The endangered maned sloth Bradypus torquatus of the Brazilian Atlantic forest: a review and update of geographical distribution and habitat preferences

• 1 The endemic maned sloth Bradypus torquatus is globally threatened due to deforestation, habitat fragmentation and isolation of populations. Despite increased interest in the species in recent years, information on its geographic distribution is limited and is derived mainly from a few museum species and from a survey based mostly on interviews carried out more than 20 years ago. Information on its habitat and altitudinal preferences is lacking. We therefore compile, update and review all occurrence records of this species using geographical information system (GIS) tools and produce a distribution map for the species.
• 2 Our compilation totalled 188 occurrence records, of which 130 (69%) were considered confirmed (museum specimens or individuals captured or seen in the field), most of which (62%) were restricted to the states of Bahia and central‐south Espírito Santo (29%). The species occurs predominantly in ombrophilous forest (80%), from sea level to 1290m, but mainly (79%) below 200m altitude.
• 3 We used GIS techniques and the alpha shape algorithm to calculate the extent of occurrence of the species and compared the resulting map with distribution maps available in the literature. We found a great deal of variation in distribution area shape and an almost two‐fold variation in distribution area size among sources.
• 4 We confirmed the existence of a main distribution gap between southern Bahia and northern Espírito Santo and discovered a second smaller gap between southern Espírito Santo and northern Rio de Janeiro.
• 5 We also confirmed that B. torquatus is sympatric with the common three‐toed sloth Bradypus variegatus in a large part of the Atlantic forest and that the two species are syntopic in at least 11 locations.

     

Multi‐scale effects of habitat structure and landscape context on a vertebrate with limited dispersal ability (the brown‐throated sloth,Bradypus variegatus)

As human population, food consumption, and demand for forest products continue to rise over the next century, the pressures of land‐use change on biodiversity are projected to intensify. In tropical regions, countryside habitats that retain abundant tree cover and structurally complex canopies may complement protected areas by providing suitable habitats and landscape connectivity for a significant portion of the native biota. Species with low dispersal capabilities are among the most at risk of extinction as a consequence of land‐use change. We assessed how the spatial distribution of the brown‐throated sloth (Bradypus variegatus), a model species for a vertebrate with limited dispersal ability, is shaped by differences in habitat structure and landscape patterns of countryside habitats in north‐central Costa Rica using a multi‐scale framework. We quantified the influence of local habitat characteristics and landscape context on sloth occurrence using mixed‐effects logistic regression models. We recorded 27 sloths within countryside habitats and found that both local and landscape factors significantly influenced their spatial distribution. Locally, sloths favored structurally complex habitats, with greater canopy cover and variation in tree height and basal area. At the landscape scale, sloths demonstrated a preference for habitats with high proportions of forest and nearby large tracts of forest. Although mixed‐use areas and tree plantations are not substitutes for protected forests, our results suggest they provide important supplemental habitats for sloths. To promote the conservation and long‐term viability of sloth populations in the tropical countryside, we recommend that land managers retain structurally complex vegetation and large patches of native habitat.     

Spatial ecology and conservation of two sloth species in a cacao landscape in limón,Costa Rica

The spatial ecology of sloths was studied in an agricultural landscape in Limón Province, Costa Rica. Two sloth species, the three-toed sloth (Bradypus variegatus) and the two-toed sloth (Choloepus hoffmanni), actively used and traveled through a cacao agroforest and its contiguous living fence rows and riparian forests. This agroecosystem was embedded in an agricultural landscape dominated by banana and pineapple plantations and pastures with dispersed trees. The two-toed sloth (C. hoffmanni) was found in 101 tree species and used 34 for food; the three-toed sloth (B. variegatus) was found in 71 tree species and used 15 for food. Choice of preferred species differed between the two sloth species. Trees commonly used by sloths for food and/or refuge in the cacao agroforest included Erythrina poeppigiana, Cecropia obtusifolia, Leucaena leucocephala; in the living fence rows, Cordia alliodora, Erythrina poeppigiana, Ocotea sinuata and Trophis racemosa; in the riparian forests, Coussapoa villosa, Cecropia obtusifolia, Hura crepitans, Pterocarpus officinalis and Spondias mombin; and in the pastures with dispersed trees, Cordia alliodora, Coussapoa villosa, Erythrina poeppigiana, Ocotea sinuata and Hura crepitans. This study demonstrates the importance of the cacao agroforest as well as arboreal elements in other land uses in providing resources for sloth conservation in a larger agricultural landscape.     

