Examination of the Taxonomy and Diversification of <Emphasis Type="Italic">Leontopithecus</Emphasis> using the Mitochondrial Control Region

Leontopithecus comprises 4 taxa: black lion tamarins (L. chrysopygus), golden lion tamarins (L. rosalia), black-faced lion tamarins (L. caissara), and golden-headed lion tamarins (L. chrysomelas). Endemic to the Atlantic Forest of Brazil, they are endangered (Appendix I, CITES; IUCN Critically Endangered: Leontopithecus chrysopygus, L. caissara; IUCN Endangered: L. rosalia, L. chrysomelas). The 4 taxa are differentiated morphologically and geographically and occupy different habitat types. However, it is not clear if all of them are separate species, particularly Leontopithecus caissara, or how they are related to one another evolutionarily. Therefore, we investigated lion tamarin differentiation and radiation. We sequenced the mtDNA control region and performed phylogenetic analyses, population aggregation analyses, and Mantel tests for geographic/genetic correlation. Mitochondrial genetic data suggest 3 distinct lion tamarin clades (Leontopithecus chrysomelas; L. caissara; and L. chrysopygus/L. rosalia). Phylogenetic analysis also supports: 1) the basal lion tamarin is Leontopithecus chrysomelas, and not L. chrysopygus, 2) L. caissara is not subspecific to L. chrysopygus, and 3) Quaternary forest refugia may have shaped lion tamarin diversification via a pattern that does not follow the theory of metachromism. Even though mitochondrial genetic analyses do not unequivocally support the 4 lion tamarins as separate species, one should consider the 4 lion tamarins, with equal conservation priority based on the combination of morphological, genetic, and habitat differentiation. Each of them is an extremely valuable flagship species that focuses attention on the diminishing, highly endemic Atlantic Forest of Brazil.     

Genetic variation within and among lion tamarins

The golden lion tamarin Leontopithecus rosalia rosalia, one of the rarest and most endangered of New World primates, has been the focus of an intensive research and conservation effort for two decades. During that period, managed breeding from 44 founders has brought the captive population to over 400 individuals, a number that equals or exceeds the estimated number of free-ranging golden lion tamarins. The extent of genetic variation among golden lion tamarins was estimated with an electrophoretic survey of 47 allozyme loci from 67 captive and 73 free-ranging individuals. The amount of variation was low, compared to 15 other primate species, with 4% of the loci being polymorphic (P), and with an average heterozygosity H estimate of 0.01 in these callitrichids. Electrophoretic analyses of captive and free-ranging animals (N = 31) of two allopatric morphotypes, Leontopithecus rosalia chrysopygus and L. r. chrysomelas, were similar to the L. r. rosalia findings insofar as they also revealed limited genetic polymorphism. Computation of the Nei-genetic distance measurements showed that the three morphotypes were genetically very similar, although discernible differentiation had occurred at two loci. These data are consistent with the occurrence of recent reproductive isolations of these subspecies.     

Comparative chromosome morphology in three Callitrichid genera: Cebuella, Callithrix, and Leontopithecus

A G-band karyotypic analysis was carried out in individual species groups of three Callitrichid primate genera: Cebuella, Callithrix, and Leontopithecus. Within Callithrix, the karyotypes of the morphologically distinct and geographically isolated morphotypes C. jacchus jacchus and C. jacchus penicillata were identical. Within the lion tamarin genus, Leontopithecus, the karyotypes of the three morphotypes (L. rosalia rosalia, L. rosalia chrysomelas and L. rosalia chrysopygus) were also indistinguishable from one another. These results are consistent with the taxonomic designation of subspecies rank to the different morphotypes. A comparison of type specimens among the three Callitrichid genera showed that their phyletic radiation has been paralleled by a limited number of chromosome rearrangements and a relatively high amount of karyotypic invariance. A fusion/fission event has been postulated to account for the difference in diploid number between Cebuella (2n = 44) and the other species (2n = 46). The karyotype of Callithrix jacchus was found to be more directly derived from Cebuella than was that of Leontopithecus. These findings differ from the previous proposition that Leontopithecus might have diverged from a common Callitrichid ancestor before the emergence of the genus Callithrix.     

