Produção e caracterização de polihidroxialcanoatos obtidos por fermentação da glicerina bruta residual do biodiesel

The aim of this study was to evaluate the production of polyhydroxyalkanoates (PHAs) by fermentation of Crude Glycerin, a byproduct of the biodiesel industry, by Cupriavidus necator IPT 026, 027 and 028. The influence of fermentation time and temperature in shake flasks were evaluated. The highest PHA production (2.82 g L-1) occurred at 35 ºC for 72 h of fermentation. The melting and initial thermal degradation temperatures of this PHA were 177.9 ºC and 306.33 ºC, respectively, with 55% crystallinity. FTIR spectrum was similar to those reported in literature. The polymer obtained presented three different methyl esters of hydroxyalkanoates in its composition, with molecular weight of 630 kDa. Bacteria can use Crude Glycerin as an inexpensive substrate to produce value-added biodegradable products, such as PHA.

Otimização da etapa de extração do ácido clavulânico presente no caldo de fermentação utilizando misturas de solventes

The purpose of this work was to study four different solvent mixtures intended to increase the yield of the extraction stage of clavulanic acid (CA), which is one of the steps in the purification process. Four central composite rotatable designs (CCRD) were utilized to optimize the solvent mixtures. The variables selected for the factorial design were solvent mixture ratio (mL/mL) and temperature (ºC). The results showed that the yield of CA extracted from fermentation broth with the solvent mixtures of methyl-ethyl-ketone and ethyl acetate, and methyl-isobutyl-ketone and ethyl acetate (44.7 and 50.0%, respectively) was higher than that of the individual ethyl acetate alone (36.5%).

Cytopathological characterization of Mal de Río Cuarto virus in corn, wheat and barley

The Mal de Río Cuarto disease is caused by Mal de Río Cuarto virus (MRCV) transmitted by Delphacodes kuscheli. Comparative studies were carried out on the cytopathological alterations produced by MRCV in corn (Zea mays), wheat (Triticum aestivum) and barley (Hordeum vulgare), as seen with a transmission electron microscope. Corn plants were infected with viruliferous D. kuscheli collected from the endemic disease area (i.e. Río Cuarto County, Córdoba, Argentina). For the viral transmission to small grain cereal plants, laboratory rared insects were used. In this case, the inoculum source was wheat and barley plants infected with MRCV isolate grown in a greenhouse. Leaf samples with conspicuous symptoms were collected: enations and size reduction in corn; crenatures, swelling veins and dark green color in small grain cereals. Viral infection was corroborated by DAS-ELISA. Viroplasms containing complete and incomplete virus particles and fibrillar material were found in the cytoplasm of infected cells in all species. Mature virions were between 60 and 70 nm diameter. In wheat and barley, viroplasms and dispersed particles were observed only in phloem, while in corn virions were also found in cells of the bundle sheath. Crystalline arrays of particles were detected in corn enation constitutive cells. Tubular inclusions were found only in wheat samples. The three species showed abnormalities in the chloroplasts of affected cells. The results showed that MRCV cytopathology has similarities with other viruses from the genus Fijivirus, family family Reoviridae, but slight differences depending upon the host plant.

Doenças do milho causadas por fungos do gênero Stenocarpella

Os fungos Stenocarpella macrospora e Stenocarpella maydis podem causar podridão de semente, morte de plântula, podridão da base do colmo e da espiga e mancha foliar em milho, sendo comumente detectados nos grãos ardidos quando as espigas são infectadas. Os danos no rendimento e na qualidade de grãos causados especificamente por Stenocarpella ainda não foram devidamente quantificados. Os patógenos são encontrados praticamente em todas as regiões produtoras de milho do Brasil. A doença ocorre com maior intensidade em lavouras de monocultura, principalmente em pequenas propriedades rurais, e em lavouras produtoras de semente, onde o cereal é freqüentemente cultivado na mesma área. Os restos culturais de milho e as sementes infectadas constituem-se na principal fonte de inóculo primário. O inóculo constituído de picniosporos, produzido nos resíduos culturais, é disseminado à curta distância pelo vento e respingos d'água. A disseminação à longa distância ocorre pelas sementes. A infecção das plantas pode ser sistêmica, com inóculo na forma de micélio oriundo das sementes, e/ou pela penetração direta com germinação dos conídios nos sítios de infecção, como bainha foliar, lâmina foliar, pedúnculo e palha da espiga. A temperatura ótima para crescimento do micélio e germinação dos conídios de ambas espécies ocorre na faixa de 23 ºC a 28 ºC. As principais estratégias de controle baseiam-se na eliminação e/ou redução da fonte de inóculo primário, como uso de semente sadia, tratamento de sementes com fungicida do grupo dos benzimidazóis e rotação de culturas. Pouca informação existe sobre a tolerância da doença em híbridos comerciais no Brasil. A adubação equilibrada do solo e evitar elevada densidade de plantas também podem reduzir a infecção.

