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Zeng W, Lu M, Wu L, Gao X, Chen Q, Zhang S. Nanozyme mediated Raman-NLISA dual-modal immunosensor for accurate and sensitive detection of microcystin-LR. Food Chem 2025; 485:144480. [PMID: 40311568 DOI: 10.1016/j.foodchem.2025.144480] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/16/2025] [Revised: 04/08/2025] [Accepted: 04/21/2025] [Indexed: 05/03/2025]
Abstract
A Raman scattering and nanozyme-linked immunosorbent assay (NLISA) dual modal immunosensor, was constructed by mesoporous SiO2/Au-Pt nanozymes (m-SAP) and nanobodies (A2.3-SBP). Oxidized TMB served as Raman and ELISA signals in a competitive binding assay. Under optimized conditions, an inverse correlation was established between the Microcystin-LR (MC-LR) concentration and the signals, spanning Raman and ELISA ranges of 0.1-100 μg L-1 and 1.0-500 μg L-1, with limit of detections (LODs, 3σ/S) of 0.015 μg L-1 and 0.12 μg L-1, respectively. The LODs showed over 90 times and 11 times higher sensitivity than that of traditional ELISA (t-ELISA, LOD, 1.36 μg L-1). The immunosensor exhibited excellent accuracy in practical samples, can be integrated together for the detection of MC-LR within 45 min, which greatly short the detection time of t-ELISA (>2 h). This method displayed potential for detecting other toxins by simply changing the nanobodies.
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Affiliation(s)
- Wei Zeng
- School of Food Science and Engineering, Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation, Hainan University, Haikou 570228, PR China
| | - Mingrui Lu
- School of Food Science and Engineering, Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation, Hainan University, Haikou 570228, PR China
| | - Long Wu
- School of Food Science and Engineering, Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation, Hainan University, Haikou 570228, PR China.; State Key Laboratory of Marine Food Processing and Safety Control, Dalian Polytechnic University, Dalian 116034, Liaoning, China.
| | - Xin Gao
- School of Food Science and Engineering, Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation, Hainan University, Haikou 570228, PR China
| | - Qi Chen
- National Engineering Research Center for Bioengineering Drugs and the Technologies, Institute of Translational Medicine, Jiangxi Medical College, Nanchang University, Nanchang 330031, PR China
| | - Sihang Zhang
- School of Food Science and Engineering, Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation, Hainan University, Haikou 570228, PR China..
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Villalobos T, Suárez-Isla B, Garcia C. Health and Environmental Impacts of Cyanobacteria and Cyanotoxins from Freshwater to Seawater. Toxins (Basel) 2025; 17:126. [PMID: 40137899 PMCID: PMC11945519 DOI: 10.3390/toxins17030126] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2024] [Revised: 02/27/2025] [Accepted: 03/04/2025] [Indexed: 03/29/2025] Open
Abstract
Cyanobacterial harmful algal blooms (cyanoHABs) are a natural phenomenon produced mainly by the interaction between natural and anthropogenic events. CyanoHABs are characterized by the production of cyanotoxins that can have harmful effects on different species within the food web and even affect human health. Among the most prevalent toxin groups worldwide are microcystins (MCs), anatoxins (ATXs), cylindrospermopsins (CYNs) and nodularins (NODs), which are characterized as toxins with hepatotoxic, neurotoxic, and cytotoxic effects. This review summarizes and analyzes research on the influence of cyanoHABs, the main toxin-producing cyanobacteria and the most prevalent cyanotoxins in freshwater and marine bodies, highlighting their global occurrence, toxicology, and bioaccumulation dynamics in vectors of the food web, and the main cases of acute and chronic intoxications in humans. This review is useful for understanding the dynamics of cyanoHABs' interaction with the ecosystem and their impact on human health, and how the implementation of a surveillance and management framework for cyanobacteria and cyanotoxins could generate vital information for stakeholders to establish health guidelines on the risks and hazards of cyanoHABs for the ecosystem and humans.
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Affiliation(s)
| | | | - Carlos Garcia
- Laboratory of Marine Toxins, Physiology and Biophysics Programme, Faculty of Medicine, University of Chile, Santiago 8330111, Chile; (T.V.); (B.S.-I.)
