Population genetics of the endangered Wattled Curassow (Crax globulosa,Cracidae, Aves) of the Colombian–Peruvian Amazon using DNA microsatellites and ND2 mitochondrial sequences

The Wattled Curassow (Crax globulosa, Cracidae, Aves) is a large bird living in the Western Amazon basin and a critically endangered species in the Colombian and in the Peruvian Amazon. We carried out the first population genetics analysis of this species employing six nuclear microsatellite markers and sequences of the mtND2 gene. The main results are as follows. (1) The levels of gene diversity were high for the overall population as well as for each of the three islands for both microsatellites and mtDNA. (2) A small amount of genetic differentiation among populations was found with both types of markers (F_ST = 0.027 for microsatellites and N_ST = 0.17 for mitochondrial sequences). (3) Using microsatellites, the Geneclass 2.0 software detected a low correct assignment of individuals to their respective populations. The Structure software only detected one gene pool for the entire area studied. These results are relevant for conservation efforts of this critically endangered species.     

Evaluation of DNA barcoding and identification of new haplomorphs in Canadian deerflies and horseflies

This paper reports the first tests of the suitability of the standardized mitochondrial cytochrome c oxidase subunit I (COI) barcoding system for the identification of Canadian deerflies and horseflies. Two additional mitochondrial molecular markers were used to determine whether unambiguous species recognition in tabanids can be achieved. Our 332 Canadian tabanid samples yielded 650 sequences from five genera and 42 species. Standard COI barcodes demonstrated a strong A + T bias (mean 68.1%), especially at third codon positions (mean 93.0%). Our preliminary test of this system showed that the standard COI barcode worked well for Canadian Tabanidae: the target DNA can be easily recovered from small amounts of insect tissue and aligned for all tabanid taxa. Each tabanid species possessed distinctive sets of COI haplotypes which discriminated well among species. Average conspecific Kimura two‐parameter (K2P) divergence (0.49%) was 12 times lower than the average divergence within species. Both the neighbour‐joining and the Bayesian methods produced trees with identical monophyletic species groups. Two species, Chrysops dawsoni Philip and Chrysops montanus Osten Sacken (Diptera: Tabanidae), showed relatively deep intraspecific sequence divergences (～10 times the average) for all three mitochondrial gene regions analysed. We suggest provisional differentiation of Ch. montanus into two haplotypes, namely, Ch. montanus haplomorph 1 and Ch. montanus haplomorph 2, both defined by their molecular sequences and by newly discovered differences in structural features near their ocelli.     

Phylogenetic relationships within the genus Jesogammarus (Crustacea, Amphipoda, Anisogammaridae) deduced from mitochondrial COI and 12S sequences

The genus Jesogammarus contains 16 species in two subgenera, Jesogammarus and Annanogammarus. To examine relationships among species in the genus, a molecular phylogenetic study including eight species of the former subgenus and four of the latter was conducted using partial DNA sequences of the mitochondrial COI and 12S rRNA genes. MP, NJ, and ML trees based on the combined COI and 12S data indicated monophyly of the subgenus Annanogammarus, though the monophyly of Jesogammarus was left unresolved. Consistent with few morphological differences, Jesogammarus (A.) naritai and J. (A.) suwaensis showed low genetic differentiation and did not show reciprocal monophyly, which suggests a close affinity of these taxa.     

Phylogeny of Betaphycus (Gigartinales,Rhodophyta) as inferred from COI sequences and morphological observations on B. philippinensis

The phylogeny of the three species that comprise the genus Betaphycus Doty in Silva, Basson and Moe and their phylogenetic relationships with other eucheumatoids are still unresolved. In this study, the utility of the mitochondrial cytochrome oxidase I (COI) marker in resolving their relationships was evaluated. Analyses of the COI sequences from Betaphycus philippinensis Doty and Betaphycus speciosus (Sonder) Doty ex Silva specimens collected from their type localities (Sorsogon, Philippines and Western Australia, respectively) revealed that the two species formed a well-supported clade distinct from Eucheuma J. Agardh and Kappaphycus Doty ex Silva. The genotyped specimens of B. philippinensis were observed to exhibit dorsal protuberances, a characteristic which has been regarded as a key diagnostic feature of Betaphycus gelatinus (Esper) Doty ex Silva. This observation raised the possibility that these two taxa are conspecific. In addition, B. philippinensis specimens were also observed to exhibit morphological features that could be used to distinguish the species from other eucheumatoids, such as the location tetrasporangial nemathecium in the thallus and the presence of apical or lateral pit connections in the tetraspores. The species referred to in the literature as “B. gelatinus” (as Eucheuma gelatinae) collected from northwestern Philippines was identified as a species of Eucheuma based on molecular and morphological evidence. The phylogenetic relationships of this species with other related eucheumatoid taxa were also determined.     