Feasibility study for acidic biorefinery concept

The integration a recovery process for spent sulfite liquor from a sulfite cooking process with a kraft pulp mill was studied in this thesis work. The process includes a fermentation process for the production of biofuel. The calculation for three cases was done. The three cases considered were with a spruce sulfite pulp production of 100, 250 and 1000 Adt/day corresponding to 10, 25 and 100% of the total pulp production. A kraft cooking process with prehydrolysis was taken in consideration as reference. Compared to kraft cooking with prehydrolysis a bigger amount of ethanol can be produced by sulfite cooking. In the kraft prehydrolysis case 40 t/day of ethanol and 1000 Adt/day pine pulp is produced and in the sulfite case the production is 113 t/day of ethanol and 1000 Adt/day of spruce pulp. The energy consumption in the sulfite process is assumed to be slightly higher than in the kraft prehydrolysis process. The recovery system for spent liquor mix was studied. The evaporation of the spent cooking liquors should be done separately. The approximately composition of melt was calculated. A comparison of all four cases was done and profitability was estimated.

Kuivurin pölynhallintajärjestelmä

Lämminilmakuivaus on viljan kuivaamisen menetelmistä ylivoimaisesti käytetyin. Siinä kuivurissa olevan viljakerroksen läpi puhalletaan lämmintä ilmaa, kunnes viljan kosteuspitoisuus laskee tavoitetasoon. Viljan jyvistä irtoaa aina pölyä niiden hangatessa toisiaan vasten. Viljankuivaamoiden pölyisyyttä on normaalisti pyritty vähentämään poistamalla leijuvaa pölyä viljan liikkeen epäjatkuvuuskohdista. Pölyä kulkeutuu kuitenkin kuivaamon ulkopuolelle myös kuivaukseen käytetyn ilman mukana. Poistoilman pölypitoisuus ei tavallisesti ole korkea, mutta kuivaamiseen käytetyistä suurista ilmamääristä johtuen pölyä voi kertyä poistoilmakanavan ympäristöön huomattavia määriä. Tämän diplomityön tavoitteena onkin viljankuivurin poistoilman pölypitoisuuden vähentämiseen soveltuvan pölynhallintajärjestelmän kehittäminen. Selvityksessä tarkasteltiin ensinnäkin kuivurin poistoilman pölypitoisuuteen liittyvää kansallista ja kansainvälistä lainsäädäntöä sekä raja-arvoja. Lähdemateriaalin perusteella kartoitettiin pölypitoisuuden vähentämiseen soveltuvia menetelmiä sekä kilpailijoiden toteuttamia ratkaisuja. Myös muita pölynhallintajärjestelmän edellytyksiä selvitettiin. Teoreettisessa tarkastelussa teknistaloudellisesti parhaaksi osoittautunut pölynhallintaratkaisu jalostettiin prototyypiksi. Sen toimintakykyä testattiin erotustehokkuuden mittaamiseen soveltuvan koelaitteiston avulla. Testitulosten perusteella kyseinen pölynhallintajärjestelmä todettiin toimivaksi ja pienellä jatkokehittelyllä myös kaupalliseen käyttöön soveltuvaksi. Testitulosten hyödyntämismahdollisuuksien lisäksi diplomityössä esitetään myös muutamia jatkokehitysehdotuksia.

CFD Simulation of Two-phase and Three-phase Flows in Internal-loop Airlift

Airlift reactors are pneumatically agitated reactors that have been widely used in chemical, petrochemical, and bioprocess industries, such as fermentation and wastewater treatment. Computational Fluid Dynamics (CFD) has become more popular approach for design, scale-up and performance evaluation of such reactors. In the present work numerical simulations for internal-loop airlift reactors were performed using the transient Eulerian model with CFD package, ANSYS Fluent 12.1. The turbulence in the liquid phase is described using κ- ε the model. Global hydrodynamic parameters like gas holdup, gas velocity and liquid velocity have been investigated for a range of superficial gas velocities, both with 2D and 3D simulations. Moreover, the study of geometry and scale influence on the reactor have been considered. The results suggest that both, geometry and scale have significant effects on the hydrodynamic parameters, which may have substantial effects on the reactor performance. Grid refinement and time-step size effect have been discussed. Numerical calculations with gas-liquid-solid three-phase flow system have been carried out to investigate the effect of solid loading, solid particle size and solid density on the hydrodynamic characteristics of internal loop airlift reactor with different superficial gas velocities. It was observed that averaged gas holdup is significantly decreased with increasing slurry concentration. Simulations show that the riser gas holdup decreases with increase in solid particle diameter. In addition, it was found that the averaged solid holdup increases in the riser section with the increase of solid density. These produced results reveal that CFD have excellent potential to simulate two-phase and three-phase flow system.

