Comparison between freeze and spray drying to obtain powder Rubrivivax gelatinosus biomass

The use of colorants in products of animal origin is justified by the improvement in the color of foods since this attribute is considered a quality criterion. These additives can be produced using industrial effluents as substrates and appropriate organisms, such as Rubrivivax gelatinosus. Oxycarotenoids represent a class of carotenes responsible for the pigmentation of animals and vegetables. R. gelatinosus grows in fish industry effluent with the resulting production of a bacterial biomass containing oxycarotenoids. The purpose of this study was to compare the use of two drying processes - spray and freeze drying - to obtain powder biomass in terms of the process parameters (yield, productivity, and product recovery) and the product characteristics (color, proximate composition, and oxycarotenoids). No difference was detected in the yield between these techniques, while productivity was higher using spray drying. Higher product recovery and moisture were achieved with freeze drying, while ash was higher with spray drying. The freeze dried biomass was redder, darker and less saturated than the spray dried biomass. No difference in oxycarotenoids was detected between the biomasses. Although it results in lower recovery rate, spray drying was faster and more productive, and it provided the same yield as freeze drying, which makes it the method of choice for obtaining R. gelatinosus biomass.

Estimating microalgae Synechococcus nidulans daily biomass concentration using neuro-fuzzy network

In this study, a neuro-fuzzy estimator was developed for the estimation of biomass concentration of the microalgae Synechococcus nidulans from initial batch concentrations, aiming to predict daily productivity. Nine replica experiments were performed. The growth was monitored daily through the culture medium optic density and kept constant up to the end of the exponential phase. The network training followed a full 3³ factorial design, in which the factors were the number of days in the entry vector (3,5 and 7 days), number of clusters (10, 30 and 50 clusters) and internal weight softening parameter (Sigma) (0.30, 0.45 and 0.60). These factors were confronted with the sum of the quadratic error in the validations. The validations had 24 (A) and 18 (B) days of culture growth. The validations demonstrated that in long-term experiments (Validation A) the use of a few clusters and high Sigma is necessary. However, in short-term experiments (Validation B), Sigma did not influence the result. The optimum point occurred within 3 days in the entry vector, 10 clusters and 0.60 Sigma and the mean determination coefficient was 0.95. The neuro-fuzzy estimator proved a credible alternative to predict the microalgae growth.

Spirulina platensis: process optimization to obtain biomass

Spirulina platensis is a photoautotrophic mesophilic cyanobacterium. Its main sources of nutrients are nitrate, urea, and ammonium salts. Spirulina cultivation requires temperature, light intensity, and nutrient content control. This microalgae has been studied and used commercially due to its therapeutic and antioxidant potential. In addition, several studies have reported its ability to use CO2, its immune activity, and use as an adjuvant nutritive factor in the treatment of obesity. The objective of this study is the production of biomass of S. platensis using different rates of stirring, nitrogen source, amount of micronutrients, and luminosity. A 2(4) experimental design with the following factors: stirring (120 and 140 RPM), amount of nitrogen (1.5 and 2.5 g/L), amount of micronutrients (0,25 and 0,75 mL/L) (11 and 15 W), and luminosity was used. Fermentation was performed in a 500 mL conical flask with 250 mL of culture medium and 10% inoculum in an incubator with controlled stirring and luminosity. Fermentation was monitored using a spectrophotometer (560 nm), and each fermentation lasted 15 days. Of the parameters studied, luminosity is the one with the highest significance, followed by the amount of nitrogen and the interaction between stirring and micronutrients. Maximum production of biomass for 15 days was 2.70 g/L under the following conditions: luminosity15W; stirring, 120 RPM; source of nitrogen, 1.5 g/L; and micronutrients, 0.75 mL/L.

Biomass production by Arthrospira platensis under different culture conditions

In biotechnological processes, the culture media components are responsible for high costs and exert a strong influence on the cyanobacteria behavior. The objective of this study was to evaluate the Arthrospira platensis growth potential for biomass production under different cultivation conditions using an experimental design. Three factors that are important for cyanobacteria growth were evaluated: sodium bicarbonate (9 to 18 g/l), sodium nitrate (1.25 to 2.5 g/l), and irradiance (20 to 120 µmol photons/m2.s–1). The results showed that the concentration of NaNO3 in the A. platensis medium can be reduced, resulting in increased concentrations of biomass produced. There was a higher biomass production due to the increase in the concentration of NaHCO3 and irradiance, mainly when these two factors varied tending towards the highest values studied. The results demonstrate the potential to produce Arthrospira platensis with lower costs and effluent generation without affecting cultivation performance.

