Recovery of Francisella tularensis from soil samples by filtration and detection by real-time PCR and cELISA

The aim of this study was to develop a specific and highly sensitive method able to detect very low concentrations of Francisella tularensis in soil samples by real-time PCR (qPCR) with SYBR Green I. tul4 gene, which encodes the 17-kDa protein (TUL4) in F. tularensis strains, was amplified using a LightCycler (LC) device. We achieved a detection limit of 0.69 fg of genomic DNA from F. tularensis subp. holarctica live vaccine strain (LVS), corresponding to a value less than 3.4 genome equivalents per reaction. The qPCR was shown to be specific, highly sensitive and reproducible. In addition, we evaluated 2 new methods for recovering bacteria from soil based on 1-step filtration using glass fiber filters and PVDF filters. These filtration methods enabled us to recover F. tularensis efficiently from soil samples. As few as 50 CFU per 0.5 g of soil were detected by qPCR. Capture enzyme-linked immunosorbent assay (cELISA) allowed us to detect and quantify the amount of bacteria recovered from soil by an immunological method. Although qPCR was more sensitive than cELISA, we did not observe substantial differences in the amount of bacteria quantified by both methods.     

Cytogenetic evaluation and the association with polymorphisms of the CPY1A1 and NR1I3 genes in individuals exposed to BTEX

The gas station attendants are exposed daily to chemical agents that compose gasoline, such as BTEX (benzene, toluene, ethylbenzene, and xylene), and the exposure to these agents can cause a variety of effects on the human health. Among the various possible cell alterations associated with these exposures are the formation of micronuclei and of binucleated cells which are used as indicators of clastogenic action. Benzene, the main carcinogenic agent, is metabolized to more soluble forms and easily excreted by isoenzymes of cytochrome P450, such as CYP1A1. The CYP1A1 gene is highly polymorphic and one of its allele variations can be detected by the use of restriction endonucleasis MspI and is originated by the transition of a thymine by a cytosine (3798T>C), resulting in the polymorphic allele CYP1A1*2A. The objective of this study was to evaluate the cytogenetic damage induced by the exposure to BTEX and to associate it with the polymorphisms of the CYP1A1 and NR1I3 genes. Samples of exfoliated cells from the oral mucosa of 27 gas station attendants and from a control group were collected. The results found show that the group exposed to BTEX presents significantly higher alterations than those in the control group for micronuclei (MN; 6.85?±?1.33 vs. 2.96?±?1.91, P?<?0.001) and for the total of nuclear alterations observed (MN + binucleated cells (BNC); 9.59?±?4.73 vs. 5.07?±?2.21, P?<?0.001). When comparing the cytological alterations and the genotypes among the exposed individuals for the polymorphism 3798T>C of the CYP1A1 gene, homozygotes TT present MN + BNC significantly higher than carriers of the allele C (10.88?±?5.36 vs. 5.33?±?2.52, P?=?0.028). No association was observed in the control group or for the NR1I3 gene. These results show that molecular and cytogenetic data can be used in the future as tools to monitor individuals exposed to such compounds.     

Invasion by <Emphasis Type="Italic">Artocarpus heterophyllus</Emphasis> (Moraceae) in an island in the Atlantic Forest Biome,Brazil: distribution at the landscape level,density and need for control

Invasion by alien species is one of the most critical factors affecting global biodiversity, leading to homogenization and species loss worldwide. Jackfruit (Artocarpus heterophyllus Lam., Moraceae) is one of the most conspicuous invasive plants in Brazil. We studied jackfruit invasion in the Atlantic Forest in Ilha Grande State Park, located on an island (Ilha Grande), by assessing jackfruit distribution and density along 23 trails (90 km). Presence or absence was verified in a total of 857 plots. Data showed that jackfruit is spread all over Ilha Grande, and that plant density varied significantly along and between trails. Jackfruit was registered as the dominant tree species on some of the trails. Control by park managers is an urgent need in order to ensure the conservation of local biodiversity. A decision key for jackfruit management according to jackfruit density, age class and slope is provided.     

