Phagotrophic protists preserve antibiotic-resistant opportunistic human pathogens in the vegetable phyllosphere.

Food safety of leafy greens is an emerging public health issue as they can harbor opportunistic human pathogens (OHPs) and expose OHPs to consumers. Protists are an integral part of phyllosphere microbial ecosystems. However, our understanding of protist-pathogen associations in the phyllosphere and their consequences on public health remains poor. Here, we examined phyllosphere protists, human pathogen marker genes (HPMGs), and protist endosymbionts from four species of leafy greens from major supermarkets in Xiamen, China. Our results showed that Staphylococcus aureus and Klebsiella pneumoniae were the dominant human pathogens in the vegetable phyllosphere. The distribution of HPMGs and protistan communities differed between vegetable species, of which Chinese chive possessed the most diverse protists and highest abundance of HPMGs. HPMGs abundance positively correlated with the diversity and relative abundance of phagotrophic protists. Whole genome sequencing further uncovered that most isolated phyllosphere protists harbored multiple OHPs which carried antibiotic resistance genes, virulence factors, and metal resistance genes and had the potential to HGT. Colpoda were identified as key phagotrophic protists which positively linked to OHPs and carried diverse resistance and virulence potential endosymbiont OHPs including Pseudomonas nitroreducens, Achromobacter xylosoxidans, and Stenotrophomonas maltophilia. We highlight that phyllosphere protists contribute to the transmission of resistant OHPs through internalization and thus pose risks to the food safety of leafy greens and human health. Our study provides insights into the protist-OHP interactions in the phyllosphere, which will help in food safety surveillance and human health.

The Effects of Heavy Metal Pollution on Collembola in Urban Soils and Associated Recovery Using Biochar Remediation: A Review.

Heavy metal pollution in urban soil continues to be a global issue that poses a serious hazard to invertebrates and human lives through oral ingestion and inhalation of soil particles. Though the toxicity of several heavy metals on invertebrates like Collembola has been studied, lead (Pb) and cadmium (Cd) have been extensively studied due to their high toxicity to collembolans. As a ubiquitous soil organism all over the world, collembolans have been used as a model species to study the effects of heavy metals on invertebrate communities. To reduce the effects of heavy metals on ecosystem functions, biotic and abiotic measures have been used for heavy metal remediation; biochar seems to be the most effective approach that not only increases the physical absorption of heavy metals but also indirectly benefits soil organisms. In this study, we briefly reviewed the application of biochar in Pb and Cd polluted soil and showed its potential in soil remediation. Furthermore, we outlined the potentially toxic effects of Pb- and Cd-polluted urban soil on the collembolan species. We searched peer-reviewed publications that investigated: (1) the level of Pb and Cd contamination on urban soil in different cities around the world; and (2) the different sources of Pb and Cd as well as factors influencing their toxicity to collembolan communities. The obtained information offers new perspectives on the interactions and effects between collembolans, Pb, and Cd, and their remediation in urban soils.

Spatial assortment of soil organisms supports the size-plasticity hypothesis.

The size-plasticity hypothesis posits that larger size organisms are less plastic in their metabolic rates and, therefore, are more strongly environmental-filtered than smaller organisms. Many studies have supported this hypothesis by evaluating the relative roles of environmental filtration and dispersal for different taxonomic groups of soil organisms. Most observations are made at large spatial scales, which are assumed to have a wide array of varying habitats. However, since urbanization causes habitat fragmentation at smaller regional scales, testing the size-plasticity hypothesis at this scale would help better understand the spatial assortment of urban soil organisms which, in turn, would help to develop improved management and conservation strategies for urban soil health. Here, we used DNA metabarcoding on five groups of soil biota (bacteria, fungi, protists, nematodes, and invertebrates) to assess the relative importance of dispersal and environmental filters to examine the size-plasticity hypothesis at this spatial scale in an urban environment. We observed strong distance-decay of community similarities associated with higher levels of stochastic changes in bacteria, nematode, and protist communities but not fungal or invertebrate communities. Bacterial communities occupied the widest niche followed by protists and nematodes, potentially because of their higher dispersal abilities compared to the larger soil organisms. Null deviation of communities varied with taxonomic groups where bacteria and nematodes were mainly driven by homogenizing dispersal, protists and fungi by drift, and soil invertebrates by environmental selection. We further identified a small percentage of locally-adapted taxa (2.1%) that could be focal taxa for conservation and restoration efforts by, for example, restoring their habitats and enhancing their regional connectivity. These results support the size-plasticity hypothesis at the relatively unexplored regional scale in an urbanization context, and provide new information for improving urban soil health and sustainable city models.

