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Biochar is a carbonaceous porous material that is produced through the thermal processing of biomass under oxygen-limited environment. Nevertheless, biochar is known to be an inexpensive and sustainable raw material with a wide range of possible applications. Recently, biochar has been discovered as an efficient biological catalyst for anaerobic conversion, mainly due to its highly porous structure with micro and macro channels, which procures a viable living area for attached-grown microorganisms. Whereas it is never applied to improve the biological conversion of gas substances such as C1 (e.g., CO, CO2) and H2, which is a promising research area with increasing commercial interest. However, considering that biological reaction is limited by the target water solubility of gas substrates, special attention is required when combining biochar for gas fermentation. The goal was to create a novel gas sparger where the biofilm grows on biochar, thus improving the interaction with the gaseous substrate. For this purpose, polystyrene foam and powdered biochar were compounded to form a mouldable composite, which was then cast as a porous monolith.â¢Biochar-made sparger (BS) was investigated for the homoacetogenic conversion of H2 gas via microbial mixed cultures as opposed to a control test equipped with a stone sparger.â¢BS showed a significantly better performance in terms of biological gas fixation rate (36% more than control) and productivity (8.5 gCOD L-1 d-1).
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Bioreactors are commonly used apparatuses generally equipped with several built-in specifications for the investigation of biological treatment studies. Each bioreactor test may require different types of specialty such as heating, agitation, re-circulation and some further technologies like online sensoring. Even thought, there are many ready-to-use fabricated bioreactors available in the market with a cost usually over than 1000 , it is often not possible to access those advanced (but inflexible) systems for many students, young-researchers or small-scale private R&D companies. In this work, a new low cost (≈100) packed-bed anaerobic bioreactor was developed, and all methodological details including open-source coding and 3D design files are shared with informative descriptions. Some preliminary tests were conducted to verify the developed bioreactor system's credibility in terms of leak-tightness, accurate gas monitoring, temperature controlling, and mass balance (COD-eq) coverage, which all have shown a very promising performance.â¢A consistent model bioreactor that will be called as "tetrapod" was developed for anaerobic treatment of challenging substrates such as pyrolytic liquids.â¢Coarse biochar grains were used as an organic packing material to stimulate the microbial bioconversion by increasing the active surface area for the attached-growth anaerobic mixed microbial culture (MMC).â¢An open-source Arduino based digital gasometer was developed for online monitoring of biogas change in the lab-scale system. Arduino was also used as a digital controller for maintaining pulse-mode liquid recirculation of the bioreactor.
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Dissolved organic matter (DOM) is an important organic matter fraction that plays a key role in many biological and chemical processes in soil. The effect of biochar addition on the content and composition of soil organic matter (SOM) and DOM in an agricultural soil in Italy was investigated within a two-year period. UV-Vis spectroscopy and analytical pyrolysis have been applied to study complex components in DOM soil samples. Additionally, analytical pyrolysis was used to provide qualitative information of SOM at molecular level and the properties of biochar before and one year after amendment. A method was developed to quantify biochar levels by thermogravimetric analysis that enabled to identify deviations from the amendment rate. The water-soluble organic carbon (WSOC) concentrations in the amended soils were significantly lower than those in the control soils, indicating that biochar decreased the leaching of DOM. DOM in treated soils was characterized by a higher aromatic character according to analytical pyrolysis and UV-Vis spectroscopy. Moreover, a relatively high abundance of compounds with N was observed in pyrolysates of treated soils, suggesting that biochar increased the proportion of microbial DOM. The results from thermal and spectroscopy techniques are consistent in highlighting significant changes in DOM levels and composition due to biochar application with important effects on soil carbon storage and cycling.
Assuntos
Matéria Orgânica Dissolvida , Solo , Agricultura , Carvão VegetalRESUMO
An overview of the main polyhydroxyalkanoates (PHA) recovery methods is here reported, by considering the kind of PHA-producing bacteria (single bacterial strains or mixed microbial cultures) and the chemico-physical characteristics of the extracted polymer (molecular weight and polydispersity index). Several recovery approaches are presented and categorized in two main strategies: PHA recovery with solvents (halogenated solvents, alkanes, alcohols, esters, carbonates and ketones) and PHA recovery by cellular lysis (with oxidants, acid and alkaline compounds, surfactants and enzymes). Comparative evaluations based on the recovery, purity and molecular weight of the recovered polymers as well as on the potential sustainability of the different approaches are here presented.
