No description available.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1966. / Typescript. Vita. Includes facsimile of one published journal article and texts for two articles to be submitted to journals, all by the author and M.L. Jackson. Description based on print version record. Includes bibliographical references (leaves 116-128).

Thesis (MScEng (Process Engineering))--University of Stellenbosch, 2009. / The abundance of alkane by-products formed in South Africa presents a feedstock opportunity for the production of a wide range of commercially important products, such as long-chain dioic acids and alcohols. These compounds are formed as intermediates through the biological conversion of alkanes, a route which is particularly attractive when compared with chemical conversion due to its operation under milder process conditions. Furthermore, advances in genetic manipulation, which enable the accumulation of a range of metabolic intermediates, make the biological route remarkably flexible. From the literature review Yarrowia lipolytica was identified as a promising organism for use in studying alkane bioconversion because of its ability to produce large quantities of fatty acids when grown on n-paraffins as a sole carbon source. The bioconversion of alkanes will not only depend on the genetic modification but also on the process conditions to maximise growth and bioconversion. The overall objective of this project was therefore to investigate the potential of Y. lipolytica for alkane bioconversion by defining the conditions that maximise both cell growth and bioconversion. The Y. lipolytica strains supplied (TVN348, TVN493 and WT), however, were not yet modified to the extent that accumulation of metabolic intermediates was possible. Use was therefore made of a model system in which the alkane substrate was substituted with an even chain alkylbenzene. Since Y. lipolytica is unable to metabolise the benzene ring, the alkylbenzene is converted to the metabolic intermediate, phenyl acetic acid (PAA), and the potential for bioconversion assessed through measuring the accumulation of PAA. The specific objectives of the project were therefore 1) to define and quantify the parameters for the establishment of an effective model system in shake flasks with respect to trace elements, buffering, added nitrogen, oxygen supply, glucose concentration, alkylbenzene substrate and inducer requirements 2) to use the defined model system to identify the most promising strain of Y. lipolytica TVN348, TVN493 and WT 3) to use the defined model system and selected strain for evaluation of the influence of time of substrate addition and glucose concentration on cell growth and bioconversion of Y. lipolytica under controlled conditions in an instrumented bioreactor Furthermore, since poor reproducibility in cell growth and bioconversion had been prevalent in previous studies, it was also aimed to identify and statistically quantify the reproducibility between duplicate or triplicate samples in each experiment and between sets of different experiments with respect to PAA formation and cell concentrations. Studies were conducted in shake flask cultures to define and quantify the parameters for the model system. The parameters assessed included trace elements, buffering, nitrogen concentration, oxygen supply, glucose concentration, alkylbenzene substrate type and possible inducer requirements. Trace elements, phosphate buffering and added nitrogen did not significantly affect the cell growth of Y. lipolytica TVN348. The cell concentration of Y. lipolytica TVN348 and TVN493 was increased by 65% and 43% respectively for an increase in oxygen supply by decreasing the working volume from 150ml to 50ml, while the cell concentration of Y. lipolytica WT was increased by 41% when oxygen supply was increased by switching from non-baffled to baffled flasks in 50ml cultures. Bioconversion was also increased for an increase in oxygen supply: 2.4mM to 29.0mM PAA (Y. lipolytica TVN348) and 1.2mM to 21.7mM PAA (Y. lipolytica TVN493) for a decrease in working volume; 10.5mM to 46.6mM PAA (Y. lipolytica WT) when switching from non-baffled to baffled flasks. These results indicated that adequate oxygen supply is crucial to both growth and bioconversion, and that further study should be conducted in 50ml working volumes. Cell concentrations obtained in 1.6% (wt/v) and 3.2% (wt/v) glucose cultures (3.95x108cells/ml and 4.03x108cells/ml respectively) indicated that cell growth was neither enhanced nor inhibited by 3.2% (wt/v) glucose. Of the range of substrates examined (propylbenzene, butylbenzene, sec-butylbenzene, hexylbenzene, ethyltoluene and tert-butyltoluene for Y. lipolytica TVN348 and TVN493; octylbenzene and decylbenzene for Y. lipolytica WT), hexylbenzene was regarded as the best substrate for bioconversion (14.7mM and 14.1mM PAA for TVN348 and TVN493 respectively; 42.6mM PAA for WT). Lastly, the absence of a requirement for an additional inducer such as ethanol or oleic acid was confirmed when PAA was formed from hexylbenzene in the culture containing additional glucose (25.0mM). This suggested that when using hexylbenzene as substrate, bioconversion