Frugivory and seed dispersal of golden lion tamarin (Leontopithecus rosalia (Linnaeus, 1766)) in a forest fragment in the Atlantic Forest, Brazil

The influence of the golden lion tamarin (Leontopithecus rosalia) as a seed disperser was studied by monitoring two groups of tamarins from December 1998 to December 2000 (871.9 hours of observations) in a forest fragment in south-east Brazil. The tamarins consumed fruits of 57 species from at least 17 families. They ingested the seeds of 39 species, and 23 of these were put to germinate in the laboratory and/or in the field. L. rosalia is a legitimate seed disperser because the seeds of all species tested germinated after ingestion, albeit some in low percentages. These primates do not show a consistent effect in final seed germination, because they benefit some species while damaging others. Feces were examined for seeds that had been preyed upon or digested.     

Food sharing in black lion tamarins (Leontopithecus chrysopygus)

Food sharing behavior was investigated by studying 10 captive black lion tamarin (Leontopithecus chrysopygus) infants (six litters) between the ages of four and 26 weeks. The frequency of sharing by parents in response to infant begging increased steadily from the age of five to nine weeks, but then declined gradually, although infants were still receiving food even at 26 weeks. Until the age of 15 weeks, infants received more food items by transfer from other group members than they did from self-feeding. Thereafter, self-feeding predominated. Offering of food by adults without prior infant begging occurred at low frequencies (5.3% of items transferred) and ceased at 20 weeks of age. Begging success (the proportion of infant begs which resulted in food sharing) remained relatively constant, averaging 65%. In common with other lion tamarins, black lion tamarins appear to continue to share food with infants well after weaning, and until at least six months of age.     

Isolation and identification of fungi from vaginal flora in three species of captive Leontopithecus

The ability to reproduce in captivity is an essential component of lion tamarin (Leontopithecus) conservation programs. However, infections such as vaginitis, cervicitis, and endometritis are important diseases that may influence the reproduction of these animals. Therefore, it is important to detect continuous or occasional vaginal microbial populations, and to understand their potential role as an endogenous source of infection [Collins, 1964; Blue, 1983; Pugh et al., 1986]. Vaginal swabs were collected from 25 female tamarins of the three currently available species (L. rosalia, L. chrysopygus, and L. chrysomelas) at the Center of Primatology in Rio de Janeiro, Brazil. The swabs were processed according to standard mycological protocols, and isolates were biochemically characterized. Fungal isolates were recovered from 16 animals (64.0%). The results showed that 70.6% of the isolated microorganisms consisted of yeast, including three species of Candida (mainly C. glabrata). We suggest that this species is a resident member of the normal vaginal flora in Leontopithecus. Filamentous fungi (mainly from Trichosporon, Aspergillus, and Penicilliumgenera) constituted 29.4% of the isolates, and were considered to be transitory contaminants of the genital area. We suggest that colonization of the vaginal environment is related to the endocrine pattern associated with the reproductive status of these animals, but not to parity.     

Ectoparasitic acari found on golden lion tamarins (Leontopithecus rosalia rosalia) from Brazil

Anesthetized golden lion tamarins (Leontopithecus rosalia rosalia), a highly endangered species, were examined for ectoparasites by brushing the fur and swabbing the ears. Ectoparasites recovered from the fur included four species of immature ticks (Amblyomma spp.), three species of chiggers (Euschoengastia sp., Microtrombicula brennani and Speleocola tamarina), and one species of follicular mite (Rhyncoptes anastosi). Ectoparasites do not seem to be of particular significance to the health of the golden lion tamarin; this may in part result from grooming.     

Patterns of urinary oestrogen excretion in female golden lion tamarins (Leontopithecus rosalia)

Daily urine samples were collected from 5 female golden lion tamarins (Leontopithecus rosalia) over a period of 3 or more months, and urinary oestrogen concentrations were determined by radioimmunoassay. Four females exhibited regular patterns of oestrogen excretion, with a peak-to-peak periodicity of 19.6 +/- 1.4 days. Levels of oestrogen excretion tended to vary between, but not within, individual females. Post-partum oestrogen patterns included periods of clear oestrogen cyclicity before conception, with dramatic elevations in oestrogen excretion following conception. Oestrone was the predominant urinary oestrogen excreted by female lion tamarins. Enzyme hydrolysis with Helix pomatia beta-glucuronidase/sulphatase was an efficient method of liberating conjugated oestrogens in tamarin urine. Urinary oestrogen determinations can provide useful information about reproductive status in female lion tamarins.     