Mitochondrial DNA genome evidence for the existence of a third divergent lineage in the western Atlantic Ocean for the bull shark (Carcharhinus leucas)

We report for the first time a highly divergent lineage in the Caribbean Sea for the bull shark (Carcharhinus leucas) based on the analysis of 51 mitochondrial DNA genomes of individuals collected in the western North Atlantic. When comparing the mtDNA control region obtained from the mitogenomes to sequences reported previously for Brazil, the Caribbean lineage remained highly divergent. These results support the existence of a discrete population in Central America due to a phylogeographic break separating the Caribbean Sea from the western North Atlantic, Gulf of Mexico and South America.     

Sloth Hair as a Novel Source of Fungi with Potent Anti-Parasitic,Anti-Cancer and Anti-Bacterial Bioactivity

The extraordinary biological diversity of tropical forests harbors a rich chemical diversity with enormous potential as a source of novel bioactive compounds. Of particular interest are new environments for microbial discovery. Sloths – arboreal mammals commonly found in the lowland forests of Panama – carry a wide variety of micro- and macro-organisms on their coarse outer hair. Here we report for the first time the isolation of diverse and bioactive strains of fungi from sloth hair, and their taxonomic placement. Eighty-four isolates of fungi were obtained in culture from the surface of hair that was collected from living three-toed sloths (Bradypus variegatus, Bradypodidae) in Soberanía National Park, Republic of Panama. Phylogenetic analyses revealed a diverse group of Ascomycota belonging to 28 distinct operational taxonomic units (OTUs), several of which are divergent from previously known taxa. Seventy-four isolates were cultivated in liquid broth and crude extracts were tested for bioactivity in vitro. We found a broad range of activities against strains of the parasites that cause malaria (Plasmodium falciparum) and Chagas disease (Trypanosoma cruzi), and against the human breast cancer cell line MCF-7. Fifty fungal extracts were tested for antibacterial activity in a new antibiotic profile screen called BioMAP; of these, 20 were active against at least one bacterial strain, and one had an unusual pattern of bioactivity against Gram-negative bacteria that suggests a potentially new mode of action. Together our results reveal the importance of exploring novel environments for bioactive fungi, and demonstrate for the first time the taxonomic composition and bioactivity of fungi from sloth hair.     

The Distributional Ecology of the Maned Sloth: Environmental Influences on Its Distribution and Gaps in Knowledge

The maned sloth Bradypus torquatus (Pilosa, Bradypodidae) is endemic to a small area in the Atlantic Forest of coastal Brazil. It has been listed as a threatened species because of its restricted geographic range, habitat loss and fragmentation, and declining populations. The major objectives of this study were to estimate its potential geographic distribution, the climatic conditions across its distributional range, and to identify suitable areas and potential species strongholds. We developed a model of habitat suitability for the maned sloth using two methods, Maxent and Mahalanobis Distance, based on 42 occurrence points. We evaluated environmental variable importance and the predictive ability of the generated distribution models. Our results suggest that the species distribution could be strongly influenced by environmental factors, mainly temperature seasonality. The modeled distribution of the maned sloth included known areas of occurrence in the Atlantic Forest (Sergipe, Bahia, Espírito Santo, and Rio de Janeiro), but did not match the observed distributional gaps in northern Rio de Janeiro, northern Espírito Santo or southern Bahia. Rather, the model showed that these areas are climatically suitable for the maned sloth, and thus suggests that factors other than climate might be responsible for the absence of species. Suitable areas for maned sloth were located mainly in the mountainous region of central Rio de Janeiro throughout Espírito Santo and to the coastal region of southern Bahia. We indicate 17 stronghold areas and recommended survey areas for the maned sloth. In addition, we highlight specific areas for conservation, including the current network protected areas. Our results can be applied for novel surveys and discovery of unknown populations, and help the selection of priority areas for management and conservation planning, especially of rare and relatively cryptic species directed associated with forested habitats.     

