, Brazil
3
SeSc~
ao de Aves, Museu de Zoologia da Universidade de S~
ao Paulo, S~
ao Paulo, Brazil
4
Departamento de Ci^
encias Ambientais, Universidade Federal de S~
ao Carlos, Campus de Sorocaba, Sorocaba,
S~
ao Paulo, Brazil
The survival of a number of birds rely on captive breeding and reintroduction into the wild, but captive populations are often
small and can be exposed to the negative effects of inbreeding and genetic drift. Then, managers are concerned not only with
producing as much offspring as possible, but also with the retention of the maximum genetic variability within and between
populations. The Black-fronted Piping Guan, Aburria jacutinga, is an endangered cracid endemic to the Atlantic Forest of
southeastern South America. Because of its conservation status and functional importance, a captive breeding program started
independently, mainly in three aviaries, in the decade of 1980. Although they have supplied animals for reintroductions,
genetic variability aspects have never been considered. Here we addressed levels of genetic variability within and between
these aviaries. Bayesian clustering analyses revealed two lineages. Inbreeding was not detected, although we found evidences
for a recent bottleneck in one of the aviaries. Then, our main management recommendations are: i) reintroducing the species in
areas where it has been extinct is more prudent than supplementing natural populations, as it could involve risks of disrupting
local adaptive complexes; ii) as far as inbreeding can be avoided, the captive groups should be managed separately to
minimize adaptation to captivity; iii) crossbreedings in pre-release generations could improve reintroduction success; and iv)
a studbook should be implemented. As populations of Black-fronted Piping Guan from conservation units are progressively
declining, these captive genetic repositories may gain importance in a near future. Zoo Biol. XX:XX–XX, 2016. © 2016
Wiley Periodicals, Inc.
Keywords: endangered species; Aves; conservation; management; genetic variability
INTRODUCTION
The survival of a growing number of bird species rely
on captive breeding and reintroduction into the wild, and
this conservation strategy applies not only to species that
have become extinct in the wild [Jones et al., 1995; Ralls
and Ballou, 2004; Silveira et al., 2004; Sven and Ryan,
2012], but also to those facing imminent risks of extinction
on nature [Jones et al., 2002; Earnhardt et al., 2014;
VanderWerf et al., 2014; Mounce et al., 2015]. The final
goal of any ex situ breeding program is to found new
populations or to genetically and demographically reinforce
the populations that still exist in the wild [Conway, 1980;
Grant sponsor: FAPESP; grant numbers: 2008/51197-0, 2010/08586-6,
2010/01251-9, 2011/06210-1; grant sponsor: Brazilian Council of
Research (CNPq); grant sponsor: SAVE Brasil.
Conflicts of interest: None.
Correspondence to: Mercival Roberto Francisco, Depto. de Ci^
encias
Ambientais, Universidade Federal de S~
ao Carlos, Campus de Sorocaba,
Rod. Jo~
ao Leme dos Santos, km 110, CEP 18052-780, Sorocaba, SP,
Brazil
E-mail: mercival@ufscar.br
Received 23 October 2015; Revised 13 April 2016; Accepted 11 May 2016
DOI: 10.1002/zoo.21296
Published online XX Month Year in Wiley Online Library
(wileyonlinelibrary.com).
© 2016 Wiley Periodicals, Inc.
Zoo Biology 9999 : 1–6 (2016)
Frankham et al., 2004; Witzenberger and Hochkirch, 2011].
Because captive populations are often small and founded by
a limited number of individuals, the major concerns of
managers are not only to produce as much offspring as
possible, but also to maintain the genetic integrity of the
populations [Frankham et al., 2004; Frankham, 2010;
Witzenberger and Hochkirch, 2011). Small populations are
more exposed to the negative impacts of inbreeding and
genetic drift, which may translate into reduced survival,
reduced fertility, and limited capability of the populations to
adapt to changing environments [Shaffer, 1987; Frankham
et al., 2004], threatening the success of both captive
breeding and reintroductions [Witzenberger and Hochkirch,
2011]. To tackle this problem, the maximum genetic
variability should be preserved within and between
populations during the captive stage of conservation
programs [Frankham et al., 2004; Witzenberger and
Hochkirch, 2011]. When founders are sampled from various
natural populations, reproduction should be achieved in as
many genetic lineages as possible, and when genetic
variability is low within each lineage, the exchange of
individuals between them generally leads to increased
heterozigosity levels, augmenting the chances of survival of
endangered taxa [Witzenberger and Hochkirch, 2011].
