The performance of an aerated horizontal subsurface flow treatment wetland was investigated before, during and after a simulated aeration failure. Conventional wastewater parameters (e.g., carbonaceous biological oxygen demand, total nitrogen, and Escherichia coli) as well as selected micropollutants (caffeine, ibuprofen, naproxen, benzotriazole, diclofenac, acesulfame, and carbamazepine) were investigated. Furthermore, the removal of biological effects was investigated using in vitro bioassays. The six bioassays selected covered environmentally relevant endpoints (indicative of activation of aryl hydrocarbon receptor, AhR; binding to the peroxisome proliferator-activated receptor gamma, PPARγ; activation of estrogen receptor alpha, ERα; activation of glucocorticoid receptor, GR; oxidative stress response, AREc32; combined algae test, CAT). During the aeration interruption phase, the water quality deteriorated to a degree comparable to that of a conventional (non-aerated) horizontal subsurface flow wetland. After the end of the aeration interruption, the analytical and biological parameters investigated recovered at different time periods until their initial treatment performance. Treatment efficacy for conventional parameters was recovered within a few days, but no complete recovery of treatment efficacy could be observed for bioassays AhR, AREc32 and CAT in the 21 days following re-start of the aeration system. Furthermore, the removal efficacy along the flow path for most of the chemicals and bioassays recovered as it was observed in the baseline phase. Only for the activation of AhR and AREc32 there was a shift of the internal treatment profile from 12.5% to 25% (AhR) and 50% (AREc32) of the fractional length.

No description available.

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are common contaminants of sediment, soils and biological tissues. These compounds pose a significant risk to biological and ecosystem health and functioning due to these compounds being mutagenic, carcinogenic and are known to disrupt the endocrine system. The bioaccumulation and biomagnification potential that these compounds possess mean that they are capable of affecting the entire food chain and are not limited to the organisms that are directly in contact with the compounds Even though there has been an increase in the attention on identifying the presence and impacts that these compounds may have in South Africa, the level of attention is lower than what it is in other countries around the world. Although South Africa has guidelines in place for other pollutants, such as metals, there are no such guidelines in place to monitor PAHs and PCBs.. Industries are known to release both PAHs and PCBs, mainly from incomplete burning processes and the release of oils and fuels in the case of PAHs, and from heat transfer fluids in the case of PCBs. Durban Bay and surrounding areas of eThekwini, KwaZulu-Natal are highly industrialised with many aquatic systems, in which these contaminants are likely to deposit. The aim of this research was to determine the concentrations of these compounds by means of chemical analysis and additionally biological analysis, using the H4IIE-luc bioassay and compare these levels to international guideline levels. It was found that the concentrations of the 23 analysed PAHs were 6.5–3 235.6 ng.g-1 and the concentration of the PCBs analysed were 0–113.83 ng.g-1. Many sites were found to be in exceedance of the guideline limits, particularly in the harbour. Toxic equivalency factors (TEF) were used to gauge the toxic equivalency (TEQ) of the PAHs and PCBs that were found. The TEQs were generally low, and were below any guideline levels. The assay revealed the extract containing the PCBs had a bioassay equivalence (BEQ) of 0–93.54 pgTCDDeq. g-1 and the extract containing the PAHs of 0–776.08 pgTCDD-eq.g-1. With a proportion of the sites exceeding guideline limits. The BEQ results were two to three orders of magnitude greater than the TEQs calculated from the concentrations determined by the instrumental analysis, however, followed a similar trend. Additionally chemical analysis was not performed on a full suite of compounds that are able to elicit a response from the cells, which could be a reason why the BEQ and TEQ did not follow a similar trend among some of the samples. It would have been more beneficial to have performed chemical analysis on the 