Rumen microbial diversity under influence of a polyclonal antibody preparation against lactate-producing and proteolytic bacteria in cows fed different energy sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do desempenho de um reator anaeróbio compartimentado no tratamento de esgoto doméstico

Pós-graduação em Engenharia Civil - FEIS

Estudo da variação da porcentagem e da estimativa de geração de gás metano para o aterro sanitário do município de Rio Claro - SP

Increased demand for energy and the search for alternative energy sources are remarkable and current facts. The management of domestic solid waste is also part of the issue in view of the possibility of power generation in existing landfills, which could provide financial autonomy to the system of waste management and improve the environmental balance of landfills in Brazil, currently only used for final solids disposal. Landfill gas is a viable alternative energy source has been tested in landfills in the World and in Brazil. Thus, the research aimed to evaluate the percentage of methane (CH4) present in three sinks of gases from a landfill in the municipality of Rio Claro – SP (Brazil), and to estimate the potential methane generation over its life through the application of a mathematical model. As a result it was found that in general the percentage of methane present in the gases is significant, between 50 and 62%. It was also noted that waste with disposal time between 8 and 12 months already are in the methanogenic stage. Finally, there was a tendency to increase the generation of methane in the transition periods of rain and drought

Estudo de dinâmicas populacionais aplicado a processos de tratamento biológico de efluentes

The purpose of this work is to perform studies of mathematical modeling of the relationship of interaction occurring between microrganisms participants from wastewater treatment processes aimed at understanding, through simulations, such as inter-relationships can affect the performance of such units. The methodology was the implementation in FORTRAN computer language of mathematical models of microbial interactions. The first model addresses the interaction of bacteria-forming flakes and filamentous bacteria in activated sludge systems, which seeks to strike a balance between these bacteria to improve efficiency of the process. Another model is studied the interaction between bacteria and protozoa in activated sludge systems and analyzing the efficiency of the process, observing the changes in daily load. Microbial interactions in anaerobic reactors were dealt a third model, in which there is the mutualistic interaction between acidogenic and methanogenic bacteria. In a fourth and final model was examined the relationship between the bacteria Acinetobacter sp. and Gordonia sp., which are present in activated sludge systems, showing the competitive capacity of Acinetobacter sp. can control the growth of unwanted bacteria.

Projeto de um sistema anaeróbico para estabilização de lodo e recuperação de energia

The treatment of domestic and industrial effluents through Wastewater Treatment Plants (WTPs) generates a residue termed sewage sludge, rich in organic matter, high-volume, occasionally containing pathogens and heavy metals. The sludge generation can minimize the benefits brought by the treatment of sewage, because this residue does not always receive appropriate treatment before final disposal. The disposal is another problem related to sludge. Landfills generally does not have physical space and alternatives such as the use in agriculture requires an intense treatment that could be in many cases operational or economic unfeasible. The objective of this work is the theoretical research about the processes of stabilization of the sludge by anaerobic digestion and the methanogenic activity during the process. Through analysis of each step and contemplating each relevant factor in anaerobic digestion process in order to optimize them, we proposed a theoretical model of reactors capable of stabilize the sludge, reduce its volume and eliminate pathogens. The obtained configuration consists of two anaerobic reactors connected in series. The first one operates in the range mesophilic temperature (35 ° C) and has higher hydraulic retention time (25 days) working primarily in the stabilization of organic matter present in the sludge and producing biogas, whereas the second one operates in the thermophilic range (55 ° C) in order to eliminate pathogens, and to reduce the volume. The hydraulic retention time in the second reactor is lower (10 days). Both mesophilic and thermophilic processes were efficient in what was proposed, promoting the stabilization of organic matter present in the sludge and significant reduction of pathogens. As a final step with the sludge previously digested, it is indicated a final dehydration... (Complete abstract click electronic access below)

Avaliação da auto-suficiência energética de reatores anaeróbios no tratamento de esgoto sanitário e de água preta

The purpose of this project is to verify whether anaerobic reactors applied to sewage treatment are energetically self-sufficient. This evaluation can be made by balancing the methane produced through the anaerobic transformation stages (hydrolysis, acidogenesis, acetogenesis and methanogenesis) and the reactor energy consumption requirements. The original project included methanogenic activity tests, which could not be performed due to setbacks in the installation of an analytical instrument. Scientific articles about bench- and full-scale anaerobic reactors were investigated instead. An average substrate-to-methane conversion efficiency of 58,2±18,6% was found for the bench-scale reactors and higher efficiencies (89,2%) were found for the cases which had higher Organic Loading Rates (OLRs) values. The average energy output was 0,013 kWh/Lsewage, value unable to meet the energy needs for the reactor operation, considering equipments normally used such as temperature controller. This balance can become positive if few hypotheses are made, for example (i) to eliminate the use of temperature controller (ii) to alter the operation pattern from continuous to intermittent. Based on energy balance assessment of eight bench-scale reactors, it was observed that the implementation of a system for biogas utilization is not energetically feasible. However, interesting results were found for a full-scale sewage treatment plant, ETE Ouro Verde – Foz do Iguaçu, PR, Brazil. Even though its substrate-tomethane conversion efficiency was about 10% only, the energy balance was quite positive, with energy consumption of 68 kWh/month and energy production of 660 kWh/month. This analysis leads us to conclude that energy recovery from full-scale sewage treatment plants should be practiced by other plants