Biomethanation of syngas: identification of metabolic pathways from CO in a natural anaerobic consortium

Au cours des dernières décennies, l’intérêt pour la gazéification de biomasses a considérablement augmenté, notamment en raison de la grande efficacité de recouvrement énergétique de ce procédé par rapport aux autres procédés de génération de bioénergies. Les composants majoritaires du gaz de synthèse, le monoxyde de carbone (CO) et l’hydrogène (H2) peuvent entre autres servir de substrats à divers microorganismes qui peuvent produire une variété de molécules chimiques d’intérêts, ou encore produire des biocarburants, particulièrement le méthane. Il est donc important d'étudier les consortiums méthanogènes naturels qui, en syntrophie, serait en mesure de convertir le gaz de synthèse en carburants utiles. Cette étude évalue principalement le potentiel de méthanisation du CO par un consortium microbien issu d’un réacteur de type UASB, ainsi que les voies métaboliques impliquées dans cette conversion en conditions mésophiles. Des tests d’activité ont donc été réalisés avec la boue anaérobie du réacteur sous différentes pressions partielles de CO variant de 0.1 à 1,65 atm (0.09 à 1.31 mmol CO/L), en présence ou absence de certains inhibiteurs métaboliques spécifiques. Dès le départ, la boue non acclimatée au CO présente une activité carboxidotrophique relativement intéressante et permet une croissance sur le CO. Les tests effectués avec de l’acide 2- bromoethanesulfonique (BES) ou avec de la vancomycine démontrent que le CO est majoritairement consommé par les bactéries acétogènes avant d’être converti en méthane par les méthanogènes acétotrophes. De plus, un plus grand potentiel de méthanisation a pu être atteint sous une atmosphère constituée uniquement de CO en acclimatant auparavant la boue. Cette adaptation est caractérisée par un changement dans la population microbienne désormais dominée par les méthanogènes hydrogénotrophes. Ceci suggère un potentiel de production à large échelle de biométhane à partir du gaz de synthèse avec l’aide de biofilms anaérobies.

Carboxydothermus hydrogenoformans comme catalyseur biologique pour la conversion du monoxyde de carbone en hydrogène simultanément a la minéralisation de calcium et phosphate

