Continuous Growth and Differentiation of Trypanosoma (Megatrypanum) freitasi Rego, Magalhaes & Siqueira, 1957, In Vitro

Trypanosoma (Megatrypanum) freitasi, a parasite of didelphid opossum, was known to be very difficult to cultivate in conventional media. Co-cultivation with L929 cell line in Baltz's medium at 27.5° C resulted in luxuriant growth of the trypanosome with the production of epimastigote colonies that adhered to the surface of culture flasks or tubes, and transformation into metacylics. Further transformation was stimulated by raising the incubation temperature. At 37° C the population was of the bloodstream type and resistant to lysis by complement.     

Mast cell types in the lymph nodes of the opossum Didelphis albiventris (Marsupialia,Didelphidae)

Summary Using histological and histochemical techniques, we have found a unique population of mast cells in the lymphatic sinuses of lymph nodes from different anatomical regions of the opossum. The lymphatic-sinus mast cells of the medullary sinuses were numerous, and could be easily distinguished from the connective-tissue mast cells of the dermis and lymph node capsule by their larger size and their enlarged cytoplasmic granules that were also more heterogeneous in shape and staining properties.     

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.     

Histoplasma capsulatum isolated from Didelphis albiventris (Marsupialia: Didelphidae) in the State of Minas Gerais, Brazil

We report the isolation of Histoplasma capsulatum from a culture of the viscera of Didelphis albiventris, one of the marsupial species found in Brazil. To our knowledge, this is the first report of the isolation of this fungus from this mammalian species. This finding confirms the ubiquitous presence of H. capsulatum in nature.     

Mitochondrial DNA-like sequences in the nuclear genome of the opossum genus Didelphis (Marsupialia: Didelphidae).

This study reports the occurrence of mtDNA-like sequences in the nuclear genome of the opossum genus Didelphis (Didelphidae, Marsupialia). A specific primer pair designed to amplify a region encompassing a 3' terminal 118 bp region of the cytochrome b gene, the Thr and Pro tRNA genes, and a 489 bp region of the D-loop of the D. virginiana mtDNA, was used in highly stringent PCR reactions. These PCR reactions resulted in several fragments per individual varying in size from 259 bp to 1 kb. The sequencing of some of these fragments showed the occurrence of paralogous mtDNA-like sequences among the PCR amplified fragments. Analyses of qualitative aspects of these sequences, their transition/transversion ratios, and phylogenetic relationships were conclusive in showing the occurrence of mtDNA-like sequences in the nuclear genome of the genus Didelphis. Comparisons and phylogenetic analysis of orthologous mtDNA from the four Didelphis species and paralogous nuclear sequences suggested that mtDNA migration to the nuclear genome occurred more than once in Didelphis evolution.     

Trypanosoma (Herpetosoma) leeuwenhoeki in Choloepus hoffmanni and Didelphis marsupialis of the Pacific Coast of Colombia

Trypanosoma (Herpetosoma) leeuwenhoeki, originally described in Panamanian sloths, was isolated from Didelphis marsupialis (Marsupialia) and Choloepus hoffmanni (Edentata) inhabiting the Pacific coast of Colombia. Trypanosomes were characterized by their large blood forms (total length 51-53 microns), poor infectivity for mice, and lack of development in Rhodnius prolixus. Isoenzyme studies, with either strains or clones, revealed homogeneous profiles clearly distinct from Trypanosoma cruzi and Trypanosoma rangeli reference strains. The present report extends the geographical distribution of T. leeuwenhoeki to South America and broadens its known host range to another order of mammals.     