Thermal behavior of corn starch granules modified by acid treatment at 30 and 50°C

Unprocessed native starches are structurally too weak and functionally too restricted for application in today's advanced food technologies. Processing is necessary to engender a range of functionality. Naturals or natives starches can be modified by using several methods physical, chemical, enzymatic or combined, according industrial purposes. In this work, native corn starch was hydrolyzed by hydrochloric acid solution and investigated by using thermoanalytical techniques (thermogravimetry - TG, differential thermal analysis - DTA and differential scanning calorimetry - DSC), as well as optical microscopy and X-ray diffractometry. After acid treatment at 30 and 50°C, a decrease of gelatinization enthalpy (ΔHgel) was verified. Optical microscopy and X-ray diffractometry allowed us to verify the granules contorn and rugosity typical of cereal starches.

Adsorptive removal of harmful organic compound from aqueous solutions

Adsorption is one of the most commonly used methods in water treatment processes. It is attractive due to it easy operation and the availability of a wide variety of commercial adsorbents. This doctoral thesis focuses on investigating and explaining the influence of external phase conditions (temperature, pH, ionic strength, acidity, presence of cosolutes) on adsorption phenomena. In order to cover a wide range of factors and phenomena, case studies were chosen from various fields where adsorption is applied. These include the adsorptive removal of surface active agents (used in cleaning chemicals, for example) from aqueous effluents, the removal of hormones (estradiol) from drinking water, and the adsorption of antibiotics onto silica. The latter can beused to predict the diffusion of antibiotics in the aquatic system if they are released into the environment. Also the adsorption of living cells on functionalized polymers to purify infected water streams was studied. In addition to these examples, the adsorptive separation of harmful compounds from internal water streams within a chemical process was investigated. The model system was removal of fermentation inhibitors from lignocelluloses hydrolyzates. The detoxification of the fermentation broth is an important step in the manufacture of bioethanol from wood, but has not been studied previously in connection with concentrated acid hydrolyzates. New knowledge on adsorption phenomena was generated for all of the applications investigated. In most cases, the results could be explained by combining classical theories for individual phenomena. As an example, it was demonstrated how liquid phase aggregation could explain abnormal-looking adsorption equilibrium data. In addition to the fundamental phenomena, also process performance was of interest. This aspect is often neglected in adsorption studies. It was demonstrated that adsorbents should not be selected for a target application based on their adsorption properties only, but regeneration of the spent adsorbent must be considered. It was found that using a suitable amount of organic co-solvent in the regeneration can significantly improve the productivity of the process.

Identifying and Characterizing New Ingredients in vitro for Prebiotic and Synbiotic use

The endogenous microbiota, constituting the microbes that live inside and on humans, is estimated to outnumber human cells by a factor of ten. This commensal microbial population has an important role in many physiological functions, with the densest microbiota population found in the colon. The colonic microbiota is a highly complex and diverse bacterial ecosystem, and a delicate balance exists between the gut microbiota and its host. An imbalance in the microbial ecosystem may lead to severe symptoms in and also beyond the gastrointestinal tract. Due to the important role of the gut microbiota in human health, means of its modification have been introduced in the dietary concepts of pro-, pre- and synbiotics. Prebiotics, which are usually carbohydrates, strive to selectively influence beneficial microbes resident in the colon with the aim of modifying the composition and functionality of the commensal microbial population towards a purportedly healthier one. The study of prebiotic effects on colonic micro-organisms is typically done by using human faecal material, though this provides relatively little information on bacterial populations and metabolic events in different parts of the colon. For this reason, several in vitro models have been developed to investigate the gut microbiota. The aim of this doctoral thesis was to screen through some of the promising prebiotic candidates, characterize their effects on the microbiota through the use of two in vitro methods (pure microbial cultures and a colon simulator model) and to evaluate their potential as emerging prebiotics or synbiotics when combined with the probiotic Bifidobacterium lactis . As a result of the screening work and subsequent colon simulation studies, several compounds with promising features were identified. Xylo-oligosaccharides (XOS), which have previously already shown promise as prebiotic compounds, were well fermented by several probiotic Bifidobacterium lactis strains in pure culture studies and in the following simulation studies utilizing the complex microbiota by endogenous B. lactis Another promising compound was panose, a trisaccharide belonging to isomalto-oligosaccharides (IMO) that also was also able to modify the microbiota in vitro by increasing the number of beneficial microbes investigated. Panose has not been widely studied previously and therefore, this thesis work provided the first data on panose fermentation in mixed colonic microbiota. Galacto-oligosaccharide (GOS) is an established prebiotic, and it was studied here in conjunction with another potential polygosaccharide polydextrose (PDX) and probiotic B. lactis Bi-07. In this final study, the synbiotics including GOS were more effective than the constituting pro- or prebiotics alone in modulating the microbiota composition, thus indicating a synergy resulting from the combination. The results obtained in this in vitro work can be, and have already been, utilized in product development aimed at the nutritional modification of the human colonic microbiota. Some of the compounds have entered the human clinical intervention phase to nvestigate in more detail the prebiotic and synbiotic properties seen in these in vitro studies.