Therefore, understanding the levels of population structur-
ing and levels of genetic variability within the different
lineages provide important guidelines for management
strategies [Nielsen et al., 2007; Nsubuga et al., 2010;
Mukesh et al., 2013].
The Black-fronted Piping Guan, Aburria jacutinga,isa
medium-sized (1.1–1.4 kg) representative of the family
Cracidae (Galliformes), endemic to the Atlantic Forest from
the Brazilian state of Bahia south to northeastern Argentina and
eastern Paraguay [Sick, 1997; Bernardo and Clay, 2006]. Over
the past 500 years, this biome has been extensively exploited
andonly11.4–16% of the original 150 million ha has been left.
Of the remaining forested areas, 97% are isolated fragments
smaller than 250 ha, that still experience strong anthropogenic
pressures [Ribeiro et al., 2009]. Furthermore, the remaining
continuums are embedded within the Neotropics’most
densely-populated region, and recent studies have revealed
that even some of the largest conservation units may not sustain
viable populations of Atlantic Forest medium-sized to large-
bodied vertebrates, especially due to overhunting [Marsden
et al., 2005; Galetti et al., 2009].
In this scenario, large-gape seed disperser birds
like the Black-fronted Piping Guan are especially
vulnerable and their local extinctions often imply in
cascading effects to Atlantic Forest ecosystem functions
[Bernardo et al., 2011; Galetti et al., 2013]. This once
abundant species has been a remarkable game bird for
centuries, which has resulted in its extinction in most of
its original distribution [Sick, 1997; Bernardo and Clay,
2006; Bernardo et al., 2011]. Due to past and ongoing
threats, the species is currently classified as endangered
in the IUCN Red List.
Because of the conservation status and functional
importance of the Black-fronted Piping Guan, a captive
breeding program started in the decade of 1980 mainly in
three Brazilian aviaries: Tropicus, in the state of Rio de
Janeiro; CESP/Paraibuna, in the state of S~
ao Paulo, and
Guaratuba, in the state of Paran
a, all in southeastern Brazil.
These breeding groups were founded independently from an
uncertain number of animals that were confiscated by the
police from illegal bird keepers, and by animals being raised
by rural people living nearby Atlantic Forest areas that were
pursued by the aviary owners. These breeding facilities are
collaborators of the Brazilian Action Plan for the Conserva-
tion of Endangered Galliformes [Silveira et al., 2008],
regulated by the Brazilian Federal Government, and over the
last two decades they have supplied animals for the
foundation of a number of small (one to three pairs) or
large (more than 10 breeding pairs) breeding groups in other
aviaries. A small number of animals (3–4) were exchanged
between Tropicus and Guaratuba more than one decade ago,
and although we cannot assert that individuals with different
origins are not present in other aviaries, to our knowledge
these are the three main genetic repositories of Black-fronted
Piping Guans in captivity. Reintroduction plans have been
conducted in Fazenda Maced^
onia, in the state of Minas
Gerais, and in Reserva Ecol
ogica de GuapiaSc
u, in the state of
Rio de Janeiro (where the species was extinct), using animals
from a breeding group maintained by Crax Foundation, in the
state of Minas Gerais; and in Paraibuna, S~
ao Paulo, using
animals from CESP [Silveira et al., 2008; Bernardo et al.,
2011], and further reintroductions have been planned for a
near future in at least three more areas in which the species
has been locally or ecologically extinct. However, genetic
variability issues have not been considered so far. As the
captive populations were founded decades ago, we predict
that genetic drift could have led to reduced genetic variability
within aviaries. However, if they retain signals of indepen-
dent foundation, crossbreedings could be considered for
rescuing genetic variability within both captive and
reintroduced populations.
