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A microbial community, selected by its ability to degrade triazinic herbicides was acclimatized by successive transfers in batch cultures. Initially, its ability to degrade prometryn, was evaluated using free cells or cells attached to fragments of a porous support. As carbon, nitrogen and sulfur sources, prometryn, (98.8 % purity), or Gesagard, a herbicide formulation containing 44.5 % prometryn and 65.5 % of adjuvants, were used. In batch cultures, a considerable delay in the degradation of prometryn, presumptively caused by the elevated concentration of inhibitory adjuvants, occurred. When pure prometryn was used, volumetric removal rates remarkably higher than those obtained with the herbicide formulation were estimated by fitting the raw experimental data to sigmoidal decay models, and differentiating them. When the microbial consortium was immobilized in a continuously operated biofilm reactor, the negative effect of adjuvants on the rate and removal efficiency of prometryn could not be detected. Using the herbicide formulation, the consortium showed volumetric removal rates greater than 20 g m(-3) h(-1), with prometryn removal efficiencies of 100 %. The predominant bacterial strains isolated from the microbial consortium were Microbacterium sp., Enterobacter sp., Acinetobacter sp., and Flavobacterium sp. Finally, by comparison of the prometryn removal rates with others reported in the literature, it can be concluded that the use of microbial consortia immobilized in a biofilm reactor operated in continuous regime offer better results than batch cultures of pure microbial strains.

Through selective enrichment of atrazinemetabolizing microorganisms, a microbial community was selected from agricultural soil. Bacterial isolates, identified by their closest similarity with 16S rDNA sequences stored in NCBI GeneBank, belonged to the genera: Massilia, Stenotrophomonas, Klebsiella, Sphingomonas, Ochrobactrum, Arthrobacter, Microbacterium, Xanthomonas and Ornithinimicrobium. From these strains, only the first six used atrazine as nitrogen and carbon source. The microbial community attached to a non-porous support was evaluated for its atrazine biodegradation rate and removal efficiency under aerobic conditions in two types of packed-bed biofilm reactors fed with a mineral salt medium containing glucose plus atrazine, or atrazine as the sole carbon and nitrogen source. Removal efficiencies near 100% were obtained at loading rates up to 10 mg l -1 h -1 . After long periods of continuous operation, the richness of microbial species in biofilm reactors diminished to only three bacterial strains; Stenotrophomonas sp., Ochrobactrum sp. and Arthrobacter sp. By PCR analysis of their DNA, the presence of atzABC genes codifying for the enzymes of the upper catabolic pathway of atrazine, was confirmed in the three strains. The gene atzD that encodes for the cyanuric acid amidohydrolase enzyme was detected only in Stenotrophomonas sp.

Cyanuric acid (1,3,5-triazine-2,4,6-triol [OOOT]) is a common biodegradation byproduct of triazinic herbicides, frequently accumulated in soils or water when supplementary carbon sources are absent. A binary bacterial culture able to degrade OOOT was selected through a continuous selection process accomplished in a chemostat fed with a mineral salt (MS) medium containing cyanuric acid as the sole carbon and nitrogen source. By sequence comparison of their 16S rDNA amplicons, bacterial strains were identified as Agrobacterium tumefaciens, and Acinetobacter sp. When the binary culture immobilized in a packed bed reactor (PBR) was fed with MS medium containing OOOT (50 mg L(-1)), its removal efficiencies were about 95%; when it was fed with OOOT plus glucose (120 mg L(-1)) as a supplementary carbon source, its removal efficiencies were closer to 100%. From sessile cells, attached to PBR porous support, or free cells present in the outflowing medium, DNA was extracted and used for Random Amplification of Polymorphic DNA analysis. Electrophoretic patterns obtained were compared to those of pure bacterial strains, a clear predominance of A. tumefaciens in PBR was observed. Although in continuous suspended cell culture, a stable binary community could be maintained, the attachment capability of A. tumefaciens represented a selective advantage over Acinetobacter sp. in the biofilm reactor, favoring its predominance in the porous stone support.