The effect of arsenic on the structure and composition of stream hyphomycetes assemblages.

Aquatic hyphomycetes are fungi with a fundamental ecological role in forested streams. These organisms are responsible for cycling of nutrients in aquatic environments. However, their structure and composition can be affected when exposed to certain pollutants. Arsenic (As) is a trace element with high toxicity for the aquatic biota. Here we evaluated the effects of different concentrations of Arsenite (AsIII) and Arsenate (AsV) on aquatic hyphomycetes assemblages. To test As toxicity, we conditioned Nectandra megapotamica leaves in a stream and after this period, we incubated leaf discs with stream water and different concentrations of AsIII and AsV. Species richness was negatively affected by both As form. Likewise, the hyphomycetes assemblages presented variation in the composition of species. However, the sporulation rates were not influenced by As. The As showed toxicity on species of hyphomycetes more sensitive, remaining only in species tolerant to its toxicity. In this way, As generated a change in the aquatic hyphomycetes composition. We observed that As had a negative effect on the aquatic hyphomycetes assemblages, regardless of the chemical form. Our results point to the toxicity of this element and its effects on a group that is fundamental to the streams ecosystems functioning.

Molecular inferences about the genus Hypostomus Lacépède, 1803 (Siluriformes: Loricariidae): a review.

This review compiles and discusses the use of genetic markers applied in the study of the fish genus Hypostomus Lacépède, 1803 (Siluriformes: Loricariidae). The database comprises 51 peer-review articles that were published in the last 52 years (1968-2020) and that approach analysis based on different classes of genetic markers. The use of cytogenetic and enzymatic markers was predominantly especially in population studies with the genus Hypostomus, while mitochondrial markers were the majority in phylogenetic studies. Although significant methodological advances have occurred for molecular evaluation, they are still modestly applied to the study of neotropical fish genera, in which Hypostomus is included. New perspectives, especially on integrative approaches, are needed to improve our knowledge of the genetic functionality of fishes.

Evaluation of heavy metal toxicity in eukaryotes using a simple functional assay.

Although carcinogenesis caused by metals has been intensively investigated, the mechanisms of action, especially at the molecular level, are still unclear. This work aimed to investigate Cd(2+), Cu(2+), Ni(2+), Cr(3+), and Zn(2+) mutagenicity and its relationship with oxidative stress. We have applied the Functional Assay for the Separation of Alleles in Yeast (FASAY) with only minor modifications to detect p53 defects caused by metals. In this method, human p53-coding gene (TP53) expressed in Saccharomyces cerevisiae activates transcription of the ADE2 reporter gene. Yeast cells, expressing p53, were exposed to increased concentrations of metals and, then, plated on media supplemented or not with adenine. Yeast colonies containing functional p53 grow independently of adenine supplementation and colonies containing nonfunctional p53 are dependent on this nutrient. Mutations in the TP53 are implicated in the pathogenesis of half of all human tumors. According to our results, Cd(2+) was found to be the most toxic metal and produced the highest oxidative damage to lipids and proteins. At low concentrations (40 µM), this metal decreased viability and completely inhibited cell growth, while higher concentrations were necessary to produce the same toxic effect by Cu(2+), Cr(3+), and Ni(2+). Zn(2+) showed no significant toxicity. Cd(2+) strongly induced damages and altered the function of p53, while Cu(2+), followed by Cr(3+), showed lower percentages of p53-mutant colonies. Our results point towards a relationship between the loss of functional p53 protein and oxidative stress, a mechanism that can be associated with tumor formation induced by heavy metals in mammalian cells. By this adaptation of FASAY developed by us it is possible to easily and rapidly detect mutations caused by metals or other stresses.

Trypanosoma cruzi response to the oxidative stress generated by hydrogen peroxide.

As an intracellular parasite, Trypanosoma cruzi is exposed to reactive oxygen species. The study of the proteins involved in the hydroperoxide detoxification cascade, tryparedoxin peroxidase included, may lead to the development of a more specific chemotherapy for Chagas'disease. In this work, the involvement of TcCPX in T. cruzi resistance to oxidant-mediated injury was investigated. At low concentrations of hydrogen peroxide cell proliferation was stimulated and parasites increased their resistance to sub-lethal doses of H2O2 (100 microM) if previously treated with a non-toxic concentration of H2O2 (20 microM). Incubation of cells with different H2O2 concentrations induced a dose-dependent increase in TcCPX levels, as detected by Western blotting analysis. The increase in TcCPX levels in the presence of high H2O2 concentrations possibly reflects an initial cell attempt to promote detoxification. To further demonstrate TcCPX involvement in T. cruzi response to oxidative stress, TcCPX overexpressing cells were produced. Compared to pTEX transformed cells, pTEX-TcCPX mutant cells showed a higher mRNA level (129%), without a corresponding increase in protein production (11%), suggesting that regulation of gene expression occurs at post-transcriptional levels. Furthermore, parasite treatment with 200 microM H2O2 for 30 min, led to an increase in mRNA (192%), but not in protein levels (24%). Higher mRNA levels correlated to protein levels were observed only after longer H2O2 incubation periods (1-2 h), suggesting that protein translation occurs accordingly to parasite needs. An increase in glucose-6-phosphate dehydrogenase activity was observed in pTEX-TcCPX epimastigotes that could provide cells with extra reducing power and a higher growth index.