The objective of this work was to investigate levels of
genetic structuring between and levels of genetic variability
within the three main genetic repositories of captive Black-
fronted Piping Guans. Specifically, we used nine microsatellite
loci and Bayesian clustering analysis to address the following
questions: 1) is there population genetic structuring between the
breeding groups? 2) are there evidences of inbreeding within
these lineages? 3) are there evidences of recent genetic
bottlenecks? These data will provide valuable information for
conservation planning in this endangered species.
METHODS
Blood Sampling and DNA Extraction
At variable times we have sampled all the breeding
pairs and young animals present in the three aviaries that
compose the main original genetic repositories of the
2Oliveira-Jr. et al.
Zoo Biology
Black-fronted Piping Guan in captivity: CESP/Paraibuna
(from 2007 to 2012) (n¼27), Tropicus (from 2010 to 2012)
(n¼31), and Criadouro de Aves Silvestres Guaratuba
(2011–2012) (n¼40). From each animal we obtained
approximately 20 ml of blood by venipuncture of the brachial
vein, and blood was preserved in 100% ethanol at 20°C.
DNA was extracted using a standard phenol:chloroform:
isoamilic alcohol protocol [Sambrook et al., 1989].
Genotyping
DNA samples were amplified at five unlinked
microsatellite loci isolated for Black-fronted Piping Guan
(Aburria 21, Aburria 22, Aburria 44, Aburria 48, Aburria
105) [Costa et al., 2014], and four heterologous loci
developed for Razor-billed Curassow, Pauxi tuberosa (Pauxi
1–4, Pauxi 2–2, Pauxi 2–30, and Pauxi 3–4) [Sousa et al.,
2013]. Details on loci isolation, PCR conditions, linkage
disequilibrium, and tests for null alleles are described in
Costa et al. [2014]. Forward primers were fluorescently
labeled, amplification products were analyzed on an ABI
3730 automated sequencer (Applied Biosystems), and alleles
were scored based on an internal size standard (500 ROX)
using GeneMarker 2.6.0 (SoftGenetics, State College, PA).
Population Structure Analyses
Population structuring and assignment of individuals
to their population of origin were obtained using the
Bayesian clustering method implemented in the software
STRUCTURE 2.3.4 [Pritchard et al., 2000], which assumes
that there are Kdistinct populations, or genetic clusters, each
with a different set of allele frequencies per locus. We used
an admixture model with correlated allele frequencies,
without prior information. We ran 1000,000 MCMC
iterations (discarding 10,000 as burn-in) for each K(1–6).
Each Kvalue was obtained in ten replicates, and the most
appropriate Kwas defined using the method proposed by
Evanno et al. [2005], implemented in the software
STRUCTURE HARVESTER [Earl and VonHoldt, 2012],
where the true Kis the greatest delta K(DK) averaged across
runs. To assign individuals probabilistically to a certain
cluster, those with membership (q) thresholds of q0.8
were assigned to one cluster, while those with q<0.8 were
considered as admixted [Lecis et al., 2006; Nsubuga et al.,
2010; Mukesh et al., 2013]. Levels of genetic differentiation
between clusters were estimated using the F
ST
of Weir and
Cockerham [1984], implemented in the software FSTAT
2.9.3.2 [Goudet, 1995].
Genetic Variability Within Aviaries
For each aviary, we obtained expected (H
E
) and
observed (H
O
) levels of heterozygosity, and number of
observed alleles (N
A
), using GENEPOP 4.2 [Raymond and
Rousset, 1995]. We inferred occurrence of inbreeding by
assessing levels of hetorizygosity deficit using the inbreeding
coefficient (F
IS
) of Weir and Cockerham [1984] implemented
in FSTAT 2.9.3.2 [Goudet, 1995]. The critical levels of
significance were also obtained with this software, and were
adjusted with the sequential Bonferroni correction for
multiple comparisons [Rice, 1989].
Contemporary Effective Population Sizes and
Recent Bottlenecks
Effective population sizes were estimated for each
aviary using the linkage disequilibrium method implemented
by software NeEstimator, Ver. 2, using the Monogamy
mating option, with a critical value of 0.02 to discard rare
alleles. This method has been demonstrated to perform
satisfactorily when a moderate number of microsatellite loci
is used [Do et al., 2014]. Potential recent bottlenecks were
assessed using the software BOTTLENECK [Piry et al.,
1999], with the two-phase microsatellite mutation model
(TPM), as it is suggested as the most appropriate model for
detecting recent bottlenecks using microsatellites [Piry et al.,
1999]. The statistical significance of the results were
obtained using Wilcoxon sign-rank tests.