Observations on the Endemic Pygmy Three-Toed Sloth,Bradypus pygmaeus of Isla Escudo de Veraguas,Panamá

Our objective was to ascertain the population status of the Pygmy Three-toed Sloth, Bradypus pygmaeus, an IUCN Critically Endangered species, on Isla Escudo de Veraguas, Panama. Bradypus pygmaeus are thought to be folivorous mangrove specialists; therefore we conducted a visual systematic survey of all 10 mangrove thickets on the island. The total mangrove habitat area was measured to be 1.67 ha, comprising 0.024% of the total island area. The population survey found low numbers of B. pygmaeus in the mangrove thickets and far lower numbers outside of them. The connectivity of subpopulations between these thickets on the island is not established, as B. pygmaeus movement data is still lacking. We found 79 individuals of B. pygmaeus; 70 were found in mangroves and 9 were observed just beyond the periphery of the mangroves in non-mangrove tree species. Low population number, habitat fragmentation and habitat loss could lead to inbreeding, a loss of genetic diversity, and extinction of B. pygmaeus.     

First observation on mating and reproductive seasonality in maned sloths <Emphasis Type="Italic">Bradypus torquatus</Emphasis> (Pilosa: Bradypodidae)

Information on mating season is of paramount importance for our understanding of reproductive behavior, yet little is currently known about the breeding season of Xenarhtra as a whole and sloths in particular. A pair of maned sloths was observed copulating in September 2005 in the Atlantic Forest region of south-eastern Brazil. Our studies on mating of the manned sloth as well of other hitherto unpublished records of newborn sloths seen in the wild, together with a review of previous information, indicate that B. torquatus is a seasonal breeder. Infants are born predominantly at the end of the wet season and beginning of the dry season (February–April), and copulation concentrates in the late dry and early wet seasons (August–October). This strategy allows for the energy-demanding gestation and lactation to take place during the less stressful period of the year.     

A striking lack of genetic diversity across the wide-ranging amphibian <Emphasis Type="Italic">Gastrophryne carolinensis</Emphasis> (Anura: Microhylidae)

We examine phylogeographic structure across a wide-ranging microhylid frog (Gastrophryne carolinensis) using both mitochondrial (mtDNA) and nuclear (AFLP) data. Species with similar ecological characteristics such as large range size, low vagility, or existence across known biogeographic barriers, often are comprised of multiple, cryptic lineages. Surprisingly, our analyses of both portions of the genome show very little phylogeographic or population genetic structure. The family Microhylidae is one of the largest families of anurans with over 60 genera and around 400 species distributed across much of the world (Americas, Asia, Africa, and Madagascar), but very few phylogeographic studies have assessed intraspecific genetic diversity across the mitochondrial and nuclear genomes. Our results suggest that G. carolinensis, one of only three species of microhylid native to the US, has experienced a severe population bottleneck with subsequent range expansion. Comparable molecular data from closely related microhylids, in addition to demographic and ecological analyses, will provide valuable insight into patterns of genetic diversity and the processes driving phylogeographic diversity in these wide-ranging frogs.     

Extensive hybridization in hawksbill turtles (Eretmochelys imbricata) nesting in Brazil revealed by mtDNA analyses

Bahia state hosts over 90% of hawksbill (Eretmochelys imbricata) nests registered in the main nesting sites monitored by Projeto Tamar-IBAMA in Brazil. The genetic diversity of this hawksbill population (n=119) was assayed through the analyses of 752 bp of the mitochondrial DNA control region in nesting females. Seven distinct haplotypes, defined by 125 polymorphic sites, were found. Most of the individuals (n=67) display four typical hawksbill haplotypes, 50 individuals display two haplotypes characteristic of the loggerhead turtle (Caretta caretta) and two individuals had a haplotype affiliated with the olive ridley (Lepidochelys olivacea). These results demonstrate hybridization between the hawksbills and two species that nest along the Bahia coast. Of special interest is the high occurrence of loggerhead × hawksbill hybrids (42%), which display loggerhead mtDNA haplotypes but are characterized morphologically as hawksbills. The true hawksbill haplotypes present only three variable sites and low genetic diversity values (h=0.358±0.069; π=0.0005±0.0001). The occurrence of several nesting individuals with identical mtDNA from another species may also suggest a long history of introgression between species producing likely F2 or further generation hybrids. Marine turtle hybrids have been previously reported, but the high frequency observed in Bahia is unprecedented. Such introgression may influence evolutionary pathways for all three species, or may introduce novel morphotypes that develop apart from the parental species. The presence of a unique hybrid swarm has profound conservation implications and will significantly influence the development and implementation of appropriate management strategies for these species.     

