The main focus of the current study was to quantify the chemical signals received by a blue crab (Callinectes sapidus) tracking a source in a laboratory flume. To make a direct linkage between tracking behavior and the odorant concentration signal, we developed a measurement system to quantify the instantaneous concentration field surrounding actively tracking blue crabs. A three-dimensional laser-induced fluorescence (3DLIF) system was designed and constructed to measure odorant concentrations around crabs tracking three source types: a continuous release with exit velocity matching the mean local velocity in the flume; a continuous release with a meander created by an upstream cylinder; and a pulsed release switching on and off and with the same mass flow rate as the other two plume types. The meandering and pulsed plumes were introduced to observe the effects of large-scale spatial (meandering) and temporal (pulsed) intermittency on crab tracking. Simultaneous with the chemical concentration measurements, crab position data was recorded for kinematic analysis during post-processing. In addition, concentration measurements were collected for the three plume types without crabs present in order to quantify the statistical characteristics of the plume structure The concentration signals arriving at the antennules and outer chemosensory organs, most notably the legs, were targeted due to the hypotheses that concentration bursts at the antennules mediate upstream movement and that spatial contrast at the leg chemosensors mediates turning. A sampling zone was placed in front of the crab's mouth parts and aligned with the crab carapace orientation to extract odorant bursts at the antennules. The data generally showed an increase in upstream walking speed when high concentration bursts arrive at the antennules location, which agrees with the hypothesis. Measurement of the odorant concentration at the outer chemosensors was less direct and involved placing a box upstream of the crab and sampled earlier in time in order to avoid shadowing interference. Based on the signal at the upstream sampling box, a general bias for turning was observed. Crabs casted transversely in response to the directional bias extracted from the upstream sampling box. A statistical analysis of crab behavioral response to concentrations at the antennules and outer chemosensors can be found in a (future) companion thesis written by Jennifer Page in the School of Biology. Data were also taken for the three plume types in the absence of blue crabs. The continuous plume average statistics displayed Gaussian behavior at nozzle centerline. The meandering plume data conformed to the meandering plume model of Gifford (1959), modified for an induced pseudo-periodic meander. The pulsed plume displayed characteristics intermediate between the cloud dispersion model (Townsend 1951, Chatwin and Sullivan 1979) and the Gaussian dispersion model for a continuous release. For the three plume types, the standard deviation of the concentration fluctuations was greater than the average concentrations, as time records consisted of intermittent high concentrations interspersed with concentrations close to zero.

Dissertation (M.Sc. (Chemical engineering)) -- University of Pretoria, 2008. / Includes bibliographical references (p. 42-52)

Thesis (MSc)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: Wine is a complex medium. Wine aroma, flavour and colour are important quality factors, but these can be influenced by many factors, such as grape-derived compounds that exist as free volatiles and also as glycosidically bound. The chemical composition of wine is determined by factors such as grape variety, geographic position, viticulture condition, microbial ecology of the grape and the winemaking process. The varietals aroma is determined by both the volatile and the non-volatile compounds, such as monoterpenes, norisoprenoids and benzene derivatives, which are naturally present in the wine. Monoterpenes are very important in the flavour and aroma of grapes and wine. They can be found in grapes and wine either in the free, volatile and odorous form, or in the glycosidically-bound, non-volatile and non-odorous form. The ratio of glycosidically-bound compounds to free aroma compounds is very high in the Gewürztraminer, Muscat and Riesling cultivars in particular. The glycosidic bonds can be hydrolysed either by the acid method or by using enzymes. The acid method is disadvantageous because it can modify the monoterpenes, whereas enzymatic hydrolysis has the advantage of not modifying the aroma character. The enzyme method of breaking the glycosidic bonds occurs in two successive steps: initial separation of glucose from the terminal sugar by a hydrolase (a-L-arabinofuranosidase, a-L-rhamnosidase or β-apiosidase, depending on the aglycone moiety), followed by the breaking of the bond between the aglycone and glucose by β-glucosidase. The enzyme β-glucosidase can be obtained from many plant (Vitis vinifera), bacterial, yeast or fungal sources. Most of the enzymes produced by these sources are not functional under the winemaking conditions of low pH, low temperature, high glucose and high ethanol content. However, β-glucosidases from fungal origins, particularly from Aspergillus spp., are tolerant of winemaking conditions. The idea of using the β-glucosidase gene from the fungus Aspergillus kawachii (BGLA), which is linked to the cell wall and the free β-glucosidase, was to determine if anchoring the enzyme to the cell wall will increase the activity of the enzyme compared to the free enzyme. Four plasmids, pCEL 16, pCEL 24, pDLG 97 and pDLG 98, were used in this study. BGLA that was cloned