was induced provided sufficient glucose was available for cell maintenance. Results from duplicate or triplicate flasks in each individual shake flask experiment were reproducible and conclusions were based solely on results which showed 95% confidence intervals. However, reproducibility problems were experienced with results between different sets of experiments carried out under the same conditions. The model system was therefore defined by: 1) no addition of trace elements, additional buffering or added nitrogen, 2) cultures grown in 50ml volumes to supply an adequate amount of oxygen crucial for growth and bioconversion, 3) 3.2% (wt/v) glucose and 4) addition of 1% (v/v) hexylbenzene at 24h with no inducer requirements. Use of the model system in shake flask cultures to identify the most promising of the three strains of Y. lipolytica supplied demonstrated that there was no significant difference in cell growth or bioconversion between these strains. Y. lipolytica WT (which has no genetic modifications) was therefore used for further investigation until an appropriate strain could be substituted when it became available. The growth and bioconversion of Y. lipolytica WT was further investigated under controlled conditions in a bioreactor. The influence of time of substrate addition (11h, 24h, 48h) and glucose concentration (3.2% and 6.4% (wt/v)) on growth and bioconversion was examined. When hexylbenzene was added at 48h, cell growth was increased (8.90x108cells/ml) when compared to two of the triplicate cultures with hexylbenzene addition at 24h (4.74x108cells/ml and 3.92x108cells/ml) and the culture with hexylbenzene addition at 11h (2.82x108cells/ml). The third of the triplicate cultures with hexylbenzene addition at 24h, on the other hand, exhibited the strongest growth (2.23x109cells/ml). The poor reproducibility between the triplicate cultures with hexylbenzene addition as 24h made it difficult to determine whether hexylbenzene addition at 24h or 48h maximised cell growth. Furthermore, the cell growth was not significantly improved when the glucose concentration was increased from 3.2% (wt/v) to 6.4% (wt/v) (7.47x108cells/ml for 6.4% glucose culture), however it was also not inhibited. The highest amount of specific PAA formed by Y. lipolytica WT was found when hexylbenzene was added at 11h (7.4x10-11mmol PAA/cell), however the highest accumulated PAA was produced in the culture that exhibited the strongest growth with hexylbenzene addition at 24h (41.4mM). This suggested that the bioconversion of hexylbenzene was maximised when it was added during the active growth phase. It is therefore recommended to conduct fed-batch experiments in future to maintain the active growth phase. Accumulated PAA was increased in 6.4% (wt/v) glucose culture (15.2mM PAA) when compared with two of the 3.2% (wt/v) glucose cultures (5.4mM and 4.3mM PAA). These results indicated that the increased glucose concentration did not inhibit the bioconversion. Furthermore, PAA was formed when 5% (wt/v) residual glucose was observed in the culture, suggesting that the bioconversion of hexylbenzene was not inhibited at glucose concentrations as high as 5.0% (wt/v). If future work were to be conducted in bioreactor culture where glucose is added in fed-batch operation, glucose concentrations in cultures of up to 5% (wt/v) could be considered for initial studies. During bioconversion by Y. lipolytica, the PAA measured after hexylbenzene exhaustion did not, however, correspond to 100% conversion. Further, poor reproducibility was found in the bioreactor cultures. The disappearance of hexylbenzene without a corresponding accumulation of PAA and poor reproducibility was investigated by determining whether PAA was further degraded or alternatively, whether other metabolic intermediates were being formed and accumulated from the hexylbenzene. However, substitution of the hexylbenzene with PAA as substrate confirmed that PAA could not be metabolised. Further, NMR analyses of both the aqueous and organic phases of the culture did not identify any additional metabolic intermediates. It is recommended that additional analyses be conducted on the aqueous and organic phases to further assess the possible accumulation of intermediates. The development of the model system in shake flask cultures demonstrated the importance of adequate oxygen supply for both cell growth and bioconversion. It was also shown that no inducer was needed because hexylbenzene acted as its own substrate inducer. Furthermore, comparison of Y. lipolytica strains TVN348, TVN493 and WT under the defined conditions of the model system revealed that the genetically modified strains (TVN348, TVN493) did not exhibit enhanced bioconversion. Bioreactor cultures using the model system under controlled conditions further showed that bioconversion was not inhibited at a 5% (wt/v) residual glucose concentration and suggested that bioconversion was maximised when hexylbenzene was added during active growth phase. This informs on future work, suggesting fed-batch operation in order to extend the active growth phase, where glucose concentrations in the bioreactor of up to 5% (wt/v) can be considered.

