The fungicide azoxystrobin promotes freshwater cyanobacterial dominance through altering competition.

BACKGROUND: Sharp increases in food production worldwide are attributable to agricultural intensification aided by heavy use of agrochemicals. This massive use of pesticides and fertilizers in combination with global climate change has led to collateral damage in freshwater systems, notably an increase in the frequency of harmful cyanobacterial blooms (HCBs). The precise mechanisms and magnitude of effects that pesticides exert on HCBs formation and proliferation have received little research attention and are poorly constrained. RESULTS: We found that azoxystrobin (AZ), a common strobilurin fungicide, can favor cyanobacterial growth through growth inhibition of eukaryotic competitors (Chlorophyta) and possibly by inhibiting cyanobacterial parasites (fungi) as well as pathogenic bacteria and viruses. Meta-transcriptomic analyses identified AZ-responsive genes and biochemical pathways in eukaryotic plankton and bacteria, potentially explaining the microbial effects of AZ. CONCLUSIONS: Our study provides novel mechanistic insights into the intertwined effects of a fungicide and eutrophication on microbial planktonic communities and cyanobacterial blooms in a eutrophic freshwater ecosystem. This knowledge may prove useful in mitigating cyanobacteria blooms resulting from agricultural intensification.

Effects of imazethapyr spraying on plant growth and leaf surface microbial communities in Arabidopsis thaliana.

Imazethapyr (IM) is an acetolactate synthase (ALS)-inhibiting herbicide that has been widely used in recent years. However, IM spraying can lead to the accumulation of herbicide residues in leaves. Here, we determined the effects of IM spraying on the plant growth and leaf surface microbial communities of Arabidopsis thaliana after 7 and 14 days of exposure. The results suggested that IM spraying inhibited plant growth. Fresh weight decreased to 48% and 26% of the control value after 7 and 14 days, respectively, of 0.035 kg/ha IM exposure. In addition, anthocyanin content increased 9.2-fold and 37.2-fold relative to the control content after 7 and 14 days of treatment, respectively. Furthermore, IM spraying destroyed the cell structures of the leaves, as evidenced by increases in the number of starch granules and the stomatal closure rate. Reductions in photosynthetic efficiency and antioxidant enzyme activity were observed after IM spraying, especially after 14 days of exposure. The diversity and evenness of the leaf microbiota were not affected by IM treatment, but the composition of community structure at the genus level was altered by IM spraying. Imazethapyr application increased the abundance of Pseudomonas, a genus that includes species pathogenic to plants and humans, indicating that IM potentially increased the abundance of pathogenic bacteria on leaves. Our findings increase our understanding of the relationships between herbicide application and the microbial community structures on plant leaves, and they provide a new perspective for studying the ecological safety of herbicide usage.

Evaluation of the taxonomic and functional variation of freshwater plankton communities induced by trace amounts of the antibiotic ciprofloxacin.

Ciprofloxacin (CIP), one of the most frequently detected antibiotics in water systems, has become an aquatic contaminant because of improper disposal and excretion by humans and animals. It is still unknown how trace amounts of CIP affect the aquatic microbial community diversity and function. We therefore investigated the effects of CIP on the structure and function of freshwater microbial communities via 16S/18S rRNA gene sequencing and metatranscriptomic analyses. CIP treatment (7 µg/L) did not significantly alter the physical and chemical condition of the water body as well as the composition of the main species in the community, but slightly increased the relative abundance of cyanobacteria and decreased the relative abundance of eukaryotes. Metatranscriptomic results showed that bacteria enhanced their phosphorus transport and photosynthesis after CIP exposure. The replication, transcription, translation and cell proliferation were all suppressed in eukaryotes, while the bacteria were not affected in any of these aspects. This interesting phenomenon was the exact opposite to both the antibacterial property of CIP and its safety for eukaryotes. We hypothesize that reciprocal and antagonistic interactions in the microcosm both contribute to this result: cyanobacteria may enhance their tolerance to CIP through benefiting from cross-feeding and some secreted substances that withstand bacterial CIP stress would also affect eukaryotic growth. The present study thus indicates that a detailed assessment of the aquatic ecotoxicity of CIP is essential, as the effects of CIP are much more complicated in microbial communities than in monocultures. CIP will continue to be an environmental contaminant due to its wide usage and production and more attention should be given to the negative effects of antibiotics as well as other bioactive pollutants on aquatic environments.