into the plasmids pCEL 24 and pDLG 97 was linked to CWP2, and in pDLG 98 it was linked to AGa1 anchor domains. All the plasmids were genome-integrated and expressed in the reference strain Saccharomyces cerevisiae 303-1A. All the transformants were grown in 2% cellobiose and showed higher biomass production compared to the reference strain. β-Glucosidase activity was also assayed and transformed strain W16 showed a fourfold increase in activity compared to the reference strain. There was no significant increase in the activity of the other transformed strains, W24, W97 and W98. Enzymatic characterisation for optimum pH and temperature was done – for all strains the optimum pH was 4 and the optimum temperature was 40ºC. The recombinant strains together with the reference strain were used to make wine from Gewürztraminer grapes. The levels of numerous monoterpenes were enhanced in the resultant wines. The concentration of nerol was increased fourfold, that of citronellol twofold, and geraniol was 20% higher than in the wild type. There was also an increase in the levels of linalool and a-terpinol, but this was not significant. In wines produced with W97, W98 and W24, monoterpene levels did not show a significant difference. In future, the expression of the W16 expression cassette in an industrial wine yeast strain could be performed. In combination with the production of enzymes such as a-arabinofuranosidase, a-rhamnosidase and β-apiosidase, which are involved in the first step of enzymatic hydrolysis, this wine strain could release the bound monoterpenes and enhance the aroma of the wine. / AFRIKAANSE OPSOMMING: Wyn is ‘n komplekse medium. Wynaroma, -geur en -kleur is belangrike kwaliteitsfaktore, hoewel hierdie kwaliteite deur verskeie faktore beïnvloed kan word, soos druifafgeleide verbindings wat as vry vlugtige stowwe teenwoordig kan wees of glikosidies gebind is. Die chemiese samestelling van wyn word bepaal deur faktore soos druifvariëteit, geografiese ligging, wingerdkundige toestande, mikrobiese ekologie van die druif en die wynbereidingsproses. Die variëteitsaroma word bepaal deur vlugtige en nie-vlugtige verbindings, soos monoterpene, norisoprenoïede en benseenderivate, wat natuurlik in die wyn voorkom. Monoterpene is baie belangrik vir die geur en aroma van druiwe en wyn. Monoterpene is teenwoordig in die druiwe en wyn in vry, vlugtige en geurige, of in glikosidiesgebinde, nie-vlugtige en nie-geurige vorms. Die verhouding van glikosidiesgebonde verbindings tot vry aromaverbindings is baie hoog, veral in die Gewürztraminer-, Muscat- en Riesling-kultivars. Glikosidiese verbindings kan deur óf die suurmetode óf die ensiemmetode gehidroliseer word. Die nadeel van die suurmetode is dat dit monoterpene kan modifiseer, terwyl die ensiemmetode die voordeel het dat dit nie die aromakarakter modifiseer nie. Die ensiemmetode waarmee die glikosidiese verbinding afgebreek word, vind in twee opeenvolgende stappe plaas: aanvanklike skeiding van glukose van die terminale suiker deur ‘n hidrolase (a-L-arabinofuranosidase, a-Lramnosidase of β-apiosidase, afhangende van die aglikoongedeelte), gevolg deur die verbreking van die verbinding tussen die aglikoon en glukose deur β- glukosidase. Die β-glukosidase-ensiem kan vanaf ‘n verskeidenheid plant- (Vitis vinifera), bakterie-, gis- en swambronne verkry word. Die meerderheid van die ensieme wat deur hierdie bronne geproduseer word, is nie onder die wynbereidingstoestande van lae pH, hoë temperatuur, hoë glukose en hoë etanol funksioneel nie. β- Glukosidase vanaf ‘n swamoorsprong, veral vanaf Aspergillus-spesies, kan egter wynbereidingstoestande verdra. Die idee agter die gebruik van die β-glukosidasegeen afkomstig van die swam Aspergillus kawachii (BGLA), wat aan die selwand en die vry β-glukosidase gekoppel is, was om te bepaal of die aktiwiteit van die ensiem in vergelyking met dié van die vry ensiem verhoog sou word indien die ensiem aan die selwand geanker is. Vier plasmiede, pCEL 16, pCEL 24, pDLG 97 en pDLG 98, is in hierdie studie gebruik. BGLA, wat in die plasmiede pCEL 24 en pDLG 97 gekloneer is, is gekoppel aan CWP2, en in pDLG 98 is dit aan AGa1-ankergebiede gekoppel. Al die plasmiede is in verwysingsras Saccharomyces cerevisiae 303-1A genoomgeïntegreer en uitgedruk. Al die transformante is in 2% sellobiose gegroei en het hoër biomassaproduksie as die verwysingsras getoon. β-Glukosidaseaktiwiteit is ook geëssaieer en die getransformeerde ras W16 het ‘n viervoudige verhoging in aktiwiteit in vergelyking met die verwysingsras getoon. Daar was geen noemenswaardige verhoging in die aktiwiteit van die ander getransformeerde rasse, W24, W97 en W98, nie. Ensimatiese karakterisering vir optimum-pH en - temperatuur is gedoen – vir al die rasse was die optimum-pH 4 en die optimumtemperatuur 40ºC. Die rekombinante rasse, tesame met die verwysingsras, is gebruik om wyn met Gewürtztraminer-druiwe te maak. Die vlakke van talryke monoterpene is in die gevolglike wyne verhoog. Die konsentrasie van nerol is viervoudig verhoog, dié van sitronellol tweevoudig, en geraniol was 20% hoër as in die wilde tipe. Daar was ook ‘n verhoging in die vlakke van linaloöl en a-terpinol, maar hierdie verhoging was nie noemenswaardig nie. In wyne wat met W97, W98 en W24 gemaak is, het die monoterpeenvlakke nie ‘n noemenswaardige verskil getoon nie. In die toekoms sal die uitdrukking van die W16-uitdrukkingskasset in ‘n industriële wyngisras uitgevoer kan word. In kombinasie met die produksie van ensieme soos a-arabinofuranosidase, a-ramnosidase, β-apiosidase, wat in die eerste stap van ensimatiese hidrolise betrokke is, sal hierdie wyngisras die gebonde monoterpene kan vrylaat en die aroma van die wyn kan verbeter.

Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: No single chemical constituent can be accredited with giving wine and brandy their overall aroma and flavour. The aroma and flavour of wine and brandy are rather attributed to a number of chemical constituents reacting together and it is these reactions that give the beverage its character. Certain chemicals within wine and brandy do, however, make larger contributions to the flavour. These include the esters, terpenes and volatile acids, although others also exist. Esters are a large group of volatile compounds with variable aroma and flavour characteristics, including banana-like (isoamyl acetate), apple-like (ethyl caproate) and chemical/solvent-like (ethyl acetate). Esters are produced as secondary metabolites during the conversion of sugar to ethanol and are formed when an alcohol binds with a fatty acid. Chemically, ester metabolism is well documented and understood; however, much work still needs to be done on a genetic level. The yeast strain used during fermentation is one of the most important factors contributing to the type and quantity of esters produced. This is due to differences in genetic makeup. The metabolism of esters is controlled largely on a genetic level, with numerous genes being involved. The alcohol acetyltransferase genes are involved in ester anabolism, whilst esterase genes are involved in ester catabolism. Esterases have a negative effect on the overall level of esters within an alcoholic beverage, as they are capable of reducing the number of esters and are thus capable of altering the beverage's aroma and flavour profile. The IAH1 and the TIP1 gene products are believed to encode for two such esterases. The objective of this study was to investigate the contribution of the IAH1 and TIP1 genes to the level of esters in both wine and brandy. This was accomplished by using two approaches. Firstly, the above genes were disrupted using a polymerise chain reaction (PCR)-generated disruption cassette homologous to either the IAH1 or the TIP1 gene. These cassettes were integrated into the industrial wine yeast, Saccharomyces cerevisiae strain VIN13. The integrations were verified by Southern blot analysis to produce yeasts VIN13-~IAH1 and VIN13-~TIP1; however, only a single copy of each was disrupted. Secondly, the IAH1 and the TIP1 genes were cloned from S. cerevisiae using PCR into plasmid pj between the phosphoglycerate kinase gene (PGK1) promoter and terminator, producing plasmids pJ-IOE1 and pJ-TOE1. The PGK1 promoter has previously been shown to constitutively express genes at high levels. These new constructs were then used as template for PCR to produce two overexpression cassettes, one for IAH1 and the other for TlP1. These cassettes were integrated into S. cerevisiae VIN13 and verified by Southern blot analysis to produce strains VIN13-IOE1 and VIN13-TOE1. The above yeast strains including VIN13 were used for the production of wines and base wines from Colombard must. Reverse-transcriptase (RT-PCR) confirmed that the VIN13-IOE1 and VIN13-TOE1 strains overexpressed the appropriate gene at a higher level than the control VIN13 strain. The VIN13-AIAH1 disrupted strain showed no difference in expression level to that of the control strain, whilst VIN13-ATIP1 showed lower levels of expression than that of the control strain. VIN13-IOE1 behaved as expected, with a decrease of between 30% and 60% in the total ester level in the wine and base wine respectively, a 30% decrease in the total acid level and no change in the higher alcohol level. The VIN13-AIAH1 strain showed no difference to the control wine, most likely as this strain still expressed the IAH1 gene at levels consistent with the control strain. VIN13-TOE1 behaved in an unexpected manner - instead of hydrolysing esters, it appeared to produce them. This increase in the total ester level was most noticeable during distillation, when a 20% increase took place. Another unexpected occurrence was a large decline in the total acid level, with acetic acid being the most significant contributor, decreasing by up to 78%. This is a very favourable finding, as acetic acid is a known spoilage molecule and is a cause of sluggish/stuck fermentations. VIN13-ATIP1 behaved in an opposite manner to VIN13-TOE1, with higher total acid levels and slightly decreased total ester levels, especially during distillation. Neither affected the total higher alcohol levels. Sensorially, the only significant difference in the wine samples was for the fruity flavour. A panel of judges distinguished that VIN13-TOE1 was fruitier than the other wines, with VIN13-ATIP1 being the least fruity. This study again proves the significant impact that a single gene can have on the chemical makeup of wine and brandy. The relatively simple genetic alteration of an organism can drastically change and improve not only the organoleptic properties of the organism, but its viability as well. These alterations can produce more favourable organisms with more desirable characteristics for the fermenting beverage industry to produce products of higher quality and better suitability. / AFRIKAANSE OPSOMMING: Geen chemiese komponent kan uitgesonderword as die produseerder van aroma en geur in wyn of brandewyn nie. Die aroma en geur van wyn en brandewyn word eerder toegeskryf aan die interaksie tussen 'n groot aantal chemiese komponente om aan die drank sy karakter te gee. Enkele van hierdie chemiese komponente sluit in esters, terpene en vlugtige sure, om maar 'n paar te noem. Esters is "n groot groep van vlugtige verbindings wat beskik oor 'n verskeidenheid van aroma- en geurkenmerke, soos piesangagtig (isoamielasetaat), appelagtig (etielkaproaat) en chemies/oplosmiddelagtig (etielasetaat). Esters word as sekondêre metaboliete geproduseer wanneer suikers na etanolomgeskakel word en word gevorm wanneer "n alkohol met "n vetsuur verbind. Estermetabolisme is chemies goed beskryf en verstaan, maar op "n genetiese vlak is daar nog heelwat aspekte wat nagevors moet word. Die gisras betrokke gedurende fermentasie word beskou as een van die grootse bydraes tot die tipe en die hoeveelheid esters wat geproduseer word. Dit word toegeskryf aan verskille in die genetiese saamestelling van die gisras. Ester metabolisme word grootliks deur genetiese faktore beheer en verskeie gene is betrokke. Dit is hoofsaaklik die alkoholasetieltransferasegene wat vir esterkatabolisme verantwoordelik is, terwyl die esterasegene vir esteranabolisme verantwoordelik is. Esterases het 'n negatiewe effek op die totale estervlak binne alkoholiese dranke deurdat hulle in staat is om die aantal esters drasties te verminder en sodoende die drank se aroma- en geurprofiel te verander. Daar is voorgestel dat die IAH1- en die TlP1-geen produkte is wat vir twee sulke esterases kodeer. Die doel van hierdie studie was om die IAH1- en die TIP1-gene se bydrae tot die totale estervlak in wyn en brandewyn te ondersoek. Dit is deur twee benaderings uitgevoer. Eerstens is die bogenoemde gene d.m.V. disrupsiekassette wat homoloog aan die IAH1- of die TlP1-gene was, uitgeslaan. Die disrupsiekassette is deur die polimerasekettingreaksie (PKR) geproduseer. Hierdie kassette is in die industriële wyngis, Saccharomyces cerevisiae VIN13, geïntegreer. Die