Mazama Species Research Articles

Integrative analysis ofMazama temama(Artiodactyla: Cervidae) and designation of a neotype for the species

AbstractMazama temama is the smallest red deer in Central America. It was one of the first species described for the genus, and until the end of the last century many authors considered it as a subspecies of Mazama americana due to morphological similarities. Cytogenetic studies of animals in captivity have shown different karyotypes that led to its recognition as a unique species, and phylogenetic studies sustained the monophyly of the species, with some inconsistencies due to its wide geographical distribution. The objectives of the study were to characterize a topotype of Veracruz and three other specimens from a Campeche locality, to complement the description of M. temama and to propose a neotype for the species. We described this species morphologically (cranial measurements, skin color, and body biometry), and obtained karyorypes from wild animals with known origins for cytogenetic analyses (C band, G band, Ag-NOR, and Giemsa staining). We also performed phylogenetic analyses of Cytb, D-loop, COI, and ND5 mitochondrial genes. The morphometric results separated M. temama from M. americana but not from the other small Mazama species. The phylogenetic trees of mitochondrial genes Cytb, COI, and ND5 corroborated the monophyly of the species with 100% of posterior probability. The topotype’s karyotype is significantly different from the one previously described for the species, and the other specimens from Campeche have karyotype variants whose chromosomal rearrangements are possibly the beginning of a reproductive isolation mechanism. We discussed the possibility to have more than one species of red brocket deer in Mexico and Central America.

Cytochrome b sequence of the Mazama americana jucunda Thomas, 1913 holotype reveals Mazama bororo Duarte, 1996 as its junior synonym.

The small red brocket deer, Mazama bororo Duarte, 1996 was described based on karyotypical and morphological characters. However, the original description of Mazama americana jucunda suggested that this subspecies could represent the same taxon as Mazama bororo. This assumption was based on the type locality of Mazama americana jucunda and on morphological similarities between Mazama americana jucunda and Mazama bororo. To solve this question, we obtained DNA sequences of the holotype of Mazama americana jucunda and compared it with other species of Mazama, including the holotype of M. bororo. A phylogenetic tree was obtained to verify the relationships among these taxa. The results clearly showed that M. americana jucunda and M. bororo represent the same biological entity. Therefore, the oldest name available for the small red brocket occurring in the Atlantic Forest of southern Brazil should be Mazama jucunda, remaining M. bororo as a junior synonym. We emphasise the importance of using DNA from museum specimens, especially from holotypes, in order to obtain a more accurate taxonomic identification. We also highlight the importance of application of valid names for labelling all aspects of biodiversity research, as well as for monitoring and conservation efforts.

Satellite DNA in Neotropical Deer Species

The taxonomy and phylogenetics of Neotropical deer have been mostly based on morphological criteria and needs a critical revision on the basis of new molecular and cytogenetic markers. In this study, we used the variation in the sequence, copy number, and chromosome localization of satellite I-IV DNA to evaluate evolutionary relationships among eight Neotropical deer species. Using FISH with satI-IV probes derived from Mazama gouazoubira, we proved the presence of satellite DNA blocks in peri/centromeric regions of all analyzed deer. Satellite DNA was also detected in the interstitial chromosome regions of species of the genus Mazama with highly reduced chromosome numbers. In contrast to Blastocerus dichotomus, Ozotoceros bezoarticus, and Odocoileus virginianus, Mazama species showed high abundance of satIV DNA by FISH. The phylogenetic analysis of the satellite DNA showed close relationships between O. bezoarticus and B. dichotomus. Furthermore, the Neotropical and Nearctic populations of O. virginianus formed a single clade. However, the satellite DNA phylogeny did not allow resolving the relationships within the genus Mazama. The high abundance of the satellite DNA in centromeres probably contributes to the formation of chromosomal rearrangements, thus leading to a fast and ongoing speciation in this genus, which has not yet been reflected in the satellite DNA sequence diversification.

Designation of a neotype for Mazama americana (Artiodactyla, Cervidae) reveals a cryptic new complex of brocket deer species.