Microsatellite analysis supports the existence of three cryptic species within the bumble bee <Emphasis Type="Italic">Bombus lucorum sensu lato</Emphasis>

Mitochondrial cytochrome oxidase I (COI) partial sequences are widely used in taxonomy for species identification. Increasingly, these sequence identities are combined with modelling approaches to delineate species. Yet the validity of species delineation based on such DNA ‘barcodes’ is rarely tested and may be called into question by phenomena such as ancestral polymorphisms in DNA sequences, phylogeographic divergence, mitochondrial introgression and hybridization, or distortion of mitochondrial inheritance through such factors as Wolbachia infection. The common and widespread European bumble bee Bombus lucorum s. lato contains three distinct mitochondrial DNA lineages that are assumed to represent three cryptic species, namely Bombus cryptarum, B. lucorum s. str. and B. magnus. To test whether nuclear gene pools of the three putative species were differentiated, we genotyped 304 sympatric members of the lucorum complex (54 B. cryptarum females, 168 B. lucorum s. str. females and 82 B. magnus females, as defined using mtDNA COI haplotypes) from 11 localities spread across the island of Ireland at seven nuclear microsatellite loci. Multilocus genotypes clustered into three discrete groups that largely corresponded to the three mtDNA lineages: B. cryptarum, B. lucorum s. str. and B. magnus. The good fit of mitochondrial haplotype to nuclear (microsatellite) genotypic data supports the view that these three bumble bee taxa are reproductively isolated species, as well as providing a vindication of species identity using so-called DNA barcodes.     

Genetic diversity and phylogenetic relationship among the western and the Asian honey bees based on two mitochondrial gene segments (COI and ND5)

The Asian honey bee species i.e., Apis cerana (the eastern honey bee), A. dorsata (the giant honey bee), and the western or European honey bee (A. mellifera) collected from Pakistan were studied using partial sequences from two mitochondrial genes (i) the Cytochrome c oxidase I (COI) and (ii) the mitochondrially encoded NADH dehydrogenase 5 (ND5) and then compared with other honey bees sequences (already submitted from different countries around the globe) obtained after the national center for biotechnology information (NCBI). DNA sequences were analyzed employing molecular evolutionary genetics analysis and Kimura 2-parameter model, neighbor-joining method was applied to investigate phylogenetic relationships, and DNA sequence polymorphism was applied to measure the genetic diversity within the genus Apis. The phylogenetic analyses yielded consistent results. Based on COI gene fragment in two Asian and European honey bee species from Pakistan and from other countries showed considerable genetic diversity levels and deviation among the species. While in contrast the phylogenetic analyses based on ND5 gene fragment in Asian and European honey bee species from Pakistan and other countries showed comparatively higher genetic diversity indices and variations than the COI gene. So, in the genus Apis, the mitochondrial ND5 region has shown the possibility to answer the interactions among species. A further detailed work (by linking the analysis of other genomic and mitochondrial genes) is required for good quality solution to establish the concise genetic diversity and interaction among the Apis species. The objective of this study was to explore the extent of genetic differences and phylogenetic links among the three kinds of honey bee species from Pakistan and comparing them with other bee species around the globe.     

An integrated closed system for fish-plankton aquaculture in Amazonian fresh water

A prototype of an integrated closed system for fish-plankton aquaculture was developed in Iquitos (Peruvian Amazonia) in order to cultivate the Tiger Catfish, Pseudoplatystoma punctifer (Castelnau, 1855). This freshwater recirculating system consisted of two linked sewage tanks with an intensive rearing unit (a cage) for P. punctifer placed in the first, and with a fish-plankton trophic chain replacing the filters commonly used in clear water closed systems. Detritivorous and zooplanktivorous fishes (Loricariidae and Cichlidae), maintained without external feeding in the sewage volume, mineralized organic matter and permitted the stabilization of the phytoplankton biomass. Water exchange and organic waste discharge were not necessary. In this paper we describe the processes undertaken to equilibrate this ecosystem: first the elimination of an un-adapted spiny alga, Golenkinia sp., whose proliferation was favored by the presence of a small rotifer, Trichocerca sp., and second the control of this rotifer proliferation via the introduction of two cichlid species, Acaronia nassa Heckel, 1840 and Satanoperca jurupari Heckel, 1840, in the sewage part. This favored some development of the green algae Nannochloris sp. and Chlorella sp. At that time we took the opportunity to begin a 3-month rearing test of P. punctifer. The mean specific growth rate and feed conversion ratio (FCR) of P. punctifer were 1.43 and 1.27, respectively, and the global FCR, including fish in the sewage part, was 1.08. This system has proven to be suitable for growing P. punctifer juveniles out to adult, and provides several practical advantages compared with traditional recirculating clear water systems, which use a combination of mechanical and biological filters and require periodic waste removal, leading to water and organic matter losses.     

