Response of bacterial and micro-eukaryotic communities to spatio-temporal fluctuations of wastewater in full scale constructed wetlands.

How microbial communities respond to wastewater fluctuations is poorly understood. Full-scale surface flow constructed wetlands (SFCWs) were constructed for investigating microbial communities. Results showed that influent wastewater changed sediment bacterial community composition seasonally, indicating that a single bacterial taxonomic group had low resistance (especially, Actinobacteriota and Gammaproteobacteria). However, copy numbers of 16S rRNA, ammonia oxidizing archaea, ammonia oxidizing bacteria, nirS and nirK in the first stage SFCWs were 2.49 × 1010, 3.48 × 109, 5.76 × 106, 8.77 × 108 and 9.06 × 108 g-1 dry sediment, respectively, which remained stable between seasons. Moreover, decreases in the nitrogen concentration in wastewater, changed microbial system state from heterotrophic to autotrophic. Micro-eukaryotic communities were more sensitive to wastewater fluctuations than bacterial communities. Overall, results revealed that microbial communities responded to spatio-temporal fluctuations in wastewater through state changes and species asynchrony. This highlighted complex processes of wastewater treatment by microbial components in SFCWs.

Human-induced homogenization of microbial taxa and function in a subtropical river and its impacts on community stability.

Combination of taxa and function can provide a more comprehensive picture on human-induced microbial homogenization. Here, we obtained 2.58 billion high-throughput sequencing reads and 479 high-quality metagenome-assembled genomes (MAGs) of planktonic microbial communities in a subtropical river for 5 years. We found the microbial taxa homogenization and functional homogenization were uncoupled. Although human activities in downstream sites significantly decreased the taxonomic diversity of non-abundant ASV communities (16S rRNA gene amplicon sequence variants), they did not significantly decrease the taxonomic diversity of abundant ASV and total observed MAG communities. However, the total observed MAG communities in downstream sites tended to homogenize into some specific taxa which encode human-activity-related functional genes, such as nutrient cycles, greenhouse gas emission, antibiotic and arsenic resistance. Those specific MAGs with high taxonomic diversity caused the weak heterogenization of total observed MAG communities in downstream sites. Moreover, functional homogenization promoted the synchrony among downstream MAGs, and these MAGs constructed some specific network modules might to synergistically execute or resist the human-activity-related functions. High synchrony also led to the tandem effects among MAGs and thus decreased community stability. Overall, our findings revealed the links of microbial taxa, functions and stability under human activity impacts, and provided a strong evidence to encourage us re-thinking biotic homogenization based on microbial taxa and their functional attributes.

Biogeographic patterns of micro-eukaryotic generalists and specialists and their effects on regional α-diversity at inter-oceanic scale.

Inter-oceanic scale studies allow us to understand the global spread of micro-organisms in marine ecosystems. In this study, micro-eukaryotic communities in marine surface sediment were collected from tropical to Arctic sites. We found that micro-eukaryotic generalists had much higher intraspecific variation than specialists which allow them to distribute more widely through higher spatiotemporal asynchrony and complementary niche preferences among conspecific taxa. Moreover, comparing to the host-associated protozoa and small metazoa, the algae and free-living protozoa with higher intraspecific variation allow them to have wider distribution ranges. Species abundance also played an important role in driving the distribution ranges of generalists and specialists. The generalists had important effects on regional α-diversity even at an inter-oceanic scale which led to the micro-eukaryotic species richness in polar sites to be mainly influenced by the regional generalists but not the local specialists. In particular, more than 97% of algal species in polar sites were shared with the tropical and subtropical sites (including toxic dinoflagellate). Overall, our study suggests that the effects of global change and human activities on the vulnerable high latitude habitats may lead to biotic homogenization for the whole microbial community (not only the dispersal of some harmful algae) through the potential long-distance spread of generalists.

Response of microbial communities to nutrient removal in coastal sediment by using ecological concrete.

Ecological concrete (eco-concrete) is a kind of environment-friendly material with porous characteristics. In this study, the eco-concrete was used to remove the total nitrogen (TN), total phosphorus (TP), and total organic carbon (TOC) in marine coastal sediment. The bacterial communities in sediment and on eco-concrete surface were also investigated by using high-throughput sequencing and quantitative PCR of 16S rRNA gene. We found that the mean removal efficiencies of TN, TP, and TOC in treatment group were 8.3%, 8.4%, and 12.3% after 28 days. The bacterial community composition in the treatment group was significantly different from that in the control group on day 28. In addition, the bacterial community composition on eco-concrete surface was slightly different from that in sediment, and the copy numbers of 16S rRNA gene were higher on eco-concrete surface than in sediment. The types of eco-concrete aggregates (gravel, pebble, and zeolite) also had effects on the bacterial community composition and 16S rRNA gene copy numbers. Furthermore, we found the abundant genus Sulfurovum increased significantly on eco-concrete surface in the treatment group after 28 days. Bacteria belonging to this genus were found having denitrification ability and were commonly detected in bioreactors for nitrate removal. Overall, our study expands the application scopes of eco-concrete and suggests that the bacterial communities in eco-concrete can potentially enhance the removal efficiency of nutrients in coastal sediment.

