Hybridization in Canids-A Case Study of Pampas Fox (Lycalopex gymnocercus) and Domestic Dog (Canis lupus familiaris) Hybrid.

Hybridization between species with different evolutionary trajectories can be a powerful threat to wildlife conservation. Anthropogenic activities, such as agriculture and livestock, have led to the degradation and loss of natural habitats for wildlife. Consequently, the incidence of interspecific hybridization between wild and domestic species has increased, although cases involving species of different genera are rare. In Vacaria, a Southern city in Brazil, a female canid with a strange phenotype, which had characteristics between the phenotype of the domestic dog (Canis familiaris) and that of the pampas fox (Lycalopex gymnocercus), was found. Our analysis suggests that the animal is a hybrid between a domestic dog and a pampas fox, but future studies are necessary to investigate additional cases of this hybridization in nature. This finding worries for the conservation of wild canids in South America, especially concerning Lycalopex species. Hybridization with the domestic dog may have harmful effects on pampas fox populations due to the potential for introgression and disease transmission by the domestic dog. Therefore, future studies to explore the consequences of hybridization on genetics, ecology, and behavior of wild populations will be essential to improve the conservation of this species.

Fluorosilicic acid and cotinine, separately and in combination, induce genotoxicity and telomeric reduction in human osteoblast cell line MG63.

Skeletal fluorosis is a severe case in which bone deformations and bone tissue weakening occur due to excessive fluorine deposition. Recently, data on smoking have been published that smoke constituents can indirectly influence bone mass and interfere in the metabolism of fluorides in humans. Thus, the present in vitro study aimed to assess the genetic instability in human osteoblast MG63 cells exposed to fluorosilicic acid (FA) and cotinine (COT), separately and in combination, in concentrations found in human plasma. For this, cell cytotoxicity was performed by MTT assay; DNA damage was performed by alkaline comet assay (CA), modified by repair endonucleases (+FPG); micronuclei test (MN) using CBMN-Cyt assay; and telomere length (TL) by qPCR in MG63 cells. No cytotoxicity was observed for all concentrations tested in this study. Alkaline CA results showed a significant increase in DNA damage at all FA concentrations (0.03125-0.300 mg/L), in the two highest concentrations of COT (125 and 250 ng/mL), and the highest concentration of FA+COT (0.300 mg/L+250 ng/mL). Alkaline CA+FPG test was used to detect oxidized nucleobases, which occurred at the two highest concentrations of FA, COT, and FA+COT. Micronuclei test showed an increase in the frequency of MN at all concentrations of FA (0.075-0.300 mg/L) except in the lowest concentration (0.03125 mg/L), in the two highest concentrations of COT (125 and 250 ng/mL), and all concentrations of FA+COT. There was no significant difference in nuclear division index, binucleated cells, nucleoplasmic bridge, and nuclear bud. A TL reduction was observed in cells treated with the highest concentrations of FA alone (0.300 mg/L) and FA+COT (0.300 mg/L+250 ng/mL). Finally, our study showed that FA and COT (mainly alone) at concentrations found in human plasma induced oxidative damage and genetic instability in human osteoblast cells.

Anthropogenic Effects on Natural Mammalian Populations: Correlation Between Telomere Length and Coal Exposure.

The Candiota coal mine in Rio Grande do Sul (RS) is one of the largest in Brazil. Coal is a fossil fuel that causes environmental impacts from its extraction to combustion due to the release of different agents, such as polycyclic aromatic hydrocarbons (PAH) and heavy metals. Ctenomys torquatus are herbivorous and subterranean rodents that dig tunnels with their paws and teeth and can be exposed to coal through contaminated food. Exposure to pollutants can cause DNA damage and affect different tissues, inducing alterations in the population structure and genetic diversity. Our study aimed to evaluate the effect of exposure to coal and its derivatives on the C. torquatus population and to examine the relationship of coal exposure with variations in absolute telomere length (aTL), global DNA methylation and genotoxicity. Our study showed an inverse correlation between telomere length and coal exposure in addition to an increase in DNA damage. The results indicate that coal and its byproducts can contribute to the alteration of the C. torquatus population structure, as evidenced by a reduction in the number of adults.

Isolation and characterization of mesenchymal stem/stromal cells from Ctenomys minutus.