The red brocket deer (Mazama americana) as a new intermediate host of Taenia omissa (Taeniidae)

A total of 13 metacestodes were collected from the lung and parietal pleura from a red brocket deer (Mazama americana) from the Peruvian Amazon. All metacestodes were identified as cysticerci of Taenia omissa by morphological and molecular analyzes. Cytochrome c oxidase subunit 1 sequences from the new isolate T. omissa had more than 96.8% identity with other Peruvian isolates of the species previously sequenced. Lower similarities (93.8–95.8%) were verified between Peruvian and Canadian isolates. This finding adds a new intermediate host for T. omissa and also expands its geographical distribution.     

Multigene Phylogeography of Bactrocera caudata (Insecta: Tephritidae): Distinct Genetic Lineages in Northern and Southern Hemispheres

Bactrocera caudata is a pest of pumpkin flower. Specimens of B. caudata from the northern hemisphere (mainland Asia) and southern hemisphere (Indonesia) were analysed using the partial DNA sequences of the nuclear 28S rRNA and internal transcribed spacer region 2 (ITS-2) genes, and the mitochondrial cytochrome c oxidase subunit I (COI), cytochrome c oxidase subunit II (COII) and 16S rRNA genes. The COI, COII, 16S rDNA and concatenated COI+COII+16S and COI+COII+16S+28S+ITS-2 nucleotide sequences revealed that B. caudata from the northern hemisphere (Peninsular Malaysia, East Malaysia, Thailand) was distinctly different from the southern hemisphere (Indonesia: Java, Bali and Lombok), without common haplotype between them. Phylogenetic analysis revealed two distinct clades (northern and southern hemispheres), indicating distinct genetic lineage. The uncorrected ‘p’ distance for the concatenated COI+COII+16S nucleotide sequences between the taxa from the northern and southern hemispheres (‘p’ = 4.46-4.94%) was several folds higher than the ‘p’ distance for the taxa in the northern hemisphere (‘p’ = 0.00-0.77%) and the southern hemisphere (‘p’ = 0.00%). This distinct difference was also reflected by concatenated COI+COII+16S+28S+ITS-2 nucleotide sequences with an uncorrected ''p'' distance of 2.34-2.69% between the taxa of northern and southern hemispheres. In accordance with the type locality the Indonesian taxa belong to the nominal species. Thus the taxa from the northern hemisphere, if they were to constitute a cryptic species of the B. caudata species complex based on molecular data, need to be formally described as a new species. The Thailand and Malaysian B. caudata populations in the northern hemisphere showed distinct genetic structure and phylogeographic pattern.     

Cytogenetic and molecular characteristics of Potamotrygon motoro and Potamotrygon sp. (Chondrichthyes,Myliobatiformes, Potamotrygonidae) from the Amazon basin: Implications for the taxonomy of the genus

The chromosomes of two freshwater stingrays, Potamotrygon motoro and Potamotrygon sp., from the Amazon River basin in Brazil were investigated using integrated molecular (cytochrome c oxidase subunit 1) and cytogenetic analyses. Potamotrygon motoro presented intraspecific variation in the diploid number, with 2n=66 in the females and 2n=65 in the males, while Potamotrygon sp. had a karyotype with 66 chromosomes, in both sexes. The C-banding revealed the presence of heterochromatic blocks accumulated in the centromeric region of all the chromosomes in both species. The FISH assays with 18S DNA probes highlighted the terminal region of three or four chromosome pairs in P. motoro and seven chromosomes in Potamotrygon sp. The rDNA 5S sequences were found in only one chromosomal pair in both species. The interspecific genetic distance based on the COI sequences, between P. motoro and Potamotrygon sp. from Amazon River was 10.8%, while that between the Amazonian P. motoro and Potamotrygon amandae from the Paraná River was 2.2%, and the genetic distance between Potamotrygon sp. and P. amandae was 11.8%. In addition to the new insights on the cytogenetics of the study species, the results of the present study confirmed the existence of heteromorphic sex-linked chromosomes in P. motoro.     