Aquatic ecotoxicity of an antidepressant, sertraline hydrochloride, on microbial communities.

Sertraline hydrochloride (Ser-HCl), a widely used antidepressant, becomes an aquatic contaminant via metabolic excretion and improper disposal; however, it is unknown how Ser-HCl affects aquatic microbial communities. The present study investigated the effects of Ser on the structures of aquatic microbial communities via high-throughput sequencing analyses. Ser-HCl treatment inhibited the growth of two model algae (the green alga, Chlorella vulgaris, and the cyanobacterium, Microcystis aeruginosa) and decreased the chlorophyll a (Chl-a) concentration in the microcosm to reduce the photosynthetic efficiency. High-throughput sequencing analyses showed that exposure to Ser-HCl disturbed the balance of cyanobacteria species by stimulating the growth of specific cyanobacteria. Among eukaryotes, the richness as well as the diversity indices were significantly enhanced after 5 days of Ser-HCl treatment but sharply decreased with exposure time. Nucleariida occupied an absolute majority (97.83%) within the eukaryotes, implicating that Ser-HCl disturbed the ecological equilibrium in microcosms. Ser-HCl will continue to be an environmental contaminant due to its wide usage and production. Our current study clarified the potential ecological risk of Ser-HCl to aquatic microorganisms. These findings suggest that more attention should be given to the negative effects of these bioactive pollutants on aquatic environments.

The combined toxicity effect of nanoplastics and glyphosate on Microcystis aeruginosa growth.

Waste plastics can be degraded to nanoplastics (NPs, diameter<1 µm) by natural forces. NPs not only directly affect aquatic organisms but also adsorb other pollutants, causing combined pollution. Glyphosate is one of the most widely used herbicides and is commonly monitored in freshwater systems. In this study, the effects of the combined toxicity of polystyrene cationic amino-modified nanoparticles (nPS-NH2) and glyphosate on a blue-green alga, Microcystis aeruginosa, were investigated. Our results demonstrated that 5 mg/L glyphosate had a strong inhibitory effect on M. aeruginosa (the 96-h inhibitory rate was 27%), while 5 mg/L nPS-NH2 had no apparent effect on the growth of M. aeruginosa. Interestingly, nPS-NH2 combined with glyphosate showed antagonistic effects on the inhibition of algal growth because nPS-NH2 displayed a strong adsorption capacity for glyphosate, which significantly alleviated the inhibitory effect of glyphosate on M. aeruginosa growth. However, the presence of glyphosate enhanced the stability of the dispersion system, which allowed more nPS-NH2 to adsorb on the surface of M. aeruginosa and may result in greater enrichment of nPS-NH2 in the food chain to show potential repercussions to human life. Our current study provides a new theoretical basis for the combined effects of NPs and pesticide pollution.

Interacting effect of diclofop-methyl on the rice rhizosphere microbiome and denitrification.

A better knowledge of the intertwined effects of herbicides on plant physiology and microbiome as well as nutrient biogeochemical cycles are needed for environmental management. Here we studied the influence of herbicide diclofop-methyl (DM) on the rice root microbiome and its relationship with N cycle. To do so, we exposed rice seedlings to 100 µg/L DM and studied rhizosphere microbiota using MiSeq-pyrosequencing, root exudation by GC-MS, and denitrification activity by 15N isotope-tracing and qRT-PCR. The richness and diversity of rhizosphere microorganisms, significantly increased after DM exposure combined with an increase in root exudation of amino acids, sugars, and fatty acids. Transcription of denitrification-related gene and denitrification rate increased significantly in the rice rhizosphere. Our results suggest that DM strongly influenced the root exudation of bacteria nutrients, which affected root microbiome community and potentially influenced N cycle in rice rhizosphere.

Changes in bacterial community structure and antibiotic resistance genes in soil in the vicinity of a pharmaceutical factory.