16 priority PAHs (as determined by the United States Environmental Protection Agency), the dioxin-like PCBs and polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDFs) which are all capable of eliciting a response from the cells and have TEF values. The areas that were most affected by contamination of these compounds was the harbour and surrounding canals, and there was point source contamination along the Umhlatuzana, Umbilo, and Amanzimnyana Rivers. All dl-PCBs should be chemically analysed at all the sample areas, as these cause adverse effects to biota. In addition to this, biota should be sampled to determine concentrations of the compounds to determine bioavailability and the degree of bioaccumulation in the food chain. Utilising biomarkers it would be possible to determine stresses of fish. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are common contaminants of sediment, soils and biological tissues. These compounds pose a significant risk to biological and ecosystem health and functioning due to these compounds being mutagenic, carcinogenic and are known to disrupt the endocrine system. The bioaccumulation and biomagnification potential that these compounds possess mean that they are capable of affecting the entire food chain and are not limited to the organisms that are directly in contact with the compounds Even though there has been an increase in the attention on identifying the presence and impacts that these compounds may have in South Africa, the level of attention is lower than what it is in other countries around the world. Although South Africa has guidelines in place for other pollutants, such as metals, there are no such guidelines in place to monitor PAHs and PCBs.. Industries are known to release both PAHs and PCBs, mainly from incomplete burning processes and the release of oils and fuels in the case of PAHs, and from heat transfer fluids in the case of PCBs. Durban Bay and surrounding areas of eThekwini, KwaZulu-Natal are highly industrialised with many aquatic systems, in which these contaminants are likely to deposit. The aim of this research was to determine the concentrations of these compounds by means of chemical analysis and additionally biological analysis, using the H4IIE-luc bioassay and compare these levels to international guideline levels. It was found that the concentrations of the 23 analysed PAHs were 6.5–3 235.6 ng.g-1 and the concentration of the PCBs analysed were 0–113.83 ng.g-1. Many sites were found to be in exceedance of the guideline limits, particularly in the harbour. Toxic equivalency factors (TEF) were used to gauge the toxic equivalency (TEQ) of the PAHs and PCBs that were found. The TEQs were generally low, and were below any guideline levels. The assay revealed the extract containing the PCBs had a bioassay equivalence (BEQ) of 0–93.54 pgTCDDeq. g-1 and the extract containing the PAHs of 0–776.08 pgTCDD-eq.g-1. With a proportion of the sites exceeding guideline limits. The BEQ results were two to three orders of magnitude greater than the TEQs calculated from the concentrations determined by the instrumental analysis, however, followed a similar trend. Additionally chemical analysis was not performed on a full suite of compounds that are able to elicit a response from the cells, which could be a reason why the BEQ and TEQ did not follow a similar trend among some of the samples. It would have been more beneficial to have performed chemical analysis on the 16 priority PAHs (as determined by the United States Environmental Protection Agency), the dioxin-like PCBs and polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDFs) which are all capable of eliciting a response from the cells and have TEF values. The areas that were most affected by contamination of these compounds was the harbour and surrounding canals, and there was point source contamination along the Umhlatuzana, Umbilo, and Amanzimnyana Rivers. All dl-PCBs should be chemically analysed at all the sample areas, as these cause adverse effects to biota. In addition to this, biota should be sampled to determine concentrations of the compounds to determine bioavailability and the degree of bioaccumulation in the food chain. Utilising biomarkers it would be possible to determine stresses of fish. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The Southern Ocean (SO) is of significant interest in the understanding of the global carbon cycle and therefore many studies have been conducted to determine the limiting factors controlling the biological pump within the region. During photosynthesis phytoplankton require various nutrients such as NO3, PO4, inorganic carbon and the micronutrient Fe. The SO is a High-Nutrient Low-Chlorophyll region, therefore no macronutrient limitation is experienced by resident phytoplankton but instead the micronutrient Fe is a significant limiting factor within these waters due to limited inputs. Due to deep mixed layer depths, ice cover, low sun angles and cloud cover throughout parts of the year, light is also considered a limiting factor in the SO. Fe and light limitation cause a decrease in photosynthetic efficiency and therefore a decrease in carbon fixation capabilities. During this study we conducted five bioassay shipboard incubation experiments during two cruises along the Greenwich meridian between South Africa and the ice edge, SOSCEx during March and SAFePool during January to February, in which we varied Fe concentrations and light levels to determine the effects of Fe and light limitation or co-limitation within resident phytoplankton. Spatial and temporal variations in phytoplankton response were studied to determine varying effects of limitation across water masses and different stages of bloom decline within the study area. The combined addition of Fe and light gave the largest increase in biomass, photosynthetic capacity and nutrient uptake. In support of the hypotheses tested changes in the photosynthetic apparatus led to changes in the photosynthetic efficiency and growth of the SO phytoplankton, as a result of variations in Fe and light availability. Variability was also observed in the response of phytoplankton to Fe and light amendments due to spatial and temporal variation in resident phytoplankton communities. It was therefore concluded that both Fe and light are significant controls in the resident phytoplankton photosynthetic apparatus, photosynthetic capabilities, organic carbon fixation and therefore the biogeochemical cycles within the Atlantic sector of the SO. / AFRIKAANSE OPSOMMING: Die Suidelike Oseaan (SO) is van beduidende belang in die begrip van die globale koolstofsiklus en dus is baie studies gedoen om die beperkende faktore te bepaal wat die biologiese pomp in die streek beheer. Tydens fotosintese benodig fitoplankton verskillende voedingstowwe soos NO3, PO4, anorganiese koolstof en die mikrovoedingstof Fe. Die SO is 'n High-Nutrient Lae-Chlorofil streek, dus word geen makrovoedingstof beperking ervaar deur inwoner fitoplankton maar in plaas daarvan is die mikrovoedingstof Fe 'n beduidende beperkende faktor binne hierdie waters weens beperkte insette. As gevolg van diep gemengde laag dieptes, ysbedekking, lae son hoeke en wolkbedekking deur dele van die jaar, word lig ook beskou as 'n beperkende faktor in die SO. Fe en lig beperking veroorsaak 'n afname in die fotosintetiese doeltreffendheid en dus 'n afname in koolstof binding vermoëns. Tydens hierdie studie het ons vyf biotoets inkubasie eksperimente aan boord die skeep gedoen tydens twee vaarte langs die Greenwich meridiaan tussen Suid-Afrika en die ys rand, SOSCEx gedurende Maart en SAFePool gedurende Januarie tot Februarie, waarin ons Fe konsentrasies en lig vlakke gewissel het om die gevolge van Fe en lig beperking, of medebeperking, binne inwoner fitoplankton te bepaal. Ruimtelike en temporale variasies in fitoplankton reaksie was bestudeer om wisselende gevolge van die beperking oor watermassas en verskillende stadiums van bloei afname in die studie area te bepaal. Die gekombineerde byvoeging van Fe en lig het die grootste toename in biomassa, fotosintetiese kapasiteit en voedingsopname gegee. Ter ondersteuning van die getoetste hipoteses, veranderinge in die fotosintetiese apparaat het gelei tot veranderinge in die fotosintetiese doeltreffendheid en groei van die SO fitoplankton, as 'n gevolg van variasies in Fe en lig beskikbaarheid. Veranderlikheid is ook waargeneem in die reaksie van fitoplankton om Fe en lig wysigings weens die ruimtelike en tydelike variasie in inwoner fitoplankton gemeenskappe. Dus was dit by die gevolgtrekking gekom dat beide Fe en lig beduidende kontrole in die inwoner fitoplankton fotosintetiese apparaat, fotosintetiese vermoëns, organiese koolstof binding en daarom die biogeochemiese siklusse binne die Atlantiese sektor van die SO.