New molecular data for tardigrade phylogeny, with the erection of Paramacrobiotus gen. nov.

Up to few years ago, the phylogenies of tardigrade taxa have been investigated using morphological data, but relationships within and between many taxa are still unresolved. Our aim has been to verify those relationships adding molecular analysis to morphological analysis, using nearly complete 18S ribosomal DNA gene sequences (five new) of 19 species, as well as cytochrome oxidase subunit 1 (COI) mitochondrial DNA gene sequences (15 new) from 20 species, from a total of seven families. The 18S rDNA tree was calculated by minimum evolution, maximum parsimony (MP) and maximum likelihood (ML) analyses. DNA sequences coding for COI were translated to amino acid sequences and a tree was also calculated by neighbour-joining, MP and ML analyses. For both trees (18S rDNA and COI) posterior probabilities were calculated by MrBayes. Prominent findings are as follows: the molecular data on Echiniscidae (Heterotardigrada) are in line with the phylogenetic relationships identifiable by morphological analysis. Among Eutardigrada, orders Apochela and Parachela are confirmed as sister groups. Ramazzottius (Hypsibiidae) results more related to Macrobiotidae than to the genera here considered of Hypsibiidae. Macrobiotidae and Macrobiotus result not monophyletic and confirm morphological data on the presence of at least two large groups within Macrobiotus. Using 18S rDNA and COI mtDNA genes, a new phylogenetic line has been identified within Macrobiotus , corresponding to the ' richtersi-areolatus group'. Moreover, cryptic species have been identified within the Macrobiotus ' richtersi group' and within Richtersius . Some evolutionary lines of tardigrades are confirmed, but others suggest taxonomic revision. In particular, the new genus Paramacrobiotus gen. n. has been identified, corresponding to the phylogenetic line represented by the ' richtersi-areolatus group'.     

Phylogeography and Sex-Biased Dispersal across Riverine Manatee Populations (Trichechus inunguis and Trichechus manatus) in South America

Phylogeographic patterns and sex-biased dispersal were studied in riverine populations of West Indian (Trichechus manatus) and Amazonian manatees (T. inunguis) in South America, using 410bp D-loop (Control Region, Mitochondrial DNA) sequences and 15 nuclear microsatellite loci. This multi-locus approach was key to disentangle complex patterns of gene flow among populations. D-loop analyses revealed population structuring among all Colombian rivers for T. manatus, while microsatellite data suggested no structure. Two main populations of T. inunguis separating the Colombian and Peruvian Amazon were supported by analysis of the D-loop and microsatellite data. Overall, we provide molecular evidence for differences in dispersal patterns between sexes, demonstrating male-biased gene flow dispersal in riverine manatees. These results are in contrast with previously reported levels of population structure shown by microsatellite data in marine manatee populations, revealing low habitat restrictions to gene flow in riverine habitats, and more significant dispersal limitations for males in marine environments.     

Genetic structure and diversity of the Diachasmimorpha longicaudata species complex in Thailand: SSCP analysis of mitochondrial 16S rDNA and COI DNA sequences

Genetic variation among 20 populations of Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae) in Thailand was investigated using single strand conformation polymorphism (SSCP) analysis of mitochondrial DNA sequences. From a total 641 individual parasitoids, seven distinct haplotypes containing a total of 32 polymorphic sites were observed from cytochrome oxidase subunit I (COI) sequences along with five distinct haplotypes containing a total 16 polymorphic sites from 16S rDNA sequences. Values obtained through pairwise F_ST comparisons and analysis of molecular variance (AMOVA) indicated significant genetic differentiation among D. longicaudata populations in Thailand. Congruent relationships showing separation of these populations into three groups were obtained from Neighbor joining and Bayesian phylogenetic tree analyses along with the use of haplotype networks. This SSCP analysis of populations of the D. longicaudata species complex is the first report using molecular population genetic methods to analyze the structure of this parasitoid species in Thailand. This may provide useful information for release of parasitoid strains to maximize their benefit in biological control programs.     

