Trypanosoma janseni n. sp. (Trypanosomatida: Trypanosomatidae) isolated from Didelphis aurita (Mammalia: Didelphidae) in the Atlantic Rainforest of Rio de Janeiro, Brazil: integrative taxonomy and phylogeography within the Trypanosoma cruzi clade

BACKGROUND Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. OBJECTIVES We characterise a new species (Trypanosoma janseni n. sp.) isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. METHODS The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp) of the small subunit (18S) ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. FINDINGS In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. CONCLUSIONS This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently know about the T. cruzi clade.

INFECTION BY Rickettsia felis IN OPOSSUMS (Didelphis sp.) FROM YUCATAN, MEXICO.

Rickettsia felis is an emergent pathogen and the causative agent of a typhus-like rickettsiosis in the Americas. Its transmission cycle involves fleas as biological vectors (mainly Ctenocephalides felis) and multiple domestic and synanthropic mammal hosts. Nonetheless, the role of mammals in the cycle of R. felis is not well understood and many efforts are ongoing in different countries of America to clarify it. The present study describes for the first time in Mexico the infection of two species of opossum (Didelphis virginiana and D. marsupialis) by R. felis. A diagnosis was carried out from blood samples by molecular methods through the gltA and 17 kDa genes and sequence determination. Eighty-seven opossum samples were analyzed and 28 were found to be infected (32.1%) from five out of the six studied localities of Yucatan. These findings enable recognition of the potential epidemiological implications for public health of the presence of infected synanthropic Didelphis in households.

Common opossum (Didelphis marsupialis Linnaeus, 1758): food and medicine for people in the Amazon.

BACKGROUND: In the Amazon rainforest, biodiversity is a significant resource for traditional communities, as it can be used as a relevant source of protein and it has a promising zootherapeutic potential. Studies on knowledge and ways how local peoples use the fauna are still incipient. This paper presents both the knowledge on and food and medicinal uses of common opossum (Didelphis marsupialis) by riverine communities in an Amazon floodplain region. METHODS: The study was conducted with riverine communities in the municipality of Abaetetuba, Pará, Brazil. The main methods used were structured and semi-structured interviews, the "snowball" technique, and participant observation. RESULTS: The study showed that D. marsupialis has an undeniable cultural significance for the local community, both in terms of food and medicine. Its meat is prized by inhabitants as it is classified as tasty, soft and, in some cases, it is designated as the best bushmeat in the region. The interviewees have demonstrated a thorough knowledge on various aspects of the animal's biology, such as its diet, behavior, and places of occurrence. The hunting activity is practiced by men, but the preparation of meat and medicinal oil are tasks mainly performed by women. In medical terms, common opossum is used in the treatment of various diseases, such as rheumatism, asthma, sore throat, and inflammation. Given the importance of this species, its meat or live individuals are often sold in the city fair at prices that can reach R$ 40.00 (U$D 18,00) per individual. CONCLUSIONS: D. marsupialis is an important source of protein for riverine communities in the region studied. Its fat is used as a traditional medicine and it is indicated for many types of diseases. Although the species concerned is treated with hostility in various Brazilian regions, in the case of Abaetetuba this animal is strongly prized due to the good quality of its meat. However, despite the value assigned to the species, its consumption should be the subject of further studies, as this marsupial species has been described as a reservoir for parasites that cause severe diseases.

Molecular characterization of an opossum Didelphis albiventris (Marsupialia: Didelphidae) population in an urban fragment of the Brazilian Atlantic rainforest and support to species barcode identification.

We made a molecular study of 40 opossums, Didelphis albiventris, from an urban fragment of the Atlantic Rainforest in southeastern Brazil, analyzing a 653-bp sequence of cytochrome c oxidase, subunit I. We found three close connected haplotypes, with low nucleotide diversity and a haplotype diversity of 59.1% and confirmed sympatry between D. albiventris and D. aurita in this region. The clear phylogenetic separation shows the appropriateness of DNA barcode identification methodology for effectively discriminating between these opossum species.

DNA barcodes effectively identify the morphologically similar Common Opossum (Didelphis marsupialis) and Virginia Opossum (Didelphis virginiana) from areas of sympatry in Mexico.