La gazéification est aujourd'hui l'une des stratégies les plus prometteuses pour valoriser les déchets en énergie. Cette technologie thermo-chimique permet une réduction de 95 % de la masse des intrants et génère des cendres inertes ainsi que du gaz de synthèse (syngaz). Le syngaz est un combustible gazeux composé principalement de monoxyde de carbone (CO), d'hydrogène (H2) et de dioxyde de carbone (CO2). Le syngaz peut être utilisé pour produire de la chaleur et de l'électricité. Il est également la pierre angulaire d'un grand nombre de produits à haute valeur ajoutée, allant de l'éthanol à l'ammoniac et l'hydrogène pur. Les applications en aval de la production de syngaz sont dictées par son pouvoir calorifique, lui-même dépendant de la teneur du gaz en H2. L’augmentation du contenu du syngaz en H2 est rendu possible par la conversion catalytique à la vapeur d’eau, largement répandu dans le cadre du reformage du méthane pour la production d'hydrogène. Au cours de cette réaction, le CO est converti en H2 et CO2 selon : CO + H2O → CO2 + H2. Ce processus est possible grâce à des catalyseurs métalliques mis en contact avec le CO et de la vapeur. La conversion catalytique à la vapeur d’eau a jusqu'ici été réservé pour de grandes installations industrielles car elle nécessite un capital et des charges d’exploitations très importantes. Par conséquent, les installations de plus petite échelle et traitant des intrants de faible qualité (biomasse, déchets, boues ...), n'ont pas accès à cette technologie. Ainsi, la seule utilisation de leur syngaz à faible pouvoir calorifique, est limitée à la génération de chaleur ou, tout au plus, d'électricité. Afin de permettre à ces installations une gamme d’application plus vaste de leurs syngaz, une alternative économique à base de catalyseur biologique est proposée par l’utilisation de bactéries hyperthermophiles hydrogénogènes. L'objectif de cette thèse est d'utiliser Carboxydothermus hydrogenoformans, une bactérie thermophile carboxydotrophe hydrogénogène comme catalyseur biologique pour la conversion du monoxyde de carbone en hydrogène. Pour cela, l’impact d'un phénomène de biominéralisation sur la production d’H2 a été étudié. Ensuite, la faisabilité et les limites de l’utilisation de la souche dans un bioréacteur ont été évaluées. Tout d'abord, la caractérisation de la phase inorganique prédominante lorsque C. hydrogenoformans est inoculé dans le milieu DSMZ, a révélé une biominéralisation de phosphate de calcium (CaP) cristallin en deux phases. L’analyse par diffraction des rayons X et spectrométrie infrarouge à transformée de Fourier de ce matériau biphasique indique une signature caractéristique de la Mg-whitlockite, alors que les images obtenues par microscopie électronique à transmission ont montré l'existence de nanotiges cristallines s’apparentant à de l’hydroxyapatite. Dans les deux cas, le mode de biominéralisation semble être biologiquement induit plutôt que contrôlé. L'impact du précipité de CaP endogène sur le transfert de masse du CO et la production d’H2 a ensuite été étudié. Les résultats ont été comparés aux valeurs obtenues dans un milieu où aucune précipitation n'est observée. Dans le milieu DSMZ, le KLa apparent (0.22 ± 0.005 min-1) et le rendement de production d’H2 (89.11 ± 6.69 %) étaient plus élevés que ceux obtenus avec le milieu modifié (0.19 ± 0.015 min-1 et 82.60 ± 3.62% respectivement). La présence du précipité n'a eu aucune incidence sur l'activité microbienne. En somme, le précipité de CaP offre une nouvelle stratégie pour améliorer les performances de transfert de masse du CO en utilisant les propriétés hydrophobes de gaz. En second lieu, la conversion du CO en H2 par la souche Carboxydothermus hydrogenoformans fut étudiée et optimisée dans un réacteur gazosiphon de 35 L. Parmi toutes les conditions opérationnelles, le paramètre majeur fut le ratio du débit de recirculation du gaz sur le débit d'alimentation en CO (QR:Qin). Ce ratio impacte à la fois l'activité biologique et le taux de transfert de masse gaz-liquide. En effet, au dessus d’un ratio de 40, les performances de conversion du CO en H2 sont limitées par l’activité biologique alors qu’en dessous, elles sont limitées par le transfert de masse. Cela se concrétise par une efficacité de conversion maximale de 90.4 ± 0.3 % et une activité spécifique de 2.7 ± 0.4 molCO·g–1VSS·d–1. Malgré des résultats prometteurs, les performances du bioréacteur ont été limitées par une faible densité cellulaire, typique de la croissance planctonique de C. hydrogenoformans. Cette limite est le facteur le plus contraignant pour des taux de charge de CO plus élevés. Ces performances ont été comparées à celles obtenues dans un réacteur à fibres creuses (BRFC) inoculé par la souche. En dépit d’une densité cellulaire et d’une activité volumétrique plus élevées, les performances du BRFC à tout le moins cinétiquement limitées quand elles n’étaient pas impactées par le transfert de masse, l'encrassement et le vieillissement de la membrane. Afin de parer à la dégénérescence de C. hydrogenoformans en cas de pénurie de CO, la croissance de la bactérie sur pyruvate en tant que seule source de carbone a été également caractérisée. Fait intéressant, en présence simultanée de pyruvate et de CO, C. hydrogenoformans n’a amorcé la consommation de pyruvate qu’une fois le CO épuisé. Cela a été attribué à un mécanisme d'inhibition du métabolisme du pyruvate par le CO, faisant ainsi du pyruvate le candidat idéal pour un système in situ de secours.

Evaluation of cyanobacteria for biomass production and effluent treatment

The microalgae gained importance as food and feed as well as source of fine chemicals since the l960’s. Spirulina became the trend setter due to its easily culturable properties as well as nutritional composition. A rapid expansion of microalgal industry occurred in the Asia-Pacific region as microalgae came to stay as a health food supplement. Microalgae have been an integral component of oxidation ponds usually incorporated with wastewater treatment. Over the last few decades, efforts have been made to apply intensive microalgal cultures to perform biological tertiary treatment of secondary effluents. Given the limited number of species still available for commercial exploitation, it is imperative to isolate and cultivate those photosynthetic organisms with high growth rate and biomass accumulation, which could play the dual role of cleaning the wastewater and also providing useful biomass. This has been the objective of this study ie. 0 To develop pure cultures of local isolates of Cyanobacteria for extraction of biochemicals of commercial value 0 To couple biomass production with effluent treatment

“Studies on glucose tolerant β-glucosidases from a novel Byssochlamys fulva and their applications in biomass to ethanol conversion”