Microbial one­carbon and nitrogen metabolisms are beneficial to the reservoir recovery after cyanobacterial bloom.

Cyanobacterial blooms have profound effects on the structure and function of plankton communities in inland waters, but few studies have focused on the effects of microbial-based processes in one­carbon and nitrogen cycling on water quality improvement following the bloom. Here, we compared the structure and function of the bacterial community, focusing on microbial one­carbon and nitrogen metabolisms during and after a cyanobacterial Microcystis bloom in a deep subtropical reservoir. Our data showed that microbial one­carbon and nitrogen cycles were closely related to different periods of the bloom, and the changes of functional genes in microbial carbon and nitrogen cycling showed the same consistent trend as that of Methylomonas sp. With the receding of the bloom, the abundance of Methylomonas as well as the functional genes of microbial one­carbon and nitrogen cycling reached the peak and then recovered. Our results indicate that microbial one­carbon and nitrogen metabolisms were beneficial to the recovery of water quality from the cyanobacterial bloom. This study lays a foundation for a deep understanding of the cyanobacterial decomposition mediated by microbes in one­carbon and nitrogen cycles in inland freshwaters.

A Novel Algicidal Bacterium and Its Effects against the Toxic Dinoflagellate Karenia mikimotoi (Dinophyceae).

The toxic dinoflagellate Karenia mikimotoi is a harmful algal bloom-forming species in coastal areas around the world. It produces ichthyotoxins and hemolytic toxins, with deleterious effects on marine ecosystems. In this study, the bacterium Pseudoalteromonas sp. FDHY-MZ2, with high algicidal efficiency against K. mikimotoi, was isolated from a bloom event. Based on the results, it completely lysed K. mikimotoi cells within 24 h 0.5% (vol/vol), with the algicidal activity of the supernatant of the bacterium culture. Algal cell wall fragmentation occurred, leading to cell death. There was a marked decline in various photochemical traits. When treated with the supernatant, cellulase, pheophorbide a oxygenase (PAO) and cyclin B genes were significantly increased, suggesting induced cell wall deterioration, chloroplast degradation and cell cycle regulation of K. mikimotoi cells. In addition, the expression levels of reactive oxygen species (ROS) scavenging gene was significantly inhibited, indicating that the ROS removal system was damaged. The bacterial culture was dried to obtain the spray-dried powder, which showed algicidal activity rates of 92.2 and 100% against a laboratory K. mikimotoi culture and a field microcosm of Karlodinium sp. bloom within 24 h with the addition of 0.04% mass fraction powder. Our results demonstrate that FDHY-MZ2 is a suitable strain for K. mikimotoi and Karlodinium sp. blooms management. In addition, this study provides a new strategy for the anthropogenic control of harmful algal bloom-forming species in situ. IMPORTANCEK. mikimotoi is a noxious algal bloom-forming species that cause damaging of the aquaculture industry and great financial losses. Bacterium with algicidal activity is an ideal agency to inhibit the growth of harmful algae. In this approach application, the bacterium with high algicidal activity is required and the final management material is ideal for easy-to-use. The algicidal characteristics are also needed to understand the effects of the bacterium for managing strategy exploration. In this study, we isolated a novel algicidal bacterium with extremely high lysis efficiency for K. mikimotoi. The algicidal characteristics of the bacterium as well as the chemical and molecular response of K. mikimotoi with the strain challenge were examined. Finally, the algicidal powder was explored for application. The results demonstrate that FDHY-MZ2 is suitable for K. mikimotoi and Karlodinium sp. blooms controlling, and this study provides a new strategy for algicidal bacterium application.

Nutrient Removal in Eutrophic Water Promotes Stability of Planktonic Bacterial and Protist Communities.

