Changes in the composition of bacterial communities and pathogen levels during wastewater treatment.

Wastewater treatment plants have been described as a potential source of spreading pathogens to the receiving water. However, few studies are reporting the presence and concentration changes of pathogens in these matrices. High-throughput sequencing provides new insights into understanding the changes of bacterial communities throughout wastewater treatment plants (WWTPs). In this study, the changes in microbial community composition and the levels of representative pathogens of effluents during the wastewater treatment process in two municipal WWTPs (A and B) were analyzed using Illumina NovaSeq sequencing and qPCR. Proteobacteria was the most abundant phylum in all samples, accounting for 45.0-75.2% of the bacterial community, followed by Firmicutes, Bacteroidetes, Actinobacteria, and Nitrospirae. A slight difference was observed between the bacterial community compositions of WWTPs A and B. However, a significant difference in the community compositions of effluent samples at different treatment stages was observed. Nutrients had a more substantial impact on bacterial community composition than physicochemical factors. Most human-associated Bacteroides and Mycobacterium were eliminated during the wastewater treatment process in both WWTPs. The bacterial community richness in WWTP A was significantly higher than that in WWTP B. The results of this study will provide insights into the potential problems that exist in WWTPs. In turn, these insights can enable the efficient and stable operation of WWTPs and help prevent the spread of pathogens.

[Co-amorphous technology to improve dissolution and physical stability of silybin].

The present study explored the effect of co-amorphous technology in improving the dissolution rate and stability of silybin based on the puerarin-silybin co-amorphous system prepared by the spray-drying method. Solid-state characterization was carried out by powder X-ray diffraction(PXRD), polarizing microscopy(PLM), Fourier transform infrared spectroscopy(FT-IR), differential scanning calorimetry(DSC), etc. Saturated powder dissolution, intrinsic dissolution rate, moisture absorption, and stability were further investigated. The results showed that puerarin and silybin formed a co-amorphous system at a single glass transition temperature which was higher than that of any crude drug. The intrinsic dissolution rate and supersaturated powder dissolution of silybin in the co-amorphous system were higher than those of the crude drug and amorphous system. The co-amorphous system kept stable for as long as three months under the condition of 40 ℃, 75% relative humidity, which was longer than that of the single amorphous silybin. Therefore, the co-amorphous technology could significantly improve the dissolution and stability of silybin.

Dummy Template-Based Molecularly Imprinted Membrane Coating for Rapid Analysis of Malachite Green and Its Metabolic Intermediates in Shrimp and Fish.

A novel malachite green molecularly imprinted membrane (MG-MIM) with specific selectivity for malachite green (MG) and leucomalachite green (LMG) was prepared using a hydrophobic glass fiber membrane as the polymer substrate, methyl violet as a template analog, 4-vinyl benzoic acid as the functional monomer, and ethyleneglycol dimethacrylate as the crosslinking agent. MG-MIM and non-imprinted membrane (NIM) were structurally characterized using scanning electron microscopy, surface area analyzer, Fourier-transform infrared spectrometer and synchronous thermal analyzer. The results showed that MG-MIM possessed a fluffier surface, porous and looser structure, and had good thermal stability. Adsorption properties of MG-MIM were investigated under optimal conditions, and adsorption equilibrium was reached in 20 min. The saturated adsorption capacities for MG and LMG were 24.25 ng·cm-2 and 13.40 ng·cm-2, and the maximum imprinting factors were 2.41 and 3.20, respectively. Issues such as "template leakage" and "embedding" were resolved. The specific recognition ability for the targets was good and the adsorption capacity was stable even after five cycles. The proposed method was successfully applied for the detection of MG and LMG in real samples, and it showed good linear correlation in the range of 0 to 10.0 µg·L-1 (R2 = 0.9991 and 0.9982), and high detection sensitivity (detection limits of MG and LMG of 0.005 µg/kg and 0.02 µg·kg-1 in shrimp, and 0.005 µg/kg and 0.02 µg/kg in fish sample). The recoveries and relative standard deviations were in the range of 76.31-93.26% and 0.73-3.72%, respectively. The proposed method provides a simple, efficient and promising alternative for monitoring MG and LMG in aquatic products.

Signs of biofilm formation in the genome of Labrenzia sp. PO1.

