Crude glycerol as feedstock for polyhydroxyalkanoates production by mixed microbial cultures.

The increase in global biodiesel production makes imperative the development of sustainable processes for the use of its main by-product, crude glycerol. In this study the feasibility of polyhydroxyalkanoates (PHA) production by a mixed microbial community using crude glycerol as feedstock was investigated. The selected culture had the ability to consume both glycerol and methanol fraction present in the crude. However, glycerol seemed to be the only carbon source contributing for the two biopolymers stored: poly-3-hydroxybutyrate (PHB) and glucose biopolymer (GB). In this work the culture reached a maximum PHB content of 47% (cdw) and a productivity of 0.27 g X/L.d, with an aerobic mixed cultures and a real waste substrate with non-volatile fatty acids (VFA) organic matter. The overall PHA yield on total substrate obtained was in the middle range of those reported in literature. The fact that crude glycerol can be used to produce PHA without any pre-treatment step, makes the overall production process economically more competitive, reducing polymer final cost.

Biopolymers production from mixed cultures and pyrolysis by-products.

Polyhydroxyalkanoates (PHAs) production from low value substrates and/or byproducts represents an economical and environmental promising alternative to established industrial manufacture methods. Bio-oil resulting from the fast-pyrolysis of chicken beds was used as substrate to select a mixed microbial culture (MMC) able to produce PHA under feast/famine conditions. In this study a maximum PHA content of 9.2% (g/g cell dry weight) was achieved in a sequencing batch reactor (SBR) operated for culture selection. The PHA obtained with bio-oil as a carbon source was a copolymer composed by 70% of hydroxybutyrate (HB) and 30% of hydroxyvalerate (HV) monomers. Similar results have been reported by other studies that use real complex substrates for culture selection indicating that bio-oil can be a promising feedstock to produce PHAs using MMC. To the best of our knowledge this is the first study that demonstrated the use of bio-oil resulting from fast pyrolysis as a possibly feedstock to produce short chain length polyhydroxyalkanoates.

Long-term operation of a reactor enriched in Accumulibacter clade I DPAOs: performance with nitrate, nitrite and oxygen.

The microbiology of denitrifying enhanced biological phosphorus removal systems has been a subject of much debate. The question has centred on the affinities of different types of Candidatus Accumulibacter PAOs, type I and type II, towards different electron acceptors such as oxygen, nitrate and nitrite. This study used a propionate anaerobic/anoxic/aerobic lab-scale sequencing batch reactor where a microbial culture was successfully enriched in Accumulibacter type I organisms (approx. 90%). The culture was able to take up phosphorus using nitrate, nitrite and oxygen as electron acceptors, although experiments with oxygen led to the fastest P removal rate. The phosphorus uptake to nitrogen consumed ratio (P/N ratio), when using both nitrate and nitrite, was shown to be affected by pH in the range of 7-8.2, achieving higher values for lower pH values (7.0-7.5). The effect of pH on P removal seems to follow a similar trend for both nitrate and nitrite. To our knowledge, this is the first study where the impact of pH in the phosphate removal stoichiometry using the three most significant electron acceptors is shown for such a high enrichment in Accumulibacter type I.

Influence of feeding strategies of mixed microbial cultures on the chemical composition and microstructure of copolyesters P(3HB-co-3HV) analyzed by NMR and statistical analysis.

NMR spectroscopy was applied for quantitative and qualitative characterization of the chemical composition and microstructure of a series of poly(3-hydroxybutyrate-co-3-hydoxyvalerate) copolymers, P(3HB-co-3HV), synthesized by mixed microbial cultures at several different feeding strategies. The monomer sequence distribution of the bacterially synthesized P(3HB-co-3HV) was defined by analysis of their high-resolution 1D (13)C NMR and 2D (1)H/(13)C HSQC and (1)H/(13)C HMBC NMR spectra. The results were verified by employment of statistical methods and suggest a block copolymer microstructure of the P(3HB-co-3HV) copolymers studied. Definitive distinction between block copolymers or a mixture of random copolymers could not be achieved. NMR spectral analysis indicates that the chemical composition and microstructure of the copolymers can be tuned by choosing a correct feeding strategy.

Kinetic and metabolic aspects of Defluviicoccus vanus-related organisms as competitors in EBPR systems.