Exploring microbial diversity and ecological function of epiphytic and surface sediment biofilm communities in a shallow tropical lake.

Microbial communities in epiphytic biofilms and surface sediments play a vital role in the biogeochemical cycles of the major chemical elements in freshwater. However, little is known about the diversity, composition, and ecological functions of microbial communities in shallow tropical lakes dominated by aquatic macrophytes. In this study, epiphytic bacterial and eukaryotic biofilm communities on submerged and floating macrophytes and surface sediments were investigated in Lake Rumira, Rwanda in August and November 2019. High-throughput sequencing data revealed that members of the phyla, including Firmicutes, Proteobacteria, Cyanobacteria, Actinobacteria, Chloroflexi, Bacteriodetes, Verrumicrobia, and Myxomycota, dominated bacterial communities, while the microeukaryotic communities were dominated by Unclassified (uncl) SAR(Stramenopiles, Alveolata, Rhizaria), Rotifers, Ascomycota, Gastrotricha, Platyhelminthes, Chloroplastida, and Arthropoda. Interestingly, the eukaryotic OTUs (operational taxonomic units) number and Shannon indices were significantly higher in sediments and epiphytic biofilms on Eicchornia crassipes than Ceratophyllum demersum (p < 0.05), while no differences were observed in bacterial OTUs number and Shannon values among substrates. Redundancy analysis (RDA) showed that water temperature, pH, dissolved oxygen (DO), total nitrogen (TN), and electrical conductivity (EC) were the most important abiotic factors closely related to the microbial community on C. demersum and E. crassipes. Furthermore, co-occurrence networks analysis (|r| > 0.7, p < 0.05) and functional prediction revealed more complex interactions among microbes on C. demersum than on E. crassipes and sediments, and those interactions include cross-feeding, parasitism, symbiosis, and predatism among organisms in biofilms. These results suggested that substrate-type and environmental factors were the strong driving forces of microbial diversity in epiphytic biofilms and surface sediments, thus shedding new insights into microbial community diversity in epiphytic biofilms and surface sediments and its ecological role in tropical lacustrine ecosystems.

Urbanization reduces resource use efficiency of phytoplankton community by altering the environment and decreasing biodiversity.

Urbanization often exerts multiple effects on aquatic and terrestrial organisms, including changes in biodiversity, species composition and ecosystem functions. However, the impacts of urbanization on river phytoplankton in subtropical urbanizing watersheds remain largely unknown. Here, we explored the effects of urbanization on phytoplankton community structure (i.e., biomass, community composition and diversity) and function (i.e., resource use efficiency) in a subtropical river at watershed scale in southeast China over 6 years. A total of 318 phytoplankton species belonging into 120 genera and 7 phyla were identified from 108 samples. Bacillariophyta biomass showed an increasing trend with increasing urbanization level. The phytoplankton community shifted from Chlorophyta dominance in rural upstream waters to Bacillariophyta dominance in urbanized downstream waters. Furthermore, phytoplankton diversity and resource use efficiency (RUE = phytoplankton biomass/total phosphorus) were significantly decreased with increasing urbanization level from upstream to downstream. Phytoplankton RUE exhibited a significant positive correlation with species richness, but a negative correlation with phytoplankton evenness. The variation in environmental factors (turbidity, total nitrogen, NH4+-N, total phosphorus, PO43--P and percentage urbanized area) was significantly correlated with phytoplankton diversity and RUE. Overall, our results revealed the influence of urbanization on phytoplankton community structure and ecosystem function was due to its altering the environmental conditions. Therefore, human-driven urbanization may play crucial roles in shaping the structure and function of phytoplankton communities in subtropical rivers, and the mechanism of this process can provide important information for freshwater sustainable uses, watershed management and conservation.

Spatio-seasonal variation of water quality influenced by land use and land cover in Lake Muhazi.