integrasies is deur Southernkladanalise bevestig en het die giste VIN13-~IAH1 en VIN13-~TIP1 gelewer. Net 'n enkele kopie van elke geen is egter uitgeslaan. Tweedens is die IAH1- en TIP1-gene d.m.V. PKR vanaf S. cerevisiae binne in plasmied pJ gekloneer, tussen die fosfogliseraatkinasegeen (PGK1) se promotor en termineerder, om plasmiede pJ-IOE1 en pJ-TOE1 te produseer. Die PGK1-promotor is al tevore geïdentifiseer as "n hoë-vlak konstitutiewe uitdrukker van gene. Hierdie twee nuwe konstrukte het vervolgens gedien as templaat vir PKR om twee ooruitdrukkingskassette, een vir IAH1 en die ander vir TIP1, te produseer. Hierdie kassette is in S. cerevisiae VIN13 geïntegreer en bevestig deur Southernkladanalise. Hierdie integrasies het die giste VIN13-IOE1 en VIN13-TOE1 geproduseer. All die nuwe gisrasse, tesame met VIN13, is gebruik vir die produksie van wyne sowel as rebatwyne vanaf Colombard-mos. Omgekeerde-transkriptase polimerasekettingreaksie (OT-PKR) het bewys dat die VIN13-IOE1 en VIN13-TOE1 rasse die geskikte geen ooruitgedruk het, met hoêr vlakke as van die kontrole VIN13-ras. Dit het ook aangedui dat die VIN13-i\IAH1-ras, waarvan die geen uitgeslaan was, geen verskil in uitdrukking gehad het in vergelyking met die kontroleras nie, terwyl VIN13-i\TIP1 'n lae uitdrukkingsvlak getoon het. VIN13-IOE1 het teen verwagting opgetree, met 'n afname van tussen 30% en 60% in die totale estervlak in beide die wyne en rebatwyne. 'n Afname van 30% in die totale suurvlak, asook geen waarneembare verskil in die hoêr alkoholvlak, in vergelyking met die kontroleras, is ook opgemerk. Die VIN13-i\IAH1-ras het glad nie van die kontroleras verskil nie, heel waarskynlik omdat hierdie ras die IAH1-geen teen dieselfde vlak as die kontroleras kon uitdruk. Die VIN13-TOE1-ras het teen verwagting opgetree deurdat dit esters geproduseer het i.p.v. om esters te hidroliseer. Hierdie toename in die totale estervlak is die meeste waarneembaar tydens distillasie, met tot 'n 20% toename. Nog 'n onverwagte effek was die groot afname in die totale suurvlak. met asynsuur wat die betekenisvolste bydrae gelewer het deurdat dit 'n afname van tot 78% getoon het. Hierdie bevinding is baie voordelig, aangesien asynsuur, 'n bekende bederfmolekuul, veral vir slepende/gestaakte fermentasies verantwoordelik is. VIN13-i\TIP1 het op die teenoorgestelde wyse opgetree as VIN13-TOE1, met 'n hoêr totale suurvlak en 'n klein afname in die totale estervlak. Weereens is dit meer gedurende distillasie waargeneem. Beide rasse het egter geen effek op die hoêr alkoholvlak gehad nie. Die proepaneel het, met betrekking tot die vrugtige geur, een betekenisvolle geurverskil tussen die wyne gevind. VIN13-TOE1 was meer vrugtig as al die ander wyne en VIN13-i\TIP1 was die minste vrugtig. Die studie het weereens bewys dat 'n enkele geen 'n betekenisvolle effek op die chemiese samestelling van wyn en brandewyn kan hê. Die relatief eenvoudige genetiese verandering van 'n organisme kan die organoleptiese eienskappe asook die lewensvatbaarheid van "n organisme, drasties verander en verbeter.

O café solúvel aglomerado é produto da aglomeração do café solúvel em pó. A sua produção envolve etapas de processo cujas variações térmicas modificam qualitativamente e quantitativamente a fração de compostos voláteis responsáveis pelo aroma e sabor do produto acabado. A indústria de bebidas de café considera o sabor e aroma de cereal como um defeito na produção de café solúvel aglomerado. Desta forma, este trabalho objetivou aplicar o planejamento experimental para avaliação dos parâmetros de processo da produção de café solúvel aglomerado em escala industrial. Os efeitos destes parâmetros nos componentes voláteis do café e na intensidade do gosto de cereal foram avaliados através do planejamento fracionário 24-1 com resolução IV. As variáveis independentes estudadas foram temperatura do queimador (TQ), fluxo de ar de processo (FA), temperatura da seção quente do leito (TL) e pressão de vapor (PV). Os resultados do planejamento experimental deram-se pela utilização de análise sensorial conduzida por equipe treinada para a detecção do aroma e sabor indesejado. Adicionalmente, os compostos voláteis foram extraídos pela técnica microextração em fase sólida com amostragem no headspace (HS-MFES), identificados e quantificados por cromatografia gasosa acoplada a espectrometria de massa (CG-EM). O efeito dos fatores estudados foi avaliado mediante análise de variância (ANOVA) com nível de significância de 90% e os resultados expressos por gráficos de pareto. A análise de componentes principais (ACP) foi realizada com o intuito de identificar dentre os compostos que podem conferir sabor e aroma de cereais aqueles tem a maior contribuição e quais as condições de processo ideias para evitar a formação do sabor e aroma de cereal. De acordo com ACP foi constatado que piridina, 2-metoxi-4-vinilfenol, 2,3-dimetilpirazina, 2,5-dimetilpirazina, 2,6-dimetilpirazina, 4-metiltiazol, pirazina, 2-metoxifenol, 4,5-dimetiltiazol contribuem para formação do sabor e aroma de cereal. Também indicou que a aglomeração deve ocorrer conforme TQ=215ºC, FA=250rpm, TL=120ºC e PV=1,2 x100KPa. / The agglomerated instant coffee is the product of the powder instant coffee agglomeration. Its production involves stages of processes whose thermal variations change qualitatively and quantitatively the volatile compounds fractions responsible for the aroma and flavor of the finished product. The coffee beverage industry considers the cereal like flavor and aroma as a fault in the production of the agglomerated instant coffee. In this way, this study had as the aim to apply experimental plan to evaluate the parameters of the processes of the agglomerated instant coffee production on an industrial scale. The effects of those parameters in the coffee volatile components and in the intensity of the cereal taste were evaluated through the fractional 24-1 with resolution IV. The independent variables studied were the burner temperature (BT), the process air flow (AF), the temperature of the fluidized bed hot section (LT) and the steam pressure (SP). The results of the experimental plan were made by the use of sensory analysis performed by a trained team for the detection of the undesired aroma and flavor. Additionally, the volatile compounds were extracted by solid-phase microextractiontechniquewithsampling in theheadspace (HS-SPME), identified and quantified by gas chromatography coupled to mass spectrometry (GC-MS). The effect of the factors studied was evaluated by variance analysis (ANOVA) with significance level of 90% and the results showed by Pareto charts. The principal components analysis (PCA) was made with the aim to identify, among several compounds, those that implies on more cereal flavor and aroma, those which major contribution and which ideal process conditions could be used to reduce the cereal flavor and aroma development. According to PCA, it was found that pyridine, 2-metoxi-4-vinylphenol, 2,3-dimethylpyrazine, 2,5-dimetylpyrazine, 2,6-dimethylpyrazine, 4-methyltiazol, pyrazine, 2-methoxyphenol, 4,5-dimethylthiazol contribute to the development of the cereal flavor and aroma. It was also found that the agglomeration must be according to TQ = 215°C, AF=250 rpm, LT=120°C and EPV = 1,2 x 100 KPa.