Labrenzia sp. are important components of marine ecology which play a key role in biochemical cycling. In this study, we isolated the Labrenzia sp. PO1 strain capable of forming biofilm, from the A. sanguinea culture. Growth analysis revealed that strain reached a logarithmic growth period at 24 hours. The whole genome of 6.21813 Mb of Labrezia sp. PO1 was sequenced and assembled into 15 scaffolds and 16 contigs, each with minimum and maximum lengths of 644 and 1,744,114 Mb. A total of 3,566 genes were classified into five pathways and 31 pathway groups. Of them, 521 genes encoded biofilm formation proteins, quorum sensing (QS) proteins, and ABC transporters. Gene Ontology annotation identified 49,272 genes that were involved in biological processes (33,425 genes), cellular components (7,031genes), and molecular function (7,816 genes). We recognised genes involved in bacterial quorum sensing, attachment, motility, and chemotaxis to investigate bacteria's ability to interact with the diatom phycosphere. As revealed by KEGG pathway analysis, several genes encoding ABC transporters exhibited a significant role during the growth and development of Labrenzia sp. PO1, indicating that ABC transporters may be involved in signalling pathways that enhance growth and biofilm formation.

Coupled nitrification and N2 gas production as a cryptic process in oxic riverbeds.

The coupling between nitrification and N2 gas production to recycle ammonia back to the atmosphere is a key step in the nitrogen cycle that has been researched widely. An assumption for such research is that the products of nitrification (nitrite or nitrate) mix freely in the environment before reduction to N2 gas. Here we show, in oxic riverbeds, that the pattern of N2 gas production from ammonia deviates by ~3- to 16-fold from that predicted for denitrification or anammox involving nitrite or nitrate as free porewater intermediates. Rather, the patterns match that for a coupling through a cryptic pool, isolated from the porewater. A cryptic pool challenges our understanding of a key step in the nitrogen cycle and masks our ability to distinguish between sources of N2 gas that 20 years' research has sought to identify. Our reasoning suggests a new pathway or a new type of coupling between known pathways in the nitrogen cycle.

A study on the nitrogen removal efficacy of bacterium Acinetobacter tandoii MZ-5 from a contaminated river of Shenzhen, Guangdong Province, China.

Heterotrophic nitrification-aerobic denitrification (HN-AD) has advantages over the traditional nitrogen removal process when removing multiple types of nitrogen in wastewater treatment. Acinetobacter tandoii MZ-5, which is capable of HN-AD, was isolated from the sediment of a polluted river for the first time. It used NH4+-N, NO2--N and NO3--N as sole nitrogen sources with maximum removal rates of 2.28, 1.18 and 1.04 mg L-1h-1, respectively. Simultaneous nitrification and denitrification were observed when using mixed N sources and NH4+-N was preferentially utilized. High nitrogen removal efficiencies (>90%) were achieved under the following conditions: C/N ratio 11-18, pH 6-8, 25-30 °C and dissolved oxygen 7.35-7.66 mg L-1. Strain MZ-5 was effective at treating wastewater from landfill leachate treatment plants, with NH4+-N, NO3--N and total nitrogen (TN) removal efficiencies of 99.28%, 44.85% and 45.31%, respectively. Thus, strain MZ-5 may be a good candidate for wastewater treatment.

Characteristics of spatial and seasonal bacterial community structures in a river under anthropogenic disturbances.

In this study, the seasonal characteristics of microbial community compositions at different sites in a river under anthropogenic disturbances (Maozhou River) were analyzed using Illumina HiSeq sequencing. Taxonomic analysis revealed that Proteobacteria was the most abundant phylum in all sites, followed by Actinobacteria, Bacteroidetes, Chloroflexi, Acidobacteria and Firmicutes. The variations of the community diversities and compositions between the seasons were not significant. However, significant differences between sites as well as water and sediment samples were observed. These results indicated that sites under different levels of anthropogenic disturbances have selected distinct bacterial communities. pH, dissolved oxygen (DO), concentrations of total nitrogen (TN) and heavy metals were the main factors that influence the diversity and the composition of bacterial community. Specifically, the relative abundance of Proteobacteria was negatively correlated with pH and DO and positively correlated with TN, while Actinobacteria and Verrucomicrobia showed the opposite pattern. Moreover, positive correlations between the relative abundances of Firmicutes and Bacteroidetes and the concentration of heavy metals were also found. Results of functional prediction analysis showed no significant differences of the carbon, nitrogen and phosphorus metabolism across the sites and seasons. Potential pathogens such as Vibrio, Arcobacter, Acinetobacter and Pseudomonas were found in these samples, which may pose potential risks for environment and human health. This study reveals the effect of anthropogenic activities on the riverine bacterial community compositions and provides new insights into the relationships between the environmental factors and the bacterial community distributions in a freshwater ecosystem under anthropogenic disturbances.

Spatial separation of anaerobic ammonium oxidation and nitrite-dependent anaerobic methane oxidation in permeable riverbeds.