The aim of this study was to understand the contribution of Linear Alkylbenzene Sulfonates (LAS) to the decline of Mediterranean coastal forest exposed to marine aerosols. LAS are a group of synthetic anionic surfactants widely used in the composition of household or industrial detergents and agrochemicals. This study was part of a large project (RISICO) aiming the assessment of the environmental impact of the LAS at multiple levels: biodegradation in the coastal waters, sorption – desorption processes in the sediments, toxicity to the aquatic life and toxicity to the coastal forest (by aerosolisation of the sea water). Previous studies pointed out this group of surfactants as the main cause of the coastal forest decline. However, the quantification of this surfactant in the environmental samples (mainly sea water and foliar deposition) was done using non-specific analytical methods as methylene blue active substances (MBAS), leading to overestimation of the environmental concentrations of LAS. The work hypothesis was that at actual environmental concentrations, the LAS does not play a key role in the foliar uptake of the sea salt deposited on the coastal vegetation by the marine aerosols, therefore the LAS may not be the main cause of the coastal forest decline. The research involved both greenhouse experiments and field measurements. The experimental work was conducted on young Mediterranean trees (Laurus nobilis L., Quercus ilex L. and Pinus halepensis P. Mill.) and investigated the synergistic toxic effects of exposure to simulated marine aerosol contaminated with surfactants. An array of endpoints was used including photosynthetic activity, relative water content, foliar deposition and uptake of salt and LAS, and pigments analysis. The results of those experiments revealed that LAS itself did not have phytotoxic effects. Nevertheless, the surfactant was shown to enhance the foliar uptake of the salt in the tested species, especially in Pinus halepensis, confirming the conclusions of previous studies regarding the sensitivity of this species to polluted marine aerosols. The field work was conducted in San Rossore National Park (Italy) and Porquerolles Island (France) and was focused on evaluating the health status of the Mediterranean forest (Quercus ilex L., Pinus halepensis Mill. and Pinus pinaster Aiton.) and also on quantification of LAS in coastal aerosols using highly specific analytical methods, like the mass spectrometry (MS). The frequencies and extent of injuries in the coastal trees were found to be correlated to the salt but not with the LAS content of the leaves. The concentrations of LAS in the Pinus and Quercus leaves were comparable in the two studied sites but the concentrations of salt were extremely high in San Rossore, suggesting that other factors may determine the excessive salt foliar uptake. The parallel MS and MBAS carried out in the same set of aerosol samples revealed that MBAS measurements were not relevant for LAS concentrations in the marine aerosols. Projecting the experimental results to the real LAS and salt exposure of the coastal forest, we concluded that LAS may play a marginal role in coastal vegetation decline.

No description available.

The aim of this study was to understand the contribution of Linear Alkylbenzene Sulfonates (LAS) to the decline of Mediterranean coastal forest exposed to marine aerosols. LAS are a group of synthetic anionic surfactants widely used in the composition of household or industrial detergents and agrochemicals. This study was part of a large project (RISICO) aiming the assessment of the environmental impact of the LAS at multiple levels: biodegradation in the coastal waters, sorption – desorption processes in the sediments, toxicity to the aquatic life and toxicity to the coastal forest (by aerosolisation of the sea water). <p>Previous studies pointed out this group of surfactants as the main cause of the coastal forest decline. However, the quantification of this surfactant in the environmental samples (mainly sea water and foliar deposition) was done using non-specific analytical methods as methylene blue active substances (MBAS), leading to overestimation of the environmental concentrations of LAS. <p>The work hypothesis was that at actual environmental concentrations, the LAS does not play a key role in the