O presente estudo teve como objetivo avaliar a qualidade ambiental do reservatório de Salto Grande (Americana, SP), localizado em área de intensa urbanização e atividade industrial. Utilizou-se, para tanto, análises físicas e químicas na água e no sedimento, bioensaios de toxicidade e estrutura (composição e densidade) da fauna de macroinvertebrados bentônicos em quatro períodos (Maio, Agosto, Novembro de 2000 e Fevereiro de 2001). Pela análise de nutrientes na água constatou-se que as concentrações são elevadas, principalmente do íon amônio e do fosfato total, os quais estiveram acima do limite estabelecido pela resolução CONAMA 20/86 e, com base no Índice do Estado Trófico o sistema caracteriza-se como eutrófico ou hipereutrófico, dependendo do período e da localização das estações amostradas, verificando-se a redução na concentração de nutrientes e conseqüentemente, do estado trófico, no sentido do rio para a barragem. A concentração de nutrientes no sedimento também foi elevada quando comparada a outros reservatórios e a análise granulométrica demonstra que as estações centrais do reservatório possuem sedimento silte-argiloso enquanto as estações marginais e o rio Atibaia apresentam sedimentos arenosos. A análise de metais indica que a água e o sedimento do sistema encontram-se contaminados para todos os metais estudados (Cd, Cu, Cr, Fe, Mg e Mn), exceto para Zn na água e Pb no sedimento. A análise de metais nos organismos bentônicos aponta para uma bioacumulação de Cd, Cu, Fe, Mg, Mn, e Zn o mesmo não sendo observado para Pb e Cr. Nos bioensaios de toxicidade aguda verificou-se, para Chironomus xanthus, maior toxicidade no rio Atibaia, com 61% de mortalidade em Novembro/2000, enquanto que para Daphnia similis a toxicidade máxima (100% de imobilidade em Agosto/2000) foi registrada nas estações próximas à barragem. A análise da fauna de macroinvertebrados bentônicos demonstra a predominância, nas estações centrais do reservatório (mais profundas), de organismos pertencentes à Classe Oligochaeta e nas estações marginais (zona litorânea) a fauna é mais diversa, sendo composta pela Classe Oligochaeta, Filo Mollusca e Ordem Ephemeroptera (Classe Insecta). A fauna do rio Atibaia é formada, predominantemente, pelas Classes Hirudinea e Oligochaeta. O estágio avançado da eutrofização do reservatório fez com que a fauna de macroinvertebrados fosse reduzida, quando comparada à estudos anteriores, possivelmente devido ao desencadeamento de processos associados ao grande aporte de nutrientes, como a redução na concentração de oxigênio dissolvido ou devido ao aporte contínuo de diversos metais para o sistema e, provavelmente, esteja havendo uma processo de substituição de espécies por organismos da Classe Oligochaeta, como a Família Tubificidae, que são mais tolerantes aos diversos tipos de poluição. Conclui-se que as altas concentrações de nutrientes e metais, com valores fora dos padrões estabelecidos pelos órgãos ambientais, na água e no sedimento do reservatório, relacionam-se com as atividades desenvolvidas na bacia hidrográfica, e indicam os impactos decorrentes desta utilização e seu reflexo nas características físicas, químicas, toxicológicas e biológicas do sistema. / The goal of this work was to asses the environmental quality of Salto Grande reservoir (Americana, SP), located in an urban area with high industrial activities. Physical and chemical analyses were made in water and sediment, toxicity bioassays and structure (composition and density) of benthic organisms in four months (May, August, November 2000 and February 2001). The water nutrients analyses demonstrated high concentrations, mainly of ammonia ion and total phosphate, exceeding the limits from CONAMA 20/86, and based on the Trophic State Index, the reservoir was classified as eutrophic or hipereutrophic, depending on the months and the sampling stations, verified by a reduction on nutrients concentrations and in the trophic state, in the river-dam direction. The nutrients concentrations in the sediment was also elevated when compared with others reservoirs and the grain size composition demonstrated that the central sampling stations had silt-clay composition while the shore sampling stations and the Atibaia river had sand composition. The metals analysis indicates that water and sediment were contaminated by Cd, Cu, Cr, Fe, Mg, e Mn, with exception for Zn in the water and Pb in the sediment. The metals analysis in the benthic organisms indicates a bioaccumulation of Cd, Cu, Fe, Mg, Mn and Zn, with exception for Pb and Cr. The acute toxicity bioassays verified, for Chironomus xanthus, higher toxicity in the Atibaia river, with 61% of mortality in November 