Trypanosoma (Megatrypanum) megachiropterorum sp. n. from the Flying Fox, Pteropus tonganus Quoy & Gaimard

SYNOPSIS. Trypanosoma (Megatrypanum) megachiropterorum sp. n. from the flying fox, Pteropus tonganus Quoy & Gaimard, from the Kingdom of Tonga, is described. The trypomastigotes measure 32 ± 1.74 (here and below S.D.) (27–37.5) × 3.6 ± 0.23 (3–4.5) μm and has a flagellum averaging in length 6.4 ± 0.92 (4.5–10) μm. Their nuclear index is 1.2 ± 0.16 (0.7–1.6) and their kinetoplastic index equals 6.2 ± 0.32 (5.5–7.1). This is the first report of a member of the subgenus Megatrypanum from bats outside the American and African continents and also the first record of a trypanosome from bats in the Pacific area.     

Mitochondrial genetic variability of Didelphis albiventris (Didelphimorphia, Didelphidae) in Brazilian localities

Didelphis albiventris is a well-known and common marsupial. Due to its high adaptability, this very widespread generalist species occurs under various environmental conditions, this even including protected regions and disturbed urban areas. We studied a 653 bp fragment of cytochrome oxidase c (COI) from 93 biological samples from seven Brazilian localities, with linear distances ranging between 58 and about 1800 km to analyze the effects of geographic distances on variability and genetic differentiation. The haplotype network presented nine haplotypes and two genetic clusters compatible with the two most distant geographic areas of the states of Minas Gerais, in the southeast, and Rio Grande do Sul, in the extreme south. As each cluster was characterized by low nucleotide and high haplotype diversities, their populations were obviously composed of closely related haplotypes. Surprisingly, moderate to high F(ST) differentiation values and a very weak phylogeographic signal characterizes interpopulation comparisons within Minas Gerais interdemes, these being correlated with the presence of privative haplotypes. On a large rgeographic scale, a comparison between demes from Minas Gerais and Rio Grande do Sul presented high F(ST) values and a robust phylogeographic pattern. This unexpected scenario implies that mtDNA gene flow was insufficient to maintain population cohesion, reflected by the observed high differentiation.     

DNA Sequence Evidence for Placement of the Ground Cuscus, Phalanger gymnotis, in the Tribe Phalangerini (Marsupialia: Phalangeridae)

A region of mitochondrial DNA, including the 3 end of tRNA phenylalanine, the complete 12S rRNA and tRNA valine genes, and the 5 end of 16S rRNA, was sequenced for four phalangerids and one burramyid; additional marsupial sequences were extracted from GenBank. Parsimony, minimum evolution, and maximum likelihood analyses show that the ground cuscus, Phalanger gymnotis, groups with the tribe Phalangerini, not with the tribe Trichosurini as had been suggested on the basis of certain morphological characters. This result is in agreement with an earlier study using DNA hybridization and is supported by some morphological evidence as well. We conclude that the character states that link the ground cuscus with the Trichosurini are the result of convergence, and therefore the placement of several other species in the trichosurin genus Strigocuscus based on the same characters should be reconsidered. The hypothesized close relationship of two fossil taxa, Strigocuscus reidi and S. notialis, to Phalanger gymnotis is also questionable because the fossils do not share morphological synapomorphies that link the ground cuscus to the Phalangerini.     

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.     

Puberty in the male owl monkey (Aotus trivirgatus griseimembra): A study of physical and hormonal development

In the male owl monkey (Aotus trivirgatus),the onset of puberty occurs at between 211 and 337 days (median, 313) as indicated by longitudinal measurements of plasma testosterone in six subjects. Larger rises in plasma testosterone were observed between 300 and 400 days. Pubertal increases in body weight and testicular volume are not pronounced in this species. Young animals first entered the weight range for adult males (800–1080 g) when they were between 370 and 520 days of age (median, 472). Marked growth of the subcaudal scentmarking gland occurred during puberty. The first signs of development of this gland (stiffening and discoloration of overlying hairs) were noted between 282 and 370 days (median 316), and it had attained an adult structure by 336–442 days (median, 397). These changes were androgen dependent as indicated by the fact that treatment of a prepubertal male with testosterone stimulated the subcaudal gland to develop prematurely. Hormonal and physical changes during puberty were the same whether males remained with their parents or were caged alone. There was no indication that puberty was retarded in males which had remained in their natal groups.     