A reactor was successfully enriched (90% as shown by Fluorescence in situ Hybridization) in Defluviicoccus vanus-related organisms presenting a Glycogen Accumulating Organisms (GAO) phenotype. Initial batch tests were performed using anaerobic/aerobic conditions to assess the capacity of different carbon sources utilization frequently abundant in wastewater: acetate, propionate, butyrate, valerate and glucose. Acetate and propionate were totally consumed in the anaerobic phase as well as butyrate and valerate, though these last ones with a very low consumption rate. All substrates were converted to polyhydroxyalkanoates (PHA). Glucose had a very slight anaerobic consumption but failed to disclose a typical GAO phenotype. In aerobic conditions, again all carbon sources were readily consumed except for glucose, with acetate and propionate having the higher consumption rates. Therefore, glucose seems not be used by this type of organisms. Acetate and propionate consumption rates indicated that these GAOs could reveal good competition advantages in EBPR systems where these carbon sources are available, especially propionate. Volatile Fatty Acid (VFA) uptake in aerobic phase and consequential PHA production indicate these organisms as possible candidates for PHA production.

Microbial community analysis with a high PHA storage capacity.

Activated sludge was submitted to aerobic dynamic substrate feeding for the production of biodegradable plastics. Two sequencing batch reactors were operated with acetate or propionate as sole carbon substrates. With acetate a homopolymer of polyhydroxybutyrate (PHB) was obtained and with propionate a copolymer of hydroxybutyrate and hydroxyvalerate P(HB/HV) was produced. Three main morphotypes were identified in both sludges: two belong to the Alphaproteobacteria class and the third to the Betaproteobacteria class. Bacilli belonging to Betaproteobacteria were shown by FISH analysis, applied in combination with Nile Blue post-staining, to be the main responsible for PHAs storage. The latter were affiliated to Azoarcus genus within Betaproteobacteria.

Polyhydroxyalkanoates production by activated sludge in a SBR using acetate and propionate as carbon sources.

In this work, sludge was submitted to aerobic dynamic substrate feeding. Two sequencing batch reactors were operated, with acetate or propionate as carbon substrates. When acetate was used the system only produced a homopolymer of polyhydroxybutyrate (PHB). In order to maximize the PHB production, tests with different concentrations of acetate and ammonia were preformed. The best results (67.2% of PHB by cell dry weight) were obtained for 0.7 Nmmol/l of ammonia and 180 Cmmol/l of acetate. The PHB cell content was further improved by pulse addition of substrate, three times 60 Cmmol/l of acetate, reaching a value of 78.5%. Propionate can be used as a precursor for hydroxyvalerate. In conjunction with other substrates, it allows for the formation of copolymers, which present better processing properties on commercial applications. Tests with different concentrations of propionate and ammonia were performed. Under the operating conditions used, the maximum PHA accumulated inside cells was 34.8%, with 30 Cmmol/l of propionate and no ammonia.

Production of polyhydroxyalkanoates by mixed microbial cultures.

Polyhydroxyalkanoates (PHAs) are biodegradable bioplastics formed from renewable resources, like sugars, with similar characteristics of polypropylene. These bioplastics are industrially produced by pure cultures using expensive pure substrates. These factors lead to a much higher selling price of PHAs compared to petroleum-based plastics, like polypropylene. The use of mixed cultures and cheap substrates (waste materials) can reduce costs of PHA production by more than 50%. Storage of PHAs by mixed populations occurs under transient conditions mainly caused by discontinuous feeding and variation in the electron donor/acceptor presence. In the last years the mechanisms of storage, metabolism and kinetics of mixed cultures have been studied. The maximum capacity of PHA storage and production rate is dependent on the substrate and on the operating conditions used. In this paper an overview and discussion of various mechanisms and processes for PHA production by mixed cultures is presented.

Production of polyhydroxyalkanoates by a mixed culture in a sequencing batch reactor: the use of propionate as carbon source.

In this work, sludge adapted to anaerobic/aerobic conditions, AN/AE, showing a high capacity of P accumulation, was submitted to aerobic dynamic substrate feeding (ADF). The fermenter was operated as a Sequencing Batch Reactor, with propionate as carbon substrate. Propionate is an important waste product from several industrial processes that can be valued, using it as a precursor for hydroxyvalerate in PHA production. Under the operational conditions used, apart from 3-hydroxyvalerate as its major component, 3-hydroxy-2-methylvalerate, 2-hydroxyisovalerate and 4-oxovalerate were also produced. A second reactor operated under the same conditions was adapted for the use of acetate as carbon substrate. The global metabolism of the organisms involved on PHA production, utilizing acetate or propionate, was studied using in vivo 13C Nuclear Magnetic Resonance (NMR).

Effect of pH control on EBPR stability and efficiency.

Biological phosphorus removal from water surfaces is an important process for the control of eutrophication. Even though much attention has been devoted to understanding the metabolism of bacteria and the effect of operational parameters on the efficiency of the phosphorus removal process, certain aspects are still unclear. The aim of this work was to evaluate the influence of pH control on the stability and efficiency of two sequenced batch reactors (SBR): one operated with pH control (pH 7.0) and the other without pH control (pH raised from around 7.8 to 8.5). The reactor operated without pH control showed higher efficiency on phosphorus removal and stability than the reactor with pH control. Based on the kinetics of both reactors it could be inferred that a different population developed in both systems.