2000, while for Daphnia similis the maximum toxicity (100% of immobility in August 2000) was observed on the sampling stations near the dam. The analysis of benthic organisms demonstrates the predominance of Oligochaeta, on central sampling stations (with higher profundity), and a higher diversity fauna, mainly by Oligochaeta, Mollusca and Ephemeroptera (Insecta), on the shore sampling stations. The Atibaia river fauna is composed mainly by Hirudinea and Oligochaeta. The higher eutrophication process in the reservoir probably was the cause of the reduction of macroinvertebrates fauna, when compared with studies in the past. The major process of these problems are the elevated input of nutrients, reduction on the dissolved oxygen and the input of metals, resulting in the substitution of the species for Oligochaeta organisms (Tubificidae), which are tolerant to different pollutants. To conclude, the high concentrations in the water and sediment for nutrients and metals, with values that exceed the limits from Environmental Agency, were related to the activities developed in the Watershed, indicating impacts produced by this utilization and its effects on the physical, chemical toxicological and biological characteristics of the system.

A cebola comum Allium cepa L. (2n=16), devido à sua excelente condição cromossômica, tem sido largamente utilizada como um sistema de testes para se examinar os efeitos citogenéticos causados por diversos compostos químicos em material biológico. Em estudos de monitoramento de riscos ambientas, observou-se limitação no crescimento das raízes quando as cebolas são expostas a aguas residuárias provenientes de indústrias químicas. Nesse estudo, nós avaliamos este sistema de testes escolhendo alguns compostos de metais pesados, cujos efeitos em outros sistemas de testes já se encontram descritos. Além disso, foram estuados vários fatores que poderiam interferir nos resultados tais como: temperatura; luminosidade; qualidade da água e qualidade das cebolas. As concentrações mais baixas empregadas (1,5 mg/l) nos testes, de cádmio e cobre demostraram alta capacidade inibidora sobre o crescimento das raízes. O zinco apresentou toxicidade moderada a 1,5 mg/l e crescente às concentrações sucessivas mais elevadas. Efluentes industriais, contendo misturas de níquel e sulfatos ou cobre e cianeto, demonstraram-se altamente inibidores às concentrações mais baixas empregadas (0,7%), resultados que demonstram a alta sensibilidade do teste aos sais de metais pesados. Quanto às condições experimentais que levaram a esses resultados, os testes revelaram que não existe influência pronunciada dos fatores luminosidade e temperatura. Quanto à água para controle e diluição, não deve ser de torneira devido ao efeito inibidor do zinco e do cobre. Os resultados foram contraditórios em relação a um possível efeito osmótico da água destilada, sendo conveniente adicionar entre 100 a 250 mg/l de NaCl. A melhor resposta foi obtida com cebolas de origem argentina de 3,5 a 4,0 cm de diâmetro sendo preferível a manutenção de cascas para evitar murchamento. / The common onion Allium cepa. L., due to its excellent chromosome conditions, has been widely used as a test system for examining the citogenetic effects of several chemical compounds on biological material. It has been observerd, in monitoring environmental studies, some growth restriction on Allium roots when they are exposed to wastewater from chemical factories. In this study, we evaluate these test systems by choosing some hard metals which harmful effects on biological materials has been already described. Furthermore, some factors that could interfere on the results, such as: temperature; light; water quality and onion quality were studied. The lowest concentration (1.5 mg/l) for cadmium and cupper were found to have hard capacity to inhibit root growth. The zinc showed moderate toxicity at 1.5 mg/l and this effect was more evident at increased concentrations. The industrial wastewater that contains mixture of nickel and sulphate or cupper and cyanate inhibited root growth at lower concentration (0.7%). These results showed that the test system has high sensitivity for hard metals. There was no direct influence of temperature and light on the experiment conditions. With regard to the type of water, tap water should not be used because of inhibition on the root growth caused by zinc and copper. The results contradicted a potential osmotic effect of distillated water, being convenient to add 100 - 250 mg/l of NaCl. The best response was obtained with onions from Argentina with 3.5 - 4.0 cm of diameter being recommended to keep the onion peel for avoiding dryness.