Characterization of immobilized biomass by amplified rDNA restriction analysis (ARDRA) in an anaerobic sequencing-batch biofilm reactor (ASBBR) for the treatment of industrial wastewater

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Toxin-antitoxin systems and its biotechnological applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Analysis of a microbial community associated with polychlorinated biphenyl degradation in anaerobic batch reactors

The degradation of polychlorinated biphenyls (PCBs) was investigated under fermentativemethanogenic conditions for up to 60 days in the presence of anaerobic biomass from a full-scale UASB reactor. The low methane yields in the PCBs-spiked batch reactors suggested that the biomass had an inhibitory effect on the methanogenic community. Reactors containing PCBs and co-substrates (ethanol/ sodium formate) exhibited substantial PCB reductions from 0.7 to 0.2 mg mL-1 . For the Bacteria domain, the PCBs-spiked reactors were grouped with the PCB-free reactors with a similarity of 55 %, which suggested the selection of a specific population in the presence of PCBs. Three genera of bacteria were found exclusively in the PCB-spiked reactors and were identified using pyrosequencing analysis, Sedimentibacter, Tissierela and Fusibacter. Interestingly, the Sedimentibacter, which was previously correlated with the reductive dechlorination of PCBs, had the highest relative abundance in the RCS-PCB (7.4 %) and RCS-PCB-PF (12.4 %) reactors. Thus, the anaerobic sludge from the UASB reactor contains bacteria from the Firmicutes phylum that are capable of degrading PCBs.

Production of active recombinant eIF5A: reconstitution in E. coli of eukaryotic hypusine modification of eIF5A by its coexpression with modifying enzymes

Eukaryotic translation initiation factor 5A (eIF5A) is the only cellular protein that contains the polyamine-modified lysine, hypusine [Nε-(4-amino-2-hydroxybutyl)lysine]. Hypusine occurs only in eukaryotes and certain archaea, but not in eubacteria. It is formed post-translationally by two consecutive enzymatic reactions catalyzed by deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). Hypusine modification is essential for the activity of eIF5A and for eukaryotic cell proliferation. eIF5A binds to the ribosome and stimulates translation in a hypusine-dependent manner, but its mode of action in translation is not well understood. Since quantities of highly pure hypusine-modified eIF5A is desired for structural studies as well as for determination of its binding sites on the ribosome, we have used a polycistronic vector, pST39, to express eIF5A alone, or to co-express human eIF5A-1 with DHS or with both DHS and DOHH in Escherichia coli cells, to engineer recombinant proteins, unmodified eIF5A, deoxyhypusine- or hypusine-modified eIF5A. We have accomplished production of three different forms of recombinant eIF5A in high quantity and purity. The recombinant hypusine-modified eIF5A was as active in methionyl-puromycin synthesis as the native, eIF5A (hypusine form) purified from mammalian tissue. The recombinant eIF5A proteins will be useful tools in future structure/function and the mechanism studies in translation.

Estudo do papel da proteína eIF5A na elongação da tradução em S. cerevisiae: análise da relação funcional com o trna de alanina e sua participação na síntese proteica geral da célula

Pós-graduação em Biotecnologia - IQ

Attenuation of ruminal methanogenesis

Ruminal methanogens reduce carbon dioxide to methane (CH 4 ), thereby preventing hydrogen use by bacteria for VFA synthesis resulting in a 2 to 12% loss in feed gross energy. Methane is a greenhouse gas that contributes to global warming. The objectives of this work were to determine: (1) the extent to which ruminal cultures acquire resistance to a nitrofuranyl derivative of para-aminobenzoate (NFP) and an extract from the plant Yucca shidigera (Yucca); (2) the effect of distillers dried grains plus solubles (DDGS) on ruminal CH4 production; (3) the effect of brome hay-based diets, corn-based diets, and in vivo 2-bromoethansulfonate treatment on ruminal methane (CH4 ) production; and (4) the effect of the above treatments on the methanogen population. Ruminal cultures treated with NFP for 90 d maintained a diminished capacity to generate CH4 , but cultures became resistant to the inhibitory effects of Yucca treatment within 10 d. Both treatments decreased (P < 0.01) the relative abundance of total Archaea and the order Methanomicrobiales, but Yucca treatment increased (P < 0.01) the relative abundance of the order Methanobacteriales. The replacement of brome hay and corn with DDGS in lamb diets decreased (P < 0.01) and increased (P < 0.05), respectively, the amount of CH4 produced per unit of digested DM. The substitution of DDGS for brome hay increased (P < 0.01) the relative abundance of the order Methanomicrobiales. The replacement of brome hay with corn decreased (P < 0.05) the amount of CH4 produced per unit of digested DM, and also decreased (P < 0.05) the relative abundance of both Archaea and the order Methanomicrobiales. However, the abundance of the order Methanobacteriales increased (P < 0.05) as corn replaced brome hay. Intraruminal administration of 2-bromoethansulfonate decreased (P < 0.05) CH4 emissions, and decreased (P < 0.05) the relative abundance of Archaea and Methanobacteriales. In conclusion, NFP may be efficacious for chronically inhibiting ruminal methanogenesis, and the replacement of dietary forage with DDGS attenuates CH4 emissions from ruminant animals. Changes in domain- and order-specific ribosomal DNA indicators of methanogens are not consistently correlated with changes in CH4 production.

