Nitrogen Removal Efficiency and Microbial Response Mechanism of Hordeum vulgare var. coeleste L. Straw as an External Carbon Source Under Different C/N Ratios

To address the bottleneck of poor biological nitrogen removal efficiency caused by the extremely low carbon-to-nitrogen (C/N) ratio of domestic sewage in alpine plateau regions, this study used Hordeum vulgare var. coeleste L., a characteristic crop endemic to the Qinghai–Tibet Plateau, as raw material and adopted pretreated highland barley straw as an external carbon source. Three parallel experiments were carried out using the anaerobic–aerobic–anoxic sequencing batch reactor (AOA-SBR) process to investigate the nitrogen removal performance and functional succession of the microbial community in the AOA-SBR system under three C/N ratio ranges: 5~7, 7~9, and 9~11. The results showed that the addition of an external carbon source significantly improved nitrogen removal efficiency. The optimal C/N ratio range for nitrogen removal in this study was determined to be 7~9. A weakly alkaline environment was conducive to denitrification. The fermentation broth prepared by alkali pretreatment contained a large amount of readily biodegradable organic matter with low toxicity, and achieved excellent nitrogen removal performance, helping to realize cost reduction and efficiency improvement in wastewater treatment. At the optimal C/N ratio of 7~9, the average removal efficiencies of ammonia nitrogen (NH4+-N) and total nitrogen (TN) reached 94.46% and 61.32%, respectively, which were significantly improved compared with the blank control group without external carbon addition. During the experimental period, no obvious changes were observed in microbial abundance at the phylum level, whereas the community structure at the genus level responded significantly to the addition of a straw carbon source. Among them, genera with specific degradation capabilities for straw hydrolysates, such as norank_f__Chitinophagaceae and unclassified_f__Comamonadaceae, were highly sensitive to variations in the C/N ratio. These genera could partially replace the nitrification and denitrification functions of other microorganisms and played a key role in the nitrogen removal process. In contrast, Thauera, a typical conventional heterotrophic denitrifier, showed no significant response to changes in the C/N ratio, indicating that the straw-based external carbon source mainly affected microbial genera with specific hydrolysate-degrading functions.