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Whole-Genome Sequencing of Bacterial Isolates That Degrade the Cyanobacterial Toxin Microcystin-LR.

ABSTRACT: We previously demonstrated that 13 bacterial isolates from Lake Erie, when grown in groups of four to five isolates per group, degraded the cyanobacterial toxin microcystin-LR (MC-LR) into nontoxic fragments. Whole-genome sequencing of these bacteria was performed to provide genus and species information and to predict putative MC-LR-degrading genes.

SUBMITTER: McCartney AT

PROVIDER: S-EPMC7530933 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Whole-Genome Sequencing of Bacterial Isolates That Degrade the Cyanobacterial Toxin Microcystin-LR.

McCartney Alexander T AT Yeo Ji-Youn JY Blomquist Thomas M TM Huntley Jason F JF

Microbiology resource announcements 20201001 40

We previously demonstrated that 13 bacterial isolates from Lake Erie, when grown in groups of four to five isolates per group, degraded the cyanobacterial toxin microcystin-LR (MC-LR) into nontoxic fragments. Whole-genome sequencing of these bacteria was performed to provide genus and species information and to predict putative MC-LR-degrading genes. ...[more]

PMID: 33004461

Similar Datasets

Isolation and Characterization of Lake Erie Bacteria that Degrade the Cyanobacterial Microcystin Toxin MC-LR.

Project description:Microcystin-LR (MC-LR) is a cyclic hepatotoxin produced by cyanobacteria, including Microcystis sp. and Planktothrix sp. Harmful algal blooms (HABs) in Lake Erie have become a major human health concern in recent years, highlighted by the August 2014 city of Toledo, Ohio, municipal water "do not drink" order that affected nearly 500,000 residents for 3 days. Given that microcystin degrading bacteria have been reported from HAB-affected waters around the world, we hypothesized that MC-LR degrading bacteria could be isolated from Lake Erie. To test this hypothesis, 13 water samples were collected from various Lake Erie locations during the summers of 2014 and 2015, MC-LR was continuously added to each water sample for 3 to 5 weeks to enrich for MC-LR-degrading bacteria, and MC-LR was quantitated over time. Whereas MC-LR was relatively stable in sterile-filtered lake water, robust MC-LR degradation (up to 19 ppb/day) was observed in some water samples. Following the MC-LR selection process, 67 individual bacterial isolates were isolated from MC-LR degrading water samples and genotyped to exclude potential human pathogens, and MC-LR degradation by smaller groups of bacterial isolates (e.g., groups of 22 isolates, groups of 11 isolates, etc.) was examined. Of those smaller groups, selected groups of four to five bacterial isolates were found to degrade MC-LR into non-toxic forms and form robust biofilms on siliconized glass tubes. Taken together, these studies support the potential use of isolated bacterial isolates to remove MC-LR from drinking water.

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Hydrogen Sulfide Signaling Protects Chlamydomonas reinhardtii Against Allelopathic Damage From Cyanobacterial Toxin Microcystin-LR.

Project description:Cyanobacterial blooms have become more frequent and serious in recent years. Not only do massive blooms cause environmental pollution and nutrient eutrophication, but they also produce microcystins (MCs), a group of toxic cycloheptapeptides, which threaten aquatic ecosystem and human health. As such, clarifying the allelopathic interactions between cyanobacteria and other algae is critical to better understand the driving factors of blooms. To date, however, such studies remain largely insufficient. Here, we treated model alga Chlamydomonas reinhardtii with microcystin-LR (MC-LR) to determine its allelopathic effects. Results showed that MC-LR markedly suppressed C. reinhardtii cell viability. Comparative proteomic and physiological analyses revealed that MC-LR significantly up-regulated protein abundance of antioxidants ascorbate peroxidase (APX) and catalase (CAT) at the beginning stage of exposure. This was accompanied by an over-accumulation of hydrogen peroxide (H2O2), suggesting that MC-LR suppresses cell viability via oxidative damage. Furthermore, we found that MCs induced desulfhydrase (DES) activity for hydrogen sulfide (H2S) generation at the beginning stage. Additional H2S donors reactivated antioxidant enzyme activity, which reduced H2O2 accumulation and ultimately enhanced C. reinhardtii tolerance to MC-LR damage. This effect could be reserved by inhibiting H2S biosynthesis. Simultaneously, we found that H2S also suppressed MC-LR-induced cell autophagy, and thus attenuated the toxic effects of MC-LR. Our findings suggest that oxidative bursts may be the main reason for the allelopathic effects of MC-LR on C. reinhardtii viability and that H2S signaling may enhance C. reinhardtii tolerance to MC-LR through the activation of antioxidant enzyme activity and suppression of cell autophagy.