Genetic Differentiation of the Giant Honey Bee (Apis dorsata) in Thailand Analyzed by Mitochondrial Genes and Microsatellites

Genetic diversity and population differentiation of the giant honey bee (Apis dorsata) in Thailand were examined. Six PCR-RFLP mitotypes were generated from digestion of the COI-COII, Cytb-tRNA^ser, ATPase6-8, and lrRNA genes with Dra I and Hin fI. Low genetic diversity (h=0.074, π=0.032%) and a lack of genetic population differentiation between A. dorsata originating from geographically different regions were observed from mtDNA polymorphisms (P > 0.05). In contrast, microsatellite (A14, A24, and A88) polymorphisms revealed a relatively high level of genetic diversity in A. dorsata (H _o=0.68–0.74, average number of alleles per locus=6.0–9.0). Both A24 and A88 indicated significant population differentiation between bees from the north-to-central region (north, northeast, and central regions), peninsular Thailand, and Samui Island.     

Genetic diversity in the bronze gudgeon, <Emphasis Type="Italic">Coreius heterodon</Emphasis>, from the Yangtze River system based on mtDNA sequences of the control region

The bronze gudgeon, Coreius heterodon (Bleeker), is an economically important species, which only inhabits the Yangtze River. The stock declined drastically in recent years due to dam construction, over-fishing, and pollution. Little is known about its population genetic structure. In this study, the sequences of the mitochondrial DNA (mtDNA) control regions of natural bronze gudgeon was determined for fish collected from four sites in the Yangtze River (n = 102). The molecular data were used to estimate the genetic diversity and differentiation of the bronze gudgeon. The results showed that 28 haplotypes and 22 variable sites were found, and the haplotype diversity (π) and nucleotide diversity (h) were 0.849 and 0.00257, respectively. A low level of genetic diversity exists in the bronze gudgeon. Analysis of molecular variance (AMOVA) suggests that 98.8% of the genetic variability occurred within the populations; the site of collection had little influence on diversity. Future research should focus on investigating the genetic divergence of populations in different tributaries and using additional polymorphic markers, such as microsatellite DNA, to verify the results and improve interpretation.     

Predation attempt and abnormal coat coloration of the tayra (Eira barbara) in the Brazilian Central Amazon

Predation is one of the most important interactions between species and a major selection pressure in wild mammal populations. Camouflage is a common pattern of colorations resulting from predation pressure, however, atypical coloration may appear in the wild. Both predation and atypical coloration in mammal species are difficult to observe in a natural setting but contain important biological information. Here we describe a leucistic tayra (Eira barbara) attempting to prey on a sloth (Bradypus tridactylus). The event occurred in Brazilian Central Amazon as a tayra unsuccessfully tried to kill an adult sloth in the mid-canopy of a secondary forest.     

GLOBAL PHYLOGEOGRAPHY OF THE LOGGERHEAD TURTLE (CARETTA CARETTA) AS INDICATED BY MITOCHONDRIAL DNA HAPLOTYPES

Restriction-site analyses of mitochondrial DNA (mtDNA) from the loggerhead sea turtle (Caretta caretta) reveal substantial phylogeographic structure among major nesting populations in the Atlantic, Indian, and Pacific oceans and the Mediterranean sea. Based on 176 samples from eight nesting populations, most breeding colonies were distinguished from other assayed nesting locations by diagnostic and often fixed restriction-site differences, indicating a strong propensity for natal homing by nesting females. Phylogenetic analyses revealed two distinctive matrilines in the loggerhead turtle that differ by a mean estimated sequence divergence p = 0.009, a value similar in magnitude to the deepest intraspecific mtDNA node (p = 0.007) reported in a global survey of the green sea turtle Chelonia mydas. In contrast to the green turtle, where a fundamental phylogenetic split distinguished turtles in the Atlantic Ocean and the Mediterranean Sea from those in the Indian and Pacific oceans, genotypes representing the two primary loggerhead mtDNA lineages were observed in both Atlantic–Mediterranean and Indian-Pacific samples. We attribute this aspect of phylogeographic structure in Caretta caretta to recent interoceanic gene flow, probably mediated by the ability of this temperate-adapted species to utilize habitats around southern Africa. These results demonstrate how differences in the ecology and geographic ranges of marine turtle species can influence their comparative global population structures.     

