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Axenic cultures of Chiamydomonas ulvaensis [CCAP 11/58], Scenedesmus brasiliensis [CCAP 276/1 B] and Ochromonas danica [0CAP933/28] were screened for their ability to grow on and remove phenol from their incubation media. Neither C. ulvaensis nor S. brasiliensis removed the phenol substrate as rapidly as 0. danica which is a nutritionally versatile chrysophyte. 0. danica was found to grow on phenol and p-cresol as the sole C-source at concentrations up to 4mM in cultures grown in both photoheterotrophic and heterotrophic conditions. The alga would not grow on cresols or xylenols unless phenol was present. Oxygen uptake and turnover studies confirmed that the enzymes involved in phenolic catabolism were inducible and that the organism showed a decrease in activity resulting from the position and number of the methyl substituents on the aromatic ring. [U- 14C]Phenol was completely mineralised with some 65% of the 14C-label appearing as 14CO2, approximately 12% remaining in the aqueous medium and the rest accounted for in the biomass. Analysis of the biomass showed that 14C-label had been incorporated into the protein, nucleic acid and lipid fractions; phenol carbon is thus unequivocally assimilated by the alga. Phenol-grown cultures of 0. danica converted phenol to catechol which was further metabolised by the meta cleavage pathway. 2-Hydroxymuconic semialdehyde and pyruvate, characteristic products of meta cleavage, were found in incubations of catechol with cell-free extracts of phenol-grown cells together with the appropriate enzyme activities. This is , as far as I am aware, the first definitive identification of the meta cleavage pathway for aromatic ring degradation in a eukaryotic microorganism.

Effluents with high concentration of organic matter and xenobiotic compounds, especially in the case of the agrochemicals, cause environmental concern. In the present work, the adaptative ability of two filamentous fungi, which cause white rot (Geotrichum candidum Link & Pers. and Phanerochaete chrysosporium Burds.), to tolerate high concentration of xenobiotic substances was analyzed. In solid medium, P. chrysosporium and G. candidum presented excellent development under aerated and not aerated conditions (30ºC and dark), in potato-dextrose-agar and malt extract-agar, respectively. In liquid medium, the same was observed in broth-potato-dextrose and malt extract. The fungi also developed well in liquid media deficient in proteins and carbohydrates, but with high concentrations (10 and 25 mg/L) of the agrochemicals Boral, Decanote, Gliphosate, Paraquat and Plenum - all used in fields of sugar-cane, as the ones of the "S.A Usina Coruripe Açúcar e Álcool" (Coruripe - AL). These chemicals are classified as persistent, belong to the toxicological classes II and III, and have different solubilities. The fungi presented tanninolitic activity when inoculated in samples of the crude effluent with supplement of nutrients, under aeration or not. The activity of the laccase of G. candidum in the same medium was higher in the 3rd day of the growth without aeration, occurring a reduction of that in the 5th day. On the other hand, the fungus P. chrysosporium presented a higher activity of laccase in the 7th day of the growth, also without aeration. Both fungi did not interfere in the color, turbidity, concentration of nitrates and nitrites of this medium. G.candidum revealed equal efficiency in the reduction of the concentration of phosphates of the crude effluent, under aeration (78.18%) and not aeration (79.33%) conditions. However, it did not reduce the COD. P. chrysosporium was efficient in the reduction of this COD, and the difference between the aerated growth (26.77%) and the not aerated one (29.55%) was very small. It did not reduce the total tanning bark concentration, but it diminished the concentration of phosphates under aerated (55.55%) and under not aerated (61.65%) condition. The addition of nutrients in the medium of diluted effluent (2 X, 4 X and 8 X) showed that there is a direct effect of the concentration of the COD on its percentage of removal by the inoculated fungi. There is also a direct effect