Da fração hexânica do extrato metanólico das folhas de Guarea guidonia (Meliaceae), foram isolados os diterpenos: [11S-(1α,2E,6E,10α)]-3,7,11-trimetil-11-(4-metil-pent-3-en-1-i1)biciclo[8.1.0]undeca-2,6-dieno e [1S(1aβ,4aα,7aβ,7bβ)]-decaidro-1,7-dimetil-1-(4-metil-pent-3-en-1-il)-4-metilen-1H-cicloprop[e]azulen-7-ol, já descritos na literatura, além de dois novos isômeros de [1S-(1aβ,4aα,7aβ, 7aβ)]-decaidro-1,7-dimetil-1-(2-hidroxi-4-metil-pent-3-en-1-il)-4metilen-1H-cicloprop[e]azulen-7-ol. No óleo essencial obtido das folhas do mesmo espécimen foram identificados, através da análise dos dados espectroscópicos (RMN 13C, RMN 1H e EM), os sesquiterpenos: eudesma-5,7-dieno, 5α,6α-epoxi-eudesm-7-eno, eudesma-5,7-dien-2α-ol, 5α,6α-epoxi-eudesm-7-en-9-ol e guaia-6-en-10-ol, ainda não descritos na literatura, além de eudesma-4(15), 11(13)-dieno, eudesm-6-em-4α-ol, eudesma-4,11-dieno e 5α,6α,7α,8α-diepoxieudesmano. Diversos extratos vegetais assim como substâncias puras e misturas de composições conhecidas foram testadas com a finalidade de se detectarem substâncias com atividade anticancerígena. Para tal, empregou-se o bioensaio com linhagens mutantes do fungo Saccharomyces cerevisiae, utilizados pelo grupo do Prof. Dr. David Kingston, do Virgínia Tech Institute. Também foi realizado um estudo químico biomonitorado por aquele bioensaio. / From the hexanic extract of the dry leaves of Guarea guidonia (Meliaceae) two new isomers of decahydro-1,7-dimethyl-1-(2-hydroxy-4-methyl-pent-3-en-1-yl)4- methylen-1H-cycloprop[e]azulen-7-ol were isolated together with the known diterpenes [11S-(1α,2E,6E,10α)]-3,7,11-trimethyl-11-(4-methyl-pent-3-en-1yl) bicyclo[8.1.0]undeca-2,6-dien and [1S-(1aβ,4aα,7aβ,7bJ3)]-decahydro-1,7dimethyl- 1-(4-methyl-pent-3-en-1-yl)-4-methylen-1H-cycloprop[e]azulen-7-ol. The essential oil from the leaves of the same plant afforded the sesquiterpenes eudesma-5,7-diene, 5α,6α-epoxy-eudesm-7-ene, eudesma-5,7-dien-2α-ol, 5α,6α-epoxy-eudesm-7-en-9-ol and guaia-6-en-10-ol not previouly described, besides eudesma-4(15),11 (13)-diene, eudesm-6-en-4α-ol, eudesma-4,11-diene and 5α,6α,7α,8α-bisiepoxy-eudesmane. All of these compounds were identified through 13C and 1H NMR spectroscopy as well as MS. Several plant extracts and pure compounds were tested in the search for potential anticancer agents, through the mechanism-based yeast bioassay, utilizing mutants of the yeast Saccharomyces cerevisiae. The screening of those plant materials and a bioassay-guided fractionation of an active extract were both carryed out in the laboratories of Professor David G.I. Kingston at Virginia Tech Institute.