foliar uptake of the sea salt deposited on the coastal vegetation by the marine aerosols, therefore the LAS may not be the main cause of the coastal forest decline. <p>The research involved both greenhouse experiments and field measurements. The experimental work was conducted on young Mediterranean trees (Laurus nobilis L. Quercus ilex L. and Pinus halepensis P. Mill.) and investigated the synergistic toxic effects of exposure to simulated marine aerosol contaminated with surfactants. An array of endpoints was used including photosynthetic activity, relative water content, foliar deposition and uptake of salt and LAS, and pigments analysis. The results of those experiments revealed that LAS itself did not have phytotoxic effects. Nevertheless, the surfactant was shown to enhance the foliar uptake of the salt in the tested species, especially in Pinus halepensis, confirming the conclusions of previous studies regarding the sensitivity of this species to polluted marine aerosols. <p>The field work was conducted in San Rossore National Park (Italy) and Porquerolles Island (France) and was focused on evaluating the health status of the Mediterranean forest (Quercus ilex L. Pinus halepensis Mill. and Pinus pinaster Aiton.) and also on quantification of LAS in coastal aerosols using highly specific analytical methods, like the mass spectrometry (MS). The frequencies and extent of injuries in the coastal trees were found to be correlated to the salt but not with the LAS content of the leaves. The concentrations of LAS in the Pinus and Quercus leaves were comparable in the two studied sites but the concentrations of salt were extremely high in San Rossore, suggesting that other factors may determine the excessive salt foliar uptake. The parallel MS and MBAS carried out in the same set of aerosol samples revealed that MBAS measurements were not relevant for LAS concentrations in the marine aerosols. Projecting the experimental results to the real LAS and salt exposure of the coastal forest, we concluded that LAS may play a marginal role in coastal vegetation decline. <p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Made available in DSpace on 2016-06-02T19:31:40Z (GMT). No. of bitstreams: 1 1959.pdf: 1033941 bytes, checksum: 78e3218226ae4e1376a101c0d5d8d71f (MD5) Previous issue date: 2008-04-25 / Financiadora de Estudos e Projetos / The main anionic surfactants world widely used are the sodium dodecyl benzene sulfonate (LAS) and sodium dodecyl sulfate (DSS), which are mainly used in the manufacturing of domestic and personal hygiene products. As a consequence of the great and increasing consumption of LAS and DSS there is an increasing wareness regarding the adverse effects of these compounds to the organisms and environment. The present study aimed to evaluate the acute and chronic toxicity of the compounds LAS and DSS to the cladocerans Daphnia similis, Ceriodaphnia dubia and Ceriodaphnia silvestrii. It was also evaluated the toxicity of the water and sediments of four reservoirs and one stream of São Paulo State Analyses of LAS concentrations in the water of these reservoirs were also performed by Liquid chromatography. Acute toxicity tests indicated a value of CE(I)50;48h for LAS of 14.17 mg L-1 to D. similis, 11.84 mg L-1 to C. dubia and 13.51 mg L-1 to C. silvestrii. Significant changes in the viability of the cladoceran offsprings were observed for C. dubia and C. silvestrii exposed to the LAS, with values of CENO equal to 1.0 mg L-1 to C. dubia and 2.5 mg L-1 to C. silvestrii. It was concluded that the maximum permissible concentration of 0.5 mg L-1 surfactants as established by the resolution CONAMA nº. 357/2005 (Brazilian Ministry of Environment) in order to protect aquatic communities is adequate, considering the sensitivity of the native species C. silvestrii. The results of the acute toxicity tests with DSS indicated a CE(I)50;48h value of 12.82; 4.37 and 5.42 mg L-1, for D. similis, C. dubia and C. silvestrii, respectively. A CENO value of 2.0 mg L-1 was obtained in the chronic toxicity tests for C. silvestrii. The surfactant LAS was detected in all the water samples collected in the Lobo (Broa), Lagoa Dourada and Fazzari reservoirs, but at concentrations lower than 5 mg L-1 (the method limit of detection). The toxicity tests with environmental samples revealed that there is