[Diversity and PICRUSt2-based Predicted Functional Analysis of Bacterial Communities During the Start-up of ANAMMOX].

Bacterial communities are vital for efficient nitrogen removal in an anaerobic ammonium oxidation (ANAMMOX) system. However, the diversity and functional characteristics of a bacterial community during the start-up of ANAMMOX has not been reported. In this study, an up-flow anaerobic sludge bed reactor was used to start-up the ANAMMOX system, and 16S rRNA high-throughput gene sequencing, combined with PICRUSt2-based functional prediction analysis, was used to investigate the dynamic changes in diversity and function of the bacterial community at different times (d0, d30, d60, and d90) during the start-up. The results showed that 48 phyla, 111 classes, 269 orders, 457 families, 840 genera, and 1497 species were present during the start-up of ANAMMOX. Candidatus Brocadia and Candidatus_Kuenenia were the main detected ANAMMOX bacteria, and their relative abundance was significantly different at different times during the start-up of ANAMMOX (P<0.05). During the start-up, the alpha diversity indices of the bacterial community were significantly decreased (P<0.05), and the structure of the bacterial community exhibited significant spatial differentiation (R=0.846, P<0.01). Functional prediction analysis with PICRUSt2 revealed that the bacterial community was active in organic systems and metabolism at hierarchy level 1, implying abundant functional diversity. Further, the abundance of functional genes was significantly different at hierarchy level 2, during the start-up of ANAMMOX. Forty-nine functional genes involving metabolic nitrogen were detected. The abundance of functional genes, involved in nitrification, denitrification, ANAMMOX, and nitrate and nitrite assimilatory/dissimilatory reduction, changed significantly during the start-up of ANAMMOX.

[Microbial Community Structure and Diversity During the Enrichment of Anaerobic Ammonium Oxidation Bacteria].

To understand the changes in microbial community characteristics during the enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria, an ASBR reactor was used to culture the ANAMMOX bacteria. The composition, diversity, and species co-occurrence network of the microbial community were investigated under different cultivation times. The results showed that the ANAMMOX bacteria were enriched by gradually increasing the substrate concentration, with removal efficiencies for NH4+-N, NO2--N, and total nitrogen of 97.6%, 95.4%, and 84.9%, respectively. The high-throughput sequencing found that the dominant phyla (relative abundance>5%) were Proteobacteria, Bacteroidetes, Chloroflexi, Planctomycetes, Armatimonadetes, and Actinobacteria in the whole culture process. Candidatus Brocadia was the main ANAMMOX bacteria in the reactor, with its relative abundance increasing from 1.42% to 24.66%. During the cultivation process, the composition of the dominant microbial community did not change, while the relative abundance showed a significant difference (P<0.05). The alpha diversity of the microbial community significantly increased first and then decreased (P<0.05), and the beta diversity of the microbial community was significantly spatially differentiated (R=0.5672, P<0.01) during the culture process. Species network densities were 0.188, 0.068, 0.059, 0.18, and 0.0735 at different times during the culture process. Although the enrichment culture process resulted in weaker correlations between microorganisms, the related group of microorganisms in the phylum Aspergillus became the main node in the network. The enrichment process weakened the correlation between microorganisms; however, the microbial taxa related to the phylum Planctomycetes became the key node in the network.

Production of fucoxanthin, chrysolaminarin, and eicosapentaenoic acid by Odontella aurita under different nitrogen supply regimes.

Microalgae are recognized as promising producers of many bioactive products, but their utility is limited due to high production costs. We subjected the marine diatom Odontella aurita to three nitrogen supply regimes [initial low nitrogen (ILN), initial high nitrogen (IHN), and initial high nitrogen plus supplementary nitrogen (SN)] to investigate the accumulation of three high-value bioactive components: fucoxanthin, chrysolaminarin and eicosapentaenoic acid (EPA). We found that SN conditions maximized fucoxanthin accumulation: a maximum productivity of 6.01 mg L-1 d-1 was obtained, a 4.32-fold and 1.42-fold increase over production in the ILN and IHN groups, respectively. After nitrogen was depleted in the growth medium, chrysolaminarin became the dominant energy storage compound. Chrysolaminarin content rose to 60.33% of dry weight (DW) in the ILN group, and 46.27% of DW in the IHN group. Variations in fatty acid composition across the different nitrogen supply regimes indicated that EPA primarily accumulated in the glycolipids, especially when nitrogen supply was sufficient. The maximum productivity of chrysolaminarin (161.55 mg L-1 d-1) and EPA (9.37 mg L-1 d-1) was observed in the IHN group. However, IHN conditions did not maximize overall content of either compound. Our results demonstrated that O. aurita is potentially useful as a producer of a variety of bioactive products; the compounds produced by this species can be controlled by altering the nitrogen supply.

Preliminary characterization, antioxidant properties and production of chrysolaminarin from marine diatom Odontella aurita.

A new chrysolaminarin, named CL2, with a molecular mass of 7.75 kDa, was purified from the marine diatom, Odontella aurita, using DEAE-52 cellulose anion-exchange chromatography and Sephadex G-200 gel-filtration chromatography. The monosaccharide and structural analysis revealed that CL2 was a glucan mainly composed of glucose, which was linked by the ß-d-(1→3) (main chain) and ß-d-(1→6) (side chain) glycosidic bond, demonstrated by infrared spectroscopy (IR) and nuclear magnetic resonance (NMR). The antioxidant activity tests revealed that the CL2 presented stronger hydroxyl radical scavenging activity with increasing concentrations, but less was effective on reducing power analysis and scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The influences of nitrogen concentration and light intensity on chrysolaminarin production of O. aurita were further investigated in a glass column photobioreactor, and a record high chrysolaminarin productivity of 306 mg L-1 day-1 was achieved. In conclusion, the chrysolaminarin CL2 from O. aurita may be explored as a natural antioxidant agent for application in aquaculture, food and pharmaceutical areas.