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Effects of a cyanobacterial bloom sample containing microcystin-LR on the ecophysiology of Daphnia similis.

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Susceptibility of the Algal Toxin Microcystin-LR to UV/Chlorine Process: Comparison with Chlorination.

Project description:Microcystin-LR (MC-LR), an algal toxin (cyanotoxin) common in sources of drinking water, poses a major human health hazard due to its high toxicity. In this study, UV/chlorine was evaluated as a potentially practical and effective process for the degradation of MC-LR. Via mass spectrometry analysis, fewer chlorinated-MC-LR products were detected with UV/chlorine treatment than with chlorination, and a transformation pathway for MC-LR by UV/chlorine was proposed. Different degrees of rapid degradation of MC-LR were observed with varying pH (6-10.4), oxidant dosage (0.5-3 mg L-1), natural organic matter (0-7 mg L-1), and natural water sources. In contrast to the formation of primarily chloroform and dichloroacetic acid in deionized water where MC-LR serves as the only carbon source, additional chlorinated disinfection byproducts were produced when sand filtered natural water was used as a background matrix. The UV/chlorine treated samples also showed quantitatively less cytotoxicity in vitro in HepaRG human liver cell line tests than chlorination treated samples. Following 16 min (96 mJ cm-2) of UV irradiation combined with 1.5 mg L-1 chlorine treatment, the cell viability of the samples increased from 80% after exposure to 1 mg L-1 MC-LR to 90%, while chlorination treatment evidenced no reduction in cytotoxicity with the same reaction time.

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Microarray Analysis of Microcystin-LR

2014-07-25 | GSE59495 | GEO

Rapid isolation of a single-chain antibody against the cyanobacterial toxin microcystin-LR by phage display and its use in the immunoaffinity concentration of microcystins from water.

Project description:A naïve (unimmunized) human semisynthetic phage display library was employed to isolate recombinant antibody fragments against the cyanobacterial hepatotoxin microcystin-LR. Selected antibody scFv genes were cloned into a soluble expression vector and expressed in Escherichia coli for characterization against purified microcystin-LR by competition enzyme-linked immunosorbent assay (ELISA). The most sensitive single-chain antibody (scAb) isolated was capable of detecting microcystin-LR at levels below the World Health Organization limit in drinking water (1 microg liter(-1)) and cross-reacted with three other purified microcystin variants (microcystin-RR, -LW, and -LF) and the related cyanotoxin nodularin. Extracts of the cyanobacterium Microcystis aeruginosa were assayed by ELISA, and quantifications of microcystins in toxic samples showed good correlation with analysis by high-performance liquid chromatography. Immobilized scAb was also used to prepare immunoaffinity columns, which were assessed for the ability to concentrate microcystin-LR from water for subsequent analysis by high-performance liquid chromatography. Anti-microcystin-LR scAb was immobilized on columns via a hexahistidine tag, ensuring maximum exposure of antigen binding sites, and the performance of the columns was evaluated by directly applying 150 ml of distilled water spiked with 4 micro g of purified microcystin-LR. The procedure was simple, and a recovery rate of 94% was achieved following elution in 1 ml of 100% methanol. Large-scale, low-cost production of anti-microcystin-LR scAb in E. coli is an exciting prospect for the development of biosensors and on-line monitoring systems for microcystins and will also facilitate a range of immunoaffinity applications for the cleanup and concentration of these toxins from environmental samples.

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Microarray Analysis of Microcystin-LR

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