no toxicity in the water of the Lobo and Lagoa Dourada sampled, however the water of Monjolinho Reservoir was toxic to D. similis and Fazzari stream was toxic to D. similis and C. dubia. Only the sediment of Monjolinho Reservoir was not toxic to cladocerans. / Os principais surfactantes aniônicos disponíveis no mercado mundial são o dodecil benzeno sulfonato de sódio (LAS) e o dodecil sulfato de sódio (DSS), utilizados principalmente em produtos de limpeza doméstica e de higiene pessoal. Devido ao grande consumo mundial de LAS e de DSS há uma crescente preocupação sobre os efeitos adversos destes compostos no ambiente e aos organismos. Este trabalho teve por objetivo avaliar a toxicidade aguda e crônica do LAS e do DSS aos organismos-teste Daphnia similis, Ceriodaphnia dubia e Ceriodaphnia silvestrii. Foi também avaliada a toxicidade da água e dos sedimentos em quatro reservatórios e um riacho do estado de São Paulo por meio de testes de toxicidade aguda e da análise quantitativa do surfactante LAS. Os testes de toxicidade aguda indicaram uma CE(I)50;48h do LAS de 14,17 mg L-1 para D. similis, 11,84 mg L-1 para C. dubia e 13,51 mg L-1 para C. silvestrii. Nos testes crônicos realizados foi observada significativa alteração viabilidade da progênie de C. dubia e C. silvestrii exposta ao LAS, com valores de CENO igual a 1,0 mg L-1 para C. dubia e 2,5 mg L-1 para C. silvestrii. Pode-se concluir que o valor máximo permissível de surfactantes de 0,5 mg L-1, estabelecido pela Resolução CONAMA nº. 357/2005 em águas destinadas à proteção das comunidades aquáticas, é adequado para a espécie nativa C. silvestrii. Nos testes de toxicidade aguda de DSS foram obtidos valores de CE(I)50;48h de 12,82; 4,37 e 5,42 mg L-1, para D. similis, C. dubia e C. silvestrii, respectivamente. No ensaio de toxicidade crônica obteve-se valor de CENO igual a 2,0 mg L-1 para C. silvestrii. O surfactante LAS foi detectado nas amostras de água dos Reservatórios do Lobo (Broa), Lagoa Dourada e Represa do Monjolinho em concentração inferior a 5 mg L-1. Os testes revelaram que não há toxicidade aguda aos cladóceros da água da Lagoa Dourada e do Reservatório do Lobo, havendo, contudo, toxicidade da água da Represa do Monjolinho para D. similis e da água do Córrego do Fazzari para os cladóceros D. similis e C. dubia. Somente a amostra de sedimento da Represa do Monjolinho não causou toxicidade aos cladóceros.

Nesse trabalho avaliaram-se quatro hipóteses sobre a remoção do alquilbenzeno linear sulfonado (LAS) em reator EGSB (expanded granular sludge bed) alimentado ora com água residuária de lavanderia comercial, ora como meio sintético acrescido de LAS Padrão, com e sem suplementação de Fe(III) afluente. Para tanto, em todas as hipóteses utilizou-se reator EGSB (1,4 L) com tempo de detenção hidráulica (TDH) de 36h, condição mesofílica (30ºC) e carga de LAS específica aplicada (CLEA) variando de 1,0 - 2,7 mgLAS.gSTV-1.d-1. DGGE e sequenciamento massivo do gene rRNA 16S (Plataforma 454-Pirosequenciamento e Ion Torrent) foram utilizados para caracterização microbiana. Em relação à Hipótese A avaliou-se o efeito da adaptação prévia da biomassa na remoção do LAS em água residuária. Para tanto, o EGSB-BA (biomassa adaptada) teve uma etapa prévia com LAS padrão e meio sintético (Etapa I), seguida da Etapa II com água residuária; e o EGSB-BNA (biomassa não adaptada) teve etapa única e alimentação diretamente com água residuária. Para a Hipótese B avaliou-se o efeito da suplementação com meio sintético na remoção de LAS em água residuária. Para tanto, o EGSB-Ag.Lav foi alimentado apenas com água residuária e bicarbonato de sódio e duas CLE (Etapa II - 1,0 e Etapa III - 2,7 mg LAS.gSTV-1.d-1). Em relação às Hipóteses C e D, avaliou-se o efeito da suplementação de Fe(III) na remoção de LAS Padrão em meio sintético e LAS em água residuária, respectivamente. A Hipótese A foi refutada uma vez que as remoções de LAS em EGSB-BA-Etapa II (76%) e EGSB-BNA-Etapa I (78%) foram similares (ambas com água residuária). A remoção de LAS foi maior quando foi adicionada água residuária (EGSB-BA-Etapa II-76%) do que com LAS Padrão (EGSB-BA-Etapa I-63%). A Hipótese B foi aceita, uma vez que a alimentação do EGSB apenas com água residuária de lavanderia (CLE 1,0 mg LAS.gSTV-1.d-1) mais bicarbonato de sódio resultou em remoções do surfactante de 93%, ou