Change in metabolism of PHA accumulation by activated sludge modifying operating conditions.

In the present work, sludge from a stable and efficient phosphorus removal process was used to evaluate the change in the metabolism when the reactor operation was modified from anaerobic/aerobic to aerobic dynamic substrate feeding. The change in operational conditions allowed the population to modify the metabolism of phosphorus and polyhydroxyalkanoates (PHA) accumulation. Under aerobic dynamic feeding, phosphorus accumulation was almost hindered while the amount of PHA accumulation increased significantly. After ten days of operation under dynamic substrate feeding, the reactor operation turned back to anaerobic/aerobic cycles and the metabolism of phosphorus was progressively recovered.

Glucose metabolism and kinetics of phosphorus removal by the fermentative bacterium Microlunatus phosphovorus.

Phosphorus and carbon metabolism in Microlunatus phosphovorus was investigated by using a batch reactor to study the kinetics of uptake and release of extracellular compounds, in combination with (31)P and (13)C nuclear magnetic resonance (NMR) to characterize intracellular pools and to trace the fate of carbon substrates through the anaerobic and aerobic cycles. The organism was subjected to repetitive anaerobic and aerobic cycles to induce phosphorus release and uptake in a sequential batch reactor; an ultrafiltration membrane module was required since cell suspensions did not sediment. M. phosphovorus fermented glucose to acetate via an Embden-Meyerhof pathway but was unable to grow under anaerobic conditions. A remarkable time shift was observed between the uptake of glucose and excretion of acetate, resulting in an intracellular accumulation of acetate. The acetate produced was oxidized in the subsequent aerobic stage. Very high phosphorus release and uptake rates were measured, 3.34 mmol g of cell(-1) h(-1) and 1.56 mmol g of cell(-1) h(-1), respectively, values only comparable with those determined in activated sludge. In the aerobic period, growth was strictly dependent on the availability of external phosphate. Natural abundance (13)C NMR showed the presence of reserves of glutamate and trehalose in cell suspensions. Unexpectedly, [1-(13)C]glucose was not significantly channeled to the synthesis of internal reserves in the anaerobic phase, and acetate was not during the aerobic stage, although the glutamate pool became labeled via the exchange with intermediates of the tricarboxylic acid cycle at the level of glutamate dehydrogenase. The intracellular pool of glutamate increased under anaerobic conditions and decreased during the aerobic period. The contribution of M. phosphovorus for phosphorus removal in wastewater treatment plants is discussed on the basis of the metabolic features disclosed by this study.

[Coronary-cavitary fistula. Embryogenesis and report of a case surgically treated]. / Fístula coronário-cavitária. Embriogênese e relato de um caso tratado cirurgicamente.

The authors report the case of a 6-year-old female patient with congenital left coronary artery-right cardiac chamber fistula. The anomalous communication began at the left main coronary artery and extended to the top of the heart, throughout the myocardial muscle thickness. Diagnosis done by 2-D echocardiogram and cineangiography. The patient underwent to surgical treatment and both sides of the anomalous fistula was closed with pericardium bovinus patch. There was found no arterial ventricular branch originary of the fistulas course. The patient had a good post-operative outcome and there was no clinical or laboratorial evidences of myocardial ischemia.

Effect of hydrogen sulfide on growth of sulfate reducing bacteria.

A culture of sulfate reducing bacteria (SRB) growing on lactate and sulfate was incubated at different pH values in the range of 5.8-7.0. The effect of pH on growth rate was determined in this pH range; the highest growth rate was observed at pH 6.7. Hydrogen sulfide produced from sulfate reduction was found to have a direct and reversible toxicity effect on the SRB. A hydrogen sulfide Concentration of 547 mg/L (16.1 mM) completely inhibited the culture growth. Comparison between acetic acid and hydrogen sulfide inhibition is presented and the concomitant inhibition kinetics are mathematically described.

Adult respiratory distress syndrome: evidence of early fibrogenesis and absence of glucocorticoid receptors.

In five lung biopsies from patients who developed the clinical picture of Adult Respiratory Distress Syndrome (ARDS) after cardiopulmonary bypass the pulmonary alterations were studied morphologically and glucocorticoid receptors determined. The time between cardiac surgery and pulmonary biopsy was 4-7 days. The results showed severe pulmonary lesions, with marked endothelial damage and active collagen secretion in the pulmonary interstitium, along the capillary bed. The glucocorticoid receptor level of the lungs with ARDS was 4.7 +/- 7.04 fmol/mg protein (control 4.28 +/- 4.32 fmol/mg protein). The results indicate that the process of collagen secretion may begin in the early phase of ARDS and that the benefits of glucocorticoid therapy in the management of ARDS should be reassessed.