Beta-glucosidases are critical enzymes in biomass hydrolysis process and is important in creating highly efficient enzyme cocktails for the bio-ethanol industry. Among the two strategies proposed for overcoming the glucose inhibition of commercial cellulases, one is to use heavy dose of BGL in the enzyme blends and the second is to do simultaneous saccharification and fermentation where glucose is converted to alcohol as soon as it is being generated. While the former needs extremely high quantities of enzyme, the latter is inefficient since the conditions for hydrolysis and fermentation are different. This makes the process technically challenging and also in this case, the alcohol generation is lesser, making its recovery difficult. A third option is to use glucose tolerant β-glucosidases which can work at elevated glucose concentrations. However, there are very few reports on such enzymes from microbial sources especially filamentous fungi which can be cultivated on cheap biomass as raw material. There has been very less number of studies directed at this, though there is every possibility that filamentous fungi that are efficient degraders of biomass may harbor such enzymes. The study therefore aimed at isolating a fungus capable of secreting glucose tolerant β- glucosidase enzyme. Production, characterization of β-glucosidases and application of BGL for bioethanol production were attempted.

Biobutanol from lignocellulosic biomass by a novel Clostridium sporogenes BE01

In the current study, a novel non-acetone forming butanol and ethanol producer Was isolated and identified. Based on the 16s rDNA sequence BLAST and phylogenetic analyses, it was found to have high similarity with the reported hydrogen producing strains of Clostridium sporogenes. Biochemical studies revealed that it is lipase and protease positive. The lipolytic and proteolytic properties are the very important characteristics of Clostridium sporogenes. Sugar utilization profile studies were positive for glucose, saccharose, cellobiose and weakly positive result to xylose. This study demonstrated C. sporogenes BE01, an isolate from NIIST is having potential to compete with existing, well known butanol producers with the advantage of no acetone in the final solvent mixture. Rice straw hydrolysate is a potent source of substrate for butanol production by C. sporogenes BE01. Additional supplementation of vitamins and minerals were avoided by using rice straw hydrolysate as substrate. Its less growth, due to the inhibitors present in the hydrolysate and also inhibition by products resulted in less efficient conversion of sugars to butanol. Calcium carbonate played an important role in improving the butanol production, by providing the buffering action during fermentation and stimulating the electron transport mediators and redox reactions favoring butanol production. Its capability to produce acetic acid, butyric acid and hydrogen in significant quantities during butanol production adds value to the conversion process of lignocellulosic biomass to butanol. High cell density fermentation by immobilizing the cells on to ceramic particles improved the solvents and VFA production. Reduced sugar utilization from the concentrated hydrolysate could be due to accumulation of inhibitors in the hydrolysate during concentration. Two-stage fermentation was very efficient with immobilized cells and high conversions of sugars to solvents and VFAs were achieved. The information obtained from the study would be useful to develop a feasible technology for conversion of lignocellulosic biomass to biobutanol.

Effects of plant diversity on bioenergy parameters in grassland biomass

Extensive grassland biomass for bioenergy production has long been subject of scientific research. The possibility of combining nature conservation goals with a profitable management while reducing competition with food production has created a strong interest in this topic. However, the botanical composition will play a key role for solid fuel quality of grassland biomass and will have effects on the combustion process by potentially causing corrosion, emission and slagging. On the other hand, botanical composition will affect anaerobic digestibility and thereby the biogas potential. In this thesis aboveground biomass from the Jena-Experiment plots was harvested in 2008 and 2009 and analysed for the most relevant chemical constituents effecting fuel quality and anaerobic digestibility. Regarding combustion, the following parameters were of main focus: higher heating value (HHV), gross energy yield (GE), ash content, ash softening temperature (AST), K, Ca, Mg, N, Cl and S content. For biogas production the following parameters were investigated: substrate specific methane yield (CH4 sub), area specific methane yield (CH4 area), crude fibre (CF), crude protein (CP), crude lipid (CL) and nitrogen-free extract (NfE). Furthermore, an improvement of the fuel quality was investigated through applying the Integrated generation of solid Fuel and Biogas from Biomass (IFBB) procedure. Through the specific setup of the Jena-Experiment it was possible to outline the changes of these parameters along two diversity gradients: (i) species richness (SR; 1 to 60 species) and (ii) functional group (grasses, legumes, small herbs and tall herbs) presence. This was a novel approach on investigating the bioenergy characteristic of extensive grassland biomass and gave detailed insight in the sward-composition¬ - bioenergy relations such as: (i) the most relevant SR effect was the increase of energy yield for both combustion (annual GE increased by 26% from SR8→16 and by 65% from SR8→60) and anaerobic digestion (annual CH4 area increased by 22% from SR8→16 and by 49% from SR8→60) through a strong interaction of SR with biomass yield; (ii) legumes play a key role for the utilization of grassland biomass for energy production as they increase the energy content of the substrate (HHV and CH4 sub) and the energy yield (GE and CH4 area); (iii) combustion is the conversion technique that will yield the highest energy output but requires an improvement of the solid fuel quality in order to reduce the risk of corrosion, emission and slagging related problems. This was achieved through applying the IFBB-procedure, with reductions in ash (by 23%), N (28%), K (85%), Cl (56%) and S (59%) and equal levels of concentrations along the SR gradient.

