Fig. 1.
Water sample sites in constructed Sewage Treatment Ecosystem (STE) of Feng Huang He Ergou River. The source of nine samples, including IC, I.NF, I.DNF, CRIC, CRI.NF, CRI.DNF, EC, E.NF, and E.DNF. The bold thick arrows indicate the water flow direction in STE, and the bold thick hollow arrows indicate the sampling site.
Table I
Sample names and processing methods in the study.
| Sample source | Processing methods | Sample names |
|---|---|---|
| STE Influent Water | Filtration of STE influent water samples with filter membrane | IC |
| Filtration of eluate from OHN medium | I.NF | |
| Filtration of eluate from OAD medium | I.DNF | |
| CRI Effluent Water | Filtration of CRI effluent water samples with filter membrane | CRIC |
| Filtration of eluate from OHN medium | CRI.NF | |
| Filtration of eluate from OAD medium | CRI.DNF | |
| STE Effluent Water | Filtration of STE effluent water samples with filter membrane | EC |
| Filtration of eluate from OHN medium | E.NF | |
| Filtration of eluate from OAD medium | E.DNF |
Table II
Physical and chemical parameters of the STE influent, CRI effluent, and STE effluent water samples.
| Sample Name | DO (mg/l) | Turbidity (NTU) | Conductivity (S/m) | pH | CODMn (mg/l) | TN (mg/l) | no3 --n (mg/l) | nh4 +–n (mg/l) | TP (mg/l) |
|---|---|---|---|---|---|---|---|---|---|
| IC | 1.47 ± 0.0 | 16.00 ± 3.3 | 0.98 ± 0.0 | 7.83 ± 0.0 | 17.46 ± 1.6 | 19.4 ± 0.0 | 0.98 ± 0.0 | 16.6 ± 0.5 | 0.92 ± 0.0 |
| CRIC | 3.92 ± 0.1 | 0.27 ± 0.2 | 0.79 ± 0.0 | 6.96 ± 0.3 | 6.46 ± 0.1 | 2.36 ± 0.1 | 0.72 ± 0.0 | 0.64 ± 0.0 | 0.30 ± 0.0 |
| EC | 5.37 ± 0.1 | 0.27 ± 0.2 | 0.77 ± 0.0 | 7.59 ± 0.0 | 4.98 ± 0.2 | 0.31 ± 0.0 | 0.10 ± 0.0 | 0.10 ± 0.0 | 0.22 ± 0.3 |
| ANOVA | *** | ** | *** | *** | *** | *** | *** | *** | *** |
Table III
Effective reads, OTUs, and richness estimators (Chao1 and ACE) of nine samples.
| Sample Names | Effective reads | OTUs | Chao1 | ACE |
|---|---|---|---|---|
| IC | 46433 | 171 | 161.5 | 163.5 |
| I.NF | 47463 | 142 | 161.0 | 158.9 |
| I.DNF | 80100 | 137 | 139.3 | 141.9 |
| CRIC | 54424 | 131 | 132.6 | 137.9 |
| CRI.NF | 59932 | 118 | 117.8 | 126.0 |
| CRI.DNF | 64800 | 103 | 123.9 | 135.2 |
| EC | 48924 | 88 | 85.2 | 89.7 |
| E.NF | 50389 | 132 | 128.3 | 130.5 |
| E.DNF | 56999 | 125 | 121.0 | 122.4 |
3 IC – Filtration of sewage treatment ecosystem (STE) influent water samples with filter membrane; I.NF – Filtration of STE influent water samples eluate from oligotrophic heterotrophic nitrification (OHN) medium; I.DNF – Filtration of STE influent water samples eluate from oligotrophic aerobic denitrification (OAD) medium; CRIC – Filtration of constructed rapid infiltration (CRI) effluent water samples with filter membrane; CRI.NF – Filtration of CRI effluent water samples eluate from OHN medium; CRI.DNF – Filtration of CRI effluent water samples eluate from OAD medium; EC – Filtration of STE effluent water samples with filter membrane; E.NF – Filtration of STE effluent water samples eluate from OHN medium; E.DNF – Filtration of STE effluent water samples eluate from OAD medium
Fig. 2.
2a) The rarefaction curve representing bacterial diversity of IC, I.NF, I.DNF, CRIC, CRI.NF, CRI.DNF, EC, E.NF, and E.DNF; 2b) The phylum community abundance of nine samples, including IC, I.NF, I.DNF, CRIC, CRI.NF, CRI.DNF, EC, E.NF, and E.DNF; 2c) The genera community abundance of nine samples, including IC, I.NF, I.DNF, CRIC, CRI.NF, CRI.DNF, EC, E.NF, and E.DNF; 2d) Principal component analysis (PCA) of the bacterial community using an unweighted clustering method. The dashed line oval indicates the cluster of IC, EC, and CRIC. The dotted dashed line oval indicates the cluster of NF and DNF groups. The solid line oval indicates the cluster of CRI.NF, CRI.DNF and CRIC, which represent a closer cluster of community structures.