Antimicrobial activity of extracellular metabolites from antagonistic bacteria isolated from potato (Solanum phureja) crops

Microorganisms for biological control are capable of producing active compounds that inhibit the development of phytopathogens, constituting a promising tool toob tain active principles that could replace synthetic pesticides. This study evaluatedtheability of severalpotentialbiocontrol microorganismsto produce active extracellular metabolites. In vitro antagonistic capability of 50 bacterial isolates from rhizospheric soils of "criolla" potato (Solanum phureja) was tested through dual culture in this plant with different plant pathogenic fungi and bacteria. Isolates that showed significantly higher antagonistic activity were fermented in liquid media and crude extracts from the supernatants had their biological activities assessed by optical density techniques. Inhibitory effecton tested pathogens was observed for concentrations between 0.5% and 1% of crude extracts. There was a correlation between the antimicrobial activity of extracts and the use of nutrient-rich media in bacteria fermentation. Using a bioguided method, a peptidic compound, active against Fusarium oxysporum, was obtained from the 7ANT04 strain (Pyrobaculum sp.). Analysis by nuclear magnetic resonance and liquid chromatography coupled to mass detector evidenced an 11-amino acid compound. Bioinformatic software using raw mass data confirmed the presence of a cyclic peptide conformed by 11 mostly non-standard amino acids.

Lethal weapons : novel approaches for receptor-targeted cancer cell elimination

The currently used forms of cancer therapy are associated with drug resistance and toxicity to healthy tissues. Thus, more efficient methods are needed for cancer-specific induction of growth arrest and programmed cell death, also known as apoptosis. Therapeutic forms of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) are investigated in clinical trials due to the capability of TRAIL to trigger apoptosis specifically in cancer cells by activation of cell surface death receptors. Many tumors, however, have acquired resistance to TRAIL-induced apoptosis and sensitizing drugs for combinatorial treatments are, therefore, in high demand. This study demonstrates that lignans, natural polyphenols enriched in seeds and cereal, have a remarkable sensitizing effect on TRAIL-induced cell death at non-toxic lignan concentrations. In TRAIL-resistant and androgen-dependent prostate cancer cells we observe that lignans repress receptor tyrosine kinase (RTK) activity and downregulate cell survival signaling via the Akt pathway, which leads to increased TRAIL sensitivity. A structure-activity relationship analysis reveals that the γ-butyrolactone ring of the dibenzylbutyrolactone lignans is essential for the rapidly reversible TRAIL-sensitizing activity of these compounds. Furthermore, the lignan nortrachelogenin (NTG) is identified as the most efficient of the 27 tested lignans and norlignans in sensitization of androgen-deprived prostate cancer cells to TRAIL-induced apoptosis. While this combinatorial anticancer approach may leave normal cells unharmed, several efficient cancer drugs are too toxic, insoluble or unstable to be used in systemic therapy. To enable use of such drugs and to protect normal cells from cytotoxic effects, cancer-targeted drug delivery vehicles of nanometer scale have recently been generated. The newly developed nanoparticle system that we tested in vitro for cancer cell targeting combines the efficient drug-loading capacity of mesoporous silica to the versatile particle surface functionalization of hyperbranched poly(ethylene imine), PEI. The mesoporous hybrid silica nanoparticles (MSNs) were functionalized with folic acid to promote targeted internalization by folate receptor overexpressing cancer cells. The presented results demonstrate that the developed carrier system can be employed in vitro for cancer selective delivery of adsorbed or covalently conjugated molecules and furthermore, for selective induction of apoptotic cell death in folate receptor expressing cancer cells. The tested carrier system displays potential for simultaneous delivery of several anticancer agents specifically to cancer cells also in vivo.