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Coenzyme Q10 (CoQ, ubiquinone) is a redox-active lipid endogenously synthesized by the cells. The final stage of CoQ biosynthesis is performed at the mitochondrial level by the 'complex Q', where coq2 is responsible for the prenylation of the benzoquinone ring of the molecule. We report that the competitive coq2 inhibitor 4-nitrobenzoate (4-NB) decreased the cellular CoQ content and caused severe impairment of mitochondrial function in the T67 human glioma cell line. In parallel with the reduction in CoQ biosynthesis, the cholesterol level increased, leading to significant perturbation of the plasma membrane physicochemical properties. We show that 4-NB treatment did not significantly affect the cell viability, because of an adaptive metabolic rewiring toward glycolysis. Hypoxia-inducible factor 1α (HIF-1α) stabilization was detected in 4-NB-treated cells, possibly due to the contribution of both reduction in intracellular oxygen tension and ROS overproduction. Exogenous CoQ supplementation partially recovered cholesterol content, HIF-1α degradation, and ROS production, whereas only weakly improved the bioenergetic impairment induced by the CoQ depletion. Our data provide new insights on the effect of CoQ depletion and contribute to shed light on the pathogenic mechanisms of ubiquinone deficiency syndrome.
Assuntos
Metabolismo Energético , Glicólise , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Ubiquinona/análogos & derivados , Alquil e Aril Transferases/antagonistas & inibidores , Alquil e Aril Transferases/metabolismo , Ataxia/metabolismo , Linhagem Celular Tumoral , Colesterol/metabolismo , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Doenças Mitocondriais/metabolismo , Debilidade Muscular/metabolismo , Nitrobenzoatos/farmacologia , Estabilidade Proteica/efeitos dos fármacos , Ubiquinona/antagonistas & inibidores , Ubiquinona/biossíntese , Ubiquinona/deficiência , Ubiquinona/metabolismoRESUMO
In order to understand the cutaneous water loss in the desert-adapted and venomous lizard Heloderma suspectum, the microscopic structure and lipid composition of epidermal molts have been examined using microscopic, spectroscopic and chemical analysis techniques. The molt is formed by a variably thick, superficial beta-layer, an extensive mesos-region and few alpha-cells in its lowermost layers. The beta-layer contains most corneous beta proteins while the mesos-region is much richer in lipids. The proteins in the mesos-region are more unstructured than those located in the beta-layer. Most interestingly, among other lipids, high contents of cholesteryl-ß-glucoside and cholesteryl sulfate were detected, molecules absent or present in traces in other species of squamates. These cholesterol derivatives may be involved in the stabilization and compaction of the mesos-region, but present a limited permeability to water movements. The modest resistance to cutaneous water-loss of this species is compensated by adopting other physiological strategies to limit thermal damage and water transpiration as previous eco-physiological studies have indicated. The increase of steroid derivatives may also be implicated in the heat shock response, influencing the relative behavior in this desert-adapted lizard.
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Adaptação Fisiológica/fisiologia , Colesterol/metabolismo , Epiderme/metabolismo , Metabolismo dos Lipídeos/fisiologia , Lagartos/metabolismo , Muda/fisiologia , Animais , Lipídeos , Permeabilidade , Peçonhas/metabolismo , Água/metabolismoRESUMO
A new method based on solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS) was developed for the analysis of seven linear (L2 - L5) and cyclic (D3 - D5) volatile methyl siloxanes (VMS) in biogas and biomethane, directly collected into Tedlar® bags (Tedlar SPME) from anaerobic digesters and wastewater treatment plants. The method was employed to monitor VMS content in biomethane produced by biogas upgrading with a pilot-plant membrane unit and provided adequate limits of quantification (< 0.05â¯mgâ¯m-3) to detect trace siloxane impurities. Tedlar SPME was validated against a standard procedure based on indirect sampling of gas streams with sorbent tubes followed by solvent extraction and GC-MS. Method precision (RSD) on total and individual VMS concentrations was lower than 10%, while RSD values of the standard procedure were higher than 20%. Tedlar SPME suitably revealed high VMS levels, expressed as total volatile silicon (> 1â¯mgSim-3), in wastewater biogas and provided a more efficient sampling of heavier VMS in comparison to the sorbent tubes method. At low values (< 0.1â¯mgSim-3) typical of wood waste biogas and biomethane, no statistically significant differences were observed between the two methods. Overall, Tedlar SPME simplified the analytical procedure by reducing the procedural steps, avoiding the use of solvents and demonstrated its applicability for testing the quality of biomethane as advanced biofuel.