Neste trabalho foi explorado o potencial das actinobactérias como produtoras de agentes antimicrobianos contra bactérias causadoras da mastite bovina. Para este fim, foram otimizados dois diferentes bioensaios um de antagonismo e o outro de antibiose, contra 13 bactérias Gram-positivas e Gram-negativas relacionadas a mastite bovina. Através do bioensaio de antagonismo 120 actinobactérias foram triadas, sendo 55% delas consideradas ativas. Para o ensaio de antibiose foram triados 70 extratos brutos e 60% mostram a formação de halos de inibição. Os extratos brutos Caat 1-54, Caat 2-63 e Caat 8-25 foram considerados os mais promissores dentre todos os extratos estudados. Esses extratos brutos foram submetidos a estudos de desreplicação empregando LC-PAD-MS/MS e isolamento guiado por bioensaio para a identificação de seus metabolitos ativos. Os extratos Caat 1-54 e Caat 2-63 foram ativos principalmente contra as linhagens de bactérias Gram-positivas e a ação antimicrobiana observada foi atribuída a presença da lisolipina I e actinomicina D e V, respectivamente. O extrato Caat 8-25 foi ativo tanto contra bactérias Gram-positivas como para as Gram-negativas, sendo a oxitetraciclina associada a atividade contra as linhagens Gram-positivas e a herbicidina B ativo contra linhagens Gram-negativas. O estudo de desreplicação por LC-MS/MS também possibilitou a identificação de outros metabolitos secundários presentes nos diferentes extratos estudados. Para o extrato Caat 2-63 foi identificada a presença de outros peptídeos provenientes da via biossintética da actinomicina D e, no extrato bruto Caat 8-25 também foi identificado herbicidina F. Um estudo de mudança no metabolismo também foi realizado para a actinobactérias Caat 1-54 produtora de lisolipina. Através da suplementação de íons brometo no meio de cultivo em diferentes períodos da fermentação foi observada a produção de um análogo da lisolipina com bromo e da lisolipina clorada e bromada. Estudos de DESI-TLC-MS associado a bioautografia em TLC também foram aplicados para análise do extrato bruto Caat 2-63 e mostraram a potencialidade desta metodologia para acelerar a identificação de compostos ativos, pois a análise por espectrometria de massas na região da placa cromatográfica correspondente ao halo de inibição revelou a presença de actinomicina D e V, as quais já haviam sido associadas a atividade antimicrobiana deste extrato bruto. Assim os resultados obtidos nesta tese confirmam o grande potencial das actinobactérias como fonte promissoras de compostos bioativos e que a espectrometria de massas empregada nos seus diferentes modos de análise acelera o processo de identificação desses compostos. / In this work were used two different bioassays to evaluate the antimicrobial potential of some actinomycetes against bacteria that cause bovine mastitis. For this propose an antagonism and antibiose assays were optimized against 13 Gram-positive and Gram-negative bacteria related with bovine mastitis. Through antagonism bioassay 120 actinomycetes were screened and 55% were considered active. In disk diffusion bioassay, 70 crude extracts were screened and 60% showed inhibition zones. The crude extracts Caat 1-54, Caat 2-63 and Caat 8-25 were considered the most promising of all the extracts. Dereplication studies and fractionation guided by bioassay were procedure with this extracts to identify the actives metabolites. The extracts Caat 1-54 and Caat 2-63 were active against strains of Gram-positive bacteria and the antimicrobial effect was related to lisolipin I and actinomycin D and V, respectively. The extract Caat 8-25 was active against both Gram-positive and Gram-negative bacteria. Oxytetracycline were associated with activity against Gram-positives strains and a herbicidin B was active against Gram-negative bacteria. Using dereplication strategies by LC-MS/MS others compounds were characterized in different crude extracts. The extract Caat 2-63 was also identified the presence of other peptides from the same biosynthetic pathway of actinomycin D and for crude extract Caat 8-25 were identified herbicidin F. In addition, studies of the shift in secondary metabolism carried out for actinobacteria Caat 1-54 (lysolipin producer). Culture medium supplementation by bromide ions at different stages of fermentation induce the production of bromine lysolipin analog and a chloro-bromine lysolipin. DESI-MS TLC studies associated with TLC bioautography were used to analyze a crude extract Caat 2-63. This study showed how this method could be used to accelerate the identification of active compounds, because mass spectrometry analysis in situ at bioautigraphy inhibition zone showed the presence of actinomycin D and V, which had been associated with antimicrobial activity of this crude extract. Therefore, the results of this thesis confirm the potential of actinobacteria as bioactives compounds source and the importance of mass spectrometry experiments to accelerate the identification of these compounds.

