As a typical sub-deep reservoir in the upper reaches of the Yangtze River in the southwest region, Zhangjiayan Reservoir is also an important source of drinking water. Exploring the role of microorganisms in the material cycle of water bodies is of great significance for preventing the exacerbation of eutrophication in the reservoir. In this study, water samples from the overlying water of five points in the reservoir were collected four times in spring (April), summer (July), autumn (November), and winter (January) of 2022–2023 using a gas-tight water sampler. Physicochemical factors were measured, and the microbial community structure was analyzed by high-throughput MiSeq sequencing of the V3–V4 hypervariable region of 16S rRNA gene in order to explore the relationship between physicochemical factors and microbial community structure and the dominant microbial populations that affect eutrophication of the reservoir. The following results were obtained through analysis. Among the 20 overlying water samples from Zhangjiayan Reservoir, a total of 66 phyla, 202 classes, 499 orders, 835 families, 1716 genera, and 27,904 ASVs of the bacterial domain were detected. The phyla Proteobacteria and Actinobacteria were dominant in the microbial community of the overlying water in Zhangjiayan Reservoir. At the genus level, hgcI_clade and Actinobacteria had the highest abundance and was the dominant population. The microbial community in the water of Zhangjiayan Reservoir has a high level of diversity. The diversity index ranked by numerical order was winter > autumn > summer > spring. Significant differences were found in the composition and structure of the microbial community between the spring/summer and autumn/winter seasons (p < 0.05). Total phosphorus, dissolved total phosphorus, soluble reactive phosphorus, and dissolved oxygen have a significant impact on the composition and structure of the microbial community (p < 0.01). The bacterial community in the overlying water of Zhangjiayan Reservoir showed a mainly positive correlation. Sphingomonas, Brevundimonas, and Blastomonas were the central populations of the bacterial community in the overlying water of Zhangjiayan Reservoir. This study indicates that environmental factors, such as phosphorus and other nutrients, have a significant impact on the formation of the microbial community structure in different seasons. Sphingomonas, Brevundimonas, and Blastomonas are key populations that may have a significant impact on eutrophication in Zhangjiayan Reservoir.

To study the microbial community structure in sediments and its relation to eutrophication environment factors, the sediments and the overlying water of Sancha Lake were collected in the four seasons. MiSeq high-throughput sequencing was conducted for the V3–V4 hypervariable regions of the 16S rRNA gene and was used to analyze the microbial community structure in sediments. Pearson correlation and redundancy analysis (RDA) were conducted to determine the relation between microbial populations and eutrophic factors. The results demonstrated four main patterns: (1) in the 36 samples that were collected, the classification annotation suggested 64 phyla, 259 classes, 476 orders, 759 families, and 9325 OTUs; (2) The diversity indices were ordered according to their values as with summer > winter > autumn > spring; (3) The microbial populations in the four seasons belonged to two distinct characteristic groups; (4) pH, dissolved oxygen (DO), total phosphorus (TP), and total nitrogen (TN) had significant effects on the community composition and structure, which further affected the dissolved total phosphorus (DTP) significantly. The present study demonstrates that the microbial communities in Sancha Lake sediments are highly diverse, their compositions and distributions are significantly different between spring and non-spring, and Actinobacteria and Cyanobacteria may be the key populations or indicator organisms for eutrophication.

The bacterial phoD gene encoding alkaline phosphatase (ALP) plays an important role in the release of soluble reactive phosphorus (SRP) from organic phosphorus in ecosystems. However, phoD gene diversity and abundance in ecosystems is poorly understood. In the present study, we sampled the surface sediments and the overlying water of Sancha Lake at 9 different sampling sites, a typical eutrophic sub-deep freshwater lake in China, in April 15 (spring) and November 3 (autumn), 2017. High-throughput sequencing and qPCR were performed to analyze the diversity and abundance of the bacterial phoD gene in the sediments. We further discussed the relationships between the diversity and abundance of the phoD gene and environmental factors and ALP activity. A total of 881,717 valid sequences were obtained from 18 samples and were classified into 41 genera, 31 families, 23 orders, 12 classes, and 9 phyla and grouped into 477 OTUs. The dominant phyla were Proteobacteria and Actinobacteria. The phylogenetic tree based on the sequences of the phoD gene was plotted and composed of three branches. The genetic sequences were aligned predominantly with genera Pseudomonas, Streptomyces, Cupriavidus, and Paludisphaer. The phoD-harboring bacterial community structure showed a significant difference in spring and autumn, but no apparent spatial heterogeneity. The phoD gene abundances at different sampling points were significantly higher in autumn than in spring. In autumn and spring, the phoD gene abundance was significantly higher in the tail of lake and where cage culture used to be intensive. pH value, dissolved oxygen (DO), total organic carbon (TOC), ALP, and phosphorus were important environmental factors affecting the diversity of the phoD gene and the phoD-harboring bacterial community structure. Changes in phoD-harboring bacterial community structure, phoD gene abundance, and ALP activity were negatively correlated with SRP in overlying water. Our study indicated phoD-harboring bacteria in the sediments of Sancha Lake with the characteristics of high diversity and significant spatial and temporal heterogeneity in abundance and community structure, which played a important role in the release of SRP.

