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  • Mineralization of 4-fluorocinnamic acid by a Rhodococcus strain
    Publication . Amorim, Catarina L.; Ferreira, António César Silva; Carvalho, M.F.; Afonso, Carlos M. M.; Castro, Paula M.L.
    A bacterial strain capable of aerobic degradation of 4-fluorocinnamic acid (4-FCA) as the sole source of carbon and energy was isolated from a biofilm reactor operating for the treatment of 2-fluorophenol. The organism, designated as strain S2, was identified by 16S rRNA gene analysis as a member of the genus Rhodococcus. Strain S2 was able to mineralize 4-FCA as sole carbon and energy source. In the presence of a conventional carbon source (sodium acetate [SA]), growth rate of strain S2 was enhanced from 0.04 to 0.14 h−1 when the culture medium was fed with 0.5 mMof 4- FCA, and the time for complete removal of 4-FCA decreased from 216 to 50 h.When grown in SA-supplemented medium, 4-FCA concentrations up to 1 mM did not affect the length of the lag phase, and for 4-FCA concentrations up to 3 mM, strain S2 was able to completely remove the target fluorinated compound. 4-Fluorobenzoate (4-FBA) was transiently formed in the culture medium, reaching concentrations up to 1.7 mM when the cultures were supplemented with 3.5mMof 4-FCA. Trans,trans-muconate was also transiently formed as a metabolic intermediate. Compounds with molecular mass compatible with 3-carboxymuconate and 3-oxoadipate were also detected in the culture medium. Strain S2 was able to mineralize a range of other haloorganic compounds, including 2- fluorophenol, to which the biofilm reactor had been exposed. To our knowledge, this is the first time that mineralization of 4-FCA as the sole carbon source by a single bacterial culture is reported.
  • Fluoroquinolones biosorption onto microbial biomass: activated sludge and aerobic granular sludge
    Publication . Ferreira, Vanessa R.A.; Amorim, Catarina L.; Cravo, Sara M.; Tiritan, M. E.; Castro, P. M. L.; Afonso, Carlos M. M.
    Fluoroquinolones (FQs) have been reported in trace amounts in different environmental matrices. The biosorption of three most prescribed FQs, ofloxacin (OFL), norfloxacin (NOR) and ciprofloxacin (CPF) by activated sludge (AS) and aerobic granular sludge (AGS) was investigated. Biosorption assays were conducted with FQs concentrations within the range of 100e700 ng mL 1, to mimic environmental conditions. At neutral pH and at the end of 48 h, AS showed higher biosorption capacity than AGS. For AS, a maximum biosorption of 1.50 ± 0.03, 3.24 ± 0.05 and 3.39 ± 0.06 mg gTSS 1 was observed for OFL, NOR and CPF respectively, whereas for AGS the maximum amount of FQs biosorbed was 1.18 ± 0.03, 2.73 ± 0.02 and 2.94 ± 0.03 mg gTSS 1. Langmuir isotherm was more applicable for describing FQs biosorption equilibrium by AS while for AGS, the Freundlich isotherm was more adequate. Given the AGS technology innovative character, the effect of change of pH on the biosorbed FQs was evaluated. FQs could be desorbed from AGS at pH 3, pH 8 and pH 9 but at pH 4 the biosorption process was promoted. This study allows a better understanding of the FQs biosorption processes. Moreover, the data from biosorption/desorption from AGS may be useful for management and operation of AGS bioreactors
  • Biodegradation of fluoroanilines by the wild strain Labrys portucalensis
    Publication . Amorim, Catarina L.; Carvalho, Maria F.; Afonso, Carlos M. M.; Castro, Paula M. L.
    Aniline and halogenated anilines are known as widespread environmental toxic pollutants released into soil and water. In contrast to aniline, which is rapidly metabolized via catechol, halosubstituted anilines are more resistant to microbial attack. A fluorobenzene-degrading bacterium, Labrys portucalensis strain F11, was tested under different culture conditions for the degradation potential towards 2-, 3- and 4-fluoroaniline (2-, 3- and 4-FA). Strain F11 was able to use FAs as a source of carbon and nitrogen however, supplementation with a nitrogen source improved substrate consumption and its dehalogenation extent. When F11 cells were previously grown on fluorobenzene (FB), higher biodegradation rates were achieved for all isomers. Complete 2-FA biodegradation with stoichiometric fluoride release was achieved when FB-induced cells were used. On the other hand, the degradation of 3- and 4-FA was characterized by incomplete defluorination of the target compounds suggesting accumulation of fluorinated intermediates. F11 cultures simultaneously supplied with FB and the fluorinated anilines showed a concomitant degradation of both substrates, suggesting co-metabolic biodegradation. To our knowledge, this is the first time that biodegradation of 2- and 3-FA as a sole carbon and nitrogen source and co-metabolic degradation of FA isomers in the presence of a structural analogous compound is reported.
  • Bioaugmentation for treating transient 4-fluorocinnamic acid shock loads in a rotating biological contactor
    Publication . Amorim, Catarina L.; Duque, Anouk F.; Afonso, Carlos M. M.; Castro, Paula M. L.
    A rotating biological contactor (RBC) was used to treat shock loadings of 4-fluorocinnamic acid (4-FCA). Intermittent 4-FCA shocks of 35 mg L-1 were applied (ca. 3 months) with only limited mineralization occurring and accumulation of 4-fluorobenzoate (4-FBA) as an intermediate. After bioaugmentation with a degrading bacterium the RBC was able to deal with 4-FCA intermittent loading of 80 mg L-1 however, a gradual decline in RBC performance occurred, leading to 4-FBA accumulation. The degrading strain was recovered from the biofilm during 2 months but intermittent feeding may have led to diminishing strain numbers. Distinct bacterial communities in the 1st and the 5th and 10th stages of the RBC were revealed by denaturating gradient gel electrophoresis. Several isolates retrieved from the RBC transformed 4-FCA into 4-FBA but only two strains mineralized the compound. Bioaugmentation allowed removal of the fluorinated compound however intermittent feeding may have compromised the bioreactor efficiency.