O processo de eutrofização das águas, principalmente pelo aporte de fósforo e nitrogênio, favorece a ocorrência de florações de cianobactérias. Uma consequência desses eventos é a produção e emissão dos metabólitos 2-metilisoborneol (MIB) e trans-1,10-dimetil-trans-9-decalol (geosmina). Estes compostos são álcoois terciários que podem ser detectados em baixas concentrações pelos consumidores, 5 a 10 ng/l, deixando a água com “cheiro e gosto de terra e mofo”. MIB e geosmina, particularmente na forma dissolvida (extracelular), mostram-se recalcitrantes aos métodos convencionais de tratamento de água. As metodologias atualmente empregadas na remoção desses metabólitos apresentam-se pouco eficientes e de elevado custo operacional, incentivando as pesquisas relacionadas às alternativas de tratamento. O presente estudo foi realizado em duas etapas, onde a primeira buscou avaliar a remoção de MIB e geosmina por adsorção em carvão ativado granular e a segunda, a degradação dos metabólitos por atividade microbiológica. Primeiramente, os carvões ativados granulares foram caracterizados através do número de iodo e área superficial BET. Em seguida, foi determinado o tempo de equilíbrio de adsorção para cada carvão e por último, foi avaliada a capacidade adsortiva através das Isotermas de Freundlich. O estudo sobre a oxidação biológica dos compostos causadores de gosto e odor consistiu no isolamento, identificação e testes de biodegradação de bactérias que utilizam MIB e geosmina como única fonte de carbono para o seu metabolismo. No que se refere à capacidade de adsorção, foi observado que os carvões de madeira e casca de coco apresentaram os melhores desempenhos na remoção de MIB e geosmina, respectivamente. Esses carvões removeram os compostos abaixo do limite de detecção olfativa (10 ng/l), com doses de 50 mg/l e tempo de contato de duas horas. Essa caracterização mostrou-se de fundamental importância na escolha do carvão mais adequado para aplicação no tratamento de água. Uma vez que pouco se conhece a respeito da comunidade bacteriana, presente em águas naturais, ao que se sabe, este trabalho é o primeiro registro da capacidade individual de bactérias Gram positivas e negativas degradarem os compostos causadores de gosto e odor, na mesma solução teste. A remoção de cerca de 90 e 96% de MIB e geosmina ao longo de 48 e 120 horas de experimento, respectivamente, demonstrou que essas bactérias podem ser utilizadas em estratégias de purificação biológica da água. A identificação molecular dos isolados bacterianos, obtidos no Lago Guaíba, registrou quatro novos gêneros capazes de degradar MIB e geosmina: Burkholderia sp, Caulobacter sp, Klebsiella sp e Leucobacter sp. Além desses, um isolado foi classificado como Arthrobacter sp, outro como Sphingomonadaceae e três como Enterobacteriaceae. O sucesso no isolamento, manutenção de culturas puras, identificação e determinação da capacidade desses microrganismos degradarem MIB e geosmina da água sugere um potencial de utilização dessas bactérias em processos de filtração biológica, que utilizam carvão ativado granular. Esse conhecimento pode ser usado como uma nova ferramenta para a remoção de substâncias que causam gosto e odor, sendo essencial para garantir uma água de qualidade para o consumo humano. / Water eutrophication process, by phosphorous and nitrogen levels, allows cyanobacteria blooms which promotes, among other effects, the generation and release of the metabolites 2-methylisoborneol (MIB) and trans-1,10-dimethyl-trans-9- decalol (geosmin). These compounds are tertiary alcohols that may lend earthymusty tastes and odors to water, characteristics detected by consumers when the metabolites are present at concentrations as low as 5 to 10 ng/l. When present in water, mainly in the dissolved form (extracellular), MIB and geosmin are difficult to be removed by conventional treatment methods. The methodologies currently employed to remove these metabolites lack efficiency and increase operational costs, arousing the interest in research to develop alternative water treatment approaches. The present study was conducted in two steps. The first step evaluated the removal of MIB and geosmin by adsorption using different types of granular activated carbon, while the second step assessed the degradation of the metabolites by microbiological activity. Firstly, granular activated carbon was analyzed for iodine number and BET surface area. Next, adsorption equilibrium time was measured for each carbon type. Then, adsorption capacity was evaluated using the Freundlich isotherm. Biological oxidation of compounds causing taste and odor included isolation, identification and biodegradation tests using bacteria that employ MIB and geosmin as the only carbon sources. Regarding adsorption capacity, wood and coconut shell carbon performed better in MIB and geosmin removal, respectively, affording removal to levels below the odor threshold concentration (10 ng/l), using 50 mg/l solutions of each carbon and exposure times of 2 h. This characterization proves the importance of choosing the most appropriate carbon to employ in water treatment. Since little is known on the bacterial community thriving in natural waters, as far as we know this is the first study to record the capacity of Gram-positive and Gramnegative bacteria to individually degrade compounds that cause taste and odor in water, in the same test solution. The removal of 90 and 96% of MIB and geosmin within 48 and 120 h of exposure, respectively, shows that these bacteria may be used in water purification strategies. The molecular identification of the bacterial isolates obtained in Lake Guaíba revealed four new genera capable to degrade MIB and geosmin: Burkholderia sp., Caulobacter sp., Klebsiella sp., and Leucobacter sp. Apart from these isolates, one Arthrobacter sp., one Sphingomonadaceae and three Enterobacteriaceae isolates were also identified. The successful isolation, maintenance of pure cultures, identification and determination of these microorganisms’ capacities to degrade MIB and geosmin in water suggests that these bacteria may be used in biological filtration processes that utilize granular activated carbon. This knowledge may be used as a new tool to remove substances that lend taste and odor to water, and is an essential aspect to guarantee quality water for human consumption.