Parasitological aspects of Schistosoma haematobium (Iran) infection in the American opossum (Didelphis marsupialis L.)

Parasitological studies on 25 American opossums (Didelphis marsupialis) exposed to 1000 cercariae each of S. haematobium (Iran) have indicated a host-parasite situation and general parasitological conditions which show this marsupial to be a satisfactory host for experimental schistosomiasis. The return of schistosomes in relation to cercarial exposure was not excessively high, but there were egg deposits in the major visceral organs. Varied macroscopic pathology due to egg deposits in different organs was demonstrated as well as a potential for urinary bladder involvement, including tumor formation. Even though the precise nature of tumor pathology is not yet known, availability of the host, a good host-parasite compatability, and a low death rate in infected hosts favor this mammal as a model for experimental schistosomiasis haematobia.     

Morphology and chemical composition of metathoracic scent glands in Dolycoris baccarum (Linnaeus, 1758) (Heteroptera: Pentatomidae)

One of the general defining characters of the Heteroptera is the presence of metathoracic scent glands (MTG). Using scanning electron microscopy, the morphology of the MTG of Dolycoris baccarum (Linnaeus 1758) (Heteroptera: Pentatomidae) was studied. The MTG belong to the diastomian type. The two glandular pores located between the mesothoracic and metathoracic coxae are associated with 'mushroom-like' structures. The MTG are composed of a reservoir and a pair of lateral glands is connected to the reservoir by a duct. A groove-like structure extends downwards from the ostiole. While this structure is long and wide, its ostiole is oval. Extracts of the volatile fractions from male and female MTG secretions were analysed by capillary gas chromatography–mass spectrometry (GC-MS) and exhibited a typical pentatomid composition. Seventeen chemical compounds were detected in female secretions, whereas 13 chemical compounds were detected in the male secretions. Most chemical compounds were similar between the sexes but were different in their quantities. In this regard, the compounds identified were investigated and the biological functions of the glandular secretions were discussed. In the analyses of the MTG of females of D. baccarum , tridecane (50.97%) was a major odour component and (Z,Z)-4,16-octadecadien-1-ol acetate (0.02%) was a minor odour component. In males, tridecane (50.80%) was a major odour component and 1,2-benzenedicarboxylic acid (0.02%) was a minor odour component.     

Footfall patterns and interlimb co-ordination in opossums (Family Didelphidae): evidence for the evolution of diagonal-sequence walking gaits in primates

Most primates typically use a diagonal-sequence footfall pattern during walking. This footfall pattern, which is unusual for mammals, is believed to have originated in ancestral primates in association with the use of grasping extremities for movement and foraging on thin, flexible branches. This theory was tested by comparing gait parameters between the grey short-tailed opossum Monodelphis domestica and the woolly opossum Caluromys philander , two didelphid marsupials that are strongly differentiated in grasping morphology of the extremities and in their reliance on foraging strategies involving thin branches. One hundred and thirty gait cycles were analysed quantitatively from videotapes of subjects moving quadrupedally on a runway and on poles of different diameters (7 and 28 mm). Duty factor (i.e. duration of the stance phase as a percentage of the stride period) for the forelimb and hindlimb, as well as diagonality (i.e. phase relationship between the forelimb and hindlimb cycles), were calculated for each of these symmetrical gait cycles. We found that the highly terrestrial Monodelphis , like most other non-primate mammals, relies primarily on lateral-sequence walking gaits on both runway and poles, and has relatively higher forelimb duty factors. Like primates, the highly arboreal Caluromys uses primarily diagonal-sequence walking gaits on the runway and pole, with relatively higher hindlimb duty factors and diagonality. The fact that the woolly opossum, a marsupial with primate-like feet that moves and forages mainly on thin branches, uses primarily diagonal-sequence gaits when walking supports the view that primate gaits evolved to meet the demands of locomotion on narrow supports. This also demonstrates the functional role of a grasping foot, in association with relatively higher hindlimb duty factors, protraction, and substrate reaction forces, in the production of such walking gaits.