RESULTS
Population Structuring
The Bayesian clustering analyses of STRUCTURE
revealed genetic structuring between the aviaries (Fig. 1),
and STRUCTURE HARVESTER indicated two as the best
Kfor the sample, suggesting two distinct lineages. Cluster 1
(dark gray color) was composed mainly by the individuals
from CESP, and cluster 2 (light gray color) contained mostly
the animals from Tropicus and Guaratuba (Fig. 1). The
Fig. 1. Graphic representation of the two lineages of captive Black-fronted Piping Guan identified using Structure (K¼2). Colors in each
bar represent the proportional membership (q) of each individual.
Managing Black-Fronted Piping Guan 3
Zoo Biology
number of admixted individuals were: two in CESP and
Guaratuba, and six in Tropicus. One individual from
Tropicus and two from Guaratuba were assigned to CESP.
The level of structuring between these two lineages was
highly significant (F
ST
¼0.18, P<0.0001).
Genetic Variability
Across the three aviaries we found a total of 34 alleles
for the nine microsatellite loci, with the number of alleles per
loci averaging 3.78 (2–10, 1.7). Mean number of alleles
within aviaries were 2.9, 3.0, and, 3.2 for CESP, Tropicus,
and Guaratuba, respectively. None of the aviaries presented
significant overall deficit of heterozigosity after Sequential
Bonferroni correction (Table 1). All of the aviaries presented
private alleles, being three for CESP, and one for each of
Tropicus and Guaratuba.
Software BOTTLENECK revealed a significant excess
of heterozygosity only for Tropicus (P¼0.019), suggesting a
recent bottleneck in this aviary. Contemporary effective
population sizes and confidence intervals obtained with
NeEstimator were 24.8 (11.4–70) for CESP, 37.1 (18–104.7)
for Tropicus, and 33.2 (19.1–63.1) for Guaratuba.
DISCUSSION
The three main geneticrepositories of the Black-fronted
Piping Guan in captivity are significantly structured into two
lineages, each preserving a few private alleles, and these
groups of birds are not inbred. Practical applications of these
findings involve answering if these groups should still be
managed separately, and if animals to be used in reintroduc-
tions should derive from only one lineage, from a combination
of lineages, or if they should have admixted ancestry.
Captive populations may be structured because they
have derived from populations that were structured in the
wild [Nsubuga et al., 2010; McGreevy et al., 2011; Simons
et al., 2013], or because of founding effects or bottlenecks
that led to random loss of alleles in different captive groups,
even when they are derived from a unique wild population
[Forstmeier et al., 2007]. When structuring in captivity
reflects the structuring of natural populations the captive
groups should be managed separately to preserve adaptive
complexes of each region, and only captive groups derived
from each specific population should be used in reintro-
ductions or supplementation plans to avoid disrupting
potential local adaptations [Frankham et al., 2004;
Champagnon et al., 2012]. Presently, it is impossible to
know what is the genetic diversity of the Black-fronted
Piping Guan in the wild because the remaining populations
are scatteredly distributed and capture to get samples enough
for any genetic analysis is not plausible. The evidence for a
recent bottleneck in at least one of the aviaries (Tropicus)
may suggest that genetic stochasticity occurred in captivity
could explain part of the observed levels of structuring, but
the potential influence of groups that were structured in the
wild also should not be ignored. It has important implications
for releasing planning because when genetic parameters of
native populations are not known, the most prudent
management strategy is avoiding the supplementation of
pre-existing native populations with captive-born animals as
it could incur in disruption of potential local adaptive
complexes [for a review, see Champagnon et al., 2012].
Therefore, we recommend that reintroduction programs
should be differentiated from those of supplementation, and
conducted only after the identification and elimination of the
causes of disappearing of the target species. Deforestation
can be considered as a minor threaten for the Black-fronted
Piping Guan, while hunting is the main threat for the survival
of this species [Bernardo et al., 2011]. This means that in
areas where the species still occurs, other conservation
strategies, such as improving surveillance, would be more
appropriate.