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas. Programa de Pós-Graduação em Farmacologia. / Made available in DSpace on 2012-10-22T15:02:55Z (GMT). No. of bitstreams: 1 225418.pdf: 2520051 bytes, checksum: b59986e81eda2d965b91443a5dedb96d (MD5) / Estudos têm demonstrado que o odor do predador é um potente estímulo ansiogênico, fornecendo elementos para a avaliação do comportamento defensivo natural em roedores. Durante situações de estresse ou perigo potencial, a liberação de adrenalina ocasiona ativação de receptores beta-adrenérgicos nas vias ascendentes do vago para estruturas do SNC envolvidas na expressão e consolidação do medo e de suas respostas defensivas. O objetivo principal do presente estudo foi avaliar a participação do sistema beta-adrenérgico no comportamento defensivo de ratos expostos ao modelo do odor de gato. Para este propósito, realizamos inicialmente uma validação biológica e farmacológica do modelo dentro das condições experimentais de nosso laboratório. Em seguida, avaliamos o papel da neurotransmissão beta-adrenérgica central e periférica na resposta defensiva de ratos submetidos a este modelo. Nesta etapa, utilizamos em nossos estudos antagonistas beta-adrenérgicos como o propranolol, com ações farmacológicas centrais e periféricas, e o nadolol, com ações restritas à periferia. Por fim, avaliamos o envolvimento da neurotransmissão noradrenérgica do núcleo PMd - uma estrutura hipotalâmica amplamente relacionada ao comportamento defensivo inato - sobre a resposta defensiva de ratos expostos ao odor de gato, através da microinjeção de atenolol, um bloqueador beta1 seletivo, diretamente nesta estrutura. Nossos resultados demonstraram que a injeção sistêmica de propranolol, bem como as microinjeções de atenolol no PMd, foram eficazes na redução da resposta defensiva. Estes achados sugerem uma participação do sistema beta-adrenérgico central, além de reforçar o papel chave do PMd no comportamento de defesa inato frente ao odor de gato. Substantial evidences have been showing that predator odor is a powerful anxiogenic stimulus, providing insights into natural defensive behavior evaluation in rodents. During situations of stress or potential danger, epinephrine release actives beta-adrenoreceptors on the ascending vagus, projecting to central nervous system structures, which are believed to be important in the expression and consolidation of the fear and its defensive responses. The main aim of this study was to verify the beta-adrenergic system participation, in the behavior response of rats toward cat odor model. For this purpose, firstly we have realized the biological and pharmacological validation of the cat odor model in accordance to the experimental conditions of our laboratory. Next, the role of central and peripheral beta-adrenergic neurotransmission in rats submitted to this model was evaluated. Whereas propranolol was used to block beta-adrenoceptor at central and peripheral sites of action, nadolol, did it predominantly at the peripheral system. Finally, we have checked the noradrenergic involvement of the PMd - a hypothalamic structure that takes part in the innate defensive behavior - in the defensive response of rats exposed to cat odor, using atenolol, a selective beta1-blocker, microinjected directly into this structure. Our results demonstrate that either propranolol systemic injection or atenolol microinjections into PMd, were able to reduce the defensive behavior of rats exposed to cat odor. These findings suggest a central beta-adrenergic system participation in the innate defensive behavior toward cat odor strengthening the key role of the PMd on this behavior.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-Graduação em Engenharia Ambiental / Made available in DSpace on 2012-10-23T14:56:07Z (GMT). No. of bitstreams: 0 / A poluição odorífera tem gerado diversos conflitos entre as fontes de emissão e a população afetada. Odores desagradáveis são provenientes de inumeras fontes, dentre as quais a indústria petrolífera. Os parques industriais de derivados de petróleo emitem diversos compostos compostos orgânicos voláteis (COV), muitos de natureza odorífera. A biofiltração, a qual é uma tecnologia de baixo custo para desodorização, foi utilizada para reduzir odores de gasolina comercial gerados a partir de uma fonte sintética. Um biofiltro em escala piloto utilizando turfa como material suporte, foi monitorado durante 140 dias a uma taxa de aplicação superficial (TAS) de 62 m3/m2.h e um tempo de contato (TC) de 52 segundos. A umidade mantida no leito variou entre 50 e 60% e o pH médio foi de 5,4. As amostragens semanais do gás odorante bruto e do gás tratado permitiram avaliar a eficiência do sistema através de análises olfatométricas e análises físico-químicas. Através da olfatometria, um corpo de jurados composto entre seis e dez pessoas avaliaram os seguintes parâmetros relativos a um estudo de impacto de odor: concentração, intensidade, qualidade, hedonicidade e nível de incômodo. As análises físico-químicas foram realizadas por uma unidade de cromatografia gasosa acoplada a um espectrômetro de massa. Estas análises permitiram a identificação dos compostos presentes nas amostras pré e pós-tratamento, bem como a avaliação da redução da concentração e do número de compostos proporcionada pela biofiltração. Após 140 dias de operação a redução da concentração do odor foi de 87%, a intensidade do odor da amostra tratada foi fraca, a hedonicidade foi agradável e o nível de incômodo foi nulo. As concentrações de benzeno, tolueno, etilbenzeno e xilenos foram reduzidas respectivamente em :89,98%, 46,43%, 25,0% e 68,60%. A quantidade de compostos foi reduzida significativamente de 65 para a amostra contendo o gás odorante bruto para 16 compostos presentes na amostra tratada. A biofiltração mostrou-se uma alternativa promissora no combate às emissões odoríferas provenientes do setor petrolífero. Malodorous atmospheric emissions have been the cause of a number of conflicts between their emission sources and the affected population. There are many different sources of odor emissions, and petrochemical companies are an example of such. Petroleum-derived fuels are formed by a large number of volatile organic compounds (VOC), and many of those are odoriferous. The biofiltration, which is a technology of low cost for deodorization, was used to reduce commercial gasoline odors generated from a synthetic source. One biofilter in pilot-scale using turf as material support was monitored during 140 days to a tax of superficial application (TAS) of 62 m3/m2.h and a time of contact (TC) of 52 seconds. The humidity kept in the bed varied between 50 and 60% and the pH average were of 5,4. Weekly samples of raw and treated gas had allowed to evaluate the efficiency of the system through olfactometric and physicalchemical analyses. Through the olfactometry, an olfactometric jury composed between six and ten people had evaluated the following relative parameters to a study of odor impact: concentration, intensity, quality, hedonic character, and annoyance level. The analyses physical-chemical had been carried through by chromatography unit coupled with a mass spectrometer. The physical-chemical analyses were conducted in order to identify the compounds present in the samples collected before and after treatment, and to evaluate the concentration reduction of the odorous compounds through biofiltration. After 140 days of operation, the observed odor concentration reduction was of 87%. The treated gas had weak odor intensity, a pleasant hedonic value and the annoyance level was inexistent. The benzene, toluene, ethyl benzene and xylene concentrations were reduced in 89,98%, 46,43%, 25,0% and 68,60%, respectively. The amount of compounds was reduced significantly of 65 for the sample contends the raw gas for 16 compounds gifts in the treated sample. The biofiltration revealed a promising alternative in the treatment the odoriferous emissions proceeding from the petrochemical sector.