Mazama americana (red brocket deer) is the genus-type species (first species described for this genus) and the basis for the identity of other Mazama species. Mazama americana is one of the most abundant and widely distributed deer species in the neotropical forest. However, recent studies suggest that this taxon belongs to a species complex. Our goal was to collect an animal at the type locality (topotype) in French Guiana with the aim of characterizing the morphological (biometric, craniometric), cytogenetic (Giemsa, C-banding, G-banding and NOR) and molecular (mitochondrial DNA) features. The comparisons showed that the collected specimen was very similar morphologically to specimens from other South American populations, but it was cytogenetically and molecularly very different from any of the cytotypes already described for this species, corroborating the existence of a complex of cryptic species. The data suggest that the M. americana topotype is a different species from all the cytotypes already described in the literature and which occupy the southern region of the Amazon River. The characterization and designation of the M. americana neotype is the first step toward a taxonomic reorganization of the genus Mazama, with the potential identification of new species.

Acoustic parameters of courtship and human-directed friendly vocalisations in seven species of Neotropical deer in captivity

Context Polygynous deer rut vocalisations have been found to be highly conspicuous, typical of the species and important in sexual selection. Information about vocalisations is not available for Neotropical species (subfamily Odocoileinae). Aims The objective was to record courtship vocalisations from all Neotropical deer available, looking for differences among species useful for taxonomic identification, characterise the vocalisations according to acoustic parameters and to try to determine functions. Methods Four species of brocket deer (red, Mazama americana (n = 9); grey, M. gouazoubira (n = 7); Amazonian brown, M. nemorivaga (n = 3); Brazilian dwarf, M. nana (n = 1)), and one individual each of marsh (Blastocerus dichotomus), white-tailed (Odocoileus virginianus) and taruca (Hippocamellus antisensis) deer were recorded in captivity at two institutions, in conditions of courtship or human interaction. The acoustic analysis was performed with Praat. Data from the first three species of Mazama were analysed statistically with a multilevel model with two layers. Key results In the context of courting either females or humans, male deer produced low-intensity, short-duration (0.1–0.3 s) bleats with fundamental frequencies (F0) between 100 and 400 Hz; calls were similar among species. The duration of these calls for Amazonian brown brocket males was significantly longer than for males of the other two species. Females of two species had longer-duration calls than did males, in friendly interactions with humans. F0 differences among species for both friendly and courtship calls were almost significant for males of M. gouazoubira. Individual differences were highly significant for both duration and mean F0. Mean F0 for courtship calls of adult males was not correlated with body size in six of the seven species, in contrast to the theory of acoustic allometry. Conclusions The production of male courtship bleats is probably a basic feature of Odocoiline deer and is probably important in sexual selection and female choice. The differences from one species to another are not sufficient for taxonomic use but may be incipient isolating mechanisms between grey and Amazonian brown brocket deer. Implications Studies of deer vocalisations have shown their importance in sexual selection and their incredible variety in closely related species and subspecies, but they have dealt mainly with Old World deer. This study fills a gap in our knowledge, as the first on Neotropical deer, which constitute six genera and 17 species and are widely distributed in the Americas. These data are important for future studies on the function and phylogeny of deer vocalisations.

Molecular identification of Mazama species (Cervidae: Artiodactyla) from natural history collections.

Natural history museum collections constitute an invaluable patrimony of biological diversity for analysing the taxa distribution and evolution. However, it is very common to discover taxonomic misidentification in museum collections based on incorrect data. The aim of this research was to identify brocket deer species (Mazama genus) using molecular markers. We collected 199 samples, performed DNA extraction and species identification using a specific mitochondrial marker based on a fragment of cytochrome b (Cytb) for Neotropical deer. We achieved the amplification and sequencing of 77 specimens and verified that 26% of the skulls were wrongly identified. Moreover, in the museum collections 57% of the specimens were only identified as Mazama sp, and we were able to identify them by molecular methods to the species level. Our findings clearly demonstrate the importance of integrating molecular analyses to identify Mazama species, since using only external morphology can result in a high probability of errors. We recommend the selection of non-convergent morphological characters, which together with the use of DNA collected from museum specimens should contribute to more accurate taxonomic identifications.