A cebola comum Allium cepa L. (2n=16), devido à sua excelente condição cromossômica, tem sido largamente utilizada como um sistema de testes para se examinar os efeitos citogenéticos causados por diversos compostos químicos em material biológico. Em estudos de monitoramento de riscos ambientas, observou-se limitação no crescimento das raízes quando as cebolas são expostas a aguas residuárias provenientes de indústrias químicas. Nesse estudo, nós avaliamos este sistema de testes escolhendo alguns compostos de metais pesados, cujos efeitos em outros sistemas de testes já se encontram descritos. Além disso, foram estuados vários fatores que poderiam interferir nos resultados tais como: temperatura; luminosidade; qualidade da água e qualidade das cebolas. As concentrações mais baixas empregadas (1,5 mg/l) nos testes, de cádmio e cobre demostraram alta capacidade inibidora sobre o crescimento das raízes. O zinco apresentou toxicidade moderada a 1,5 mg/l e crescente às concentrações sucessivas mais elevadas. Efluentes industriais, contendo misturas de níquel e sulfatos ou cobre e cianeto, demonstraram-se altamente inibidores às concentrações mais baixas empregadas (0,7%), resultados que demonstram a alta sensibilidade do teste aos sais de metais pesados. Quanto às condições experimentais que levaram a esses resultados, os testes revelaram que não existe influência pronunciada dos fatores luminosidade e temperatura. Quanto à água para controle e diluição, não deve ser de torneira devido ao efeito inibidor do zinco e do cobre. Os resultados foram contraditórios em relação a um possível efeito osmótico da água destilada, sendo conveniente adicionar entre 100 a 250 mg/l de NaCl. A melhor resposta foi obtida com cebolas de origem argentina de 3,5 a 4,0 cm de diâmetro sendo preferível a manutenção de cascas para evitar murchamento. / The common onion Allium cepa. L., due to its excellent chromosome conditions, has been widely used as a test system for examining the citogenetic effects of several chemical compounds on biological material. It has been observerd, in monitoring environmental studies, some growth restriction on Allium roots when they are exposed to wastewater from chemical factories. In this study, we evaluate these test systems by choosing some hard metals which harmful effects on biological materials has been already described. Furthermore, some factors that could interfere on the results, such as: temperature; light; water quality and onion quality were studied. The lowest concentration (1.5 mg/l) for cadmium and cupper were found to have hard capacity to inhibit root growth. The zinc showed moderate toxicity at 1.5 mg/l and this effect was more evident at increased concentrations. The industrial wastewater that contains mixture of nickel and sulphate or cupper and cyanate inhibited root growth at lower concentration (0.7%). These results showed that the test system has high sensitivity for hard metals. There was no direct influence of temperature and light on the experiment conditions. With regard to the type of water, tap water should not be used because of inhibition on the root growth caused by zinc and copper. The results contradicted a potential osmotic effect of distillated water, being convenient to add 100 - 250 mg/l of NaCl. The best response was obtained with onions from Argentina with 3.5 - 4.0 cm of diameter being recommended to keep the onion peel for avoiding dryness.