Calcium Superphosphate-Mediated Reshaping of Denitrifying Bacteria Community Contributed to N2O Mitigation in Pig Manure Windrow Composting

Composting is recognized as an effective strategy for the sustainable use of organic wastes, but also as an important emission source of nitrous oxide (N2O) contributing to global warming. The effects of calcium superphosphate (CaSSP) on N2O production during composting are reported to be controversial, and the intrinsic microbial mechanism remains unclear. Here, a pig manure windrow composting experiment lasting for ~60 days was performed to evaluate the effects of CaSSP amendment (5%, w/w) on N2O fluxes in situ, and to determine the denitrifiers’ response, and their driving factors. Results indicated that CaSSP amendment significantly reduced N2O emissions as compared to the control pile (maximum N2O emission rate reduced by 64.5% and total emission decreased by 49.8%). CaSSP amendment reduced the abundance of nirK gene encoding for nitrite reductase, while the abundance of nosZ gene (N2O reductase) was enriched. Finally, we built a schematic model and indicated that the abundance of nirK gene was likely to play a key role in mediating N2O production, which were correlated with NH4+-N and NO3−-N changing responsive to CaSSP. Our finding implicates that CaSSP application could be a potential strategy for N2O mitigation in manure windrow composting, and the revealed microbial mechanism is helpful for deepening the understanding of the interaction among N-cycle functional genes, physicochemical factors, and greenhouse gases (GHG) emissions.

Isolation and Characterization of an Aerobic Denitrifier Bacillus sp. SC16 from an Intensive Aquaculture Pond

Overloading of ammonia and nitrite nitrogen in aquaculture can result in toxicity to aquatic animals. In order to eliminate the hazardous substances, a highly efficient denitrifying bacterium, Bacillus sp. SC16, was identified in a fishery pond and isolated subsequently. The strain SC16 could remove nitrate up to 97%, ammonia up to 36.6%, and nitrite up to 99.99% when incubated with nitrate at an initial concentration of 306.9 mg·L−1 for 72 h, ammonia at 165.49 mg·L−1 for 48 h, and nitrite at 200 mg·L−1 for 24 h under aerobic conditions. The nitrite reductase gene was identified as the nirK gene. The transcriptional levels of the nirK gene in strain SC16 incubated with ammonia, nitrate, and nitrite showed similar expression patterns. When the strain SC16 was used to treat the aquaculture water, the concentration of ammonia decreased significantly, from 8.35 mg·L−1 to 4.56 mg·L−1, and there was almost no accumulation of nitrite by the end of experiment. Therefore, the results indicated that Bacillus sp. SC16 could be a promising candidate for aquaculture water treatment.

Predominance of ammonia-oxidizing archaea andnirK-gene-bearing denitrifiers among ammonia-oxidizing and denitrifying populations in sediments of a large urban eutrophic lake (Lake Donghu)

The coupled nitrification–denitrification process plays a pivotal role in cycling and removal of nitrogen in aquatic ecosystems. In the present study, the communities of ammonia oxidizers and denitrifiers in the sediments of 2 basins (Guozhenghu Basin and Tuanhu Basin) of a large urban eutrophic lake (Lake Donghu) were determined using the ammonia monooxygenase subunit A (amoA) gene and the nitrite reductase gene. At all sites of this study, the archaeal amoA gene predominated over the bacterial amoA gene, whereas the functional gene for denitrification nirK gene far outnumbered the nirS gene. Spatially, compared with the Tuanhu Basin, the Guozhenghu Basin showed a significantly greater abundance of the archaeal amoA gene but less abundance of the nirK and nirS genes, while there was no significant difference of bacterial amoA gene copy numbers between the 2 basins. Unlike the archaeal amoA gene, the nirK gene showed a significant difference in community structure between the 2 basins. Archaeal amoA diversity was limited to the water–sediment cluster of Crenarchaeota, in sharp contrast with nirK for which 22 distinct operational taxonomic units were found. Accumulation of organic substances were found to be positively related to nirK and nirS gene copy numbers but negatively related to archaeal amoA gene copy numbers, whereas the abundance of the bacterial amoA gene was related to ammonia concentration.

Function of the Rhizobium etli CFN42 nirK gene in nitrite metabolism

Rhizobium etli CFN42 is not capable of growing anaerobically with nitrate but it grows with nitrite as a terminal electron acceptor. This bacterium contains the nirK gene encoding the copper-containing Nir (nitrite reductase), which is located on the cryptic plasmid pCFN42f. Mutational analysis has demonstrated that a nirK deficient mutant was not capable of growing under nitrite-respiring conditions. Moreover, microaerobic growth of this mutant was inhibited by the presence of nitrite. Nir activity and nitrite uptake were highly diminished in a nirK mutant, compared with the wild-type levels after incubation under anaerobic conditions. Our results suggest that the copper-containing Nir may have both a respiratory and a nitrite-detoxifying role in R. etli.