In the face of the lack of information on structuring
of natural populations, we suggest that releasing programs
TABLE 1. Number of alleles (N
A
), observed (H
O
) and expected (H
E
) heterozygosities, inbreeding coefficient (F
IS
), and probability
that F
IS
has differed significantly from zero (P) for each of the three aviaries that hold the main genetic repositories of the Black-
fronted Piping Guan in captivity
CESP TROPICUS GUARATUBA
Locus N
A
H
O
H
E
F
IS
PN
A
H
O
H
E
F
IS
PN
A
H
O
H
E
F
IS
P
Aburria21 4 0.89 0.70 0.279 0.993 4 0.45 0.43 0.057 0.778 4 0.77 0.74 0.045 0.754
Aburria22 2 0.52 0.50 0.031 0.704 2 0.52 0.46 0.111 0.839 2 0.37 0.37 0.023 0.711
Aburria44 3 0.07 0.07 0.010 1.000 3 0.26 0.54 0.529 0.002 3 0.25 0.55 0.550 0.002
Aburria48 4 0.67 0.55 0.209 0.981 2 0.32 0.27 0.176 1.000 3 0.12 0.14 0.129 0.167
Aburria105 2 0.37 0.45 0.185 0.291 2 0.35 0.50 0.293 0.093 2 0.47 0.48 0.012 0.580
Pauxi1-4 2 0.18 0.17 0.083 1.000 3 0.55 0.66 0.171 0.146 3 0.72 0.67 0.084 0.806
Pauxi2-2 5 0.78 0.68 0.154 0.909 6 0.93 0.82 0.147 0.989 7 0.80 0.72 0.107 0.944
Pauxi2-30 2 0.18 0.17 0.083 1.000 3 0.35 0.31 0.142 1.000 3 0.62 0.57 0.098 0.817
Pauxi3-4 2 0.11 0.48 0.774 0.002 2 0.00 0.32 1.000 0.002 2 0.10 0.49 0.796 0.002
Average 2.89 0.42 0.42 0.002 0.493 3.00 0.41 0.48 0.135 0.009 3.22 0.47 0.53 0.103 0.011
SD 0.99 0.19 0.21 ––0.89 0.18 0.13 ––1.01 0.23 0.14
Critical value after Sequential Bonferroni correction ¼0.002.
4Oliveira-Jr. et al.
Zoo Biology
should give priority to areas where the species has been
locally extinct beyond any reasonable doubt. In these sites,
however, as local adaptations are not known, and often the
habitat has changed e.g. due to deterioration or fragmenta-
tion, a combination of individuals from various captive
lineages, or admixtured animals with high levels of
heterozigosity can increase reintroduction success, as it
improves the chances that at least some of the genetic
complexes are adaptive [Frankham et al., 2004; Frankham,
2008; Robert, 2009; Zeisset and Beebee, 2013]. This seems
to be the case of the three areas in which releasing has
already occurred: Paraibuna and Fazenda Maced^
onia are
forest fragments largely isolated by eucalyptus plantations
[personal observation]. Although GuapiaSc
u is very close to
the forest continuums of Serra do Mar, the species vanished
from this region decades ago. However, animals used in
reintroductions in Paraibuna were exclusively from CESP,
and those reintroduced in Fazenda Maced^
onia and
GuapiaSc
u were all derived from Crax Foundation, whose
lineage was founded mainly by animals from Tropicus and
Guaratuba [R. Azeredo, pers. comm.]. As a consequence,
these reintroduced populations are certainly not represen-
tative of all the genetic variability existing in captivity.
Although the presence of second generation animals
(unmarked animals) at least in Fazenda Maced^
onia
[personal observation] is an indicative of success,
thoroughly monitoring programs have never been con-
ducted, and we suggest that mixing the captive lineages
could increase the long term viability of these and future
reintroduced populations.
Despite population structuring issues, when the differ-
ent captive lineages are inbred crossbreedings are indicated to
rescue the genetic variability and to avoid inbreeding
depression within each lineage [Frankham et al., 2004;
Witzenberger and Hochkirch, 2011]. As we did not find
evidences for inbreeding, crossbreedings between aviaries do
not seem to be an urgent need. Indeed, maintaining these
lineages separately may contribute to avoiding genetic
adaptations to captivity [Margan et al., 1998; Frankham,
2008). After a number of generations, captive populations
adapt to captive environment, which implies in reduced
reintroduction success [Williams and Hoffman, 2009].
