The aim of this study was to investigate the microbial community of three tannery wastewater treatment plants (WWTPs) involved in nitrification by Illumina MiSeq sequencing. The results showed that highly diverse communities were present in tannery wastewater. A total of six phyla, including Proteob...
The aim of this study was to investigate the microbial community of three tannery wastewater treatment plants (WWTPs) involved in nitrification by Illumina MiSeq sequencing. The results showed that highly diverse communities were present in tannery wastewater. A total of six phyla, including Proteobacteria (37-41%), Bacteroidetes (6.04-16.80), Planctomycetes (3.65-16.55), Chloroflexi (2.51-11.48), Actinobacteria (1.91-9.21), and Acidobacteria (3.04-6.20), were identified as the main phyla, and Proteobacteria dominated in all the samples. Within Proteobacteria, Beta-proteobacteria was the most abundant class, with the sequence percentages ranging from 9.66% to 17.44%. Analysis of the community at the genus level suggested that Thauera, Gp4, Ignavibacterium, Phycisphaera, and Arenimonas were the core genera shared by at least two tannery WWTPs. A detailed analysis of the abundance of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) indicated that Nitrosospira, Nitrosomonas, and Nitrospira were the main AOB and NOB in tannery wastewater, respectively, which exhibited relatively high abundance in all samples. In addition, real-time quantitative PCR was conducted to validate the results by quantifying the abundance of the AOB and total bacteria, and similar results were obtained. Overall, the results presented in this study may provide new insights into our understanding of key microorganisms and the entire community of tannery wastewater and contribute to improving the nitrogen removal efficiency.
The aim of this study was to investigate the microbial community of three tannery wastewater treatment plants (WWTPs) involved in nitrification by Illumina MiSeq sequencing. The results showed that highly diverse communities were present in tannery wastewater. A total of six phyla, including Proteobacteria (37-41%), Bacteroidetes (6.04-16.80), Planctomycetes (3.65-16.55), Chloroflexi (2.51-11.48), Actinobacteria (1.91-9.21), and Acidobacteria (3.04-6.20), were identified as the main phyla, and Proteobacteria dominated in all the samples. Within Proteobacteria, Beta-proteobacteria was the most abundant class, with the sequence percentages ranging from 9.66% to 17.44%. Analysis of the community at the genus level suggested that Thauera, Gp4, Ignavibacterium, Phycisphaera, and Arenimonas were the core genera shared by at least two tannery WWTPs. A detailed analysis of the abundance of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) indicated that Nitrosospira, Nitrosomonas, and Nitrospira were the main AOB and NOB in tannery wastewater, respectively, which exhibited relatively high abundance in all samples. In addition, real-time quantitative PCR was conducted to validate the results by quantifying the abundance of the AOB and total bacteria, and similar results were obtained. Overall, the results presented in this study may provide new insights into our understanding of key microorganisms and the entire community of tannery wastewater and contribute to improving the nitrogen removal efficiency.
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제안 방법
In this study, the microbial community of tannery wastewater involved in nitrification was investigated by using Illumina MiSeq sequencing and the results were further validated by qPCR analysis. This research may further elucidate the microbial community structure and aid in developing promising strategies and proper management technologies for tannery wastewater treatment.
Operational taxonomic units (OTUs) were categorized using CD-HIT at a 97% sequence similarity threshold, and the taxonomic assignment of OTUs was performed by the RDP classifier with 50% confidence [25]. Rarefaction curves were generated, and Shannon, Chao 1, and abundance-based coverage estimator (ACE) indexes were calculated using the Mothur program to compare the microbial diversity and richness between these sludge samples. Hierarchical clustering analysis was performed using C LUSTER and visualized using TREEVIEW.
To validate the sequencing result, qPCR assay was conducted to quantify the abundance of AOA, AOB, and total bacteria in the three samples (Fig. 7). As shown in Fig.
데이터처리
Rarefaction curves were generated, and Shannon, Chao 1, and abundance-based coverage estimator (ACE) indexes were calculated using the Mothur program to compare the microbial diversity and richness between these sludge samples. Hierarchical clustering analysis was performed using C LUSTER and visualized using TREEVIEW.
이론/모형
In this study, the microbial community of activated sludge involved in nitrification from tannery WWTPs was investigated by the Illumina MiSeq sequencing approach. The results showed that the diversity and richness of the tannery wastewater were similar to those of municipal wastewater and significantly higher than those of other industrial wastewaters.
After removing low-quality sequences and chimeras, at least 25,714 (eubacteria) and 34,658 (archaea) effective sequences were obtained for each sample. The sequence number of each sample was normalized and tags with 97% similarity were then grouped into OTUs by the CD-HIT clustering method. At least 2,035 and 364 OTUs were generated from the eubacterial and archaeal sequences, respectively (Table 1).
To validate the abundance of the total bacteria AOB and AOA, qPCR was conducted with the fluorescent SYBR-Green method. Genomic DNA was extracted using the Mag-Bind Soil DNA Kit (Omega Biotek) according to the manufacturer’s instructions.