Effects of grassland conversion and tillage intensities on soil microbial biomass, residues and community structure

Agricultural intensification has a strong impact on level of soil organic matter (SOM), microbial biomass stocks and microbial community structure in agro-ecosystems. The size of the microbial necromass C pool could be about 40 times that of the living microbial biomass C pool in soils. Due to the specificity, amino sugar analysis gives more important information on the relative contribution of fungal and bacterial residues to C sequestration potential of soils. Meanwhile, the relationship between microbial biomass and microbial necromass in soil and its ecological significance on SOM are not fully understood and likely to be very complex in grassland soils. This thesis focuses on the effects of tillage, grassland conversion intensities and fertilisation on microbial biomass, residues and community structure. The combined analyses of microbial biomass and residue formation of both fungi and bacteria provided a unique opportunity to study the effect of tillage, grassland conversion and fertilisation on soil microbial dynamics. In top soil at 0-30 cm layer, a reduction in tillage intensity by the GRT and NT treatments increased the accumulation of saprotrophic fungi in comparison with the MBT treatment. In contrast, the GRT and NT treatments promoted AMF at the expense of saprotrophic fungi in the bottom soil layer at 30-40 cm depth. The negative relationship between the ergosterol to microbial biomass C ratio and the fungal C to bacterial C ratio points to the importance of the relationship between saprotrophic fungi and biotrophic AMF for tillage-induced changes in microbial turnover of SOC. One-season cultivation of winter wheat with two tillage events led to a significant loss in SOC and microbial biomass C stocks at 0-40 cm depth in comparison with the permanent grassland, even 5 years after the tillage event. However, the tillage induced loss in microbial biomass C was roughly 40% less in the long-term than in the short-term of the current experiment, indicating a recovery process during grassland restoration. In general, mould board tillage and grassland conversion to maize monoculture promoted saprotrophic fungi at the expense of biotrophic AMF and bacteria compared to undisturbed grassland soils. Slurry application promoted bacterial residues as indicated by the decreases in both, the ergosterol to microbial biomass C ratio and the fungal C to bacterial C ratio. In addition, the lost microbial functional diversity due to tillage and maize monoculture was restored by slurry application both in arable and grassland soils. I conclude that the microbial biomass C/S ratio can be used as an additional indicator for a shift in microbial community. The strong relationships between microbial biomass and necromass indices points to the importance of saprotrophic fungi and biotrophic AMF for agricultural management induced effects on microbial turnover and ecosystem C storage. Quantitative information on exact biomass estimates of these two important fungal groups in soil is inevitably necessary to understand their different roles in SOM dynamics.

Modeled and observed atmospheric radiation balance during the West African dry season: role of mineral dust, biomass burning aerosol, and surface albedo

The global radiation balance of the atmosphere is still poorly observed, particularly at the surface. We investigate the observed radiation balance at (1) the surface using the ARM Mobile Facility in Niamey, Niger, and (2) the top of the atmosphere (TOA) over West Africa using data from the Geostationary Earth Radiation Budget (GERB) instrument on board Meteosat-8. Observed radiative fluxes are compared with predictions from the global numerical weather prediction (NWP) version of the Met Office Unified Model (MetUM). The evaluation points to major shortcomings in the NWP model's radiative fluxes during the dry season (December 2005 to April 2006) arising from (1) a lack of absorbing aerosol in the model (mineral dust and biomass burning aerosol) and (2) a poor specification of the surface albedo. A case study of the major Saharan dust outbreak of 6–12 March 2006 is used to evaluate a parameterization of mineral dust for use in the NWP models. The model shows good predictability of the large-scale flow out to 4–5 days with the dust parameterization providing reasonable dust uplift, spatial distribution, and temporal evolution for this strongly forced dust event. The direct radiative impact of the dust reduces net downward shortwave (SW) flux at the surface (TOA) by a maximum of 200 W m−2 (150 W m−2), with a SW heating of the atmospheric column. The impacts of dust on terrestrial radiation are smaller. Comparisons of TOA (surface) radiation balance with GERB (ARM) show the “dusty” forecasts reduce biases in the radiative fluxes and improve surface temperatures and vertical thermodynamic structure.