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Volatile fatty acids (VFAs) are among the most important parameters in process monitoring of anaerobic digestion plants for biogas production. The concentration of single VFA species is typically determined by direct injection of the acidified aqueous phase of digestate samples into GC-FID. Analysis of dimethyl carbonate extracts was investigated as an alternative method consisting of a simple and rapid in-vial procedure of acidification and solvent extraction of the sample, followed by centrifugation and GC-MS analysis. The principal figures of merit resulting from internal standard calibration were comparable to those proposed for the direct analysis of aqueous digestate, while the analysis of real samples did not provide statistically significant differences between the two methods according to parametric and non-parametric tests. Procedural aspects including sample amount and solid removal improved with dimethyl carbonate, while GC contamination was reduced. The method was applied to seventeen samples from fully operating anaerobic digesters fed with various feedstocks and enabled the individuation of high probability of system stress through the values of total VFA, propanoic acid, longer chained VFA concentrations and the ratio between acetic and propanoic acid concentrations. The use of dimethyl carbonate allowed the detection of alicyclic and aromatic acids that could represent new molecular markers in assessing the origin of feed and process conditions.
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Ácidos Graxos Voláteis/análise , Formiatos/química , Ácidos Graxos Voláteis/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Solventes/químicaRESUMO
Polyhydroxyalkanoates (PHA) are a key constituent of excess sludge produced by Aerobic Sewage Sludge Treatment plants. The accumulation of significant amount of PHA inside aerobic microbial cells occurs when a surplus of an easily degradable carbon source (e.g., volatile fatty acids, VFA) is found in combination with other nutrients limitation. Herein, hydrothermal treatment (HT) of PHA-containing sludge at 300 and 375 °C was demonstrated to be effective in converting most (>70% w/w) of the bacterial PHA stored inside microbial cells into alkene/CO2 gas mixtures. Simultaneously, most of non-PHA biomass was converted into water-soluble compounds (50% carbon yield) that were acidogenic fermented to produce volatile fatty acids, ideal substrate to feed aerobic bacteria and produce more PHA. According to results here presented, HT of excess sludge with moderate (13%) PHA content can produce about 50 kg of alkenes per tonne of suspended solids treated, with a significant reduction of sludge mass (80% reduction of wet sludge volume) and consequent disposal cost.
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Reatores Biológicos , Poli-Hidroxialcanoatos , Alcenos , Ácidos Graxos Voláteis , EsgotosRESUMO
The study evaluated the growth of Desmodesmus communis on column photobioreactor and its thermochemical treatment by catalytic pyrolysis using HZSM-5 zeolite. D. communis showed good results in terms of growth (0.05gL-1d-1). Analytical pyrolysis of original algae and derived bio-oil mixed with zeolite was used as a screening method in order to gather information on the cracking process. Preparative pyrolysis on bench scale reactor was performed on algae biomass over a zeolite bed at 1:10 ratio (wt/wt). Py-GC-MS of biomass/catalyst mixture showed that the denitrogenation/deoxygenation increased with increasing zeolite load from 1:5 to 1:20 ratio and became significant at 1:10 ratio. The composition observed by analytical pyrolysis was featured by the predominance of alkylated monoaromatic hydrocarbons. The scaling-up to bench scale confirmed the results obtained with analytical pyrolysis in terms of monoaromatic hydrocarbons. However, low yield of catalytic oil (8% by weight) was observed.