Limiting the number of generations in captivity, e.g. up to
three generations, can minimize genetic adaptation [Araki
et al., 2007; Frankham, 2008], but like the Black-fronted
Piping Guan, many captive breeding programs have
accumulated a greater number of generations. We estimate
that younger Black-fronted Piping Guans have been through
at least six generations of captive breeding, indicating that
adaptation to captivity may have occurred. Then, a plausible
alternative strategy for minimizing this problem involves
maintaining a number of populations separately, so that each
small group retains less genetic variability than the whole
captive population, limiting the capability of each group to
adapt to captivity [Margan et al., 1998; Frankham, 2008]. As
far as levels of inbreeding can be constantly monitored
[Frankham, 2008], this strategy seems to be adequate for the
groups of Black-fronted Piping Guans analyzed here. It is
important to note that the theoretical loss of heterozygosity in
finite populations, assuming constant population size and
random mating, is (1/2N
e
)
t
,wheretis the number of
generations [Wright, 1931; Frankham et al., 2004]. Then,
with a generation time of 3 years for the Black-fronted Piping
Guan, the studied populations might lose from 37 to 50% of
their heterozygosity in 100 years, which is above the
acceptable level of 10% proposed by Frankham et al.
[2004]. It suggests that the implementation of relatedness
analyses and of genealogical recordings to guide pairing
would be essential to minimize future losses of
heterozygosity.
Then, in summary, we propose four main management
guidelines:
Reintroducing the species in areas where it has been extinct is
more prudent than supplementing natural populations;
As far as inbreeding can be avoided within these three captive
groups, they should be managed separately to minimize
adaptation to captivity;
Crossbreedings, however, are indicated for pre-release
generations. Then, pre-release pairs should be formed and
managed separately, e.g. in other aviaries, to avoid
homogenizing the original lineages;
Relatedness analyses and a studbook should be implemented
to avoid future losses of genetic variability.
The lack of inbreeding is surprising after so many
generations in captivity. Because of the limited financial
support, these private aviaries have tended to reduce the
number of breedingpairs, and in the more dramatic cases, they
can be closed, which may imply in more bottlenecks. The
creation of governmental mechanisms of financial support for
breeding facilities involved in the conservation of endangered
species is urgently needed in Brazil, as captive breeding has
become the last reduct for a growing number of species
[Silveira et al., 2008]. Today, the management of Brazilian
endangered species has relied mostly on researchers, private
institutions, and non-governmental organizations, with lim-
ited governmental participation. While the populations from
the large Atlantic Forests conservation units are declining
progressively [Bernardo et al., 2011], the genetic repositories
of Black-fronted Piping Guans maintained in captivity can
become very important in a near future.
ACKNOWLEDGMENTS
To Instituto Chico Mendes para a ConservaSc~
ao da
Biodiversidade (ICMBio) for authorizing sample collec-
tions and to the owners/keepers of Criadouro Tropicus,
Criadouro Guaratuba, and CESP/Paraibuna for providing
the samples and logistical support. This research is part of
the recommendations of the Brazilian Action Plan for the
Conservation of Endangered Galliformes, ruled by the
Brazilian Federal government.
Managing Black-Fronted Piping Guan 5
Zoo Biology
REFERENCES
Araki H, Arden WR, Olsen EM, Cooper B, Blouin MS. 2007. Reproductive
success of captive-bred steelhead trout in the wild: evaluation of three
hatchery programs in the Hood River. Conserv Biol 21:181–190.
Bernardo CSS, Clay RP. 2006. Black-fronted piping guan (Aburria jacutin
ga). In: Brooks DM, editor. Conserving cracids: the most threatened
family of birds in the americas. Houston, TX: Miscellaneous Publications
of the Houston Museum of Natural Science. p 53–56.
Bernardo CSS, Rubim P, Bueno RS, et al. 2011. Density estimates of the
black-fronted piping guan in the brazilian atlantic forest. Wilson J Ornithol
123:690–698.
Champagnon J, Elmberg J, Guillemain M, Gauthier-Clerc M, Lebreton JD.