Anaerobic ammonium oxidation (anammox) and nitrite-dependent anaerobic methane oxidation (n-damo) play important roles in nitrogen and carbon cycling in fresh waters but we do not know how these two processes compete for their common electron acceptor, nitrite. Here, we investigated the spatial distribution of anammox and n-damo across a range of permeable riverbed sediments. Anammox activity and gene abundance were detected in both gravel and sandy riverbeds and showed a simple, common vertical distribution pattern, while the patterns in n-damo were more complex and n-damo activity was confined to the more reduced, sandy riverbeds. Anammox was most active in surficial sediment (0-2 cm), coincident with a peak in hzsA gene abundance and nitrite. In contrast, n-damo activity peaked deeper down (4-8 cm) in the sandy riverbeds, coincident with a peak in n-damo 16S rRNA gene abundance and higher methane concentration. Pore water nitrite, methane and oxygen were key factors influencing the distribution of these two processes in permeable riverbeds. Furthermore, both anammox- and n-damo- activity were positively correlated with denitrification activity, suggesting a role for denitrification in supplying both processes with nitrite. Our data reveal spatial separation between anammox and n-damo in permeable riverbed sediments that potentially avoids them competing for nitrite.

Active pathways of anaerobic methane oxidation across contrasting riverbeds.

Anaerobic oxidation of methane (AOM) reduces methane emissions from marine ecosystems but we know little about AOM in rivers, whose role in the global carbon cycle is increasingly recognized. We measured AOM potentials driven by different electron acceptors, including nitrite, nitrate, sulfate, and ferric iron, and identified microorganisms involved across contrasting riverbeds. AOM activity was confined to the more reduced, sandy riverbeds, whereas no activity was measured in the less reduced, gravel riverbeds where there were few anaerobic methanotrophs. Nitrite-dependent and nitrate-dependent AOM occurred in all sandy riverbeds, with the maximum rates of 61.0 and 20.0 nmol CO2 g-1 (dry sediment) d-1, respectively, while sulfate-dependent and ferric iron-dependent AOM occurred only where methane concentration was highest and the diversity of AOM pathways greatest. Diverse Candidatus Methylomirabilis oxyfera (M. oxyfera)-like bacteria and Candidatus Methanoperedens nitroreducens (M. nitroreducens)-like archaea were detected in the sandy riverbeds (16S rRNA gene abundance of 9.3 × 105 to 1.5 × 107 and 2.1 × 104 to 2.5 × 105 copies g-1 dry sediment, respectively) but no other known anaerobic methanotrophs. Further, we found M. oxyfera-like bacteria and M. nitroreducens-like archaea to be actively involved in nitrite- and nitrate/ferric iron-dependent AOM, respectively. Hence, we demonstrate multiple pathways of AOM in relation to methane, though the activities of M. oxyfera-like bacteria and M. nitroreducens-like archaea are dominant.

[Isolation, identification and characterization of a microcystin-degrading bacterium Paucibacter sp. strain CH].

A bacterium capable of degrading microcystin (MC), strain CH, was isolated from the sediment of Lake Chaohu, China. Strain CH was tentatively identified as Paucibacter sp. based on the analysis of 16S rRNA gene sequences. Paucibacter sp. strain CH can use microcystin LR (MCLR) as the sole carbon and energy sources, and 11.6 microg x mL(-1) of MCLR was degraded to below the detection limit within 10 hours with the first-order reaction rate constant of 0.242 h(-1). The optimum temperature and initial pH for MC degradation were 25-30 degrees C and pH 6-9, respectively. A novel intermediate product containing the Adda residue was detected during the degradation of MCLR, which is different from those produced by strain ACM-3962, and Adda was recognized as the final product of the degradation process. Furthermore, no homologue to any of the four genes, mlrA, mlrB, mlrC and mlrD previously associated with the degradation of MCLR by strain ACM-3962 was found in strain CH. These findings suggest that Paucibacter sp. strain CH mighe degrade MC through a different pathway from that of strain ACM-3962.

Fates of Microcystis aeruginosa cells and associated microcystins in sediment and the effect of coagulation process on them.

During toxic Microcystis aeruginosa blooms, large amounts of cells can enter sediment through natural settlement, and coagulation treatment used to control water blooms can enhance the accumulation of cells. However, the current understanding of the fates of these cells and associated microcystins (MCs), as well as the effect of coagulation treatment on these factors, is limited. The results of the present study show that Microcystis aeruginosa cells in sediment were steadily decomposed under experimental conditions, and that they completely disappeared within 28 days. The major MCs released from settled cells were immediately degraded in sediment, and microbial degradation may be the main mechanism involved in this process. Coagulation treatment with PAC (polyaluminium chloride) + sepiolite can efficiently remove Microcystis aeruginosa cells from the water column and prevent their re-invasion. Furthermore, coagulation treatment with PAC + sepiolite had no significant effect on the release and decomposition of MCs and, thus, will not enhance the MCs pollution. However, coagulation treatment can accelerate the nutrient cycle by enhancing the settlement of cells. More attention should be paid to the effect on nutrient cycle when coagulation treatment is used for restoration of aquatic ecosystems.