First phylogenetic analysis of Mesoamerican brocket deer Mazama pandora and Mazama temama (Cetartiodactyla: Cervidae) based on mitochondrial sequences: Implications for Neotropical deer evolution

Several recent studies have tried to unravel the complex evolutionary history of Neotropical cervids. However, the two Mesoamerican brocket deer Mazama pandora and M. temama have not been included in these studies and their relationship to other Neotropical cervids remains unclear. Here, we present analyses that included concatenated sequences from the mitochondrial genes ND2, Cytb, and tRNA-Pro-Control region. Our results suggest that both Mesoamerican brocket deer fall within the “red” clade, more closely related to genus Odocoileus and red brocket deer species, such as South American Mazama americana, M. bororo, M. nana, and M. rufina. We estimated that M. temama diverged from other red brocket deer ca. two MYA. Two hypothesis emerge regarding the relationship of the Yucatan brocket deer M. pandora with respect to the other Neotropical deer. The first one, based on analyses of concatenated sequences from all three genes, implies an early split from a larger clade that includes Odocoileus and other red Mazama species, with an estimated divergence time dating back to 2.7 MYA. This would suggest isolation on the Yucatan peninsula, limiting the crossing of this species into South America. The second one, based only on Cytb, places M. pandora more closely related to Odocoileus and with an estimated divergence time of ca. 1.9 MYA. This would suggest that the humid tropical forests of Mexico and Central America could have been the most likely place of origin of members of the red brocket deer clade. Deer colonization into South America occurred after the complete formation of the Panamanian land bridge in the late Pliocene, more than one ancestor of the gray brocket deer clade crossed into South America, as well as a red brocket deer ancestor and Odocoileus dispersed southward. A posterior event probably occurred when divergent ancestors of M. temama and M. pandora respectively moved northward after the split with South American red brocket deer. Our results add to the growing body of evidence calling for an extensive taxonomic revision of this group, and we concur with previous recommendations that the generic taxonomic designation of Mazama should be applied not only to the red brocket deer but also to all of the species currently recognized under the genus Odocoileus.

Phylogenetic signal in the circadian rhythm of morphologically convergent species of Neotropical deer

Deer species included in the genus Mazama descend from two different clades that experienced a strong evolutionary convergence in morphology and behaviour when they adapted to Neotropical forests. We would expect that circadian activity rhythms also converged according to habitat features or responded to temporal niche segregation in sympatric species. We used camera trapping in four study areas, representing three main biomes in Brazil, together with data taken from the literature, to analyse activity patterns of five Mazama species in four biomes in South America. Our results show that clade assignment was the main predictor of diurnal versus nocturnal activity, thus suggesting a phylogenetic constraint rather than any other ecological influence on circadian activity. We discuss how the evolutionary history of both lineages may have influenced their activity patterns.

Differentiation of deer species of the genus Mazama by track morphometry

We analyzed morphometric variables of tracks of the five species in the genus Mazama known from Brazil to determine if footprint morphometry can distinguish these species. Tracks from adult M. americana, M. bororo, M. gouazoubira, M. nemorivaga and M. nana maintained at the Deer Research and Conservation Center (NUPECCE) were recorded on a carpet of fine wet sand. Discriminant analysis was successful (99.28%) at classifying the Amazon Rainforest species M. americana and M. nemorivaga whereas for the Atlantic Forest species it was not possible. The analysis of variance (ANOVA) analysis showed longer and wider tracks of M. americana in comparison with other Mazama species, probably due to its larger body size. We do not recommend species identification based on the morphometry of tracks, except for the Amazonian species when imprints have been formed under similar conditions.

Meiotic pairing of B chromosomes, multiple sexual system, and Robertsonian fusion in the red brocket deer Mazama americana (Mammalia, Cervidae)

Deer species of the genus Mazama show significant inter- and intraspecific chromosomal variation due to the occurrence of rearrangements and B chromosomes. Given that carriers of aneuploidies and structural rearrangements often show anomalous chromosome pairings, we here performed a synaptonemal complex analysis to study chromosome pairing behavior in a red brocket deer (Mazama americana) individual that is heterozygous for a Robertsonian translocation, is a B chromosome carrier, and has a multiple sex chromosome system (XY₁Y₂). The synaptonemal complex in spermatocytes showed normal chromosome pairings for all chromosomes, including the autosomal and sex trivalents. The electromicrographs showed homology among B chromosomes since they formed bivalents, but they also appeared as univalents, indicating their anomalous behavior and non-Mendelian segregation. Thus, synaptonemal complex analysis is a useful tool to evaluate the role of B chromosomes and rearrangements during meiosis on the intraspecific chromosomal variation that is observed in the majority of Mazama species.