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Wang YT, Wu QH, Chen L, Giesy JP, Xu LL, Xu WL, He J, Shi T, Liu YQ, Xiao SM, Wang YK, Chen F, Chen Y, Xu NH, Ge YL, Chu L, Yan YZ, Chen J, Xie P. Effects of sub-chronic exposure to microcystin-LR on the endocrine system of male rats. THE SCIENCE OF THE TOTAL ENVIRONMENT 2024; 906:166839. [PMID: 37690761 DOI: 10.1016/j.scitotenv.2023.166839] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/04/2023] [Revised: 08/14/2023] [Accepted: 09/02/2023] [Indexed: 09/12/2023]
Abstract
Microcystins (MCs) can cause reproductive and developmental toxicity and disrupt endocrine homeostasis in mammals. In the present study, male, Sprague-Dawley (SD) rats were administrated 3 or 30 μg MC-LR/kg, body mass (bm) per day via intraperitoneal (i.p.) injections for 6 weeks. Effects of MC-LR on histology, hormone concentrations, gene transcriptional profiles and protein expressions along the hypothalamic-pituitary-adrenal (HPA), -gonad (HPG) and -thyroid (HPT) axes were assessed. Sub-chronic administration with MC-LR caused histological damage to hypothalamus, pituitary, adrenal, testes and thyroid and affected relative masses of pituitary, adrenal and testes. The HPA axis was activated and serum concentrations of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were significantly augmented. Along the HPG axis, serum concentrations of gonadotropin-releasing hormone (GnRH) and dihydrotestosterone (DHT) were diminished, while concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T) and estradiol (E2) were augmented. For the HPT axis, only concentrations of free tetra-iodothyronine (fT4) were significantly diminished, while concentrations of thyrotropin-releasing hormone (TRH), thyroid-stimulating hormone (TSH) or free tri-iodothyronine (fT3) were not significantly changed. Also, several genes and proteins related to synthesis of steroid hormones were significantly altered. Findings of the present study illustrate that MC-LR can cause endocrine-disrupting effects through the disruption of synthesis and secretion of hormones along the HPA, HPG and HPT axes and negative feedback regulation. Also, there could be crosstalk among HPA, HPG and HPT axes. These findings elucidate mechanisms of endocrine-disrupting effects of MCs.
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Affiliation(s)
- Yu-Ting Wang
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China; Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
| | - Qian-Hui Wu
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China; Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
| | - Liang Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China; Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
| | - John P Giesy
- Department of Veterinary Biomedical Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B3, Canada; Department of Integrative Biology and Center for Integrative Toxicology, Michigan State University, 1129 Farm Lane Road, East Lansing, MI, USA; Department of Environmental Sciences, Baylor University, Waco, TX 76706, USA
| | - Lin-Lin Xu
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Wen-Li Xu
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Jun He
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Ting Shi
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Yi-Qing Liu
- Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
| | - Shi-Man Xiao
- Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
| | - Ye-Ke Wang
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Feng Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Yang Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China
| | - Ning-Hui Xu
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China; Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
| | - Ya-Li Ge
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China
| | - Ling Chu
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China
| | - Yun-Zhi Yan
- Collaborative Innovation Center of Recovery and Reconstruction of Degraded Ecosystem in Wanjiang Basin Co-founded by Anhui Province and Ministry of Education, School of Ecology and Environment, Anhui Normal University, Wuhu 241002, China.
| | - Jun Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China.
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China; Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
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He J, Chen Y, Dai S, Chen F, Wang Y, Shi T, Chen L, Liu Y, Chen J, Xie P. First insights into region-specific lipidome alterations of prefrontal cortex and hippocampus of mice exposed chronically to microcystins. ENVIRONMENT INTERNATIONAL 2023; 177:108018. [PMID: 37329758 DOI: 10.1016/j.envint.2023.108018] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/15/2023] [Revised: 05/31/2023] [Accepted: 06/01/2023] [Indexed: 06/19/2023]
Abstract
Microcystins (MCs), a group of most widespread freshwater cyanotoxins that possess strong neurotoxicity, can adversely affect brain structures and functions and are linked to neurodegenerative diseases. Despite the essential role of lipids in brain structures and functions, the brain lipidome profile of mammals exposed to MCs remains unexplored, hindering a clear understanding of the neurotoxic effects of MCs and underlying mechanisms. In this study, we performed untargeted lipidomic profiling using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) on the prefrontal cortex and hippocampus of mice orally exposed to 30 and 300 μg/kg body mass/day of microcystin-leucine arginine (MC-LR) for 180 days to evaluate the impacts of MC-LR on the brain lipidome profile and functions. Our results show that MC-LR resulted in a decline in cognitive parameters, as assessed by the Morris water maze test. Interestingly, apparent neurodegenerative changes were observed in the prefrontal cortex, but not in the hippocampus. Comprehensive lipidomic analyses uncovered profound, region-specific changes in the phospholipid and sphingolipid profile at the levels of lipid subclasses, lipid species, and fatty acyl composition. These changes showed overall decrease trends of lipid content in the prefrontal cortex yet increasing trends in the hippocampus. We identified distinct transcriptional regulations of lipid metabolism and apoptosis by MC-LR in the two regions, which appeared to underlie the neurodegenerative changes. Collectively, this study uncovers region-specific changes in the brain lipidome profile and functions induced by MCs, shedding light on the role of lipid dysfunction in neurotoxicity mechanism of MCs.