Understanding the influence of land use/land cover (LULC) on water quality is pertinent to sustainable water management. This study aimed at assessing the spatio-seasonal variation of water quality in relation to land use types in Lake Muhazi, Rwanda. The National Sanitation Foundation Water Quality Index (NSF-WQI) was used to evaluate the anthropogenically-induced water quality changes. In addition to Principal Components Analysis (PCA), a Cluster Analysis (CA) was applied on 12-clustered sampling sites and the obtained NSF-WQI. Lastly, the Partial Least Squares Path Modelling (PLS-PM) was used to estimate the nexus between LULC, water quality parameters, and the obtained NSF-WQI. The results revealed a poor water quality status at the Mugorore and Butimba sites in the rainy season, then at Mugorore and Bwimiyange sites in the dry season. Furthermore, PCA displayed a sample dispersion based on seasonality while NSF-WQI's CA hierarchy grouped the samples corresponding to LULC types. Finally, the PLS-PM returned a strong positive correlation (+ 0.831) between LULCs and water quality parameters in the rainy season but a negative correlation coefficient (- 0.542) in the dry season, with great influences of cropland on the water quality parameters. Overall, this study concludes that the lake is seasonally influenced by anthropogenic activities, suggesting sustainable land-use management decisions, such as the establishment and safeguarding protection belts in the lake vicinity.

On the use of chemotaxonomy, a phytoplankton identification and quantification method based on pigment for quick surveys of subtropical reservoirs.

Traditionally, composition and biomass of phytoplankton communities are determined by microscopy, but this method is time-consuming and so does not allow for high-frequency data acquisition across space and time. Pigment-based chemotaxonomy (CHEMTAX) is now widely applied to study of phytoplankton community structure on broader spatial and temporal scales of oceans, but the ability of this approach to provide estimates of phytoplankton assemblage in freshwater ecosystems is yet underdeveloped. To investigate the efficiency of the high-performance liquid chromatography (HPLC)-CHEMTAX in quantifying the different phytoplankton groups in inland freshwater, we present a comparison between phytoplankton pigment analyses by HPLC with CHEMTAX and microscopic counting of phytoplankton samples from four subtropical reservoirs in January and July 2014, respectively. The correlation between pigment and phytoplankton abundance detected by microscopy was stronger than that between pigment and phytoplankton biomass. The published marker pigments and their revised ratios can be used to describe phytoplankton abundances in a mixed community of freshwater phytoplankton, and pigment-based CHEMTAX can successfully describe the overall pattern of phytoplankton community dynamics during different seasons. The use of pigment-based CHEMTAX for quick surveys of phytoplankton communities can be recommended as a useful supplement or alternative tool to microscopy for freshwater ecosystem management.

Urbanization drives riverine bacterial antibiotic resistome more than taxonomic community at watershed scale.

Although the occurrence and distribution of antibiotic resistance genes (ARGs) in various aquatic ecosystems are well explored, understanding of the ecological processes and mechanisms governing the composition and dynamics of bacterial ARGs still remains limited across space and time. Here, we used high-throughput approaches to detect spatial patterns of bacterial ARGs and operational taxonomic units (OTUs) in an urbanizing subtropical watershed, Xiamen, southeast China over a five-year period. At watershed scale, the OTU profiles were undergoing a directional change, but the ARG profiles showed a high stability or stochastic change over time. Compared with the upstream and midstream, the richness, absolute abundance, normalized abundance and diversity of ARGs were significantly higher in the downstream waters. Our results revealed a clear rural-urban disparity in ARG and OTU profiles which were mainly governed by deterministic and stochastic assembly processes, respectively. With the increase of urban building area along the river, the ecological processes of ARG profiles shifted from stochastic to deterministic. In downstream waters, the bacterial ARG profiles were much more stable than bacterial OTUs. Further, our results indicated that both human-dominated environment (e.g., land use) and mobile genetic elements (MGEs) played an important role in shaping the ARG profiles and dynamics. Overall, this was a response to spatially extensive human-landscape interactions that included urban development in the river downstream region, which were common across subtropical coastal cities of China and can alter the ARG profile dynamics along rural-urban gradient. Therefore, watershed management actions aiming at reducing threats posed by ARGs in urbanizing watershed should first consider the surrounding urbanization level and the mode and intensity of human activity. Our findings also imply that due to the decoupling of bacterial function and taxonomy, both aspects should be studied separately.

Correction to: Stochastic processes shape microeukaryotic community assembly in a subtropical river across wet and dry seasons.

