Cerradopsora, a new genus of neotropical rust fungi.

Two Cerrado rust fungi, Phakopsora rossmaniae and Aplopsora hennenii, described in 1993 and 1995 and originally assigned to families Phakopsoraceae and Ochropsoraceae, respectively, were subjected to molecular phylogenetic analyses using fragments of the nuc 28S and 18S rDNA and mitochondrial cytochrome c oxidase subunit 3 (CO3) gene. Although both taxa were morphologically well placed in their original genera, they were shown to belong in a strongly supported new lineage within the Raveneliineae distant from the Phakopsoraceae and Ochropsoraceae. Therefore, we properly treated this lineage as the new genus Cerradopsora now harboring C. rossmaniae (type species) and C. hennenii. However, this novel phakopsoroid genus remains in uncertain familial position without support to be included in any of the families that share space within the Raveneliineae.

Raveneliopsis, a new genus of ravenelioid rust fungi on Cenostigma (Caesalpinioideae) from the Brazilian Cerrado and Caatinga.

The multicellular discoid convex teliospore heads represent a prominent generic feature of the genus Ravenelia. However, recent molecular phylogenetic work has shown that this is a convergent trait, and that this genus does not represent a natural group. In 2000, a rust fungus infecting the Caesalpinioid species Cenostigma macrophyllum (= C. gardnerianum) was described as Ravenelia cenostigmatis. This species shows some rare features, such as an extra layer of sterile cells between the cysts and the fertile teliospores, spirally ornamented urediniospores, as well as strongly incurved paraphyses giving the telia and uredinia a basket-like appearance. Using freshly collected specimens of Rav. cenostigmatis and Rav. spiralis on C. macrophyllum, our phylogenetic analyses based on the nuc 28S, nuc 18S, and mt CO3 (cytochrome c oxidase subunit 3) gene sequences demonstrated that these two rust fungi belong in a lineage within the Raveneliineae that is distinct from Ravenelia s. str. Besides proposing their recombination into the new genus Raveneliopsis (type species R. cenostigmatis) and briefly discussing their potentially close phylogenetic affiliations, we suggest that five other Ravenelia species that are morphologically and ecologically close to the type species of Raveneliopsis, i.e., Rav. corbula, Rav. corbuloides, Rav. parahybana, Rav. pileolarioides, and Rav. Striatiformis, may be recombined pending new collections and confirmation through molecular phylogenetic analyses.

Freezing for the Future: Obtaining Fibroblast Samples from Deceased Wild Mammals for the Brazilian Cerrado Germplasm Bank.

We isolated and further characterized fibroblasts obtained from postmortem skin biopsies of three different Brazilian wild species (Chrysocyon brachyurus-maned wolf, Cerdocyon thous-crab-eating fox, Mazama gouazoubira-brown brocket deer). The effects of two cryoprotectants, 10% dimethyl sulfoxide (DMSO) and 5% dimethylformamide (DMF), were assessed to determine the most efficient cryopreservation protocol. Such an investigation promotes the creation of germplasm banks, using samples that would otherwise be rejected and permanently lost following the death of the animals. We utilized animal corpses that were involved in highway accidents, found dead in the natural environment, or referred to us from the veterinary hospital at the Brasília Zoo. Fibroblasts from C. brachyurus specimens presented a delay in cell growth in Dulbecco's modified Eagle's medium in relation to other species. This observation is a limiting factor for the future storage of cells from this species. Differences in cellular morphology were observed between C. brachyurus, C. thous, and M. gouazoubira, presenting branched, fusiform, and spherical forms, respectively. The cryoprotective solution containing 10% DMSO was more efficient than 5% DMF medium in preserving the viability of fibroblasts of the three species (p < 0.05). After defining the best cryopreservation solution, a germplasm bank was successfully formed. This biological reservoir is configured as the first germplasm bank containing somatic cells and gametes of wild mammals of the Cerrado biome of Brazil. This material will be used for future characterization of the species and multiplication by means of nuclear transfer cloning.

Cellular Characterization and Effects of Cryoprotectant Solutions on the Viability of Fibroblasts from Three Brazilian Wild Cats.

Preserving genetic material in cryogenic conditions presents a viable alternative for the protection of species' gene variability. However, there is an enormous need to establish and test cryopreservation protocols that are suitable for each diverse cell type to guarantee technical success in the long run. Considering this, fibroblasts from jaguar (Panthera onca), oncilla (Leopardus tigrinus), and pampas cat (Leopardus colocolo) were subjected to cell characterization and then cryopreservation in different cryoprotectant solutions (2.5%, 10% dimethyl sulfoxide [DMSO] or CryoSOfree™). Further testing was conducted to determine each solution's performance in preserving cell viability. In culture, a growth curve to assess cellular growth potential showed that exponential proliferation lasts for about the first 50 hours of in vitro culturing, declining in pace afterward. L. colocolo and L. tigrinus presented no difference in cell viability while using 2.5% DMSO protocols. P. onca cells did not present difference on viability for both concentrations of DMSO. Protocols using CryoSOfree resulted in a decreased viability of P. onca fibroblasts. Morphological differences between fibroblasts among the species were noted under bright field microscopy and scanning electron microscopy. L. colocolo and P. onca cells are fusiform, and L. tigrinus are spherical. All cells presented cytoplasmic projections. Transmission electron microscopy revealed vacuoles and secretion granules, indicating intense cell activity after thawing. Differences found in the efficiency of cryopreservation protocols according to the type of cryoprotectant indicate that species react differently to freezing and thawing processes. This research evaluates key aspects of in vitro protocols for cryopreservation of wild animals, which need to be optimized to guarantee successful cell culturing. More suitable protocols lead to increased efficiency in establishing fibroblast cryobanks and also facilitating the use of wild cats' cells in cloning techniques, contributing directly to preserving wild fauna.