seja, 15-17% maior que nos reatores suplementados com meio sintético (EGSB-BA Etapa II e EGSB-BNA Etapa I). Na Etapa III verificou-se diminuição da remoção em 30%. A Hipótese C foi aceita uma vez que se notou 20% de aumento na remoção de LAS quando comparado com reator não suplementado com Fe(III) (EGSB-Fe - 84,3% e EGSB-BA Etapa I - 63,5%). A Hipótese D foi refutada, uma vez que embora tenha sido obtida alta remoção de LAS (91,2%), esta não foi acompanhada pela redução férrica. Por meio do DGGE (domínio Bactéria) notou-se estratificação microbiana ao longo do reator na Etapa III (Hipótese B), provavelmente, em função do tamanho do grânulo que variou ao longo do reator. Por meio do sequenciamento massivo identificou-se bactérias semelhantes à Geobacter na amostra proveniente do reator EGSBFe da Hipótese C (17% da abundância relativa), portanto, as condições impostas favoreceram esse gênero. Fato este não observado para o reator EGSB-Fe-Ag.Lav. da Hipótese D. A comparação da análise filogenética das bactérias para os diferentes reatores permitiu identificar gêneros em comum relacionados com a degradação de LAS, a saber: Desulfobulbus, Geobacter, Syntrophorhabdus, Sporomusa, Comamonas, Holophaga, Mycobacterium, Pseudomonas, Stenotrophomonas e Synergistes. / This study evaluated four hypotheses about the removal of linear alkylbenzene sulfonate (LAS) in EGSB reactor (expanded granular sludge bed) fed sometimes with commercial laundry wastewater, sometimes with synthetic medium more Standard LAS, with and without Fe(III) influent supplementation. Therefore, in all hypotheses were used an EGSB reactor (1.4 L) with a hydraulic retention time (HRT) of 36 h, mesophilic condition (30°C) and load specific LAS (CLE) ranging from 1,0 to 2,7 mgLAS.gSTV-1.d-1. DGGE and massive sequencing of 16S rRNA gene (454-pyrosequencing and Ion Torrent platform) were used for microbial characterization. Regarding the Hypothesis A, it was evaluated the effect of biomass pre-adaptation for removal of LAS in wastewater. Then, the EGSBBA (adapted biomass) had a previous step with standard LAS and synthetic medium (Phase I), followed by Stage II with wastewater; and EGSB-BNA (not adapted biomass) had single step and feeding directly with wastewater. Regarding the Hypothesis B, it was evaluated the effect of synthetic medium supplementation in the removal of LAS in wastewater. Then, the EGSB-Ag.Lav was fed only with wastewater and sodium bicarbonate with two CLE (Stage II - 1,0 e Stage III - 2,7 mg LAS.gSTV-1.d-1). Regarding the Hypothesis C and D, it was evaluated the effect of Fe(III) supplementation in the removal of standard LAS and LAS in wastewater, respectively. The Hypothesis A was refuted since the LAS removal in EGSB-BA-Stage II (76%) and EGSB-BNA-Step I (78%) were similar (both with wastewater). The LAS removal was highest when wastewater was added (EGSB-BA-Stage-II 76%) than with standard LAS (EGSB-BAStage- I 63%). The Hypothesis B was accepted, since the feed of the EGSB only with wastewater from laundry (CLE 1,0 mg LAS.gSTV-1.d-1) more sodium bicarbonate resulted in removal of 93% of surfactant, in other words, 15-17% higher than in the reactors supplemented with synthetic medium (EGSB-BA Stage II e EGSB-BNA Stage I). In the Stage III, there was a decrease by 30% of LAS removal. The Hypothesis C was accepted, since there was an increase of 20% in the removal of LAS as compared to unsupplemented reactor with Fe (III) (EGSB-Fe - 84,3% e EGSB-BA Stage I - 63,5%). The Hypothesis D was refuted since although high LAS removal was obtained (91,2%), this was not accompanied by ferric reduction. By means of DGGE (Bacteria domain) was noted a microbial stratification along the reactor in the Stage III (Hypothesis B), probably in function of granule size along the reactor. By means of massive sequencing were identified bacteria similar to Geobacter in the sample from the reactor EGSB-Fe Hypothesis C (relative abundance 17%), therefore, the conditions favored this genre. This fact was not observed in the reactor EGSB-Fe-Ag.Lav. hypothesis D. A comparison of phylogenetic analysis of bacteria for different reactors allowed to identify common genera related to LAS degradation, namely: Desulfobulbus, Geobacter, Syntrophorhabdus, Sporomusa, Comamonas, Holophaga, Mycobacterium, Pseudomonas, Stenotrophomonas and Synergistes.