China is the largest global producer of antibiotics. With the demand for antibiotics increasing every year, it is necessary to assess potential environmental risks and the spread of antibiotic resistance genes (ARGs) associated with antibiotic production. Here, we investigated the occurrence and distribution of ARGs in soil in the vicinity of a pharmaceutical factory. The results showed that antibiotic concentrations were under the detection limit; however, ARGs were present in soil and tended to be enriched near the factory. A significant correlation between the relative abundance of intI-1 and tetracycline ARGs implied that horizontal gene transfer might play an important role in the spread of ARGs. The occurrence of these ARGs could be the results of previous antibiotic contamination. However, the soil bacterial community structure seemed to be more affected by nutrients or other factors than by antibiotics. Overall, this study supports the viewpoint that long-term pharmaceutical activity might have a negative effect on environmental health, thus, underscoring the need to regulate antibiotic production and management.

Investigation of Rhizospheric Microbial Communities in Wheat, Barley, and Two Rice Varieties at the Seedling Stage.

The plant rhizosphere microbiota plays multiple roles in plant growth. We investigated the taxonomic and functional variations in the rhizosphere microbial community, examining both prokaryotes and eukaryotes, of four crops at the seedling stage: wheat, barley, and two rice varieties ( indica and japonica) seeded in paddy soil. The diversity of rhizosphere communities in these four species was determined. Results showed that wheat and barley had much stronger selection effects than rice for the rhizosphere microbial community. Functional metagenomic profiling indicated that a series of sequences related to glycan, limonene, and pinene degradation pathways as well as some relatively rare functions related to N or S metabolism were enriched in the rhizosphere soil. We conclude that the four tested crops induced the formation of the microbial community with specific features that may influence the plant growth but stochastic processes also appreciably influenced the functional selection.

Reproductive and endocrine-disrupting toxicity of Microcystis aeruginosa in female zebrafish.

Microcystis aeruginosa, a primary species in cyanobacterial blooms, is ubiquitously distributed in water. Microcystins (MCs) purified from M. aeruginosa can exert reproductive toxicity in fish. However, the effects of M. aeruginosa at environmentally relevant levels on the reproductive and endocrine systems of zebrafish are still unknown. The present study investigated the reproductive and endocrine-disrupting toxicity of M. aeruginosa on female zebrafish (Danio rerio) by short-term exposure (96 h). After exposure, marked histological lesions in the liver or gonads, such as nuclear pyknosis and deformation, were observed, and the fertilization rate and hatchability of eggs spawned from treated females were both significantly lower than they were in females in the control group, suggesting the possibility of transgenerational effects of M. aeruginosa exposure. Moreover, M. aeruginosa exposure decreased the concentration of 17ß-estradiol (E2) and testosterone (T) in female zebrafish. Interestingly, the vtg1 transcriptional level significantly decreased in the liver, whereas plasma vitellogenin (VTG) protein levels increased. The present findings indicate that M. aeruginosa could modulate endocrine function by disrupting transcription of hypothalamic-pituitary-gonadal-liver (HPGL) axis-related genes, and impair the reproductive capacity of female zebrafish, suggesting that M. aeruginosa causes potential adverse effects on fish reproduction in Microcystis bloom-contaminated aquatic environments.

Evaluation of the toxic response induced by azoxystrobin in the non-target green alga Chlorella pyrenoidosa.

The top-selling strobilurin, azoxystrobin (AZ), is a broad-spectrum fungicide that protects against many kinds of pathogenic fungi by preventing their ATP production. The extensive use of AZ can have negative consequences on non-target species and its effects and toxic mechanisms on algae are still poorly understood. In this work, Chlorella pyrenoidosa that had been grown in BG-11 medium was exposed to AZ (0.5-10 mg L-1) for 10 d. The physiological and molecular responses of the algae to AZ treatment, including photosynthetic efficiency, lipid peroxidation level, antioxidant enzyme activities, as well as transcriptome-based analysis of gene expression, were examined to investigate the potential toxic mechanism. Results shows that the photosynthetic pigment (per cell) increased slightly after AZ treatments, indicating that the photosystem of C. pyrenoidosa may have been strengthened. Glutathione and ascorbate contents were increased, and antioxidant enzyme activities were induced to relieve oxidative damage (e.g., from lipid peroxidation) in algae after AZ treatment. Transcriptome-based analysis of gene expression combined with physiological verification suggested that the 5 mg L-1 AZ treatment did not inhibit ATP generation in C. pyrenoidosa, but did significantly alter amino acid metabolism, especially in aspartate- and glutamine-related reactions. Moreover, perturbation of ascorbate synthesis, fat acid metabolism, and RNA translation was also observed, suggesting that AZ inhibits algal cell growth through multiple pathways. The identification of AZ-responsive genes in the eukaryotic alga C. pyrenoidosa provides new insight into AZ stress responses in a non-target organism.

