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Oligotrophic Nitrification and Denitrification Bacterial Communities in a Constructed Sewage Treatment Ecosystem and Nitrogen Removal of Delftia tsuruhatensis NF4

RUILAN YANG
RUILAN YANG
, 
JING LI
JING LI
, 
LUYAO WEI-XIE
LUYAO WEI-XIE
 and 
LIN SHAO
LIN SHAO
   | Mar 11, 2020
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Article Category: original-paper
Published Online: Mar 11, 2020
Page range: 99 - 108
Received: Nov 27, 2019
Accepted: Feb 15, 2020
DOI: https://doi.org/10.33073/pjm-2020-013
Keywords
© 2020 RuiLan Yang et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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.
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.

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
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

Fig. 3.

Nitrate and TN removal efficiency with Delftia tsuruhatensis NF4 in sterilized STE influent water.
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.
Nitrate and TN removal efficiency with Delftia tsuruhatensis NF4 in the oligotrophic aerobic denitrification medium.

j.pjm-2020-013.tab.006

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)

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

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 *** ** *** *** *** *** *** *** ***

j.pjm-2020-013.tab.005

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)

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

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)
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