Stereology of the myocardium in Leontopithecus (Lesson, 1840) callitrichidae - primates

Rare morphological features of the Leontopithecus cardiovascular system have been reported in the literature. The samples analyzed in this study came from 33 specimens of Leontopithecus from the collection of the Center of Primatology of Rio de Janeiro-FEEMA (CPRJ-FEEMA). Morphometry and stereological data were obtained from all animals. Adult body weights of L. rosalia were the lowest, the greatest being those of L. chrysopygus caissara; body weights of L. chrysomelas and L. c. chrysopygus were similar and in between those of the two former species. Cardiomyocytes (left ventricular myocardium) were bigger in adults than in infants. The myocardium of L. rosalia showed focal fibrosis, fatty vacuoles, and hyalinization. In L. chrysomelas the myocardium showed areas of fibrosis and presence of mononuclear cells. Fibrosis and areas of congestion were observed in L. c. chrysopygus; areas of disorganization and vascular congestion were found in L. c. caissara. In L. rosalia infants, a greater density of vessels per myocardial area and a greater length density of vessels were observed as compared with those of L. chrysomelas. In adults, L. chrysomelas showed greater density of connective tissue in the myocardium than L. c. chrysopygus and L. c. caissara did. In L. rosalia, cardiomyocyte nuclei had a greater area density than those of the other forms of Leontopithecus. These characteristics may explain the faster development of L. rosalia infants as compared with that of L. chrysomelas and L. c. chrysopygus kept under the same handling conditions at the CPRJ-FEEMA.     

Color Vision in <Emphasis Type="Italic">Leontopithecus chrysomelas</Emphasis>: A Behavioral Study

We assessed the color discriminations by golden-headed lion tamarins (Leontopithecus chrysomelas) via a series of tasks involving a behavioral paradigm that maximizes the naturalness of the stimuli. The stimuli were pairs of Munsell color chips used in earlier experiments with human dichromats. We tested 3 male and 3 female monkeys with stimuli of random brightness values in order to assure that discriminations were based on color rather than brightness cues. Results indicate that each male and one female presented random performances for stimuli resembling hue conditions under which tamarins forage: oranges vs. greens. Two females exhibited discriminations consistent with allelic trichromacy. Findings indicate the presence of an M/L cone opsin polymorphism, a condition of most platyrrhines that is characterized by dichromatic and/or trichromatic females and dichromatic males. Interspecific differences in allelic frequency among lion tamarins raises the possibility that habitat fragmentation is affecting heterozygous frequencies, a trend that could impact tamarin foraging efficiency.     

Histology of the sternal and suprapubic skin areas in lion tamarins (Leontopithecus sp. Callitrichidae-primates)

Though knowledge regarding the biology and morphology of lion tamarins is scarce in the literature, it is very important for their conservation. This paper focuses on the anatomical and histological aspects of the glands involved in the scent-marking behavior of lion tamarins. It examines the histological aspects of sternal and suprapubic skin sections of specimens that were preserved in formaldehyde and were the property of the Rio de Janeiro Primatology Center Museum. Eighteen specimens from three lion tamarin (Leontopithecus sp.) species (L. rosalia, L. chrysomelas, and L. chrysopygus) were analyzed. Both sexes were represented, and macroscopic hypertrophy was quantified by direct observation of the tegument on the sternal area and classified as discrete, moderate, or accentuated for each specimen. The skin of both sexes had a high degree of histological resemblance to that of other primates, including humans. The epidermis presented stratified squamous keratinous epithelia, with a few cellular layers and dermis with cutaneous appendages (i.e., hair follicles and both sebaceous and sweat glands). The dermal papillae were short, and the sebaceous and apocrine sweat glands resembled those of humans. These glands were present in the dermis of the analyzed skin fragments of both sternal and suprapubic regions in great numbers. Furthermore, we were able to establish a relationship between the macroscopic appearance of the sternal tegument and the degree of microscopic gland hyperplasia.     

Cross-generic Food Sharing in Tamarins

Adult-infant food-sharing behavior is a major component of the infant care strategies of callitrichids (marmosets and tamarins). It is particularly well-developed in cotton-top tamarins (Saguinus Oedipus) and lion tamarins (Leontopithecus spp), which show frequent adult-initiated food offering, as well as sharing of food in response to begging by infants. This report documents a case of cross-generic food sharing, in which a male golden-headed lion tamarin (Leontopithecus chrysomelas) shared food with an infant cotton-top tamarin. The lion tamarin provided more food to the infant than its mother did. This emphasizes the importance of this behavior in the reproductive strategies of the communally-rearing Callitrichidae and raises questions about mechanisms that underly it.     