Amazonian Anthrosols Support Similar Microbial Communities that Differ Distinctly from Those Extant in Adjacent, Unmodified Soils of the Same Mineralogy

We compared the microbial community composition in soils from the Brazilian Amazon with two contrasting histories; anthrosols and their adjacent non-anthrosol soils of the same mineralogy. The anthrosols, also known as the Amazonian Dark Earths or terra preta, were managed by the indigenous pre-Colombian Indians between 500 and 8,700 years before present and are characterized by unusually high cation exchange capacity, phosphorus (P), and calcium (Ca) contents, and soil carbon pools that contain a high proportion of incompletely combusted biomass as biochar or black carbon (BC). We sampled paired anthrosol and unmodified soils from four locations in the Manaus, Brazil, region that differed in their current land use and soil type. Community DNA was extracted from sampled soils and characterized by use of denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism. DNA bands of interest from Bacteria and Archaea DGGE gels were cloned and sequenced. In cluster analyses of the DNA fingerprints, microbial communities from the anthrosols grouped together regardless of current land use or soil type and were distinct from those in their respective, paired adjacent soils. For the Archaea, the anthrosol communities diverged from the adjacent soils by over 90%. A greater overall richness was observed for Bacteria sequences as compared with those of the Archaea. Most of the sequences obtained were novel and matched those in databases at less than 98% similarity. Several sequences obtained only from the anthrosols grouped at 93% similarity with the Verrucomicrobia, a genus commonly found in rice paddies in the tropics. Sequences closely related to Proteobacteria and Cyanobacteria sp. were recovered only from adjacent soil samples. Sequences related to Pseudomonas, Acidobacteria, and Flexibacter sp. were recovered from both anthrosols and adjacent soils. The strong similarities among the microbial communities present in the anthrosols for both the Bacteria and Archaea suggests that the microbial community composition in these soils is controlled more strongly by their historical soil management than by soil type or current land use. The anthrosols had consistently higher concentrations of incompletely combusted organic black carbon material (BC), higher soil pH, and higher concentrations of P and Ca compared to their respective adjacent soils. Such characteristics may help to explain the longevity and distinctiveness of the anthrosols in the Amazonian landscape and guide us in recreating soils with sustained high fertility in otherwise nutrient-poor soils in modern times.

Spatial and Temporal Variation of Archaeal, Bacterial and Fungal Communities in Agricultural Soils

Background: Soil microbial communities are in constant change at many different temporal and spatial scales. However, the importance of these changes to the turnover of the soil microbial communities has been rarely studied simultaneously in space and time. Methodology/Principal Findings: In this study, we explored the temporal and spatial responses of soil bacterial, archaeal and fungal beta-diversities to abiotic parameters. Taking into account data from a 3-year sampling period, we analyzed the abundances and community structures of Archaea, Bacteria and Fungi along with key soil chemical parameters. We questioned how these abiotic variables influence the turnover of bacterial, archaeal and fungal communities and how they impact the long-term patterns of changes of the aforementioned soil communities. Interestingly, we found that the bacterial and fungal b-diversities are quite stable over time, whereas archaeal diversity showed significantly higher fluctuations. These fluctuations were reflected in temporal turnover caused by soil management through addition of N-fertilizers. Conclusions: Our study showed that management practices applied to agricultural soils might not significantly affect the bacterial and fungal communities, but cause slow and long-term changes in the abundance and structure of the archaeal community. Moreover, the results suggest that, to different extents, abiotic and biotic factors determine the community assembly of archaeal, bacterial and fungal communities.

Characterization of Immobilized Biomass by Amplified rDNA Restriction Analysis (ARDRA) in an Anaerobic Sequencing-Batch Biofilm Reactor (ASBBR) for the Treatment of Industrial Wastewater

The performance of an anaerobic sequencing-batch biofilm reactor (ASBBR-laboratory scale- 14L) containing biomass immobilized on coal was evaluated for the removal of elevated concentrations of sulfate (between 200 and 3,000 mg SO4-2.L-1) from industrial wastewater effluents. The ASBBR was shown to be efficient for removal of organic material (between 90% and 45%) and sulfate (between 95% and 85%). The microbiota adhering to the support medium was analyzed by amplified ribosomal DNA restriction analysis (ARDRA). The ARDRA profiles for the Bacteria and Archaea domains proved to be sensitive for the determination of microbial diversity and were consistent with the physical-chemical monitoring analysis of the reactor. At 3,000 mg SO4-2.L-1, there was a reduction in the microbial diversity of both domains and also in the removal efficiencies of organic material and sulfate.