Following publication of the original article [1], the authors reported an error in Figure 1 and a text on page 13. In Figure 1, some areas are missing and the correct figure is presented here. On page 13, the sentence should be updated to the following (change has been indicated in bold italics).

Stochastic processes shape microeukaryotic community assembly in a subtropical river across wet and dry seasons.

BACKGROUND: The deep mechanisms (deterministic and/or stochastic processes) underlying community assembly are a central challenge in microbial ecology. However, the relative importance of these processes in shaping riverine microeukaryotic biogeography is still poorly understood. Here, we compared the spatiotemporal and biogeographical patterns of microeukaryotic community using high-throughput sequencing of 18S rRNA gene and multivariate statistical analyses from a subtropical river during wet and dry seasons. RESULTS: Our results provide the first description of biogeographical patterns of microeukaryotic communities in the Tingjiang River, the largest river in the west of Fujian province, southeastern China. The results showed that microeukaryotes from both wet and dry seasons exhibited contrasting community compositions, which might be owing to planktonic microeukaryotes having seasonal succession patterns. Further, all components of the microeukaryotic communities (including total, dominant, always rare, and conditionally rare taxa) exhibited a significant distance-decay pattern in both seasons, and these communities had a stronger distance-decay relationship during the dry season, especially for the conditionally rare taxa. Although several variables had a significant influence on the microeukaryotic communities, the environmental and spatial factors showed minor roles in shaping the communities. Importantly, these microeukaryotic communities were strongly driven by stochastic processes, with 89.9%, 88.5%, and 89.6% of the community variation explained by neutral community model during wet, dry, and both seasons, respectively. The neutral community model also explained a large fraction of the community variation across different taxonomic groups and levels. Additionally, the microeukaryotic taxa, which were above and below the neutral prediction, were ecologically and taxonomically distinct groups, which might be interactively structured by deterministic and stochastic processes. CONCLUSIONS: This study demonstrated that stochastic processes are sufficient in shaping substantial variation in river microeukaryotic metacommunity across different hydrographic regimes, thereby providing a better understanding of spatiotemporal patterns, processes, and mechanisms of microeukaryotic community in waters.

An extensively shared antibiotic resistome among four seasons suggests management prioritization in a subtropical riverine ecosystem.

Although seasonality is a key driver of environmental fluctuation in aquatic ecosystems, there exists limited knowledge on the factors controlling the distribution of antibiotic resistance genes (ARGs) across seasons at a watershed scale. Here we used high-throughput quantitative PCR to quantify 285 ARGs conferring resistance to most major classes of antibiotics, reveal their spatial and seasonal distribution patterns, and depict the underlying mechanisms in a subtropical riverine ecosystem under low and high human pressures, in Xiamen city, southeast China. Our results showed that spatial differences in ARG richness and abundance overwhelmed their seasonal variations, with only ARGs that confer resistance to sulfonamide and vancomycin being significantly different across seasons. Only a few abundant ARGs (19 ARGs) could contribute to >70% of the total ARGs abundance and were found in all seasons. The significantly higher number of ARGs in the summer rainy period than other seasons coincided with high number of significant edges in ARG co-occurrence networks. Summer rainfall had strong dilution effect on ARGs in upstream waters and enrichment effect in downstream waters. The variance partitioning analysis indicated that the environment explained larger variance of ARG profiles than mobile genetic elements (MGEs), spatial predictors and the rainfall. Nevertheless, strong and significant correlations between transposase gene absolute abundance and aminoglycoside, chloramphenicol, MLS, multidrug and tetracycline classes of resistance genes inferred the role of MGEs on ARG distribution. Overall, our results imply that the modelling and management of ARGs in highly dynamic ecosystems could be better implemented by considering priority genes that dominate at spatial and seasonal gradients.

Spartina alterniflora invasion alters soil bacterial communities and enhances soil N2O emissions by stimulating soil denitrification in mangrove wetland.