Mesenchymal stem/stromal cells (MSCs) are multipotent cells distributed in all tissues and characterized by adherence, morphology, immunophenotype and trilineage differentiation potential. The present study aimed to isolate and characterize adherent MSC-like populations from different tissues of Ctenomys minutus, a threatened wildlife rodent popularly known as tuco-tuco. Adherent cells were isolated from bone marrow, brain, liver, pancreas and adipose tissue of three adult animals collect in southern Brazil. Cultures showed typical morphology and proliferation potential. Adipose-derived MSCs showed trilineage potential. Cultures derived from adipose tissue, bone marrow and brain were immunophenotyped with negative results for CD31, CD44, CD45, CD106, and MHC class II, as well as strong positive results for CD29. Low fluorescence levels were seen for CD49d, CD90.2 and CD117. Cultures were negative for CD49e, except for brain-derived cultures that were weakly positive. CD11b was negative in adipose-derived MSCs, but positive in brain and bone marrow-derived cultures. The scratch assay showed high migration potential for pancreas and adipose tissue-derived cells. This study represents the first report of isolation and characterization of cultures having characteristics of MSCs from Ctenomys minutus. The collection of biological information for biobanks represents an important contribution to the creation of strategies for prevention of loss of genetic diversity.

DNA damage induced by coal dust, fly and bottom ash from coal combustion evaluated using the micronucleus test and comet assay in vitro.

Coal mining and combustion generating huge amounts of bottom and fly ash are major causes of environmental pollution and health hazards due to the release of polycyclic aromatic hydrocarbons (PAH) and heavy metals. The Candiota coalfield in Rio Grande do Sul, is one of the largest open-cast coal mines in Brazil. The aim of this study was to evaluate genotoxic and mutagenic effects of coal, bottom ash and fly ash samples from Candiota with the comet assay (alkaline and modified version) and micronucleus test using the lung fibroblast cell line (V79). Qualitative and quantitative analysis of PAH and inorganic elements was carried out by High Performance Liquid Chromatography (HPLC) and by Particle-Induced X-ray Emission (PIXE) techniques respectively. The samples demonstrated genotoxic and mutagenic effects. The comet assay modified using DNA-glicosilase formamidopirimidina (FPG) endonuclease showed damage related to oxidative stress mechanisms. The amount of PAHs was higher in fly ash followed by pulverized coal. The amount of inorganic elements was highest in fly ash, followed by bottom ash. It is concluded that the samples induce DNA damage by mechanisms that include oxidative stress, due to their complex composition, and that protective measures have to be taken regarding occupational and environmental hazards.

Genotoxicity induced by water and sediment samples from a river under the influence of brewery effluent.

Brewery effluents contain complex mixtures that are discharged into rivers. Therefore, it is necessary to evaluate the genotoxic potential of these effluents. The study evaluated the genotoxicity of surface water and sediment samples from the Jacuí River in the state of Rio Grande do Sul, Brazil, which received effluents discharged from a brewery. The Salmonella/microsome test, Comet Assay and Micronucleus test on V79 cells, as well as the element profile (PIXE) and PAHs levels were used for this purpose. The surface water and sediment samples were collected in summer at three sites: 1 km upstream from the brewery discharge site (Site A); in front of the effluent discharge site, after chemical and biological treatment (Site B); about 1 km downstream from the discharge site (Site C). Only a sediment sample from Site A induced a mutagenic effect using the Salmonella/microsoma test (TA97a). All three sites presented genotoxicity (A, B and C), both for water and sediments using comet assay, and mutagenicity in the samples from Site B (surface water) and Site A and Site C (sediments) using the micronuclei tests. The results of PIXE and PAHs showed higher levels of elements for samples obtained from sites upstream and downstream from the effluent discharge. Environmental samples consist of complex mixtures of chemicals, and it is difficult to associate DNA damage with a specific element. This study showed that brewery effluent contains metals and PAHs that can induce in vitro genotoxicity under the conditions of this study.

Comparative genotoxicity of airborne particulate matter (PM2.5) using Salmonella, plants and mammalian cells.

This study compared genotoxicity in bacteria, plants and cell cultures in areas at risk of exposure to airborne pollution. Genotoxicity of moderately polar organic extracts of PM2.5 from areas with urban airborne pollution (Site 1) and urban-industrial pollution (Site 2) was evaluated using microsuspension assays in Salmonella/microsome, micronucleus test with Tradescantia pallida (Trad-MN) with acute exposure, and in V79 (V79-MN) cells, Comet assay in V79 and human lymphocyte, besides Trad-MN in situ at Site 1. In the Salmonella/microsome assay all samples presented frameshift mutagenic activity (-/+S9), most intense at Site 2 (rev/m(3)). The presence of nitro-PAHs and hydroxylamines in PM2.5 was shown by positive mutagenic responses with YG1021 and YG1024. In tests with Trad-MN, no significant genotoxic responses were found (MN %). In V79-MN a genotoxic response was not found. The Comet assay damages were found in the DNA at Site 1 in both cell systems. Non-detection of genotoxicity with Trad-MN at sites or in environmental samples from polluted areas detected using other biomarkers suggests the need for careful evaluation when biomonitoring genotoxic compounds using plants. The microsuspension assay in Salmonella/microsome was sensitive to detect and identify different classes of airborne mutagenic compounds present in fine particulate matter in Porto Alegre city, showing that monitoring air quality with PM2.5 using this methodology is relevant.