Two morphologically similar species of opossum from the genus Didelphis-Didelphis virginiana and Didelphis marsupialis-cooccur sympatrically in Mexico. High intraspecific variation complicates their morphological discrimination, under both field and museum conditions. This study aims to evaluate the utility and reliability of using DNA barcodes (short standardized genome fragments used for DNA-based identification) to distinguish these two species. Sequences of the cytochrome c oxidase subunit I (Cox1) mitochondrial gene were obtained from 12 D. marsupialis and 29 D. virginiana individuals and were compared using the neighbor-joining (NJ) algorithm with Kimura's two-parameter (K2P) model of nucleotide substitution. Average K2P distances were 1.56% within D. virginiana and 1.65% in D. marsupialis. Interspecific distances between D. virginiana and D. marsupialis varied from 7.8 to 9.3% and their barcode sequences formed distinct non-overlapping clusters on NJ trees. All sympatric specimens of both species were effectively discriminated, confirming the utility of Cox1 barcoding as a tool for taxonomic identification of these morphologically similar taxa.

Body size of commom opossum Didelphis aurita Wied-Neuwied 1826 (Didelphimorphia: Didelphidae) on southern brazilian islands.

The body size of vertebrates isolated on islands can undergo changes due to ecological features of these environments. This study aimed to compare the body size of the common opossum, Didelphis aurita, from different insular populations within the same archipelago in southern Brazil. The opossum populations showed corporal variation and different hypotheses were raised to understand the results. This study constitutes the most detailed body size comparison of a marsupial within different insular populations in the Neotropical zone and the data gathered represents an initial contribution for regional fauna biometric knowledge.

Body size of commom opossum Didelphis aurita Wied-Neuwied 1826 (Didelphimorphia: Didelphidae) on southern Brazilian islands / Tamanho corpóreo de gambás Didelphis aurita Wied-Neuwied 1826 (Didelphimorphia: Didelphidae) em ilhas no sul do Brasil

The body size of vertebrates isolated on islands can undergo changes due to ecological features of these environments. This study aimed to compare the body size of the common opossum, Didelphis aurita, from different insular populations within the same archipelago in southern Brazil. The opossum populations showed corporal variation and different hypotheses were raised to understand the results. This study constitutes the most detailed body size comparison of a marsupial within different insular populations in the Neotropical zone and the data gathered represents an initial contribution for regional fauna biometric knowledge.

O tamanho corpóreo de vertebrados isolados em ilhas pode sofrer alterações em função das particularidades ecológicas desses ambientes. O presente trabalho comparou o tamanho corpóreo de gambás, Didelphis aurita, em ilhas costeiras de um mesmo arquipélago no Sul do Brasil e encontrou diferenças de tamanho entre as localidades. Os gambás apresentaram variações para as medidas corporais e diferentes hipóteses foram consideradas para entender os resultados. Esta é a comparação mais detalhada de tamanho corpóreo entre populações insulares de um marsupial neotropical e uma contribuição inicial para o conhecimento da biometria da fauna regional.

Different patterns of selection on the nuclear genes IRBP and DMP-1 affect the efficiency but not the outcome of phylogeny estimation for didelphid marsupials.

Selection at the protein-level can influence nucleotide substitution patterns for protein-coding genes, which in turn can affect their performance as phylogenetic characters. In this study, we compare two protein-coding nuclear genes that appear to have evolved under markedly different selective constraints and evaluate how selection has shaped their phylogenetic signal. We sequenced 1,100+ bp of exon 6 of the gene encoding dentin matrix protein 1 (DMP1) from most of the currently recognized genera of New World opossums (family: Didelphidae) and compared these data to an existing matrix of sequences from the interphotoreceptor retinoid-binding protein gene (IRBP) and morphological characters. In comparison to IRBP, DMP1 has far fewer sites under strong purifying selection and exhibits a number of sites under positive directional selection. Furthermore, selection on the DMP1 protein appears to conserve short, acidic, serine-rich domains rather than primary amino acid sequence; as a result, DMP1 has significantly different nucleotide substitution patterns from IRBP. Using Bayesian methods, we determined that DMP1 evolves almost 30% faster than IRBP, has 2.5 times more variable sites, has less among-site rate heterogeneity, is skewed toward A and away from CT (IRBP has relatively even base frequencies), and has a significantly lower rate of change between adenine and any other nucleotide. Despite these different nucleotide substitution patterns, estimates of didelphid relationships based on separate phylogenetic analyses of these genes are remarkably congruent whether patterns of nucleotide substitution are explicitly modeled or not. Nonetheless, DMP1 contains more phylogenetically informative characters per unit sequence and resolves more nodes with higher support than does IRBP. Thus, for these two genes, relaxed functional constraints and positive selection appear to improve the efficiency of phylogenetic estimation without compromising its accuracy.