Carlos R. Ruiz-Miranda

Food transfers in wild and reintroduced golden lion tamarins, Leontopithecus rosalia

We collected data from wild and reintroduced golden lion tamarins (Leontopithecus rosalia) to describe the behavior of donor and recipient during food transfers, evaluate the effect of supplemental feeding on food transfer behavior, and examine various hypotheses concerning the function of food transfers in primates. Behavioral observations were conducted on 12 groups of tamarins with young (N = 30) between the ages of 1 week and 1 year old. Results show that food transfers involve various behaviors, from steals by recipients to offers by donors; transfers mostly derive from adults and are directed at immature weaned young (between 3 and 9 months old); and that most items transferred were prey or fruits that require skill to process. Eleven percent of food transfers were preceded by an adult vocalization specific to that context, whereas 86% were preceded by conspicuous infant vocalizations and begging behavior. The most common vocalizations were loud and atonal (rasps) and broad banded frequency modulated (trills). Infants born to reintroduced parents vocalized less, whereas reintroduced adults vocalized more before transferring food than their wild counterparts. Reintroduced adults and young received more food transfers (4.4 per hr) than did wild‐born adults and young (2.2 per hr). Our findings suggest that food transfer in golden lion tamarins is best understood as provisioning of young that have not fully developed foraging skills to ensure they get the necessary resources for growth and survival. Am. J. Primatol. 48:305–320, 1999.

The Risks of Assisted Colonization

There is a growing debate over whether species should be translocated outside their historic ranges to deal with extinction risks as habitats shift due to climate change. This idea of taking preemptive action to avert predicted extinction risks has been given emphasis by the recent International Union for Conservation of Nature (IUCN) assessment of species susceptibility to climate-change impacts (Foden et al. 2008), prompting suggestions that “more aggressive measures, such as so-called ‘assisted migration’” be considered (Marris 2008). Hoegh-Guldberg et al. (2008) provide a decision framework for identifying scenarios in which what they term “assisted colonization” (AC) is justified. We see problems with the impact of these articles, despite their conservative approach. First, there are current international translocation guidelines (IUCN 1998) that provide a strong rationale against the early adoption of AC as a conservation tool. The Reintroduction Specialist Group (RSG) was created in 1988 to address the proliferation of ill-conceived translocations that had been taking place, including many releases of species outside historic ranges (Stanley Price & Soorae 2003). The RSG formulated the guidelines for translocation planning to ensure that conservation benefits accrue. “Benign introduction” (BI)—the translocation of species to suitable habitat outside their historic range as a conservation measure—was considered appropriate only when there was no habitat left within the original species range (IUCN 1998). Although AC appears to fall within the definition of BI, the two differ in that AC aims proactively to establish species outside their historic range to preempt predicted climate-driven changes in habitat suitability. Calls to take proactive conservation measures need to consider that there are currently huge uncertainties involved, not only in climatechange predictions and consequent species responses (Araújo et al. 2005; Hulme 2005; Sekercioglu et al. 2008) but also in our understanding of the habitat requirements of species (Stamps & Swaisgood 2007) and the effects of translocations on ecosystem function (Armstrong & Seddon 2008). At a recent conference (First International Wildlife Reintroduction Conference, Lincoln Park Zoo, Chicago, Illinois,April2008, http://www. reintroduction.org/), RSG members discussed climate-change implications for translocations, acknowledging the need for the integration of reintroduction biology and restoration ecology, and the updating of translocation guidelines to consider issues such as the mitigation of climate-driven habitat change and overcoming barriers to natural dispersal of species. Given current uncertainty, however, there is substantial risk that prematurely embracing the undeniably sexy AC concept will initiate a new era of ill-conceived species translocations. Philip J. Seddon,∗ Doug P. Armstrong,† Pritpal Soorae,‡ Frederic Launay,§ Sally Walker,∗∗ Carlos R. Ruiz-Miranda,†† Sanjay Molur,∗∗ Heather Koldewey,‡‡ and Devra G. Kleiman§§ ∗RSG Bird Section Chair, Department of Zoology, University of Otago, Dunedin, New Zealand, email philip.seddon@stonebow. otago.ac.nz †RSG Australasia Chair, Massey University, Palmerston North, New Zealand ‡RSG Programme Officer, Environment Agency, Abu Dhabi, United Arab Emirates §RSG Chair, Environment Agency, Abu Dhabi, United Arab Emirates ∗∗RSG South Asia co-Chair, Zoo Outreach, Coimbatore, India ††RSG Meso-South America Chair, Environmental Sciences Laboratory, Universidade Estadual do Norte Fluminense, Rio de Janeiro, Brazil ‡‡RSG Fish Section Chair, Zoological Society of London, London, United Kingdom §§RSG North America Chair, Zoo-Logic, Chevy Chase, MD 20815, U.S.A.