O objetivo desse trabalho foi avaliar a degradação de alquilbenzeno linear sulfonado (LAS) em condições anaeróbias. Os primeiros experimentos foram realizados em reatores em batelada alimentados com diferentes substratos e concentrações de LAS. Apesar do surfactante ficar adsorvido no lodo, não foram observadas interferências no metabolismo de microrganismos anaeróbios, pois dessa forma o LAS tornou-se indisponível para a degradação celular. Reatores anaeróbios horizontais de leito fixo (RAHLF) foram avaliados quanto à remoção de LAS e inoculados com lodos anaeróbios provenientes de reatores UASB usados respectivamente no tratamento de esgoto sanitário (R1) e tratamento de dejetos suinocultura (R2) imobilizados em espuma de poliuretano. A adição de LAS não influenciou na estabilidade do reator. O LAS começou a ser degradado após 108 dias da sua adição no afluente dos reatores. Porcentagens de remoção, considerando adsorção e degradação de LAS, com 313 dias de operação foram iguais a 50% e 91% para o R1 e R2, respectivamente, quando foram alimentados com esgoto sintético e 14 mg/L de LAS (reator - R1) e somente LAS a 14 mg/L (reator - R2). Em relação ao balanço de massa de LAS, os reatores apresentaram degradações muito semelhantes, sendo 35% para o reator R1 e 34% para o reator - R2. A diversidade microbiana referente aos domínios Bacteria e Archaea e ao grupo BRS foi avaliada utilizando a técnica de PCR/DGGE. Para o domínio Archaea, foram observadas diferenças significativas nas populações quando os reatores foram alimentados com LAS. Diferenças foram observadas no domínio Bactéria e grupo das BRS, para concentrações de LAS de 14 mg/L. A alteração na diversidade microbiana pode ter ocorrido devido à seleção dos microrganismos pela presença do surfactante. A biomassa presente no final da operação foi submetida à técnica de clonagem e seqüenciamento do fragmento do RNAr 16S para o domínio Bacteria. Observou-se que os reatores que apresentaram maior número de clones relacionados ao filo Firmicutes, classe Clostridia, ordem Clostridiales. Provavelmente os microrganismos pertencentes a esse grupo estejam envolvidos com a degradação do LAS / The objective of this work was to evaluate the degradation of linear alkylbenzene sulfonate (LAS) in anaerobic conditions. The first experiments were accomplished in reactors in batch fed with different substrates and concentration of LAS. In spite of the surfactant to be adsorbed in the sludge interferences was not observed in the metabolism of anaerobic microorganisms, because in that way LAS became unavailable for the cellular degradation. Horizontal anaerobic immobilized biomass (HAIB) reactors were appraised as for the removal of LAS and inoculated with coming anaerobic slugde of reactors UASB used respectively in the treatment of sanitary sewage (R1) and treatment of wastewater swine (R2) immobilized polyurethane foam. The addition of LAS didn’t influence in the stability of the reactor. LAS began to be degraded after 108 days of its addition in the tributary of the reactors. Removal percentages, considering adsorption and degradation of LAS, with 313 days of operation was same to 50% and 91% for R1 and R2, respectively, when they were fed with synthetic sewage and 14 mg/L of LAS (reactor – R1) and only LAS to 14 mg/L (reactor – R2). In relation to the balance of mass of LAS, the reactors presented very similar degradations, being 35% for the reactor R1 and 34% for the reactor R2. The microbial diversity regarding the Bacteria and Archaea domain and to the group BRS was evaluated using the technique of PCR/DGGE. The alteration in the microbial diversity might have happened due to the selection of the microorganisms for the presence of the surfactant. The biomass present in the end of the operation was submitted the cloning technique and sequencing of the fragment of 16S rRNA for the bacteria domain. It was observed that the reactors presented larger number of clones related to the phylum Firmicutes, Clostridia, Clostridiales. Probably the microorganisms belonging to that group are involved with the degradation of LAS