Integrative taxonomy of Pseudolepeophtheirus longicauda (Crustacea: Copepoda: Caligidae) parasitic on Platichthys stellatus (Actinopterygii: Pleuronectidae)

Based on specimens from the gill cavities of one Platichthys stellatus individual collected in the Sea of Japan, we investigated the taxonomic status of the enigmatic caligid genus Pseudolepeophtheirus and its type species, Pseudolepeophtheirus longicauda. In a maximum likelihood (ML) tree based on 18S rRNA gene sequences, the sequence from our sample was nested in a well-supported Lepeophtheirus clade, along with the type species, confirming that Pseudolepeophtheirus should be considered a junior synonym of Lepeophtheirus; our morphological data support this synonymy. Although a previous study had synonymized Pseudolepeophtheirus longicauda with Lepeophtheirus parvicruris, we found that the former differs morphologically from the latter in having a short leg-4 exopod, with the articulation between the first and second segments not evident (the shape of the posterior striated membrane on the leg-2 intercoxal sclerite also differs between two species), and detected slight differences in 18S rRNA sequences between two taxa. We thus concluded that this synonymy is invalid, and reinstate Lepeophtheirus longicauda as a valid species. A ML analysis of COI sequences from Pl. stellatus (the host fish for both L. longicauda and L. parvicruris) showed the host species to comprise distinct northwestern- and northeastern-Pacific clades. Lepeophtheirus longicauda is distributed in the northwestern Pacific and L. parvicruris in the northeastern Pacific, indicating co-divergence of the two copepod species with the host lineages.     

From species to individuals: combining barcoding and microsatellite analyses from non-invasive samples in plant ecology studies

By means of DNA barcoding and microsatellite analyses, we studied the species and individuals of legitimate seed dispersers of the Mediterranean shrub Pistacia lentiscus, a keystone species that represents the main source of food in winter for frugivorous birds. We collected dropping of birds containing seeds, and after DNA extraction we amplified and sequenced a fragment of the mitochondrial COI gene. Through BLASTN queries of the sequenced fragments against registered sequences in the GenBank database we identified the bird species that are currently dispersing P. lentiscus seeds. Further, through the amplification of specific nuclear microsatellite loci we calculated standard genetic diversity parameters of the population of birds from the genus Sylvia (the blackcap and Sardinian warbler), the most important dispersers of P. lentiscus. Five bird species were identified as seed dispersers through their barcode match. Further, we found that S. melanocephala displayed slightly lower levels of genetic diversity than S. atricapilla. In this study we show how the genetic analyses of environmental faecal samples can be a useful and convenient tool for the study of plant-frugivore interactions through the ascertainment of the identity of the species involved and through the analyses of genetic variability of their populations.     

The Praon dorsale–yomenae s.str. complex (Hymenoptera,Braconidae, Aphidiinae): Species discrimination using geometric morphometrics and molecular markers with description of a new species

In this paper we apply an integrative approach combining morphometric and molecular analyses to explore parasitoids of the Praon dorsale–yomenae s.str. complex. These parasitoids occur in natural and agricultural ecosystems throughout the Palaearctic and parasitize aphid hosts belonging to the tribe Macrosiphini. The P. dorsale–yomenae species complex represents a morphologically cryptic group, consisting of several hidden taxa with specific host adaptations and distributions. For the morphometric analyses we used a large dataset comprising 230 female specimens that emerged from 30 different species of aphid hosts throughout the Palaearctic. The molecular analysis included a reduced dataset of 44 specimens that emerged from 19 aphid hosts. The mitochondrial cytochrome oxidase subunit I (COI) and the nuclear second expansion segment of the 28S rRNA gene (28SD2) were used to estimate a genetic diversity within this complex. Although all Praon species clustered closely together in the 28SD2 tree, confirming their close relatedness, the molecular characterization based on COI identified five clearly separate taxa with sequence divergences in the range of 4.7–8.9%. These taxa also exhibited significant differences in forewing shape as revealed by geometric morphometric analyses. Classical morphometric analyses revealed the length of m-cu vein as a new taxonomic character, but suggested that one commonly used trait, the color pattern of the Rs + M and m-cu veins, cannot be used for species distinction as it was highly variable in one of the taxa. Based on the combined morphometric and genetic data, we confirm the species status of Praon dorsale, P. yomenae, P. longicorne and P. volucre, and describe and illustrate a new parasitoid species of the “dorsale–yomenae” group, i.e. Praon staticobii n.sp. associated with Staticobium limonii on Limonium angustifolium.     

