ABSTRACT Anthropogenic activity, such as accidental oil spills, are typical sources of urban mangrove pollution that may affect mangrove bacterial communities as well as their mobile genetic elements. To evaluate remediation strategies, we followed over the time the effects of a petroleum hydrocarbon degrading consortium inoculated on mangrove tree Avicennia schaueriana against artificial petroleum contamination in a phytoremediation greenhouse experiment. Interestingly, despite plant protection due to the inoculation, denaturing gradient gel electrophoresis of the bacterial 16S rRNA gene fragments amplified from the total community DNA indicated that the different treatments did not significantly affect the bacterial community composition. However, while the bacterial community was rather stable, pronounced shifts were observed in the abundance of bacteria carrying plasmids. A PCR-Southern blot hybridization analysis indicated an increase in the abundance of IncP-9 catabolic plasmids. Denaturing gradient gel electrophoresis of naphthalene dioxygenase (ndo) genes amplified from cDNA (RNA) indicated the dominance of a specific ndo gene in the inoculated petroleum amendment treatment. The petroleum hydrocarbon degrading consortium characterization indicated the prevalence of bacteria assigned to Pseudomonas spp., Comamonas spp. and Ochrobactrum spp. IncP-9 plasmids were detected for the first time in Comamonas sp. and Ochrobactrum spp., which is a novelty of this study.

Uma cultura mixta e uma linhagem bacteriana pura foram isoladas de um bioreator para tratamento de tiocianato. As culturas removeram 5mM de tiocianato do meio em 36 horas. A cultura mixta foi capaz de tolerar concentrações superiores a 60mM. A eficiência da degradação de tiocianato diminuiu quando as células foram imobilizadas.

A mixed culture and a pure bacterial strain (BMV8) were isolated from a bioreactor for thiocyanate treatment. Both cultures removed 5 mM of thiocyanate from the medium in 36 hours. The mixed culture was able to tolerate concentrations up to 60 mM. The efficiency of thiocyanate degradation decreased when the cells were immobilized.

Nesse trabalho descrevemos um protocolo otimizado para extração simultânea de DNA e RNA de solo. O tratamento das amostras de solo com etanol e posterior agitação com partículas foi uma estratégia bem sucedida para lise das células sem degradação significativa dos ácidos nucléicos, resultando em bom rendimento de DNA e RNA íntegros. O RNA transcrito pode ser amplificado com iniciadores com alvo no fragmento do gene da glutamina sintetase (glnA). Os fragmentos 16S rDNA, tanto do DNA como do cDNA, foram amplificados e analisados por DGGE. O método foi aplicado para amostras de solo e rizosfera (morango e canola). Dois outros protocolos para extração de ácidos nucléicos de solo foram aplicados para o mesmo lote de amostras, de forma a comparar os métodos quanto à eficiência e reprodutibilidade. Os perfis de DGGE mostraram não haver diferença relevante nos padrões obtidos. O método descrito é apropriado para o processamento rápido de muitas amostras e, conseqüentemente, adequado para estudos ecológicos.

In this work we report an optimized protocol for simultaneous extraction of DNA and RNA from soil matrices. Treatment of soil matrices with ethanol followed by bead-beating worked as a successful strategy to lyse the cells without considerable degradation of nucleic acids, resulting in DNA and RNA of good yield and integrity. The reverse transcribed RNA could be amplified with primers targeting a glutamine synthetase (glnA) gene fragment. From both DNA and cDNA, 16S rDNA fragments were amplified and analyzed by Denaturing Gradient Gel Electrophoresis (DGGE). The method was applied to soil and rhizosphere (strawberry and oilseed rape) samples. Two other protocols for the extraction of nucleic acids from soil were applied to the same set of samples in order to compare the methods in terms of efficiency and reproducibility. The DGGE profiles indicated no relevant differences between the patterns obtained. The method described here is suitable for rapid processing of many samples and therefore appropriate for ecological studies.

Quatro linhagens de Rhodotorula mucilaginosa e uma linhagem padrão dessa mesma espécie foram investigadas quanto a habilidade de acumular íons de prata livres (Ag+) e complexados [Ag (CN)2-], através de processos dependentes e independentes do metabolismo. A habilidade de acumular Ag+ foi observada em ambas as células, vivas e mortas. Contudo, o acúmulo de Ag (CN)2- foi um processo estritamente dependente de energia. Durante os estudos que avaliaram a dependência de uma fonte energética para o acumulo de íons de prata livres (Ag+) foi observado que a adição de glicose resultou na diminuição do acúmulo de Ag+ por três linhagens (UFMG -- Y02, 27 and 35). Ao passo que as linhagens CBS 316 e UFMG -- Y01 apresentaram aumento do acúmulo desse metal. A analise dos resultados de biossorção através do modelo de Langmuir, sugerem que a linhagem UFMG -- Y27 possui a maior capacidade de acúmulo de prata (carga). Enquanto que a linhagem padrão CBS 316 apresentou a menor capacidade de carga e a maior afinidade por ions de prata. Analises de infra vermelho da biomassa fúngica, com e sem prata, mostraram que os grupamentos C=O foram os principais sítios de ligação desse metal pela linhagem UFMG -- Y27.

Five strains of Rhodotorula mucilaginosa were tested for the ability to accumulate free and complexed silver ions by metabolism-dependent and -independent processes. The ability to take up Ag+ was observed in both live and dead biomass, whereas silver dicyanide [Ag (CN)2-] uptake was strictly glucose dependent. In contrast to Ag (CN)2-, glucose addition inhibited by 16 to 25% the Ag+ uptake rate of living UFMG - Y02, Y27, and Y35 cells, while strains CBS 316 and UFMG-Y01 showed an improved uptake rate of about 115% and 13%, respectively. The Langmuir sorption model was used to evaluate the silver sorption capability of the R. mucilaginosa strains. The calculated q max value suggested that R. mucilaginosa strains UFMG-Y27 had the highest loading capacity. The type strain CBS 316 had the lowest q max but showed the highest affinity for silver ions. The results provided by the Fourier Transform Infra Red analysis (FTIR) suggest that C=O groups represent the main reactive site for silver uptake by the strain UFMG-Y27.