성능/효과
In addition, previous studies also revealed that AOA amoA is absent in aerobic sludge of different WWTPs and it was detected only in the anaerobic sludge [26, 47]. Analysis of the abundance of AOB and total bacteria showed that the log copy numbers of the AOB amoA gene and bacterial 16S rRNA in the three samples were 5.4-6.0 and 6.3-6.6 copies per nanogram of DNA, respectively (Fig. 7). It was shown that the qPCR quantification results were in agreement with the sequencing ones.
44% of the total identified eubacterial sequences, respectively. As for sample AS-B, Phycisphaera, Nitrosomonas, Gp4, and Truepera were the main genera, with 15.63%, 3.94%, 3.38%, and 3.46% of the sequences on average. In sample AS-C, Phycisphaera (8.
As for the archaea, the most abundant phyla were Euryarchaeota (56.87-94.92%), Thaumarchaeota (1.47-26.28%), and Crenarchaeota (0.06-14.99%), which accounted for approximately 98% of all the archaeal sequences (Fig. 2B). Among them, Euryarchaeota and Thaumarchaeota were also detected previously as the predominant phyla in tannery WWTPs by a combined PCR-DGGE and highthroughput sequencing method [28].
For the archaea, the largest genus was Methanosaeta (10.08-27.23%) in all the three samples, which was affiliated to phylum Euryarchaeota. Previous research demonstrated that genus Methanosaeta generally existed in the anaerobic digestion of organic compounds, converting acetate to methane [41], and further research is needed to elucidate the role of genus Methanosaeta during nitrification.
NOB included the four groups Nitrobacter, Nitrospina, Nitrococcus, and Nitrospira, belonging to Alpha-proteobacteria, Gamma-proteobacteria, Beta-proteobacteria, and Nitrospirae, respectively [44]. In this study, only Nitrospira was detected in the three samples, with high distribution in AS-A and AS-C, suggesting that Nitrospira is the major NOB in tannery WWTPs. This result was in agreement with that obtained in other activated sludge [25, 44].
These results showed that samples AS-A and AS-C had higher eubacterial and archaeal diversities, respectively. In this study, relatively high Shannon indexes were obtained for all samples expect archaeal community in sample AS-B, corresponding to high OTU values. The diversity and richness of the tannery wastewater were similar to those of municipal wastewater plants and significantly higher than those of other industrial wastewaters [15, 18, 25].
5, there was a distinct difference in the microbial composition in each sample. Thauera, Gp4, Ignavibacterium, and Nitrospira were dominant genera in sample AS-A, which accounted for 4.74%, 3.38%, 4.57%, and 4.44% of the total identified eubacterial sequences, respectively. As for sample AS-B, Phycisphaera, Nitrosomonas, Gp4, and Truepera were the main genera, with 15.
08%) were the predominant genera. The genus Thauera, of the class Beta-proteobacteria, was the primary genus in AS-A, and it was detected in all the three samples (Fig. 5A). Similar results were also reported in aerobic sludge samples collected from tannery WWTPs, and extremely high abundance was detected (4.
3B). The largest family in AS-A and AS-B were Nitrososphaeraceae, whereas in sample AS-C, Methanomicrobiale sincertae-sedis was the abundant family (Fig. 4B). Although a relatively low abundance of archaea was detected, they may play important roles during nitrification.
In this study, the microbial community of activated sludge involved in nitrification from tannery WWTPs was investigated by the Illumina MiSeq sequencing approach. The results showed that the diversity and richness of the tannery wastewater were similar to those of municipal wastewater and significantly higher than those of other industrial wastewaters. The taxonomic diversity of the microbial community of tannery wastewater was identified from the phylum to genus levels, and the core genera were further discussed in the present manuscript.
09) for the eubacteria and archaeal community, respectively. These results showed that samples AS-A and AS-C had higher eubacterial and archaeal diversities, respectively. In this study, relatively high Shannon indexes were obtained for all samples expect archaeal community in sample AS-B, corresponding to high OTU values.
후속연구
23%) in all the three samples, which was affiliated to phylum Euryarchaeota. Previous research demonstrated that genus Methanosaeta generally existed in the anaerobic digestion of organic compounds, converting acetate to methane [41], and further research is needed to elucidate the role of genus Methanosaeta during nitrification.
qPCR was performed to quantify the abundance of the amoA gene of AOB and total bacteria, and the results were in agreement with the sequencing ones. The results obtained in this study can extend our knowledge about nitrifiers in tannery WWTPs, and might be practically useful in efficiently removing nitrogen from industrial wastewaters.
In this study, the microbial community of tannery wastewater involved in nitrification was investigated by using Illumina MiSeq sequencing and the results were further validated by qPCR analysis. This research may further elucidate the microbial community structure and aid in developing promising strategies and proper management technologies for tannery wastewater treatment.
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