Desmophyllum dianthus (Esper, 1794) in the Scleractinian Phylogeny and Its Intraspecific Diversity

The cosmopolitan solitary deep-water scleractinian coral Desmophyllum dianthus (Esper, 1794) was selected as a representative model species of the polyphyletic Caryophylliidae family to (1) examine phylogenetic relationships with respect to the principal Scleractinia taxa, (2) check population structure, (3) test the widespread connectivity hypothesis and (4) assess the utility of different nuclear and mitochondrial markers currently in use. To carry out these goals, DNA sequence data from nuclear (ITS and 28S) and mitochondrial (16S and COI) markers were analyzed for several coral species and for Mediterranean populations of D. dianthus. Three phylogenetic methodologies (ML, MP and BI), based on data from the four molecular markers, all supported D. dianthus as clearly belonging to the “robust” clade, in which the species Lophelia pertusa and D. dianthus not only grouped together, but also shared haplotypes for some DNA markers. Molecular results also showed shared haplotypes among D. dianthus populations distributed in regions separated by several thousands of kilometers and by clear geographic barriers. These results could reflect limited molecular and morphological taxonomic resolution rather than real widespread connectivity. Additional studies are needed in order to find molecular markers and morphological features able to disentangle the complex phylogenetic relationship in the Order Scleractinia and to differentiate isolated populations, thus avoiding the homoplasy found in some morphological characters that are still considered in the literature.     

DNA barcodes and phylogenetic affinities of the terrestrial slugs Arion gilvus and A. ponsi (Gastropoda,?Pulmonata,Arionidae)

The Iberian Peninsula is a region with a high endemicity of species of the terrestrial slug subgenus Mesarion. Many of these species have been described mainly on subtle differences in their proximal genitalia. It therefore remains to be investigated 1) whether these locally diverged taxa also represent different species under a phylogenetic species concept as has been shown for other Mesarion species outside the Iberian Peninsula, and 2) how these taxa are phylogenetically related. Here, we analysed DNA sequence data of two mitochondrial (COI and 16S) genes, and of the nuclear ITS1 region, to explore the phylogenetic affinities of two of these endemic taxa, viz. Arion gilvus Torres Mínguez, 1925 and A. ponsi Quintana Cardona, 2007. We also evaluated the use of these DNA sequence data as DNA barcodes for both species. Our results showed that ITS did not allow to differentiate among most of the Mesarion molecular operational taxonomic units (MOTUs) / morphospecies in Mesarion. Yet, the overall mean p-distance among the Mesarion MOTUs / morphospecies for both mtDNA fragments (16.7% for COI, 13% for 16S) was comparable to that between A. ponsi and its closest relative A. molinae (COI: 14.2%; 16S: 16.2%) and to that between A. gilvus and its closest relative A. urbiae (COI: 14.4%; 16S: 13.4%). Hence, with respect to mtDNA divergence, both A. ponsi and A. gilvus, behave as other Mesarion species or putative species-level MOTUs and thus are confirmed as distinct ‘species’.     

Evaluation of the utility of COI and ITS markers as tools for population genetic studies of temperate gorgonians

The aim of this study was to analyse the polymorphism of two molecular markers, mitochondrial COI and nuclear ITS 2, to determine the genetic structure of Mediterranean gorgonian populations. Four gorgonians, Eunicella cavolinii, Eunicella singularis, Paramuricea clavata and Corallium rubrum were analysed for COI, while only E. cavolinii was analysed for ITS. These species have been recently affected by mass mortality events probably linked to climate change. In the context of these strong disturbances, reliable data on the genetic structure of affected populations could be critical in order to propose management and conservation strategies for these species. A hierarchical sampling plan encompassing different spatial scales (from tenths to hundreds of kilometres) was designed to test these markers. Both markers failed to make evident any genetic structure of these populations. The evolutionary features and utility of these markers are critically discussed and future research directions are proposed.     

Variations of chromosomal structures in Caluromys philander (Didelphimorphia: Didelphidae) from the Amazon region

Caluromys is considered to be one of the most ancient genera of extant marsupials and is positioned among the basal taxa of the family Didelphidae. At least two species occur in Brazil, C. philander and C. lanatus, both of which have 2n = 14 chromosomes. For the first time, we present evidence of an intrapopulation polymorphism of the sexual chromosome pair in C. philander females from the Central Amazon region. Detailed cytogenetic results of animals from three localities on the Amazon region were analyzed using classical cytogenetics (NOR, C-Band and G-Band) and molecular techniques (18S rDNA and telomere probes). Similar to other conspecific individuals, the diploid number of these animals is 2n = 14, and their fundamental number is 24, with NOR present on the 6th autosomal pair. The X chromosome presented variation detectable by G banding, suggesting a pericentric inversion.