Roal Oy:n käyttöhyödykejärjestelmän arviointi, optimointi ja raportointi

Prosessiteollisuudessa tarvitaan usein erilaisia apujärjestelmiä pääprosessin tueksi. Tyypillisiä tällaisia järjestelmiä ovat jäähdytys-, höyry- ja ilmajärjestelmät. Hyödykejärjestelmien kehitys jää helposti pääprosessin varjoon, joka usein johtaa tarpeettoman suuriin hyödykekustannuksiin ja järjestelmien teknisen tilan laskuun. Työn kirjallisuusosassa käsitellään Roal Oy:n fermentointiprosessin kannalta olennaisimpia hyödykevaatimuksia, niiden laskennallista ilmaisua ja vuorovaikutuksia sekä PINCH-menetelmää lämpöenergian hyötykäyttöön. Jäähdytysjärjestelmän osalta käydään läpi merkittävimmät laitetekniset ratkaisut, jäähdytystorni ja lämpöpumppu, toimintaperiaatteineen sekä luonnonvesien käyttö jäähdytykseen. Työn soveltavassa osassa seurattiin Aspergillus, Trichoderma ja Bacillus fermentointeja, joiden pohjalta luotiin kasvatuskohtainen empiirinen malli jäähdytystarpeen arviointiin perustuen sekoitustehoon, kasvatuksen hiilidioksidituottoon ja haihtumisen vaikutukseen. Kasvatuksien aikana seurattiin myös tilavuusperusteista lämmönsiirtokerrointa. Mitattujen lämmönsiirtokertoimien perustella laskettiin ominaislämmöntuottoon perustuva maksimilämpötila käytettävälle jäähdytysvedelle ja fermentorien maksimitilavuudet tunnetuilla kasvatusparametreilla eri lämpöisille jäähdytysvesille. Soveltavassa osassa käydään myös läpi Roalin höyry- ja kuumavesikulutukset ja tärkeimmät käyttökohteet. Mittaustulosten ja mallien perusteella on tehtiin kehitysehdotukset hyödykejärjestelmän optimoimiseksi.

Demanda energética nas operações mecanizadas na silagem de milho no sistema de "silo bag"

Foi avaliado o consumo energético das operações mecanizadas envolvidas na produção de silagem de planta inteira e silagem de "grão úmido" de milho, tendo como referência o processamento seco desse cereal. O ensaio foi conduzido na Fazenda Experimental Lageado, pertencente à Faculdade de Ciências Agronômicas, e nas instalações da Faculdade de Medicina Veterinária e Zootecnia - UNESP, localizada no município de Botucatu - SP. O delineamento experimental foi em blocos ao acaso, com parcelas subdivididas no tempo (três épocas de colheita: silagem de planta inteira, silagem de "grão úmido" e colheita de grãos secos), com 10 repetições. As análises estatísticas foram realizadas por meio do programa ESTAT, pelo teste de média de Tukey, a 5% de probabilidade. A silagem de planta inteira teve o maior consumo de combustível por área. A secagem dos grãos de 15,5% para 13% foi responsável por 87% do gasto de energia por área. A silagem de "grão úmido" demandou o menor uso de energia por área nas operações mecanizadas.

Identification of croplands of winter cereals in Rio Grande do Sul state, Brazil, through unsupervised classification of normalized difference vegetation index images

This study aimed to propose methods to identify croplands cultivated with winter cereals in the northern region of Rio Grande do Sul State, Brazil. Thus, temporal profiles of Normalized Difference Vegetation Index (NDVI) from MODIS sensor, from April to December of the 2000 to 2008, were analyzed. Firstly, crop masks were elaborated by subtracting the minimum NDVI image (April to May) from the maximum NDVI image (June to October). Then, an unsupervised classification of NDVI images was carried out (Isodata), considering the crop mask areas. According to the results, crop masks allowed the identification of pixels with greatest green biomass variation. This variation might be associated or not with winter cereals areas established to grain production. The unsupervised classification generated classes in which NDVI temporal profiles were associated with water bodies, pastures, winter cereals for grain production and for soil cover. Temporal NDVI profiles of the class winter cereals for grain production were in agree with crop patterns in the region (developmental stage, management standard and sowing dates). Therefore, unsupervised classification based on crop masks allows distinguishing and monitoring winter cereal crops, which were similar in terms of morphology and phenology.