Low genetic diversity in the endangered great Indian bustard (Ardeotis nigriceps) across India and implications for conservation

The great Indian bustard (Ardeotis nigriceps) is an endemic endangered bird of the Indian subcontinent with a declining population, as a result of hunting and continuing habitat loss. In this first genetic study of this little-known species, we investigate the diversity of the mitochondrial DNA (hypervariable control region II and cytochrome b gene) among samples (n = 63) from five states within the current distribution range of great Indian bustards in India. We find just three haplotypes defined by three variable sites, a comparatively low genetic diversity of π = 0.0021 ± 0.0012 for cytochrome b, 0.0008 ± 0.0007 for the control region (CR), and 0.0017 ± 0.0069 for combined regions and no phylogeographic structure between populations. We provide evidence for a bottleneck event, estimate an effective population size (Ne) that is roughly concordant with recent population size estimates based on field surveys (~200 to 400), but extremely low for a widely distributed species. We also discuss the conservation implications. Based on our findings, we strongly recommend upgrading the IUCN threat status from Endangered to Critically Endangered.     

Rate of digesta passage in the Philippine flying lemur,Cynocephalus volans

The rate of digesta passage was measured in five captive Philippine flying lemurs (Cynocephalus volans). These animals were force fed capsules containing known quantities of either particulate or soluble markers. The volumes of the gastrointestinal tracts of three flying lemurs were determined based on the wet weight of the contents of each section of the gut. The mean rate of digesta passage was 14.37±3.31 h when determined using the particulate marker and 21.9±0.03 h when determined using the soluble marker. The values based on the particulate marker are between 2% and 10% of similar values for other arboreal folivores. The morphology of the gastrointestinal system of the Philippine flying lemur is similar to that of other hindgut fermenters. Flying lemurs have a simple stomach and a large caecum. The total gut capacity of the Philippine flying lemur is similar to that of other herbivores, but is slightly smaller than that of either the koala (Phascolarctos cinereus), a hindgut fermenter, or the three-toed sloth (Bradypus variegatus), a foregut fermenter. These data suggest that flying lemurs deal with the problems of a folivorous diet very differently than some other arboreal mammals. Phascolarctos cinereus and Bradypus variegatus may represent one extreme with Cynocephalus volans representing the other extreme along a continuum of foraging strategies that are compatible with the arboreal folivore lifestyle.Abbreviations bw body weight - EDTA ethylenediaminetetraaceticacid - MR metabolic rate - i mean rate of passage - t _exp expected time to appearance - t _o time to first appearance     

Patterns of genetic diversity and differentiation in the tsetse fly <Emphasis Type="Italic">Glossina morsitans morsitans</Emphasis> Westwood populations in East and southern Africa

Genetic diversity and differentiation within and among nine G. morsitans morsitans populations from East and southern Africa was assessed by examining variation at seven microsatellite loci and a mitochondrial locus, cytochrome oxidase (COI). Mean COI diversity within populations was 0.63 ± 0.33 and 0.81 taken over all populations. Diversities averaged over microsatellite loci were high (mean number of alleles/locus ≥7.4; mean H _E ≥ 65%) in all populations. Diversities averaged across populations were greater in East Africa (mean number of alleles = 22 ± 2.6; mean h _e = 0.773 ± 0.033) than in southern Africa (mean number of alleles = 18.7 ± 4.0; mean h _e = 0.713 ± 0.072). Differentiation among all populations was highly significant (R _ST = 0.25, F _ST = 0.132). Nei’s G _ij statistics were 0.09 and 0.19 within regions for microsatellites and mitochondria, respectively; between regions, G _ij was 0.14 for microsatellites and 0.23 for mitochondria. G _ST among populations was 0.23 for microsatellite loci and 0.40 for mitochondria. The F, G and R statistics indicate highly restricted gene flow among G. m. morsitans populations separated over geographic scales of 12–917 km.     