Long-call structure and its relation to taxonomy in lion tamarins

Recent evidence on craniodental morphology suggests the acceptance of three species of lion tamarins (Leontopithecus). Confirmatory evidence is presented here using the morphology of long-call vocalizations recorded from several individuals of each type of lion tamarin. Recordings were made of Leontopicthecus rosalia, Leontopithecus chrysopygus, and Leontopithecus chrysomelas at the Centro de Primatologia do Rio de Janeiro (FEEMA) and of L. rosalia at Monkey Jungle in Florida. Thirty separate parameters were measured, and 17 of them differed significantly between populations. In general, L. chrysomelas had higher pitched calls with shorter note duration, while L. chrysopygus, the larger of the animals, had lower pitched calls with longer note duration. L. rosalia was either intermediate to the other two populations or resembled L. chrysopygus. Thus, the results from the analysis of vocal structures closely paralleled the results obtained with more traditional taxonomic methods and suggests that the quantitative analysis of vocal strucures can be a useful adjunct in taxonomy.     

Infection with Trypanosoma cruzi TcII and TcI in free-ranging population of lion tamarins (Leontopithecus spp): an 11-year follow-up

Here, we present a review of the dataset resulting from the 11-years follow-up of Trypanosoma cruzi infection in free-ranging populations of Leontopithecus rosalia (golden lion tamarin) and Leontopithecus chrysomelas (golden-headed lion tamarin) from distinct forest fragments in Atlantic Coastal Rainforest. Additionally, we present new data regarding T. cruzi infection of small mammals (rodents and marsupials) that live in the same areas as golden lion tamarins and characterisation at discrete typing unit (DTU) level of 77 of these isolates. DTU TcII was found to exclusively infect primates, while TcI infected Didelphis aurita and lion tamarins. The majority of T. cruzi isolates derived from L. rosalia were shown to be TcII (33 out 42) Nine T. cruzi isolates displayed a TcI profile. Golden-headed lion tamarins demonstrated to be excellent reservoirs of TcII, as 24 of 26 T. cruzi isolates exhibited the TcII profile. We concluded the following: (i) the transmission cycle of T. cruzi in a same host species and forest fragment is modified over time, (ii) the infectivity competence of the golden lion tamarin population fluctuates in waves that peak every other year and (iii) both golden and golden-headed lion tamarins are able to maintain long-lasting infections by TcII and TcI.     

Description and analysis of the arch display in the golden lion tamarin, Leontopithecus rosalia rosalia.

Three different arch displays are described in captive lion tamarins (Leontopithecus rosalia rosalia) at the National Zoological Park. Analysis of social behaviours including huddling, sexual activity, territorial encounters and arch displays suggests that arch displaying is motivated by conflicting tendencies of a nonaggressive nature and may function as a regulator of social contact, thus maintaining homeostasis within and between a social group.     

Dinucleotide microsatellite primers designed for a critically endangered primate,the black lion tamarin (Leontopithecus chrysopygus)

Black lion tamarin (BLT) monkeys (Leontopithecus chrysopygus) have suffered a severe reduction in their natural range and are consequently critically endangered. Because allozyme data showed very low levels of variation, it was not clear if these monkeys had much genetic diversity. We designed microsatellite primers for BLTs, and from them we identified nine polymorphic loci, seven of which were tested on golden lion tamarins (GLTs) (Leontopithecus rosalia). All of the seven polymorphic loci and two other monomorphic BLT loci were polymorphic in GLTs. The microsatellite markers identified here are directly applicable to ongoing lion tamarin population and conservation genetics studies.     

Morphological and morphometric age-related changes of the upper thoracic aorta in Leontopithecus (Lesson, 1840) (callitrichidae--primates)

Few morphologic aspects of Leontopithecus are listed in the literature, which does not provide sufficient knowledge of those simians. However, such results are important to evaluate the potential of those species as a model for studying comparative pathology. In this study, the material came from the collection of the Museu de Primatologia do Centro de Primatologia do Rio de Janeiro, fixed in 10% formalin pH 7.2. The samples were 33 individuals (five infant and five adult L. rosalia, L. chrysomelas, L. chrysopygus chrysopygus, and three adult L. chrysopygus caissara). The sections of the thoracic aorta were prepared for histological study and measurements were obtained. In the infant Leontopithecus, the aorta presents endothelial line and subendothelial space, the tunica media is not comparatively very thick. In the adults, the aorta showed enlarged tunica intima with consequent thick arterial wall, which increased with the aging process. The atheromatous plates were seen in the aortic wall of all adult L. chrysopygus chrysopygus, in two adult L. rosalia and only one adult L. chrysomelas. Differences in the aortic internal and external diameters were not significant in infants. In the adults, no significant difference was found among L. rosalia, L. chrysomelas and L. c. caissara, although the average of the aortic internal diameter in L. chrysomelas was larger than in L. c. caissara. In infant and adults L. c. chrysopygus, the thickness of the thoracic aorta is larger than in the other forms of Leontopithecus. These findings suggest a morphometric and morphologic aortic difference among the species of Leontopithecus, which could have biological significance that needs major investigation in the future.     