Human predisposition to cognitive impairment and its relation with environmental exposure to potentially toxic elements

New lines of evidence suggest that less than 10% of neurodegenerative diseases have a strict genetic aetiology and other factors may be prevalent. Environmental exposures to potentially toxic elements appear to be a risk factor for Parkinson’s, Alzheimer’s and sclerosis diseases. This study proposes a multidisciplinary approach combining neurosciences, psychology and environmental sciences while integrating socio-economic, neuropsychological, environmental and health data. We present the preliminary results of a neuropsychological assessment carried out in elderly residents of the industrial city of Estarreja. A battery of cognitive tests and a personal questionnaire were administered to the participants. Multivariate analysis and multiple linear regression analysis were used to identify potential relationships between the cognitive status of the participants and environmental exposure to potentially toxic elements. The results suggest a relationship between urinary PTEs levels and the incidence of cognitive disorders. They also point towards water consumption habits and profession as relevant factors of exposure. Linear regression models show that aluminium (R ² = 38%), cadmium (R ² = 11%) and zinc (R ² = 6%) are good predictors of the scores of the Mini-Mental State Examination cognitive test. Median contents (µg/l) in groundwater are above admissible levels for drinking water for aluminium (371), iron (860), manganese (250), and zinc (305). While the World Health Organization does not provide health-based reference values for aluminium, results obtained from this study suggest that it may have an important role in the cognitive status of the elderly. Urine proved to be a suitable biomarker of exposure both to elements with low and high excretion rates.     

Changes in glucose uptake rate and in the energy status of PC-12 cells acutely exposed to hexavalent chromium,an established human carcinogen

In the 1920s, Otto Warburg reported a striking metabolic shift in solid tumors: contrary to their normal counter parts, which exhibited a nearly pure respiratory metabolism, where cancer cells relied strongly on lactic fermentation for energy production, even in the presence of ample oxygen. This metabolic shift, later named the Warburg effect, is now viewed as a nearly universal cancer phenotype. To investigate whether it is operating in hexavalent chromium (Cr(VI))-induced carcinogenesis, PC-12 cells were exposed to low Cr(VI) concentrations and effects determined on the rates of glucose uptake, lactate production and oxygen consumption, critical indicators of the type of energy metabolism adopted by the cells. Further, the influence on the cells’ energy charge, an important parameter in the evaluation of the cellular physiological state was assessed. In the presence of ample oxygen, concentration-dependent, statistically significant decreases in the energy charge were detected, which were accompanied by an increased glucose uptake rate. This enhanced uptake may constitute the first step in a compensatory mechanism aimed at counteracting the decrease in energy charge. Although these changes may be too small to exert an impact in the cellular functions, they may provide insight into the initial steps of Cr(VI)-induced carcinogenesis.     

Effect of temperature on biogeochemistry of marine organic-enriched systems: implications in a global warming scenario

Coastal biogeochemical cycles are expected to be affected by global warming. By means of a mesocosm experiment, the effect of increased water temperature on the biogeochemical cycles of coastal sediments affected by organic-matter enrichment was tested, focusing on the carbon, sulfur, and iron cycles. Nereis diversicolor was used as a model species to simulate macrofaunal bioirrigation activity in natural sediments. Although bioirrigation rates of N. diversicolor were not temperature dependent, temperature did have a major effect on the sediment metabolism. Under organic-enrichment conditions, the increase in sediment metabolism was greater than expected and occurred through the enhancement of anaerobic metabolic pathway rates, mainly sulfate reduction. There was a twofold increase in sediment metabolism and the accumulation of reduced sulfur. The increase in the benthic metabolism was maintained by the supply of electron acceptors through bioirrigation and as a result of the availability of iron in the sediment. As long as the sediment buffering capacity toward sulfides is not surpassed, an increase in temperature might promote the recovery of organic-enriched sediments by decreasing the time for mineralization of excess organic matter.     