O Alquilbenzeno Linear Sulfonado (LAS) é um surfactante aniônico considerado prioridade de risco ambiental entre os contaminantes emergentes atuais. Estudos anteriores reportaram a degradação anaeróbia de LAS por co-digestão da água residuária de lavanderia comercial e co-substratos sintéticos (extrato de levedura e etanol), em reatores em escala de bancada. Todavia, na literatura ainda são limitados os estudos envolvendo co-substratos economicamente viáveis, como o esgoto doméstico. O objetivo deste estudo foi avaliar a eficiência de remoção do LAS, de água residuária de lavanderia comercial em co-digestão com esgoto doméstico. O reator de leito granular expandido (EGSB) escala piloto de 62,0 L foi operado em condição mesofílica, em TDH de 40 h, por 480 dias. O reator foi inoculado com 12,0 L de lodo proveniente de reator UASB usado no tratamento de água residuária de abatedouro de aves. Na primeira etapa, de adaptação, com duração de 60 dias, o reator foi alimentado apenas com esgoto doméstico. Em seguida, o reator foi alimentado com água residuária de lavanderia comercial diluída em esgoto doméstico para obtenção de concentrações de LAS afluente de 5 ± 4; 19 ± 10 e 36 ± 19 mg L-1, nas etapas II, III e IV, respectivamente. Nestas etapas o reator foi monitorado por 60, 270 e 90 dias. Verificou-se diminuição da eficiência de remoção de matéria orgânica de 73 ± 23 % na etapa de adaptação, para 61 ± 25%, 50 ± 15% e 44 ± 15%, respectivamente, para as etapas II, III e IV. Assim como, verificou-se diminuição da eficiência de remoção do LAS de 73 ± 31% para 55 ± 29% e 32 ± 17% nas três últimas etapas. Verificou-se aumento significativo de sulfeto da etapa II para a etapa III e IV, respectivamente de 8 ± 2 mg L-1 para 29 ± 12 mg L-1 e 31 ± 5 mg L-1, respectivamente. Foi realizada a caracterização taxonômica da biomassa do leito do lodo do reator EGSB ao final de cada etapa experimental, sendo identificadas bactérias semelhantes a Bellinea (domínio Bacteria) e Methanosaeta (domínio Archaea), as quais foram as de maior abundância relativa ao longo de toda operação. A tecnologia empregada para remoção do surfactante foi adequada para concentrações afluente de até 20 mgLAS L-1. Aclimatação da biomassa microbiana foi observada, após longo período e exposição aos compostos recalcitrantes da água residuária de lavanderia. / Classified as an emergent contaminant, Alkylbenzene Linear Sulfonate (LAS) is an anionic surfactant of high priority given its environmental risk. Previous studies reported LAS anaerobic degradation from laundry wastewater in co-digestion with synthetic substrates (such as methanol, ethanol and yeast extract). However, studies with more economic viable substrates as domestic sewage still lacking. This study assessed LAS removal from laundry wastewater diluted in domestic sewage, using an Expanded Bed Granular Sludge (EGSB) pilot scale reactor. The reactor was kept in mesophilic temperature, with 62,0 L total volume and it was operated with a hydraulic retention time (HRT) of 40 h. The inoculum was 12,0 L of sludge from an UASB used on a poultry slaughterhouse wastewater treatment plant. On the first stage, of adaptation, the reactor was fed only with domestic sewage for 60 days. Then, commercial laundry wastewater was diluted in domestic sewage to obtain LAS affluent concentrations of 5 ± 4; 19 ± 10 and 36 ± 19 mg L-1, on stages II, III and IV, respectively. These stages lasted for 60, 270 and 90 days. The increase of commercial laundry wastewater proportion on fed resulted in lower organic matter removal efficiencies. These parameter decreased from 73 ± 23% on adaptation, to 61 ± 25%, 50 ± 15% and 39 ± 23% on following stages. As well as LAS removal efficiencies that dropped from 73 ± 31% to 55 ± 29% and 44 ± 15 % on three last stages. Sulphide production enhanced from stage II (8 ± 2 mg L-1) to stage III (29 ± 12 mg L-1) and remained 31 ± 5 mg L-1 on stage IV. Taxonomic characterization of granular sludge biomass in the end of each stage was performed and genders similar to Bellinea (Bacteria domain) and Methanosaeta (Archae domain) were the ones with higher relative abundance in all stages. The technology applied was suitable for anaerobic surfactant removal with LAS affluent concentration of 20 mg L-1. Acclimation of microbial biomass was observed after long exposure of microbial community to recalcitrant compounds from laundry wastewater.