2012. Conspecific can be aliens too: a review of effects of practices in
vertebrates. J Nat Conserv 20:231–241.
Conway WG. 1980. An overview of captive propagation. In: Soul
eME,
Wilcox BA, editors. Conservation biology an evolutionary-
ecological perspective. Massachusetts: Sinauer Associates INC. p
199–208.
Costa MC, Camargo C, Laganaro NM, et al. 2014. A suite of microsatellite
markers for genetic management of captive cracids (Aves, Galliformes).
Genet Mol Res 13:9867–9873.
Do C, Waples RS, Peel D, et al. 2014. NeEstimator v2: re-implementation of
software for the estimation of contemporary effective population size (N
e
)
from genetic data. Mol Ecol Resour 14:209–214.
Earl DA, VonHoldt BM. 2012. STRUCTURE HARVESTER: a website and
program for visualizing STRUCTURE output and implementing the
Evanno method. Conserv Genet Resour 4:359–361.
Earnhardt J, V
elez-Valent
ın J, Valentin R, et al. 2014. The Puerto Rican
parrot reintroduction program: sustainable management of the aviary
population. Zoo Biol 33:89–98.
Evanno G, Regnaut S, Goudet J. 2005. Detecting the number of clusters of
individuals using the software STRUCTURE: a simulation study. Mol
Ecol 14:2611–2620.
Forstmeier W, Segelbacher G, Mueller JC, Kempenaers B. 2007. Genetic
variation and differentiation in captive and wild zebra finches
(Taeniopygia guttata). Mol Ecol 16:4039–4050.
Frankham R, Ballou JD, Briscoe DA. 2004. A primer of conservation
genetics. Cambridge, UK: Cambridge University Press. p 236.
Frankham R. 2008. Genetic adaptation to captivity in species conservation
programs. Mol Ecol 17:325–333.
Frankham R. 2010. Challenges and opportunities of genetic approaches to
biological conservation. Biol Conserv 143:1919–1927.
Galetti M, Giacomini HC, Bueno RS, et al. 2009. Priority areas for the
conservation of Atlantic forest large mammals. Biol Conserv 142:
1229–1241.
Galetti M, Guevara R, C^
ortes MC, et al. 2013. Functional extinction of birds
drives rapid evolutionary changes in seed size. Science 340:1086–1090.
Goudet J. 1995. FSTAT (version 1.2): a computer program to calculate F-
statistics. J Hered 86:485–486.
Jones CG, Heck W, Lewis RE, et al. 1995. The restoration of the Mauritius
Kestrel Falco punctatus population. Ibis 137:S173–S180.
Jones KL, Glenn TC, Lacy RC, et al. 2002. Refining the Whooping Crane
studbook by incorporating microsatellite DNA and leg-banding analyses.
Conserv Biol 16:789–799.
Lecis R, Pierpaoli M, Bir
o S, et al. 2006. Bayesian analyses of admixture in
wild and domestic cats (Felis silvestris) using linked microsatellite loci.
Mol Ecol 15:119–131.
Margan SH, Nurthen RK, Montgomery ME. 1998. Single large or several
small? Population fragmentation in the captive management of endan-
gered species. Zoo Biol 17:467–480.
Marsden SJ, Whiffin M, Galetti M, Fielding AH. 2005. How well will
Brazil’s system of Atlantic forest reserves maintain viable bird
populations?. Biodivers Conserv 14:2835–2853.
McGreevy TJ, Jr., Dabek L, Husband TP. 2011. Genetic evaluation of the
Association of Zoos and Aquariums Matschie’s tree kangoroo
(Dendrolagus matschiei) captive breeding program. Zoo Biol 30:
636–646.
Mounce HL, Raisin C, Leonard DL, et al. 2015. Spatial genetic architecture
of the critically-endangered Maui Parrotbill (Pseudonestor xanthophrys):
management considerations for reintroduction strategies. Conserv Genet
16:71–84.
Mukesh, Fernandes M, Han J, Sathyakumar S. 2013. Genetics driven
interventions for ex situ conservation of red junglefowl (Gallus gallus
murghi) populations in India. Zoo Biol 32:476–483.