Behavioral and ecological interactions between reintroduced golden lion tamarins (Leontopithecus rosalia Linnaeus, 1766) and introduced marmosets (Callithrix spp, Linnaeus, 1758) in Brazil's Atlantic Coast forest fragments

Marmosets (Callithrix spp.) have been introduced widely in areas within Rio de Janeiro state assigned for the reintroduction of the endangered golden lion tamarin (Leontopithecus rosalia). The objetives of this study were to estimate the marmoset (CM) population in two fragments with reintroduced golden lion tamarin to quantify the association and characterize the interactions between species. The CM population density (0,09 ind/ha) was higher than that of the golden lion tamarin (0,06 ind/ha). The mean association index between tamarins and marmosets varied among groups and seasons (winter=62% and summer=35%). During the winter, competition resulted in increases in territorial and foraging behavior when associated with marmosets. Evidence of benefits during the summer was reduced adult vigilance while associated to marmosets. Golden lion tamarins were also observed feeding on gums obtained from tree gouges made by the marmosets. Marmosets represented a threat for the conservation of golden lion tamarins.

Natural and Anthropogenic Hybridization in Two Species of Eastern Brazilian Marmosets (Callithrix jacchus and C. penicillata).

Animal hybridization is well documented, but evolutionary outcomes and conservation priorities often differ for natural and anthropogenic hybrids. Among primates, an order with many endangered species, the two contexts can be hard to disentangle from one another, which carries important conservation implications. Callithrix marmosets give us a unique glimpse of genetic hybridization effects under distinct natural and human-induced contexts. Here, we use a 44 autosomal microsatellite marker panel to examine genome-wide admixture levels and introgression at a natural C. jacchus and C. penicillata species border along the São Francisco River in NE Brazil and in an area of Rio de Janeiro state where humans introduced these species exotically. Additionally, we describe for the first time autosomal genetic diversity in wild C. penicillata and expand previous C. jacchus genetic data. We characterize admixture within the natural zone as bimodal where hybrid ancestry is biased toward one parental species or the other. We also show evidence that São Francisco River islands are gateways for bidirectional gene flow across the species border. In the anthropogenic zone, marmosets essentially form a hybrid swarm with intermediate levels of admixture, likely from the absence of strong physical barriers to interspecific breeding. Our data show that while hybridization can occur naturally, the presence of physical, even if leaky, barriers to hybridization is important for maintaining species genetic integrity. Thus, we suggest further study of hybridization under different contexts to set well informed conservation guidelines for hybrid populations that often fit somewhere between “natural” and “man-made.”

Hybridization effects and genetic diversity of the common and black-tufted marmoset (Callithrix jacchus and Callithrix penicillata) mitochondrial control region

Hybridization is continually documented in primates, but effects of natural and anthropogenic hybridization on biodiversity are still unclear and differentiating between these contexts remains challenging in regards to primate evolution and conservation. Here, we examine hybridization effects on the mitochondrial DNA (mtDNA) control region of Callithrix marmosets, which provide a unique glimpse into interspecific mating under distinct anthropogenic and natural conditions. DNA was sampled from 40 marmosets along a 50-km transect from a previously uncharacterized hybrid zone in NE Brazil between the ranges of Callithrix jacchus and Callithrix penicillata. DNA was also collected from 46 marmosets along a 30-km transect in a hybrid zone in Rio de Janeiro state, Brazil, where exotic marmosets appeared in the 1980s. Combining Callithrix DNA sampled inside and outside of these hybrid zones, phylogenetic and network analyses show C. jacchus and C. penicillata being parental species to sampled hybrids. We expand limited Callithrix population genetics work by describing mtDNA diversity and demographic history of these parental species. We show ancient population expansion in C. jacchus and historically constant population size in C. penicillata, with the latter being more genetically diverse than the former. The natural hybrid zone contained higher genetic diversity relative to the anthropogenic zone. While our data suggest hybrid swarm formation within the anthropogenic zone due to removed physical reproductive barriers, this pattern is not seen in the natural hybrid zone. These results suggest different genetic dynamics within natural and anthropogenic hybridization contexts that carry important implications for primate evolution and conservation.

Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

Temporal genetic dynamics of reintroduced and translocated populations of the endangered golden lion tamarin ( Leontopithecus rosalia )

Reintroductions—captive-born animals introduced into the species’ original distribution area—and translocations—free-living animals transferred to another location within the historical distribution area—are important conservation strategies for endangered species. Genetic analyses of 239 individuals from unmanaged, translocated and reintroduced populations of Leontopithecus rosalia were performed using 14 microsatellites. These samples were collected during two periods: (a) 1996–1997 (historic), when individuals were translocated and reintroduced into forest fragments in the lowland Atlantic Forest, and (b) 2007–09 (recent). We hypothesized that effective population size and genetic diversity would increase over time and that these management strategies would affect the resulting population genetic structure. We found trends indicating that the effective population size at the translocation site increased while that at the reintroduction sites diminished over time. The inbreeding coefficient of the translocated population diminished over time (from 0.38 to 0.03) and was much lower than that of the native (0.29) and reintroduced (0.13) recent populations. We observed a greater genetic admixture among the reintroduced sites on the historic sampling, as well as a strong genetic structure at the translocation site. In the recent sampling, the population structuring became more site-related suggesting low or inconsistent gene flow between sampling sites. This research highlights how conservation management decisions have an important influence on the genetic outcome of translocations and reintroductions. Future conservation planning should consider population genetic monitoring before and after management measures and maintain population connectivity thereafter to avoid the negative effects of a population size reduction.

Acoustic differences between spontaneous and induced long calls of golden lion tamarins, Leontopithecus rosalia

Long calls are emitted by several primate taxa and appear to have multiple functions related to spacing mechanisms and reproduction. Yet, in some species the spontaneous rate of emission of these calls is low, and playbacks are used to elicit long calls to augment the sample size of studies. The objective of this study was to compare the acoustic structure of spontaneously emitted and experimentally induced long calls of golden lion tamarins. Results from 6 adult tamarins show that spontaneously emitted long calls differ acoustically from experimentally induced long calls in several frequency measures. Several explanations for these differences are discussed. We conclude that regardless of the underlying mechanisms, playbacks elicit responses that may be different from naturally occurring calls.

Application of PE-RADSeq to the study of genomic diversity and divergence of two Brazilian marmoset species (Callithrix jacchus and C. penicillata).

Callithrix jacchus and C. penicillata are among the smallest anthropoid primates, are highly specialized tree gougers, and largely occupy Brazils most extreme, semi‐arid biomes. However, the underlying genomic factors that underpin the evolution of these species and their unique traits are under‐investigated. Additionally, exotic populations of these two species are widely established throughout Brazil and hybridize with threatened native congers. Thus, both genomic and conservation factors call for a better understanding of C. jacchus and C. penicillata evolution. Here, we applied PE‐RADseq to characterize genomic variation in these two species, using six C. jacchus and seven C. penicillata individuals. We identified an average of 7,463 and 5,180u2009SNPs/individual in C. penicillata and C. jacchus, respectively, and also found 1,395 variable sites that were represented in both species. C. penicillata showed overall higher levels of genetic diversity than C. jacchus at the variable sites present in both species. Additionally, among these variable sites, 106 showed relative interspecific divergence levels that were significantly higher than the genome‐wide average. We further compared relative and absolute divergence for C. penicillata and C. jacchus between RAD loci associated with the 106 significantly diverged variable sites and all other RAD loci present in both species. The former RAD loci set showed significantly elevated relative and absolute divergence measures in comparison to the latter set. This convergence suggests that C. jacchus and C. penicillata may have diverged under a scenario of gene flow under secondary contact. Here, we demonstrate that RADseq is an efficient method to simultaneously discover and genotype a large number of markers and validate the utility of RADseq for examining Callithrix evolution.

Population Genetic Structure of an Endangered Endemic Primate ( Leontopithecus chrysomelas ) in a Highly Fragmented Atlantic Coastal Rain Forest

This study evaluated the genetic structure of wild populations of the endangered primate, Leontopithecus chrysomelas. We tested the assumption that populations of L. chrysomelas, given their larger population size and a higher degree of habitat continuity, would have higher genetic diversity and less genetic structuring than other lion tamarins. We used 11 microsatellites and 122 hair samples from different locations to assess their genetic diversity and genetic structure, and to make inferences about the isolation by distance. The overall expected heterozygosity (0.51 ± 0.03) and the average number of alleles (3.6 ± 0.2) were relatively low, as is the case in other endangered lion tamarins. Genetic clustering analyses indicated two main clusters, whereas the statistical analyses based on genotype similarities and Fst suggested further substructure. A Mantel test showed that only 34% of this genetic differentiation was explained by the linear distance. In addition to linear distance, structural differences in the landscape, physical barriers and behavioural factors may be causing significant genetic structuring. Overall, this study suggests that these populations have a relatively low genetic diversity and a relatively high population genetic structure, putting in question whether the presence of agroforest systems (known locally as cabruca) is enough to fully re-establish functional landscape connectivity.