Chinese mangrove, an important ecosystem in coastal wetlands, is sensitive to the invasive alien species Spartina alterniflora. However, the effects of the S. alterniflora invasion on mangrove soil N2O emissions and the underlying mechanisms by which emissions are affected have not been well studied. In this study, the N2O emitted from soils dominated by two typical native mangroves (i.e. Kandelia obovata: KO; Avicennia marina: AM), one invaded by S. alterniflora (SA), and one bare mudflat (Mud) were monitored at Zhangjiang Mangrove Estuary (where S. alterniflora is exotic). Together with soil biogeochemical properties, the potential denitrification rate and the composition of soil bacterial communities were determined simultaneously by 15NO3- tracer and high-throughput sequencing techniques, respectively. Our results showed that S. alterniflora invasion significantly (p < 0.05) increases soil N2O emissions by 15-28-fold. In addition, isotope results revealed that the soil potential denitrification rate was significantly (p < 0.05) enhanced after S. alterniflora invasion. Moreover, the S. alterniflora invasion significantly (p < 0.05) decreased soil bacterial α-diversity and strongly modified soil bacterial communities. Indicator groups strongly associated with S. alterniflora were Chloroflexia, Alphaproteobacteria, and Bacilli, each of which was abundant and acts as connector in the co-occurrence network. FAPROTAX analysis implied that the S. alterniflora invasion stimulated soil denitrification and nitrification while depressing anaerobic ammonium oxidation (anammox) and dissimilatory nitrate reduction to ammonium (DNRA). Redundancy analysis (RDA) found that soil organic matter (SOM) and pH were the most important environmental factors in altering soil bacterial communities. Taken together, our results imply that the S. alterniflora invasion in mangrove wetlands significantly stimulates soil denitrification and N2O emissions, thereby contributing N2O to the atmosphere and contributing to global climate change.

Community assembly processes underlying phytoplankton and bacterioplankton across a hydrologic change in a human-impacted river.

Although the influence of microbial community assembly processes on aquatic ecosystem function and biodiversity is well known, the processes that govern planktonic communities in human-impacted rivers remain largely unstudied. Here, we used multivariate statistics and a null model approach to test the hypothesis that environmental conditions and obstructed dispersal opportunities, dictate a deterministic community assembly for phytoplankton and bacterioplankton across contrasting hydrographic conditions in a subtropical mid-sized river (Jiulong River, southeast China). Variation partitioning analysis showed that the explanatory power of local environmental variables was larger than that of the spatial variables for both plankton communities during the dry season. During the wet season, phytoplankton community variation was mainly explained by local environmental variables, whereas the variance in bacterioplankton was explained by both environmental and spatial predictors. The null model based on Raup-Crick coefficients for both planktonic groups suggested little evidences of the stochastic processes involving dispersal and random distribution. Our results showed that hydrological change and landscape structure act together to cause divergence in communities along the river channel, thereby dictating a deterministic assembly and that selection exceeds dispersal limitation during the dry season. Therefore, to protect the ecological integrity of human-impacted rivers, watershed managers should not only consider local environmental conditions but also dispersal routes to account for the effect of regional species pool on local communities.

Disturbance-induced phytoplankton regime shifts and recovery of cyanobacteria dominance in two subtropical reservoirs.

Many countries in the world still suffer from high toxic cyanobacterial blooms in inland waters used for human consumption. Regional climate change and human activities within watersheds exert a complex and diverse influence on aquatic ecosystem structure and function across space and time. However, the degree to which these factors may contribute to the long-term dynamics of plankton communities is still not well understood. Here, we explore the impacts of multiple disturbance events (e.g. human-resettlement, temperature change, rainfall, water level fluctuations), including six combined disturbances, on phytoplankton and cyanobacteria in two subtropical reservoirs over six years. Our data showed that combined environmental disturbances triggered two apparent and abrupt switches between cyanobacteria-dominated state and non-cyanobacterial taxa-dominated state. In late 2010, the combined effect of human-resettlement (emigration) and natural disturbances (e.g. cooling, rainfall, water level fluctuations) lead to a 60-90% decrease in cyanobacteria biomass accompanied by the disappearance of cyanobacterial blooms, in tandem with an abrupt and persistent shift in phytoplankton community. After summer 2014, however, combined weather and hydrological disturbances (e.g. warming, rainfall, water level fluctuations) occurred leading to an abrupt and marked increase of cyanobacteria biomass, associated with a return to cyanobacteria dominance. These changes in phytoplankton community were strongly related to the nutrient concentrations and water level fluctuations, as well as water temperature and rainfall. As both extreme weather events and human disturbances are predicted to become more frequent and severe during the twenty-first century, prudent sustainable management will require consideration of the background limnologic conditions and the frequency of disturbance events when assessing the potential impacts on reservoir biodiversity and ecosystem functioning and services.