IC – Filtration of sewage treatment ecosystem (STE) influent water samples with filter membrane; I.NF – Filtration of STE influent water samples eluate from oligotrophic heterotrophic nitrification (OHN) medium; I.DNF – Filtration of STE influent water samples eluate from oligotrophic aerobic denitrification (OAD) medium; CRIC – Filtration of constructed rapid infiltration (CRI) effluent water samples with filter membrane; CRI.NF – Filtration of CRI effluent water samples eluate from OHN medium; CRI.DNF – Filtration of CRI effluent water samples eluate from OAD medium; EC – Filtration of STE effluent water samples with filter membrane; E.NF – Filtration of STE effluent water samples eluate from OHN medium; E.DNF – Filtration of STE effluent water samples eluate from OAD medium
Table IV
The relative abundance of the top three populations at different levels of the taxonomy.
| Taxonomy | IC | CRIC | EC | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | |
| Phylum | Proteobacteria (96.6) | Firmicutes (17.4) | Bacteroidetes (0.96) | Proteobacteria (99.3) | Firmicutes (0.17) | Bacteroidetes (0.16) | Proteobacteria (99.7) | Bacteroidetes (0.14) | Firmicutes (0.09) |
| Class | Gammaproteobacteria (83.8) | Alphaproteobacteria (12.8) | Clostridia (0.97) | Gammaproteobacteria (83.7) | Alphaproteobacteria (15.6) | Bacteroidia (0.16) | Gammaproteobacteria (81.7) | Alphaproteobacteria (18.0) | Bacteroidia (0.13) |
| Order | Xanthomonadales (78.0) | Sphingomonadales (6.1) | Rhizobiales (5.2) | Xanthomonadales (72.7) | Rhizobiales (7.8) | Pseudomonadales (5.8) | Xanthomonadales (72.7) | Rhizobiales (7.8) | Sphingomonadales (5.8) |
| Family | Xanthomonadaceae (78.0) | Rhizobiaceae (4.9) | Others (11.4) | Xanthomonadaceae (72.6) | Others (8.5) | Rhizobiaceae (7.8) | Xanthomonadaceae (72.7) | Rhizobiaceae (7.8) | Others (8.5) |
| Genus | Stenotrophomonas (78.0) | Sphingomomas (12.1) | Phyllobacterium (4.7) | Stenotrophomonas (72.7) | Phyllobacterium (10.0) | Sphingomomas (7.5) | Stenotrophomonas (72.6) | Sphingomomas (9.5) | Delftia (9.1) |
| Taxonomy | I.NF | CRI.NF | E.NF | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | |
| Phylum | Proteobacteria (79.0) | Bacteroidetes (20.6) | Firmicutes (0.09) | Proteobacteria (96.6) | Bacteroidetes (2.9) | Actinobacteria (0.12) | Proteobacteria (75.4) | Bacteroidetes (24.5) | Firmicutes (0.06) |
| Class | Gammaproteobacteria (77.0) | Bacteroidia (20.6) | Alphaproteobacteria (2.0) | Gammaproteobacteria (66.2) | Alphaproteobacteria (30.3) | Bacteroidia (2.9) | Gammaproteobacteria (73.8) | Bacteroidia (24.5) | Alphaproteobacteria (1.6) |
| Order | Pseudomonadales (61.4) | unidentified_Gammaproteobacteria (6.7) | Aeromonadales (5.7) | Pseudomonadales (42.7) | Rhizobiales (28.6) | Aeromonadales (11.5) | Pseudomonadales (35.6) | Flavobacteriales (24.0) | unidentified_Gammaproteobacteria (18.3) |
| Family | Moraxellaceae (45.7) | Flavobacteriaceae (19.1) | Pseudomonadaceae (15.8) | Moraxellaceae (32.8) | Rhizobiaceae (28.7) | Xanthomonadaceae (10.5) | Moraxellaceae (31.9) | unidentified_Flavobacteriales (20.6) | Burkholderiaceae (9.9) |
| Genus | Acinetobacter (46.7) | Flavobacterium (20.5) | Pseudomonas (15.8) | Acinetobacter (32.1) | unidentified_Rhizobiaceae (28.5) | Pseudomonas (10.5) | Acinetobacter (31.9) | unidentified_Rhizobiaceae (22.5) | Aeromonas (20.6) |
| Taxonomy | I.DNF | CRI.DNF | E.DNF | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | Top1 (%) | Top2 (%) | Top3 (%) | |
| Phylum | Proteobacteria (81.7) | Bacteroidetes (18.1) | Firmicutes (0.05) | Proteobacteria (99.5) | Bacteroidetes (0.4) | Actinobacteria (0.07) | Proteobacteria (74.9) | Bacteroidetes (25.0) | Firmicutes (0.05) |
| Class | Gammaproteobacteria (80.4) | Bacteroidia (18.1) | Alphaproteobacteria (1.2) | Alphaproteobacteria (60.9) | Gammaproteobacteria (38.6) | Bacteroidia (0.39) | Gammaproteobacteria (72.9) | Bacteroidia (25.0) | Alphaproteobacteria (1.9) |
| Order | Pseudomonadales (56.5) | Flavobacteriales (17.0) | Aeromonadales (11.6) | Rhizobiales (60.5) | Pseudomonadales (24.5) | Aeromonadales (7.7) | Aeromonadales (29.6) | Flavobacteriales (24.1) | Pseudomonadales (17.1) |
| Family | Pseudomonadaceae (35.1) | Moraxellaceae (21.4) | Flavobacteriaceae (16.7) | Rhizobiaceae (60.5) | Moraxellaceae (14.3) | Pseudomonadaceae (10.2) | Aeromonadaceae (25.1) | unidentified_Flavobacteriales (21.5) | Moraxellaceae (14.3) |
| Genus | Pseudomonas (34.8) | Acinetobacter (20.3) | Flavobacterium (16.7) | unidentified_Rhizobiaceae (61.4) | Acinetobacter (12.7) | Pseudomonas (6.1) | Aeromonas (22.5) | Cloacibacterium (20.9) | Acinetobacter (16.8) |
Fig. 3.
Nitrate and TN removal efficiency with Delftia tsuruhatensis NF4 in sterilized STE influent water.
Fig. 4.
Nitrate and TN removal efficiency with Delftia tsuruhatensis NF4 in the oligotrophic aerobic denitrification medium.