Low Phylogeographic Structure in a Wide Spread Endangered Australian Frog <Emphasis Type="Italic">Litoria aurea</Emphasis> (Anura: Hylidae)

The green and golden bell frog (Litoria aurea) has a widespread distribution along the south-east coast of Australia. The species range, however, is highly fragmented and remaining populations are predominately isolated and restricted to the coastline. Previously, the range extended further inland and the species was considered common. Here we report a study designed to identify the phylogeographic and conservation genetic parameters of L. aurea. Mitochondrial DNA sequences were examined from 263 individuals sampled from 26 locations using both phylogenetic and population analyses. Despite a general consensus that amphibians are highly structured we found no phylogeographic divisions within the species, however, there was significant structure amongst extant populations (F _ST=0.385). Patterns of haplotype relatedness, high haplotypic diversity (mean h=0.547) relative to low nucleotide diversity (mean π=0.003) and mismatch distribution analysis supported a Pleistocene expansion hypothesis with continued restricted dispersal and gene flow. We conclude that the genetic structure of the species may permit ‘well managed’ intervention to mediate gene flow amongst isolated populations and provide some guidelines for the implementation of such conservation strategies.     

Evolutionary and demographic processes shaping geographic patterns of genetic diversity in a keystone species,the African forest elephant (Loxodonta cyclotis)

The past processes that have shaped geographic patterns of genetic diversity may be difficult to infer from current patterns. However, in species with sex differences in dispersal, differing phylogeographic patterns between mitochondrial (mt) and nuclear (nu) DNA may provide contrasting insights into past events. Forest elephants (Loxodonta cyclotis) were impacted by climate and habitat change during the Pleistocene, which likely shaped phylogeographic patterns in mitochondrial (mt) DNA that have persisted due to limited female dispersal. By contrast, the nuclear (nu) DNA phylogeography of forest elephants in Central Africa has not been determined. We therefore examined the population structure of Central African forest elephants by genotyping 94 individuals from six localities at 21 microsatellite loci. Between forest elephants in western and eastern Congolian forests, there was only modest genetic differentiation, a pattern highly discordant with that of mtDNA. Nuclear genetic patterns are consistent with isolation by distance. Alternatively, male‐mediated gene flow may have reduced the previous regional differentiation in Central Africa suggested by mtDNA patterns, which likely reflect forest fragmentation during the Pleistocene. In species like elephants, male‐mediated gene flow erases the nuclear genetic signatures of past climate and habitat changes, but these continue to persist as patterns in mtDNA because females do not disperse. Conservation implications of these results are discussed.     

Stocking may increase mitochondrial DNA diversity but fails to halt the decline of endangered Atlantic salmon populations

Over the last 50 years, Spanish Atlantic salmon (Salmo salar) populations have been in decline. In order to bolster these populations, rivers were stocked with fish of northern European origin during the period 1974–1996, probably also introducing the furunculosis-inducing pathogen, Aeromonas salmonicida. Here we assess the relative importance of processes influencing mitochondrial (mt)DNA variability in these populations from 1948 to 2002. Genetic material collected over this period from four rivers in northern Spain (Cantabria) was used to detect variability at the mtDNA ND1 gene. Before stocking, a single haplotype was found at high frequency (0.980). Following stocking, haplotype diversity (h) increased in all rivers (mean h before stocking was 0.041, and 0.245 afterwards). These increases were due principally to the dramatic increase in frequency of a previously very low frequency haplotype, reported at higher frequencies in northern European populations proximate to those used to stock Cantabrian rivers. Genetic structuring increased after stocking: among-river differentiation was low before stocking (1950s/1960s Φ _ST = –0.00296–0.00284), increasing considerably at the height of stocking (1980s Φ _ST = 0.18932) and decreasing post-stocking (1990s/2002 Φ _ST = 0.04934–0.03852). Gene flow from stocked fish therefore seems to have had a substantial role in increasing mtDNA variability. Additionally, we found significant differentiation between individuals that had probably died from infectious disease and apparently healthy, angled fish, suggesting a possible role for pathogen-driven selection of mtDNA variation. Our results suggest that stocking with non-native fish may increase genetic diversity in the short term, but may not reverse population declines.