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Biotecnologia/métodos , Clorófitas/metabolismo , Hidrocarbonetos/metabolismo , Microalgas/metabolismo , Biocombustíveis/análise , Biomassa , Biotecnologia/instrumentação , Catálise , Clorófitas/química , Clorófitas/crescimento & desenvolvimento , Cromatografia Gasosa-Espectrometria de Massas/métodos , Hidrocarbonetos/análise , Hidrocarbonetos/química , Microalgas/química , Microalgas/crescimento & desenvolvimento , Fotobiorreatores , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Zeolitas/químicaRESUMO
In view of the potential use of pyrolysis-based technologies, it is crucial to understand the environmental hazards of pyrolysis-derived products, in particular bio-oils. Here, three bio-oils were produced from fast pyrolysis of pine wood and intermediate pyrolysis of corn stalk and poultry litter. They were fully characterized by chemical analysis and tested for their biodegradability and their ecotoxicity on the crustacean Daphnia magna and the green alga Raphidocelis subcapitata. These tests were chosen as required by the European REACH regulation. These three bio-oils were biodegradable, with 40-60% of biodegradation after 28 days, and had EC50 values above 100mgL(-1) for the crustacean and above 10mgL(-1) for the alga, showing low toxicity to the aquatic life. The toxic unit approach was applied to verify whether the observed toxicity could be predicted from the data available for the substances detected in the bio-oils. The predicted values largely underestimated the experimental values.
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Biomassa , Óleos , Animais , Biodegradação Ambiental , Galinhas , Clorófitas/efeitos dos fármacos , Daphnia/efeitos dos fármacos , Ecotoxicologia , Esterco , Óleos/metabolismo , Óleos/toxicidade , Pinus , Zea maysRESUMO
A Life Cycle Assessment is conducted on pyrolysis coupled to anaerobic digestion to treat corn stovers and to obtain bioenergy and biochar. The analysis takes into account the feedstock treatment process, the fate of products and the indirect effects due to crop residue removal. The biochar is considered to be used as solid fuel for coal power plants or as soil conditioner. All results are compared with a corresponding fossil-fuel-based scenario. It is shown that the proposed system always enables relevant primary energy savings of non-renewable sources and a strong reduction of greenhouse gases emissions without worsening the abiotic resources depletion. Conversely, the study points out that the use of corn stovers for mulch is critical when considering acidification and eutrophication impacts. Therefore, removal of corn stovers from the fields must be planned carefully.
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Carvão Vegetal , Meio Ambiente , Lignina , Eliminação de Resíduos/métodos , Zea mays , Anaerobiose , Biocombustíveis , Biomassa , Biotecnologia/métodos , Carvão Vegetal/química , Carvão Mineral , Lignina/química , Centrais Elétricas , Solo , Zea mays/químicaRESUMO
Biogas is a mixture of CO2 and CH4 produced by a consortia of Bacteria and Archeae operating in anaerobic digestion (AD) plants. Biogas can be burnt as such in engines to produce electricity and heat or upgraded into biomethane. Biomethane is a drop-in fuel that can be injected in the natural gas grid or utilised as a transport fuel. While a wide array of biomass feedstock can be degraded into biogas, unconverted lignin, hemicellulose and cellulose end up in the co-product digestate leaving a large portion of chemical energy unutilised. Pyrolysis (Py) transforms in a single step and without chemical reagents the lignocellulose matrix into gaseous (syngas), liquid (bio-oil, pyrolysis oil) and solid (biochar) fractions for the development of renewable fuels and materials. The Py route applied downstream to AD is actively investigated in order to valorise the solid digestate presently destined only for soil applications. Coupling Py upstream to AD is an emerging field of research aimed at expanding the feedstock towards biologically recalcitrant substrates (wood, paper, sludge). The biomethanation potential was demonstrated for gaseous (H2/CO) and water soluble pyrolysis products, while the influence of insoluble pyrolytic lignin remains fairly unexplored. Biochar can promote the production of biomethane by acting as a support for microorganism colonisation, conductor for direct interspecies electron transfer, sorbent for hydrophobic inhibitors, and reactant for in situ biogas upgrading. Enhancing the advantages (carbon source) over the side effects (toxicity) of Py fractions represents the main challenge of Py-AD. This can be addressed by increasing the selectivity of the thermochemical process or improving the ecological flexibility of mixed bacterial consortia towards chemically complex environments.