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Affiliation(s)
- Jun He
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Yang Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Shiming Dai
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Feng Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Yeke Wang
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Ting Shi
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China
| | - Liang Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China; Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
| | - Ying Liu
- Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China
| | - Jun Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China.
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences (UCAS), Beijing 100049, China; Institute for Ecological Research and Pollution Control of Plateau Lakes, School of Ecology and Environmental Science, Yunnan University, Kunming 650500, China.
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Davidović P, Blagojević D, Meriluoto J, Simeunović J, Svirčev Z. Biotests in Cyanobacterial Toxicity Assessment-Efficient Enough or Not? BIOLOGY 2023; 12:biology12050711. [PMID: 37237524 DOI: 10.3390/biology12050711] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/23/2023] [Revised: 04/27/2023] [Accepted: 05/09/2023] [Indexed: 05/28/2023]
Abstract
Cyanobacteria are a diverse group of organisms known for producing highly potent cyanotoxins that pose a threat to human, animal, and environmental health. These toxins have varying chemical structures and toxicity mechanisms and several toxin classes can be present simultaneously, making it difficult to assess their toxic effects using physico-chemical methods, even when the producing organism and its abundance are identified. To address these challenges, alternative organisms among aquatic vertebrates and invertebrates are being explored as more assays evolve and diverge from the initially established and routinely used mouse bioassay. However, detecting cyanotoxins in complex environmental samples and characterizing their toxic modes of action remain major challenges. This review provides a systematic overview of the use of some of these alternative models and their responses to harmful cyanobacterial metabolites. It also assesses the general usefulness, sensitivity, and efficiency of these models in investigating the mechanisms of cyanotoxicity expressed at different levels of biological organization. From the reported findings, it is clear that cyanotoxin testing requires a multi-level approach. While studying changes at the whole-organism level is essential, as the complexities of whole organisms are still beyond the reach of in vitro methodologies, understanding cyanotoxicity at the molecular and biochemical levels is necessary for meaningful toxicity evaluations. Further research is needed to refine and optimize bioassays for cyanotoxicity testing, which includes developing standardized protocols and identifying novel model organisms for improved understanding of the mechanisms with fewer ethical concerns. In vitro models and computational modeling can complement vertebrate bioassays and reduce animal use, leading to better risk assessment and characterization of cyanotoxins.
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Affiliation(s)
- Petar Davidović
- Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 2, 21000 Novi Sad, Serbia
| | - Dajana Blagojević
- Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 2, 21000 Novi Sad, Serbia
| | - Jussi Meriluoto
- Faculty of Science and Engineering, Biochemistry, Åbo Akademi, Tykistökatu 6 A, 20520 Turku, Finland
| | - Jelica Simeunović
- Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 2, 21000 Novi Sad, Serbia
| | - Zorica Svirčev
- Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 2, 21000 Novi Sad, Serbia
- Faculty of Science and Engineering, Biochemistry, Åbo Akademi, Tykistökatu 6 A, 20520 Turku, Finland
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Varriale F, Tartaglione L, Zervou SK, Miles CO, Mazur-Marzec H, Triantis TM, Kaloudis T, Hiskia A, Dell'Aversano C. Untargeted and targeted LC-MS and data processing workflow for the comprehensive analysis of oligopeptides from cyanobacteria. CHEMOSPHERE 2023; 311:137012. [PMID: 36397634 DOI: 10.1016/j.chemosphere.2022.137012] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/28/2022] [Revised: 09/26/2022] [Accepted: 10/23/2022] [Indexed: 06/16/2023]
Abstract
Cyanobacteria produce a plethora of structurally diverse bioactive secondary metabolites, including cyanotoxins which pose a serious threat to humans and other living organisms worldwide. Currently, a wide variety of mass spectrometry-based methods for determination of microcystins (MCs), the most commonly occurring and studied class of cyanotoxins, have been developed and employed for research and monitoring purposes. The scarcity of commercially available reference materials, together with the ever-growing range of mass spectrometers and analytical approaches, make the accuracy of quantitative analyses a critical point to be carefully investigated in view of a reliable risk evaluation. This study reports, a comparative investigation of the qualitative and quantitative MCs profile obtained using targeted and untargeted liquid chromatography-mass spectrometry approaches for the analyses of cyanobacterial biomass from Lake Kastoria, Greece. Comparison of the total MCs content measured by the two approaches showed good correlation, with variations in the range of 3.8-13.2%. In addition, the implementation of an analytical workflow on a hybrid linear ion trap/orbitrap mass spectrometer is described, based on combining data-dependent acquisition and a powerful database of cyanobacterial metabolites (CyanoMetDB) for the annotation of known and discovery of new cyanopeptides. This untargeted strategy proved highly effective for the identification of MCs, microginins, anabaenopeptins, and micropeptins. The systematic interpretation of the acquired fragmentation patterns allowed the elucidation of two new MC structural variants, MC-PrhcysR and MC-Prhcys(O)R, and proposal of structures for two new microginins, isomeric cyanostatin B and MG 821A, and three isomeric micropeptins at m/z 846.4715, 846.4711 and 846.4723.