Nielsen RK, Pertoldi C, Loeschcke V. 2007. Genetic evaluation of the
captive breeding program of the Persian wild ass. J Zool 272:349–357.
Nsubuga AM, Holzman J, Chemnick LG, Ryder OA. 2010. The cryptic
genetic structure of the North American captive gorilla population.
Conserv Genet 11:161–172.
Piry S, Luikart G, Cornuet JM. 1999. Bottleneck: a computer program for
detecting recent reductions in the effective size using allele frequency
data. J Hered 90:502–503.
Pritchard JK, Stephens M, Donnelly P. 2000. Inference of population
structure using multilocus genotype data. Genetics 155:945–959.
Ralls K, Ballou JD. 2004. Genetic status and management of California
Condors. Condor 106:215–228.
Raymond M, Rousset F. 1995. GENEPOP (version 1.2): population genetics
software for exact tests and ecumenicism. J Hered 86:9–10.
Ribeiro MC, Metzger JP, Martensen AC, Ponzoni FJ, Hirota MM. 2009. The
Brazilian Atlantic Forest: how much is left, and how is the remaining
forest distributed? Implications for conservation. Biol Conserv 142:
1141–1153.
Rice W. 1989. Analyzing tables of statistical tests. Evolution 43:223–225.
Robert A. 2009. Captive breeding genetics and reintroduction success. Biol
Conserv 142:2915–2922.
Sambrook J, Fritsch E, Maniatis T. 1989. Molecular cloning: a laboratory
manual. New York, USA: Cold Spring Harbor Laboratory Press.
Shaffer ML. 1987. Minimum viable population: coping with uncertainty. In:
Soul
e ME, editor. Viable populations conservation. New York:
Cambridge University Press. p 69–85.
Sick H. 1997. Ordem galliformes. In: Sick H, editor. Ornitologia Brasileira.
Rio de Janeiro, Brazil: Nova Fronteira. p 270–282.
Silveira LF, Olmos F, Long AJ. 2004. Taxonomy, history, and status of
Alagoas Curassow Mitu mitu (Linnaeus, 1766), the world’s most
threatened cracid. Ararajuba 12:43–50.
Silveira LF, Soares ES, Bianchi CA. 2008. Plano de ASc~
ao Nacional para a
conservaSc~
ao dos Galliformes ameaSc ados de extinSc~
ao (aracu~
as, jacus,
jacutingas, mutuns e urus). Bras
ılia, DF, Brazil: Instituto Chico Mendes de
ConservaSc~
ao da Biodiversidade.
Simons ND, Wagner RS, Lorenz JG. 2013. Genetic diversity of North
American captive-born gorillas (Gorilla gorilla gorilla). Ecol Evol 3:
80–88.
Sousa LMS, Laganaro NM, Camargo C, et al. 2013. Microsatellite markers
for detecting hybrids between the extinct in the wild Alagoas Curassow
(Pauxi mitu) and Razor-billed Curassow (P. tuberosa) (Aves, Galli-
formes). Conserv Genet Res 5:181–183.
Sven H, Ryan W. 2012. The challenge of managing Spix Macaws
(Cyanopsitta spixii) at Qatar an eleven-year retrospection. Zool Garten
81:81–95.
VanderWerf EA, Crampton LH, Diegmann JS, Atkinson CT, Leonard DL.
2014. Survival estimates of wild and captive-bred released Puaiohi, and
endangered Hawaiian thrush. Condor 116:609–618.
Weir B, Cockerham C. 1984. Estimating F-statistics for the analysis of
population sctructure. Evolution 38:1358–1370.
Williams SE, Hoffman EA. 2009. Minimizing genetic adaptation in captive
breeding programs: a review. Biol Conserv 142:2388–2400.
Witzenberger KA, Hochkirch A. 2011. Ex situ conservation genetics: a
review of molecular studies on the genetic consequences of captive
breeding programs for endangered animal species. Biodivers Conserv
20:1843–1861.
Wright S. 1931. Evolution in mendelian populations. Genetics 16:97–159.
Zeisset I, Beebee TJC. 2013. Donor population size rather than local
adaptation can be a key determinant of amphibian tanslocation success.
Anim Conserv 16:359–366.
6Oliveira-Jr. et al.
Zoo Biology