O processo de eutrofização das águas, principalmente pelo aporte de fósforo e nitrogênio, favorece a ocorrência de florações de cianobactérias. Uma consequência desses eventos é a produção e emissão dos metabólitos 2-metilisoborneol (MIB) e trans-1,10-dimetil-trans-9-decalol (geosmina). Estes compostos são álcoois terciários que podem ser detectados em baixas concentrações pelos consumidores, 5 a 10 ng/l, deixando a água com “cheiro e gosto de terra e mofo”. MIB e geosmina, particularmente na forma dissolvida (extracelular), mostram-se recalcitrantes aos métodos convencionais de tratamento de água. As metodologias atualmente empregadas na remoção desses metabólitos apresentam-se pouco eficientes e de elevado custo operacional, incentivando as pesquisas relacionadas às alternativas de tratamento. O presente estudo foi realizado em duas etapas, onde a primeira buscou avaliar a remoção de MIB e geosmina por adsorção em carvão ativado granular e a segunda, a degradação dos metabólitos por atividade microbiológica. Primeiramente, os carvões ativados granulares foram caracterizados através do número de iodo e área superficial BET. Em seguida, foi determinado o tempo de equilíbrio de adsorção para cada carvão e por último, foi avaliada a capacidade adsortiva através das Isotermas de Freundlich. O estudo sobre a oxidação biológica dos compostos causadores de gosto e odor consistiu no isolamento, identificação e testes de biodegradação de bactérias que utilizam MIB e geosmina como única fonte de carbono para o seu metabolismo. No que se refere à capacidade de adsorção, foi observado que os carvões de madeira e casca de coco apresentaram os melhores desempenhos na remoção de MIB e geosmina, respectivamente. Esses carvões removeram os compostos abaixo do limite de detecção olfativa (10 ng/l), com doses de 50 mg/l e tempo de contato de duas horas. Essa caracterização mostrou-se de fundamental importância na escolha do carvão mais adequado para aplicação no tratamento de água. Uma vez que pouco se conhece a respeito da comunidade bacteriana, presente em águas naturais, ao que se sabe, este trabalho é o primeiro registro da capacidade individual de bactérias Gram positivas e negativas degradarem os compostos causadores de gosto e odor, na mesma solução teste. A remoção de cerca de 90 e 96% de MIB e geosmina ao longo de 48 e 120 horas de experimento, respectivamente, demonstrou que essas bactérias podem ser utilizadas em estratégias de purificação biológica da água. A identificação molecular dos isolados bacterianos, obtidos no Lago Guaíba, registrou quatro novos gêneros capazes de degradar MIB e geosmina: Burkholderia sp, Caulobacter sp, Klebsiella sp e Leucobacter sp. Além desses, um isolado foi classificado como Arthrobacter sp, outro como Sphingomonadaceae e três como Enterobacteriaceae. O sucesso no isolamento, manutenção de culturas puras, identificação e determinação da capacidade desses microrganismos degradarem MIB e geosmina da água sugere um potencial de utilização dessas bactérias em processos de filtração biológica, que utilizam carvão ativado granular. Esse conhecimento pode ser usado como uma nova ferramenta para a remoção de substâncias que causam gosto e odor, sendo essencial para garantir uma água de qualidade para o consumo humano. / Water eutrophication process, by phosphorous and nitrogen levels, allows cyanobacteria blooms which promotes, among other effects, the generation and release of the metabolites 2-methylisoborneol (MIB) and trans-1,10-dimethyl-trans-9- decalol (geosmin). These compounds are tertiary alcohols that may lend earthymusty tastes and odors to water, characteristics detected by consumers when the metabolites are present at concentrations as low as 5 to 10 ng/l. When present in water, mainly in the dissolved form (extracellular), MIB and geosmin are difficult to be removed by conventional treatment methods. The methodologies currently employed to remove these metabolites lack efficiency and increase operational costs, arousing the interest in research to develop alternative water treatment approaches. The present study was conducted in two steps. The first step evaluated the removal of MIB and geosmin by adsorption using different types of granular activated carbon, while the second step assessed the degradation of the metabolites by microbiological activity. Firstly, granular activated carbon was analyzed for iodine number and BET surface area. Next, adsorption equilibrium time was measured for each carbon type. Then, adsorption capacity was evaluated using the Freundlich isotherm. Biological oxidation of compounds causing taste and odor included isolation, identification and biodegradation tests using bacteria that employ MIB and geosmin as the only carbon sources. Regarding adsorption capacity, wood and coconut shell carbon performed better in MIB and geosmin removal, respectively, affording removal to levels below the odor threshold concentration (10 ng/l), using 50 mg/l solutions of each carbon and exposure times of 2 h. This characterization proves the importance of choosing the most appropriate carbon to employ in water treatment. Since little is known on the bacterial community thriving in natural waters, as far as we know this is the first study to record the capacity of Gram-positive and Gramnegative bacteria to individually degrade compounds that cause taste and odor in water, in the same test solution. The removal of 90 and 96% of MIB and geosmin within 48 and 120 h of exposure, respectively, shows that these bacteria may be used in water purification strategies. The molecular identification of the bacterial isolates obtained in Lake Guaíba revealed four new genera capable to degrade MIB and geosmin: Burkholderia sp., Caulobacter sp., Klebsiella sp., and Leucobacter sp. Apart from these isolates, one Arthrobacter sp., one Sphingomonadaceae and three Enterobacteriaceae isolates were also identified. The successful isolation, maintenance of pure cultures, identification and determination of these microorganisms’ capacities to degrade MIB and geosmin in water suggests that these bacteria may be used in biological filtration processes that utilize granular activated carbon. This knowledge may be used as a new tool to remove substances that lend taste and odor to water, and is an essential aspect to guarantee quality water for human consumption.