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Biomassa , Lignina/metabolismo , Anaerobiose , Animais , Temperatura Alta , Humanos , Lignina/químicaRESUMO
Three biochars were prepared by intermediate pyrolysis from poultry litter at different temperatures (400, 500, and 600 °C with decreasing residence times) and compared with biochars from corn stalk prepared under the same pyrolysis conditions. The phytotoxicity of these biochars was estimated by means of seed germination tests on cress (Lepidium sativum L.) conducted in water suspensions (at 2, 5, and 40 g/L) and on biochars wetted according to their water-holding capacity. Whereas the seeds germinated after 72 h in water suspensions with corn stalk biochar were similar to the control (water only), significant inhibition was observed with poultry litter biochars. In comparison to corn stalk, poultry litter generated biochars with higher contents of ash, ammonium, nitrogen, and volatile fatty acids (VFAs) and a similar concentration of polycyclic aromatic hydrocarbons (PAHs). Results from analytical pyrolysis (Py-GC-MS) indicated that nitrogen-containing organic compounds (NCCs) and aliphatic components were distinctive constituents of the thermally labile fraction of poultry litter biochar. The inhibition of germination due to poultry litter biochar produced at 400 °C (PL400) was suppressed after solvent extraction or treatment with active sludge. A novel method based on solid-phase microextraction (SPME) enabled the identification of mobile organic compounds in PL400 capable of being released in air and water, including VFAs and NCCs. The higher phytotoxicity of poultry litter than corn biochars was tentatively attributed to hydrophilic biodegradable substances derived from lipids or proteins removable by water leaching or microbial treatments.
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Carvão Vegetal/química , Carvão Vegetal/toxicidade , Lepidium sativum/efeitos dos fármacos , Esterco/análise , Animais , Hidrólise , Lepidium sativum/crescimento & desenvolvimento , Aves Domésticas , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimentoRESUMO
Polyhydroxyalkanoates (PHAs) can be extracted from mixed microbial cultures (MMCs) by means of dimethyl carbonate (DMC) or combination of DMC and sodium hypochlorite (NaClO). The protocol based on DMC, a green solvent never used before for the extraction of PHAs from MMC, allows an overall polymer recovery of 63%; also the purity and the molecular weight of the recovered polymers are good (98% and 1.2 MDa, respectively). The use of NaClO pretreatment before DMC extraction increases the overall PHA recovery (82%) but lowers the mean molecular weight to 0.6-0.2 MDa. A double extraction with DMC results to be the method of choice for the recovery of high quality PHAs from attractive but challenging MMCs.
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Bactérias/metabolismo , Poli-Hidroxialcanoatos/isolamento & purificação , Bactérias/crescimento & desenvolvimento , Técnicas de Cultura Celular por Lotes , Biomassa , Reatores Biológicos/microbiologia , SolventesRESUMO
Intermediate pyrolysis produces a two-phase liquid whose aqueous phase is characterized by low heating value and high water content (aqueous pyrolysis liquid, APL). Anaerobic digestion can be the straightest way to produce a fuel (methane) from this material. Batch tests showed poor performance in anaerobic digestion of APL, which underlined the inhibition of biological process. Nutrient supplementation was ineffective, whereas biochar addition increased yield of methane (60±15% of theoretical) with respect to pure APL (34±6% of theoretical) and improved the reaction rate. On the basis of batch results, a semi-continuous biomethanation test was set up, by adding an increasingly amount of APL in a 30ml reactor preloaded with biochar (0.8gml(-1)). With a daily input of 5gd(-1)l(-1) of APL (corresponding to overall amount of 0.1kgl(-1) added before the end of the study) the yield of methane was 65±5% of the theoretical.