Disseminated lethal Encephalitozoon cuniculi (genotype III) infections in cotton-top tamarins (Oedipomidas oedipus)--a case report

For the first time, Encephalitozoon (E.) cuniculi genotype III ('dog strain') was verified in two cotton-top tamarins (Oedipomidas oedipus) by light microscopy, immunohistochemistry, electron microscopy, PCR and sequencing. The animals had a disseminated lethal infection with this protist. In earlier reports, genotype III had been found only in domestic dogs, man, emperor tamarins (Saguinus imperator) and golden lion tamarins (Leontopithecus rosalia). This investigation establishes now that the 'dog strain' can occur in cotton-top tamarins too. This is further evidence for the zoonotic potential of E. cuniculi. Furthermore, free E. cuniculi spores were identified also in blood vessels of several tissues. These findings indicate that during a disseminated infection E. cuniculi spores can occur in peripheral blood, too. We propose that blood should also be included in the investigations for the detection of microsporidia, so that a possible disseminated course of an infection can be detected.     

Diet, foraging, and use of space in wild golden-headed lion tamarins

Lion tamarins (Callitrichidae: Leontopithecus) are small frugi-faunivores that defend large home ranges. We describe results from the first long-term investigation of wild golden-headed lion tamarins (L. chrysomelas; GHLTs). We present data about activity budgets, daily activity cycles, diet, daily path length, home range size, home range overlap, and territorial encounters for three groups of GHLTs that were studied for 1.5-2.5 years in Una Biological Reserve, Bahia State, Brazil, an area characterized by aseasonal rainfall. We compare our results to those from other studies of lion tamarins to identify factors that may influence foraging and ranging patterns in this genus. Ripe fruit, nectar, insects, and small vertebrates were the primary components of the GHLT diet, and gums were rarely eaten. Fruit comprised the majority of plant feeding bouts, and the GHLTs ate at least 79 different species of plants from 32 families. The most common foraging sites for animal prey were epiphytic bromeliads. The GHLTs defended large home ranges averaging 123 ha, but showed strong affinities for core areas, spending 50% of their time in approximately 11% of their home range. Encounters with neighboring groups averaged two encounters every 9 days, and they were always aggressive. Data about time budgets and daily activity cycles reveal that the GHLTs spent most of their time foraging for resources or traveling between foraging sites distributed throughout their home ranges. The GHLTs spent much less time consuming exudates compared to lion tamarins in more seasonal environments. Additionally, the GHLTs had much larger home ranges than golden lion tamarins (L. rosalia), and did not engage in territorial encounters as frequently as L. rosalia. GHLT ranging patterns appear to be strongly influenced by resource acquisition and, to a lesser extent, by resource defense.     

Trypanosoma cruzi infection in Leontopithecus rosalia at the Reserva Biológica de Poco das Antas, Rio de Janeiro, Brazil

Wild golden lion tamarins (Leontopithecus rosalia) - endangered primates that are native to the Brazilian Atlantic coastal forest - were surveyed for the presence of Trypanosoma cruzi with the use of Giemsa-stained blood smears, hemocultures and an indirect immunofluorescence assay (IFAT). Positive IFAT with titers ranging from 1:20 to 1:1280 were observed in 52% of the 118 wild tamarins examined and the parasite was isolated from 38 tamarins. No patent parasitemia was observed among the tamarins from which T. cruzi was isolated. Serum conversion and positive hemoculture was observed for three animals that had yielded negative results some months earlier, which indicates that T. cruzi is actively transmitted among tamarins. In contrast to observations with other sylvatic isolates, those from the tamarins were significantly more virulent and most of them produced mortality in experimentally infected Swiss mice. Some variation in the kDNA restriction profiles among the isolates was observed. Electrophoresis with GPI, G6PDH, IDH, MDH and ME enzymes showed a Z2 profile.