Nutrient (nitrogen and phosphorus) removal by using bioremediation technologies in eutrophic water alters bacterial and protist community structure and function, but how it changes the stability of community remains unclear. To fill this gap, in this study, bacterial and protist communities were investigated using 16S and 18S rRNA gene high-throughput sequencing during the nutrient removal by using ecological floating beds of Canna indica L. Our results showed that both bacterial and protist community compositions in the treatment group were similar to those in the control group at the beginning of the experiment (day 1 to day 11), but then bacterial and protist community compositions became more stable with the removal of nutrients in the treatment group than those in the control group (day 12 to day 18). We further explored the mechanisms for this increased stability and found that the contribution of the stochastic process to bacterial and protist community variations was higher in the control group than that in the treatment group. This suggests that the high nutrient concentration in the control group might increase the random colonization or extinction, and therefore resulted in the high temporal variability (i.e., unstable) of bacterial and protist communities. Our findings suggest that bioremediation for eutrophication can promote the stability of aquatic communities, and therefore potentially maintain aquatic ecosystem functions and services to humanity.

Anthropogenic activities change the relationship between microbial community taxonomic composition and functional attributes.

Functional redundancy is considered common in microbial systems, but recent studies have challenged this idea. The mechanism for this contradictory result is not clear. However, in this study, we hypothesize that strong environmental filtering which links to the anthropogenic activities is able to weaken microbial functional redundancy. We used metagenome and 16S rRNA gene high-throughput sequencing to investigate planktonic microbial communities in a subtropical river. We found that the weak anthropogenic activities might result in a low selection pressure in the river upstream area. Therefore, the microbial community functional attributes were stable although the community composition changed with the water temperature and NO3 -N in upstream area (this indicates functional redundancy). However, the strong anthropogenic activities in river downstream area selected pollutant-degraded functions (e.g. nitrogen metabolism, toluene, xylenes and ethylbenzene degradation) and potentially pollutant-degraded (tolerant) microbes, and therefore caused the microbial community composition synchronously changed with the variation of community functional attributes. Our results reveal that strong environmental filtering which associates with the anthropogenic activities not only has effects on microbial community composition and community functional attributes but also on their relationships. These results provide a new insight to refine the functional redundancy idea.

Transformations from specialists to generalists cause bacterial communities are more stable than micro-eukaryotic communities under anthropogenic activity disturbance.

Different microbial components have different responses to environmental disturbances. Here, we found that the planktonic bacterial and micro-eukaryotic communities had different responses to anthropogenic activity disturbance in a subtropical river, because they had different survival strategies (generalist and specialist). We used nutrients (nitrogen and phosphorus) as indicators of anthropogenic activities. We found that river stretch 1 showed low nutrient concentrations from October 2018 to September 2019. However, a nutrient disturbance was observed in river stretch 2. The nutrient concentrations increased largely in December and January but recovered to low values in June. Bacterial communities had higher resilience under this disturbance than micro-eukaryotic communities in river stretch 2. The bacterial community composition were quite different between the two river stretches in December and January but were similar in June and July. However, the differences of micro-eukaryotic community composition between the two river stretches were always high during the study period. The bacterial communities in river stretch 2 contained more generalists and nutrient tolerant specialists. The bacterial nutrient tolerant specialists rapidly decreased in the low nutrient months and were replaced by the generalists. Bacteria which were involved in this shifts accounted for 29.3% of the total abundance. However, the micro-eukaryotic communities in river stretch 2 contained more moderate generalists. These moderate generalists were insensitive to the variation of nutrients and only 19.56% of the micro-eukaryotes had significant responses to the disturbance. The survival strategies caused bacterial communities had higher adaptability than eukaryotes to environmental fluctuation.

Transcriptome responses of the dinoflagellate Karenia mikimotoi driven by nitrogen deficiency.

The availability of ambient N nutrient is often correlated with the occurrences of harmful algal bloom formed by certain dinoflagellates, making it important to understand how these species might be responding to such conditions. Here, transcriptome sequencing of Karenia mikimotoi was conducted to understand the underlying molecular mechanisms by which this dinoflagellate copes with nitrogen (N) deficiency. Transcriptomic analysis revealed 8802 unigenes (3.56%) that were differentially expressed with ≥ 2-fold change. Under N-depleted conditions, genes involved in glycolysis, fatty acid metabolism, and the tricarboxylic acid (TCA) cycle as well as lipid accumulation were significantly upregulated. The elevated expression of enzymes used in protein degradation and turnover suggests possible metabolic reconfiguration towards accelerated N recycling. Moreover, a significant increase in urea transporter was observed, indicating increased assimilation of organic nitrogen resources as an alternative in N-depleted cultures of K. mikimotoi. The down-regulated glutamate synthase genes were also identified under N deficiency, suggesting suppression of primary amino acid synthesis to save N resource. Taken together, results of this study show enhanced multiple N resource acquisition and reuse of multiple N resources constitute a comprehensive strategy to cope with N deficiency in a dinoflagellate.