Global mismatches in aboveground and belowground biodiversity

Human activities are accelerating global biodiversity change and have resulted in severely threatened ecosystem services. A large proportion of terrestrial biodiversity is harbored by soil, but soil biodiversity has been omitted from many global biodiversity assessments and conservation actions, and understanding of global patterns of soil biodiversity remains limited. In particular, the extent to which hotspots and coldspots of aboveground and soil biodiversity overlap is not clear. We examined global patterns of these overlaps by mapping indices of aboveground (mammals, birds, amphibians, vascular plants) and soil (bacteria, fungi, macrofauna) biodiversity that we created using previously published data on species richness. Areas of mismatch between aboveground and soil biodiversity covered 27% of Earth's terrestrial surface. The temperate broadleaf and mixed forests biome had the highest proportion of grid cells with high aboveground biodiversity but low soil biodiversity, whereas the boreal and tundra biomes had intermediate soil biodiversity but low aboveground biodiversity. While more data on soil biodiversity are needed, both to cover geographic gaps and to include additional taxa, our results suggest that protecting aboveground biodiversity may not sufficiently reduce threats to soil biodiversity. Given the functional importance of soil biodiversity and the role of soils in human well-being, soil biodiversity should be considered further in policy agendas and conservation actions by adapting management practices to sustain soil biodiversity and considering soil biodiversity when designing protected areas.     

Determinants of population persistence and abundance of terrestrial and arboreal vertebrates stranded in tropical forest land-bridge islands

Megadams are among the key modern drivers of habitat and biodiversity loss in emerging economies. The Balbina Hydroelectric Dam of Central Brazilian Amazonia inundated 312,900 ha of primary forests and created approximately 3500 variable-sized islands that still harbor vertebrate populations after nearly 3 decades after isolation. We estimated the species richness, abundance, biomass, composition, and group size of medium- to large-bodied forest vertebrates in response to patch, landscape, and habitat-quality metrics across 37 islands and 3 continuous forest sites throughout the Balbina archipelago. We conducted 1168 km of diurnal censuses and had 12,420 camera-trapping days along 81 transects with 207 camera stations. We determined the number of individuals (or groups) detected per 10 km walked and the number of independent photographs per 10 camera-trapping days, respectively, for each species. We recorded 34 species, and patch area was the most significant predictor of vertebrate population relative abundance and aggregate biomass. The maximum group size of several group-living species was consistently larger on large islands and in continuous patches than on small islands. Most vertebrate populations were extirpated after inundation. Remaining populations are unlikely to survive further ecological disruptions. If all vertebrate species were once widely distributed before inundation, we estimated that approximately 75% of all individual vertebrates were lost from all 3546 islands and 7.4% of the animals in all persisting insular populations are highly likely to be extirpated. Our results demonstrate that population abundance estimates should be factored into predictions of community disassembly on small islands to robustly predict biodiversity outcomes. Given the rapidly escalating hydropower infrastructure projects in developing counties, we suggest that faunal abundance and biomass estimates be considered in environmental impact assessments and large strictly protected reserves be established to minimize detrimental effects of dams on biodiversity. Conserving large tracts of continuous forests represents the most critical conservation measure to ensure that animal populations can persist at natural densities in Amazonian forests.     

Agro-industrial wastes as effective amendments for ecotoxicity reduction and soil health improvement in aided phytostabilization

Aided phytostabilization is a technology that uses metal tolerant plants and organic and/or inorganic amendments to reduce soil metal bioavailability, while improving soil health. Our objective was to determine the effects of the application of amendments [sheep manure (SHEEP), poultry litter (POULTRY), cow slurry (COW), and paper mill sludge mixed with poultry litter (PAPER)], together with the growth of a metallicolous Festuca rubra L. population, on (i) chemical and microbial indicators of soil health and (ii) soil ecotoxicity, during the aided phytostabilization of a Zn/Pb contaminated mine soil. Amendment application led to an increase in soil pH, organic matter content, and inorganic salts, resulting in a decrease in Pb and Zn CaCl₂-extractable concentrations in soil, which, in turn, contributed to lower ecotoxicity and a stimulation of plant growth and soil microbial communities. The factor most affecting the metal extractability was probably soil pH. POULTRY was the best amendment in terms of increasing plant growth, chlorophylls content, and soil microbial biomass and activity, but resulted in higher levels of phytoavailable Pb and Zn. SHEEP and PAPER were more effective at reducing metal CaCl₂-extractability and, consequently, led to lower values of metal accumulation in plant tissues, thereby reducing the risk of metals entering into the food chain. When combined with the application of organic amendments, the metallicolous F. rubra population studied here appears an excellent candidate for aided phytostabilization. Our results indicate that the application of organic amendments is essential for the short-term recovery of highly contaminated metalliferous soils during aided phytostabilization.