The organophosphate-mineralizing bacteria (OPB) convert environmental organic phosphorus (P) into soluble P that can be directly absorbed and utilized by organisms. OPB is an important group of microorganisms in lake sediments. The P decomposed and released from the sediments by OPB is an important P-source in eutrophic water bodies. In this study, the egg-yolk organophosphate medium was used to isolate and screen OPB strains from the sediments of Sancha Lake. Furthermore, the obtained OPB strains were classified based on their 16S rDNA sequence. Both the solid and liquid lecithin hydrolyzing experiments were conducted to investigate the P-solubilizing characteristics of the obtained OPB strains. Microcosm experimentsiwere performed to study the P-release ability of OPB strains from sediments. A total of 39 OPB strains were isolated from the sediments of Sancha Lake. They belonged to three phyla, five families, and five genera, and contained two potentially new species. Bacillus and Pseudomonas were the dominant genera. On the solid lecithin plate, 35 of the 39 OPB strains produced visible phosphate halos, and 24 strains showed a high ratio of P halo diameter (HD)/colony diameter (CD). In the liquid lecithin medium, all 39 OPB strains demonstrated P-solubilizing ability, but with significant differences. The Pseudomonas strain demonstrated the strongest P-solubilizing ability, at 70.91 mg·L⁻¹. There was no significant correlation between the amount of released phosphorus by OPB strains and pH. The P-solubilizing characteristics of OPB were affected by the interaction of dissolved inorganic phosphate and alkaline phosphatase. In the microcosm experiments, the added OPB strains significantly promoted the decomposition and release of organic phosphorus (OP) in the sediments. OPB in the sediments of Sancha Lake is rich in diversity and had a strong ability to release OP in the sediments.

The aerobic denitrifying phosphate accumulating bacteria (ADPB) use NO₃⁻ as an electron acceptor and remove nitrate by denitrification and concomitant uptake of excessive phosphorus in aerobic conditions. Activated sludge was collected from the A²O aerobic biological pool of the sewage treatment plant at Hezuo Town, Chengdu City. The candidate ADPB strains were obtained by cultivation in the enriched denitrification media, followed by repeated isolation and purification on bromothymol blue (BTB) solid plates. The obtained candidates were further screened for ADPB strains by phosphorus uptake experiment, nitrate reduction test, metachromatic granules staining, and poly-β-hydroxybutyrate (PHB) staining. The 16 sedimentation ribosome deoxyribonucleic acid (16 S rDNA) molecular technique was used to determine their taxonomy. Further, the denitrification and dephosphorization capacities of ADPB strains were ascertained through their growth characteristics in nitrogen-phosphorus-rich liquid media. The results revealed a total of 25 ADPB strains screened from the activated sludge of the A²O aerobic biological pool of the sewage treatment plant at Hezuo Town. These strains belonged to two classes, four orders, and five genera. Among them, the strain SW18NP2 was a potentially new species in the Acinetobacter genus, while the strain SW18NP24 was a potential new species in the Pseudomonas genus at the time of their characterization. The Acinetobacter was the dominant genus. The obtained ADPB strains demonstrated a rich diversity. The ADPB strains had significant variations in denitrification and dephosphorization capacities. Twenty-three strains exhibited a total phosphorus removal rate of above 50%, and 19 strains exhibited a total nitrogen removal rate of above 50%. The strain SW18NP2 showed the best denitrifying phosphorus removal (DPR) capacity, with a dephosphorization rate of 82.32% and a denitrification rate of 73.73%. The ADPB in the A²O aerobic biological pool of the sewage treatment plant at Hezuo Town demonstrated a rich diversity and a strong DPR capacity.

The activated sludge was collected from the Membrane BioReactor (MBR) pool of the sewage treatment system of Sanxing Town, Jintang County, Chengdu, to obtain a good population of heterotrophic nitrifying/aerobic denitrifying (HNAD) bacteria. After undergoing enrichment, isolation, and purification, the HNAD bacteria were selected using the pure culture method. The 16S rDNA molecular technology was used to determine the taxonomy of bacteria. The heterophic nitrifying ability and denitrification capacity of HNAD strains was ascertained through their growth characteristics in heterotrophic nitrification and denitrification media. The results showed that 53 HNAD strains selected from the MBR pool belonged to 2 phyla, 3 classes, 6 orders, 6 families, and 7 genera, with 26 species. Acinetobacter was the largest and dominant genus. Among these, strains numbered (bacterial strain) SW21HD14, SW21HD17, and SW21HD18 were potentially new species in the Acinetobacter genus. Each HNAD strain showed a significant heterotrophic nitrifying and aerobic denitrifying efficiency compared with the control strain (P < 0.05). Specifically, 10 strains demonstrated ammonia nitrogen degradation of greater than 70 mg·L^-1 and 9 strains demonstrated nitrate nitrogen degradation above 150 mg·L^-1. The HNAD bacteria, which were selected from the MBR pool of sewage treatment system of the Sanxing Town sewage treatment plant, exhibited rich diversity and strong nitrogen removal ability. These findings offered an effective strain source and theoretical basis for implementing biological denitrification technology that involves synchronous nitrification and denitrification.