Wide biosurfactant application on biorremediation is limited by its high production cost. The search for cheaper biossurfactant production alternatives has guided our study. The use of selective media containing sucrose (10 g.L -1 ) and Arabian Light oil (2 g.L -1 ) as carbon sources showed to be effective to screen and maintain biosurfactant-producing consortia isolated from mangrove hydrocarbon-contaminated sediment. The biosurfactant production was assayed by kerosene, gasoline and Arabian Light Emulsification activity and the bacterial growth curve was determined by bacterial quantification. The parameters analyzed for biosurfactant production were the growth curve, salinity concentration, flask shape and oxygenation. All bacteria consortia screened were able to emulsify the petroleum derivatives tested. Biosurfactant production increased according to the incubation time; however the type of emulsification (non-aqueous phase or aqueous phase) did not change with time but with the compound tested. The methodology was able to isolate biosurfactant-producing consortia from superficial mangrove sediment contaminated by petroleum hydrocarbons and was recommended for selection of biosurfactant producing bacteria in tropical countries with low financial resources.Keywords: bacteria consortia, bioremediation, petroleum, sediment.
Metodologia alternativa para isolamento de bactérias produtoras de biossurfactante ResumoA ampla aplicação de biossurfactantes em biorremediação é limitada pelo seu alto custo de produção. A procura de alternativas de produção mais baratas motivou nosso estudo. O uso de meio seletivo, contendo sacarose (10 g.L ), mostrou-se eficiente na triagem e manutenção de consórcios bacterianos isolados de sedimento de mangue contaminado com hidrocarboneto. A produção de biossurfactante foi avaliada pela Atividade Emulsificante do querosene, gasolina e Árabe Leve e a curva de crescimento foi determinada pela quantificação bacteriana. Os parâ-metros analisados para a produção de biossurfactante foram: curva de crescimento, concentração da salinidade, forma do frasco e oxigenação. Todos os consórcios bacterianos pesquisados foram capazes de emulsificar os derivados de petróleo testados. A produção de biossurfactante aumentou de acordo com o tempo de incubação, entretanto, o tipo de emulsificação aquosa e não-aquosa não mudou com o tempo, mas com o composto testado. A metodologia permitiu o isolamento de consórcios bacterianos produtores de biossurfactante de sedimento superficial de mangue contaminado com hidrocarbonetos do petróleo, sendo indicada para seleção de bactéria produtora de biossurfactante em países tropicais com baixos recursos financeiros.Palavras-chave: consórcio bacteriano, biorremediação, petróleo, sedimento.
The cell surface hydrophobicity of 60 isolates and three reference strains of Staphylococcus epidermidis was assayed by means of bacterial aggregation in liquid broth, phosphate-buffered saline, and in ammonium sulfate, as well as by affinity of the bacteria to n-hexadecane and polystyrene surfaces. In order to better characterize the isolates, the influence of bacterial growth time and enzyme treatment on cell hydrophobicity and the analysis of the slime production were also investigated. The strains presented the following profiles when assayed by the ammonium sulfate aggregation test (SAT): SAT < 1M, SAT 1M - <2M, SAT 2M - <4M, and SAT >or=4M. When SAT < 1M, the strains showed positive results for most of the cell surface hydrophobicity tests. None of the strains belonging to the groups with SAT >or= 1M showed spontaneous aggregation (SA), auto-aggregation (AA), or glass adherence, albeit 32 (62.7%) strains were polystyrene adherent and 42 (82.3%) presented weak adherence to n-hexadecane (>20%). The best correlation of the results was found among the AA and glass adherence tests (100%), followed by SA/ glass adherence (98%) and SA/ AA test (98%). The polystyrene adherence test and microbial adherence to n-hexadecane test (MATH) showed 78% correlation. Proteinase K treatment reduced bacterial adherence to polystyrene, but did not influence the SAT values. Three distinct groups of strains were distinguished by the polystyrene micromethod and glass tube adherence assay: 0.0-0.4 O.D. group, including non-glass adherent isolates; 0.5-0.7 O.D. group, including strains with variable profiles (adherent or non-adherent); and 0.8-1.3 O.D. group, composed of glass-adherent strains. Evaluation by a single method seemed not to reliably determine the surface hydrophobicity characteristics of S. epidermidis clinical isolates. Auto-aggregation properties of the strains that adhered to glass seemed related to slime expression, rather than cell surface hydrophobicity. Data also suggested involvement of protein components in adherence to polystyrene, but not in auto-aggregation properties assayed by SAT.
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