Anammox Granular Sludge Of Uncultured And Culture Of Bacterial Diversity In A Preliminary Study And Identification Of New Species

Environmental contamination was tending to become worse in terms of the industrial development. Our strategy must be taken to control of the pollution while pollutants are produced. The nitrogen compounds were very important pollutant in the wastewater and landfill leachate, of which ammonia was the major pollutant. Anaerobic ammonia oxidation (Anammox) was a promising biological nitrogen removal process and had a lot of merits such as none of additional organics and aeration, and less sludge production. More and more scientists have joined in the research of anammox.The anammox granules sludge of a UASB (upflow anaerobic sludge blanket) reactor was selected to survey the characterization of bacterial communities using the culture-independent and-dependent methods. Culture-independent method in view of 16S rRNA gene phylogenetic analysis indicated that there were abundant bacterial diversity in the granules sludge. There were 22 OTUs within 29 clones, belonged to 8 phyla, which they accounted for one third of 27 phyla in bacterial taxonomy of RDP (3% cutoff). Culture-independent method in view of hzo gene phylogenetic analysis stated that there were two groups of anammox bacteria such as Candidatus Kuenenia and Candidatus Jettenia. The bacteria of nitrogen cycling such as nitrogen-fixing bacteria, ammonia oxidization bacteria, nitrite oxidization bacteria, anammox bacteria and denitrifying bacteria were distributed in the granules sludge, they formed a completed nitrogen cycling.Thirty-four distinct isolates were obtained from culture-dependent method. They were belonged to 6 genera of 3 phyla based on the phylogenetic analysis of 16S rRNA gene sequences. Most of them were belonged to denitrifying bacteria. The isolates of Bacillus were not detected in the culture-independent method. It illustrated that the culture-dependent method should be a supplement to the culture-independent method.Two strains HY-14T and HY-24, of which the similarity to all known typical bacteria were blow 97%, were selected to classify and identify. Phylogenetic trees based on the neighbour-joining and maximum-parsimony methods indicated that two strains formed a monophyletic clade within the genus Pseudomonas. For the large genic difference, two strains had the large different physiological and biochemical characteristics, G+C contents and fatty acid profiles with the type strains of the genus Pseudomonas. Strains HY-14T and HY-24 were classified as a novel species of the genus Pseudomonas, for which the name Pseudomonas caeni sp. nov. is proposed (HY-14T=KCTC 22292T=CCTCCAB 208156T).