Multiwall carbon nanotubes modulate paraquat toxicity in Arabidopsis thaliana.

Carbon nanotubes can be either toxic or beneficial to plant growth and can also modulate toxicity of organic contaminants through surface sorption. The complex interacting toxic effects of carbon nanotubes and organic contaminants in plants have received little attention in the literature to date. In this study, the toxicity of multiwall carbon nanotubes (MWCNT, 50 mg/L) and paraquat (MV, 0.82 mg/L), separately or in combination, were evaluated at the physiological and the proteomic level in Arabidopsis thaliana for 7-14 days. The results revealed that the exposure to MWCNT had no inhibitory effect on the growth of shoots and leaves. Rather, MWCNT stimulated the relative electron transport rate and the effective photochemical quantum yield of PSII value as compared to the control by around 12% and lateral root production up to nearly 4-fold as compared to the control. The protective effect of MWCNT on MV toxicity on the root surface area could be quantitatively explained by the extent of MV adsorption on MWCNT and was related to stimulation of photosynthesis, antioxidant protection and number and area of lateral roots which in turn helped nutrient assimilation. The influence of MWCNT and MV on photosynthesis and oxidative stress at the physiological level was consistent with the proteomics analysis, with various over-expressed photosynthesis-related proteins (by more than 2 folds) and various under-expressed oxidative stress related proteins (by about 2-3 folds). This study brings new insights into the interactive effects of two xenobiotics (MWCNT and MV) on the physiology of a model plant.

Physiological and Molecular Response of Arabidopsis thaliana to CuO Nanoparticle (nCuO) Exposure.

The widespread application of copper oxide nanoparticles (nCuO) results in ecological risk when nanoparticles enter the environment. This study clarifies the mechanism of nCuO toxicity in Arabidopsis thaliana seedlings via comparison with copper (Cu) ion bioeffects. Under the same culture conditions, Cu2+ ion exposure exerted a stronger inhibitory effect on plant fresh weight and growth and caused stronger oxidative disruption (measured by malondialdehyde, MDA) than nCuO exposure. The Cu2+ ions also showed a stronger induction effect than did nCuO on the activity of antioxidant enzymes and the transcription of antioxidant-related genes. Dissolved Cu2+ ions contributed a minority of the toxicity of nCuO, implying that nCuO itself showed relative strong phytotoxicity. The work presented here will help increase our understanding of the toxicity of metal nanoparticles in plants.

A comparison of the effects of copper nanoparticles and copper sulfate on Phaeodactylum tricornutum physiology and transcription.

Copper nanoparticles (CuNPs) have been used in a broad range of applications. However, they are inevitably released into the marine environment, making it necessary to evaluate their potential effects on marine phytoplankton. In this study, the short-term (96h) effects of CuNPs and CuSO4 on Phacodactylum tricornutum growth, photosynthesis, reactive oxygen species production and transcription were assessed. It was found that high concentrations (40µM) of CuNPs and CuSO4 significantly inhibited the growth, photosynthesis and induced oxidative stress of P. tricornutum, while lower concentrations caused a hormetic response as indicated by a slight stimulation in algal growth. The high percentage of dissolved Cu (78-100%) in culture medium suggested that the dissolved Cu was the main driver of toxicity during CuNPs treatment. The algal cells upregulated electron transport chain-related genes to produce more energy and restore photosynthesis after 96h of treatment with CuNPs and CuSO4. This study delineates the cellular mechanism behind the toxicity of CuNPs and CuSO4 on marine diatoms.