Transferability of microsatellite loci from exotic Cervidae to Brazilian brocket deer (Mazama spp, Mammalia: Cervidae)

Transferability of microsatellite loci between closely related species has been reported in several species. This helps reduce costs involved with the development of primers for newly investigated species. Fifteen microsatellite primers developed for Rangifer tarandus, Cervus elaphus, C. axis, and Moschus berezovskii were tested on five species of Brazilian brocket deer of the genus Mazama (M. americana, M. bororo, M. gouazoubira, M. nana, and M. nemorivaga). These primers were tested with DNA extracted from blood samples of two individuals of each species obtained from the Núcleo de Pesquisa e Conservação de Cervídeos (NUPECCE) of Universidade Estadual Paulista (UNESP). Fourteen of the 15 primers tested amplified microsatellite regions of all five species of Mazama, confirmed by sequencing of the amplified fragments. We conclude that these primers could be used for population studies of brocket deer.

Identification of the endangered small red brocket deer (Mazama bororo) using noninvasive genetic techniques (Mammalia; Cervidae)

The small red brocket deer Mazama bororo is one of the most endangered deer in the Neotropics. The great morphological similarities with three other sympatric brocket deer species, coupled with the fact that they inhabit densely forested habitats complicate detection and prevent the use of traditional methodologies for accurate identification of species. The ability to determine the presence of this endangered species in an area is crucial for estimating its distribution range, and is critical for establishing conservation management strategies. Here we describe a fast and reliable noninvasive genetic method for species identification of Mazama species from faeces. We designed a primer set that amplifies a short 224-bp fragment of the cytochrome b and demonstrate its effectiveness in successful amplification of DNA isolated from both tissue and faecal samples. This fragment contains a BSTNI/ECORII digestion site that is unique to the endangered M. bororo. The digested polymerase chain reaction products yielded a 160-bp fragment that is clearly visible in a 2% agarose gel. Two other diagnostic sites were identified to differentiate the other three sympatric species, SspI (M. gouazoubira) and AflIII (M. americana, and M. nana).

Latin American deer diversity and conservation: A review of status and distribution

We reviewed the scientific literature published from 1940 to 2002 on Latin American (Mexican and Central and South American) deer (Mammalia: Cervidae), focusing on conservation status and prediction of geographic distributions. The extant Latin American deer are the most diverse deer assemblage in the world (Cervinae: one genus, one species: Elk [Cervus elaphus]; Odocoilineae: six genera, 18 species: White-tailed deer [Odocoileus virginianus], mule deer [O. hemionus], marsh deer [Blastocerus dochotomus], pampas deer [Ozotoceros bezoarticus], huemul [Hippocamelus bisulcus], taruca [Hippocamelus antisensis], greater pudu [Pudu mephistophiles], lesser pudu [Pudu pudu], and six recognized species of brocket deer [Mazama americana, M. gouazoubira, M. nana, M. bricenni, M. chunyi, and M. rufina] as well as three recently proposed species [M. pandora, M. rondoni, and M. bororo]). Within this group, three genera (Odocoileus, Hippocamelus, and Blastocerus) have been well studied, whereas the genus Mazama, P. puda, P. mephistophiles, and H. antisensis have received little attention. The marsh deer, pampas deer, taruca, huemul, lesser pudu, and two Mazama species (Mazama nana and M. bororo) are the most endangered species. They are each listed as Convention International for the Trade of Endangered Species Appendix I and are considered by the International Union for Nature Conservation to be near threatened or endangered. We updated or created distribution maps for these species. Current research predominantly centres on easily available species (e.g., white-tailed deer in Mexico) or endangered species (e.g., marsh deer in Brazil). In contrast, the ecology and conservation status of the pudu, taruca, and Mazama sp. remain largely unknown.