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Bactérias Anaeróbias/fisiologia , Carvão Vegetal/metabolismo , Temperatura Alta , Metano/metabolismo , Componentes Aéreos da Planta/microbiologia , Água/metabolismo , Zea mays/microbiologia , Reatores Biológicos/microbiologia , Transição de Fase , Componentes Aéreos da Planta/química , Esgotos , Água/química , Zea mays/químicaRESUMO
Poly(hydroxyalkanoates) (PHAs) are polyesters formed by saturated short chain hydroxyacids, among which 3-hydroxybutanoic (HB) and 3-hydroxypentanoic (3-hydroxyvalerate, HV) are the most common monomers of homopolymers (e.g. poly(3-hydroxybutyrate), PHB) and copolymers (e.g. poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), PHB-HC). The most widely used approach for their determination is the polymer methanolysis followed by gas chromatography-mass spectrometry (GC-MS) analysis of the methylated monomers; this procedure generally requires the use of additional reagents (e.g. sulfuric acid) and is performed with harmful chlorinated solvents, such as chloroform. The development of fast routine solventless methods for the quantitative determination of PHAs and their monomeric composition is highly desirable to reduce sample pretreatment, speed up the analysis and decrease overall costs. It has been reported that under thermal treatment (e.g. pyrolysis, Py), PHAs are degraded in high yield (>40%, w/wPHA) into the corresponding 2-alkenoic acid (e.g. crotonic acid from PHB). This work aimed at investigating this reaction for direct analysis of PHAs in bacterial cells. The sample was directly subjected to pyrolysis and trapped pyrolysis products were analyzed by GC-FID. Off-line Py/GC-FID was first optimized on pure polymers with different monomer composition (PHB, PHB-HV, PHB-HC) and then applied to bacterial samples deriving from both mixed microbial cultures or selected strains, containing various types and amounts of PHAs. The Py/GC-FID method provided RSD <15% range, limit of detection of 100µg (1% PHAs in biomass), and results comparable to that of methanolysis (R(2)=0.9855), but with minimal sample pretreatment.
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Ácido 3-Hidroxibutírico/química , Biopolímeros/química , Caproatos/química , Cupriavidus necator/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Hidroxibutiratos/química , Poliésteres/química , Ácido 3-Hidroxibutírico/metabolismo , Biomassa , Biopolímeros/metabolismo , Caproatos/metabolismo , Cupriavidus necator/crescimento & desenvolvimento , Cupriavidus necator/metabolismo , Ionização de Chama , Hidroxibutiratos/metabolismo , Poliésteres/metabolismoRESUMO
The role of lipids in controlling water exchange is fundamentally a matter of molecular organization. In the present study we have observed that in snake molt the water permeability drastically varies among species living in different climates and habitats. The analysis of molts from four snake species: tiger snake, Notechis scutatus, gabon viper, Bitis gabonica, rattle snake, Crotalus atrox, and grass snake, Natrix natrix, revealed correlations between the molecular composition and the structural organization of the lipid-rich mesos layer with control in water exchange as a function of temperature. It was discovered, merging data from micro-diffraction and micro-spectroscopy with those from thermal, NMR and chromatographic analyses, that this control is generated from a sophisticated structural organization that changes size and phase distribution of crystalline domains of specific lipid molecules as a function of temperature. Thus, the results of this research on four snake species suggest that in snake skins different structured lipid layers have evolved and adapted to different climates. Moreover, these lipid structures can protect, "safety", the snakes from water lost even at temperatures higher than those of their usual habitat.
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Lipídeos/fisiologia , Muda/fisiologia , Pele/metabolismo , Pele/fisiopatologia , Serpentes/metabolismo , Serpentes/fisiologia , Água/metabolismo , Animais , Ecossistema , Permeabilidade , TemperaturaRESUMO
Bio-oils were produced from bench-scale slow-pyrolysis of three different biomass samples (corn stalks, poplar and switchgrass). Experimental protocols were developed and applied in order to screen their chemical composition. Several hazardous compounds were detected in the bio-oil samples analysed, including phenols, furans and polycyclic aromatic hydrocarbons. A procedure was outlined and applied to the assessment of toxicological and carcinogenic hazards of the bio-oils. The following hazardous properties were considered: acute toxicity; ecotoxicity; chronic toxicity; carcinogenicity. Parameters related to these properties were quantified for each component identified in the bio-oils and overall values were estimated for the bio-oils. The hazard screening carried out for the three bio-oils considered suggested that: (i) hazards to human health could be associated with chronic exposures to the bio-oils; (ii) acute toxic effects on humans and eco-toxic effects on aquatic ecosystems could also be possible in the case of loss of containment; and (iii) bio-oils may present a marginal potential carcinogenicity. The approach outlined allows the collection of screening information on the potential hazards posed by the bio-oils. This can be particularly useful when limited time and analytical resources reduce the possibility to obtain detailed specific experimental data.