Made available in DSpace on 2016-08-29T15:09:36Z (GMT). No. of bitstreams: 1 tese_2643_Dissertação - kenyo.pdf: 1538823 bytes, checksum: 5f5d581d8ef6f3456294c46a2498b203 (MD5) Previous issue date: 2007-09-28 / Algumas Estações de Tratamento de Esgotos já possuem equipamentos e processos que reduzem as emissões de gases odorantes, entretanto, a população circunvizinha a esse tipo de empreendimento ainda pode ser afetada, visto que o sistema olfativo humano é capaz de perceber a presença de odores em concentrações extremamente baixas e em curtos intervalos de tempo. A estimativa da emissão de gases odorantes em ETE pode ser obtida através de modelos matemáticos que envolvem um balanço de massa em cada unidade da ETE, onde equações empíricas simulam os fenômenos físicos, químicos e biológicos (volatilização, stripping, adsorção, absorção, biodegradação e oxidação química) responsáveis pela remoção dos gases odorantes dissolvidos na massa de esgoto. Lopes (2005) adaptou um modelo de emissão para estimar a concentração e as taxas de remoção e emissão de H2S a partir de uma ETE do tipo compacta, composta por digestor anaeróbio de fluxo ascendente (UASB) associado a um biofiltro aerado submerso (BF). Apesar dos bons resultados apresentados pelo modelo de emissão adaptado, foi verificada a necessidade de sua calibração devido ao uso de parâmetros empíricos obtidos a partir de outros trabalhos que utilizaram outros tipos de ETE. O objetivo deste trabalho consiste na calibração do modelo de emissão adaptado e avaliado por Lopes (2005) através do uso da técnica de algoritmo genético (AG), responsável por reestimar alguns parâmetros associados às equações empíricas do modelo. Os resultados obtidos revelaram que os parâmetros associados à transferência global de massa entre a fase líquida e a fase gasosa diminuíram em torno de 25% após a calibração; o parâmetro associado à correção do oxigênio para H2S aumentou em 66.6%; e o parâmetro associado à biodegradação (remoção de H2S na fase líquida do BF) não sofreu alteração significativa após a calibração do modelo. As novas estimativas da concentração de H2S nas unidades estação elevatória (EE), caixa de areia (CA), UASB e BF, aproximaram-se das concentrações medidas por Sá (2004), comparando com as concentrações estimadas antes da calibração do modelo de emissão, em 0.51%, 0.25%, 0.29% e 1.96%, respectivamente. Em relação as taxas de emissão de H2S estimadas após a calibração do modelo de emissão, houve uma redução de 23% na EE, 11.6% na CA, 1.2% no UASB, 10.5% na CDV e 12.6% no BF, comparando com as taxas de emissão estimadas antes da calibração do modelo. / Some wastewater treatment plants already have equipments and processes that reduce emissions of odorant gases, however, the population surrounding this type of plant can still be affected by it, due to the fact that human olfactory system is capable of perceiving the presence of odors in extremely low concentrations and short time interval.. The estimative of the odorant gases emission in WWTP can be obtained through mathematical models which involve a balance of mass in each of the WWTP units, where empirical equations simulate the physical, chemical and biological phenomena (volatilization, stripping, adsorption, absorption, biodegradation and chemical oxidation) responsible for the removal of the odorant gases dissolved in the wastewater mass. Lopes (2005) adapted an emission model in order to estimate concentration, removal and emission rates of H2S from a compact type of WWTP, composed by upward flow anaerobic digester (UASB) associated with a submerged aired biofilter (BF). In spite of the good results showed by adapted emission model, it was detected a need to calibrate it due to the use of empirical parameters obtained from other works that used other types of WWTP. The goal of this work is to calibrate the adapted and evaluated emission model by Lopes (2005) through the use of the genetic algorithm (AG) technique, responsible for reestimate some of the parameters associated with the empirical equations of the model. The results obtained showed that the parameters associated with the overall mass transfer between the gas phase and liquid phase decreased by around 25% after calibration; the parameter associated with the correction of oxygen to H2S increased by 66.6%, and the parameter associated with biodegradation (H2S removal of the liquid phase of BF) suffered no significant changes after the model calibration. The new estimates of the concentration of H2S in pumping station (EE) units, grid chamber (CA), UASB and BF were similar to the ones measured by Sá (2004), comparing to the concentrations estimated before the calibration of the emissions model, 0.51%, 0.25%, 0.29% and 1.96%, respectively. Regarding the estimated emission rates of H2S after the calibration of the emissions model, there was a reduction of 23% in EE, 11.6% in CA, 1.2% in the UASB, 10.5% in the CDV and 12.6% in BF, compared to the estimated emission rates before the calibration of the model.