MICRORGANISMS WITH ANAMMOX ACTIVITY IN SLUDGE ACCLIMATED FROM A SWINE MANURE TREATMENT LAGOON

Aline Viancelli (UFSC); Airton Kunz (CNPSA); Paulo Augusto Esteves (CNPSA); T. Fujii (Dept SLS); Matias Vanotti (ARS/USDA); T Nishiyama, (Dept SLS); Regina Vasconcellos Antônio (UFSC)

Introduction: ANAMMOX (anaerobic ammonium oxidation) process is an alternative technology in nitrogen removal from wastewater and consists on the oxidation of NH₄ to N₂. The advantages of ANAMMOX over conventional nitrification/denitrification process are less oxygen and no organic carbon requirement. The bacteria responsible for ANAMMOX process belong to phylum Planctomycetes and have a widespread distribution. There are five uncultured genus C. Brocadia; C. Kuenenia; C. Scalindua; C. Anammoxoglobus and C. Jettenia. The present study aimed to analyze the bacterial community from an ANAMMOX reactor by molecular methods. Material and methods: Reactor was inoculated with sludge from an anaerobic lagoon in experimental swine manure treatment at EMBRAPA - Swine and Poultry. After 20 months operating on ANAMMOX activity, samples were collected; DNA was extracted and treated with phenol/chloroform isoamylalcohol. Amplification of 16S rDNA gene was performed with KOD -plus- DNA polymerase using primers 6F 5’-GGAGAGTTAGATCTTGGCTCAG-3’ and 1492R 5’-GGTTACCTTGTTACGACT-3’. PCR was carried out according to: 2 min at 94°C, 25 cycles (15 sec at 94°C, 30 sec at 58°C, 30 sec at 68°C). Amplified fragments were inserted into pBluescript II KS+ and transformed with Escherichia coli DH10B. Plasmidial DNA extracted was sequenced with BigDye terminator v3.1 methodology and compared with sequences from nr-database using BLAST. Results and Discussion: Seventeen clones were obtained. Three of these clones had 97% homology with organisms belonging to Anammox group (Planctomycete KSU-1 and Candidatus Jettenia asiatica). One clone, showed homology with phylum Chlorobium (93%) that lives in anoxic aquatic settings and in phototrophic consortiums with â-proteobacteria. Twelve clones had 99% homology with Janthinobacterium. Conclusion: Bacteria with ANAMMOX activity in a laboratory reactor was confirmed by the present study. Were identified organisms homologous to Candidatus Jettenia asiatica, Planctomycete KSU-1, Chlorobium and Janthinobacterium.