on the remotion of total phenolic compounds and reduced glycids. The conversion of COD in the diluted effluent (8X) was higher in the growth of P. chrysosporium (65.66%) than in the one of G.candidum (57%). The same it can be said regarding to the best rates of phenol degradation (59.66% and 50.33%, respectively). The pH measured in the media supplemented with nutrients was always higher (7,56-7,8) than the initial one (5,5). In the higher dilution of the effluent (8X) plus the supplement of nutrients, the maximum removal of COD and total phenols after the treatment occurred in 7th day of incubation of both fungi. In all the cases of diluted samples plus supplement of nutrients, the discoloration drastically increased after the inoculation of the fungi. Therefore, the use of these microorganisms in the bioremediation of the effluent of sugar-alcohol industry is recommended only after a correction of the ratio C: N: P to about 100:1,6:1. / Efluentes com alta concentração de matéria orgânica e compostos xenobióticos, especialmente no caso dos agroquímicos, causam preocupação ambiental. No presente trabalho, dois fungos filamentosos causadores de podridão branca (Geotrichum candidum Link e Pers. e Phanerochaete chrysosporium Burds.) mostraram-se capazes de se adaptar a meios com elevados teores de matéria orgânica xenobiótica ou não. Em meio sólido, verificou-se que P. chrysosporium e G. candidum, apresentaram ótimo desenvolvimento em condições aeradas e não aeradas, a 30ºC e escuro, em meios batata-dextrose-ágar e extrato de malte-ágar, respectivamente. Em meio líquido, o mesmo foi observado em caldo-batatadextrose e meio extrato de malte. Os fungos também desenvolveram-se bem em cultivos líquidos deficientes em proteínas e carboidratos, e contendo altas concentrações (10 e 25 mg/L) dos agroquímicos Boral, Decanote, Glifosato, Paraquat e Plenum, todos utilizados nas lavouras de cana de açúcar da S.A Usina Coruripe Açúcar e Álcool (Coruripe AL), classificados como persistentes, de classes toxicológicas II e III, e com diferentes solubilidades. Ambos os fungos apresentaram atividade taninolítica quando inoculados em amostras de efluente de agroindústria sucro-alcooleira não diluídas e não acrescida de nutrientes, sob aeração ou não. A atividade da lacase do fungo G. candidum, nesse mesmo meio, foi maior no 3o dia de cultivo sem aeração, ocorrendo uma diminuição no 5o dia. Já o fungo P. chrysosporium, apresentou maior atividade da lacase no 7o dia de cultivo sem aeração. Em relação à cor, turbidez, concentração de nitratos e nitritos, não houve variação desses parâmetros nas amostras inoculadas por ambos os fungos. G. candidum mostrou-se igualmente eficiente na redução da concentração de fosfatos, em cultivos aerados (78,18%) e não aerados (79,33%) do efluente bruto. Todavia, não reduziu a DQO do mesmo. P. chrysosporium mostrou-se eficiente na redução da DQO, sendo pequena a diferença entre os cultivos aerados (26,77%) com relação aos não aerados (29,55%), e não reduziu o teor de taninos totais, mas diminuiu a concentração de fosfatos sob aeração (55,55%) e não aerado (61,65%). A adição de nutrientes no cultivo dos fungos em efluente agroindustrial diluído (2X, 4X e 8X), mostrou que houve um efeito direto da concentração da DQO sobre sua porcentagem de remoção e sobre a redução de fenóis totais e glicídeos redutores. A conversão da DQO em efluente diluído (8X) foi maior no cultivo de P. chrysosporium (65,66%) do que no de G. candidum (57%). O mesmo pode ser dito quanto às melhores taxas de degradação de fenóis (59,66% e 50,33%, respectivamente). Quanto ao pH mensurado nos cultivos suplementados com nutrientes, foi sempre maior (7,56-7,8) que o inicial (5,5). Já nos cultivos utilizando a maior diluição (8X) e o suplemento de nutrientes, a máxima remoção de DQO e de fenóis totais após o tratamento ocorreu no 7º dia do cultivo para ambos os fungos. Em todos os casos de amostras diluídas e acrescidas de nutrientes, a descoloração aumentou drasticamente após o cultivo dos fungos estudados. Portanto, recomenda-se a utilização desses microrganismos na biorremediação de efluentes sucro-alcooleiros desde que haja uma correção da proporção C:N:P de cerca de 100:1,6:1.