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Affiliation(s)
- Fabio Varriale
- University of Napoli Federico II, Department of Pharmacy, School of Medicine and Surgery, Via D. Montesano 49, 80131, Napoli, Italy
| | - Luciana Tartaglione
- University of Napoli Federico II, Department of Pharmacy, School of Medicine and Surgery, Via D. Montesano 49, 80131, Napoli, Italy; CoNISMa - National Inter-University Consortium for Marine Sciences, Piazzale Flaminio 9, 00196, Rome, Italy.
| | - Sevasti-Kiriaki Zervou
- Laboratory of Photo-Catalytic Processes and Environmental Chemistry, Institute of Nanoscience and Nanotechnology (INN), National Center for Scientific Research "Demokritos", Patriarchou Grigoriou E & 27 Neapoleos Str, 15341 Agia Paraskevi, Athens, Greece
| | - Christopher O Miles
- Biotoxin Metrology, National Research Council Canada, 1411 Oxford Street, Nova Scotia, Canada
| | - Hanna Mazur-Marzec
- Division of Marine Biotechnology, Faculty of Oceanography and Geography, University of Gdańsk, Poland
| | - Theodoros M Triantis
- Laboratory of Photo-Catalytic Processes and Environmental Chemistry, Institute of Nanoscience and Nanotechnology (INN), National Center for Scientific Research "Demokritos", Patriarchou Grigoriou E & 27 Neapoleos Str, 15341 Agia Paraskevi, Athens, Greece
| | - Triantafyllos Kaloudis
- Laboratory of Photo-Catalytic Processes and Environmental Chemistry, Institute of Nanoscience and Nanotechnology (INN), National Center for Scientific Research "Demokritos", Patriarchou Grigoriou E & 27 Neapoleos Str, 15341 Agia Paraskevi, Athens, Greece
| | - Anastasia Hiskia
- Laboratory of Photo-Catalytic Processes and Environmental Chemistry, Institute of Nanoscience and Nanotechnology (INN), National Center for Scientific Research "Demokritos", Patriarchou Grigoriou E & 27 Neapoleos Str, 15341 Agia Paraskevi, Athens, Greece
| | - Carmela Dell'Aversano
- University of Napoli Federico II, Department of Pharmacy, School of Medicine and Surgery, Via D. Montesano 49, 80131, Napoli, Italy; CoNISMa - National Inter-University Consortium for Marine Sciences, Piazzale Flaminio 9, 00196, Rome, Italy
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Rattner BA, Wazniak CE, Lankton JS, McGowan PC, Drovetski SV, Egerton TA. Review of harmful algal bloom effects on birds with implications for avian wildlife in the Chesapeake Bay region. HARMFUL ALGAE 2022; 120:102319. [PMID: 36470599 DOI: 10.1016/j.hal.2022.102319] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/25/2022] [Revised: 09/16/2022] [Accepted: 09/18/2022] [Indexed: 06/17/2023]
Abstract
The Chesapeake Bay, along the mid-Atlantic coast of North America, is the largest estuary in the United States and provides critical habitat for wildlife. In contrast to point and non-point source release of pesticides, metals, and industrial, personal care and household use chemicals on biota in this watershed, there has only been scant attention to potential exposure and effects of algal toxins on wildlife in the Chesapeake Bay region. As background, we first review the scientific literature on algal toxins and harmful algal bloom (HAB) events in various regions of the world that principally affected birds, and to a lesser degree other wildlife. To examine the situation for the Chesapeake, we compiled information from government reports and databases summarizing wildlife mortality events for 2000 through 2020 that were associated with potentially toxic algae and HAB events. Summary findings indicate that there have been few wildlife mortality incidents definitively linked to HABs, other mortality events that were suspected to be related to HABs, and more instances in which HABs may have indirectly contributed to or occurred coincident with wildlife mortality. The dominant toxins found in the Chesapeake Bay drainage that could potentially affect wildlife are microcystins, with concentrations in water approaching or exceeding human-based thresholds for ceasing recreational use and drinking water at a number of locations. As an increasing trend in HAB events in the U.S. and in the Chesapeake Bay have been reported, additional information on HAB toxin exposure routes, comparative sensitivity among species, consequences of sublethal exposure, and better diagnostic and risk criteria would greatly assist in predicting algal toxin hazard and risks to wildlife.
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Affiliation(s)
- Barnett A Rattner
- U.S. Geological Survey, Eastern Ecological Science Center at the Patuxent Research Refuge, Beltsville, MD 20705, USA.
| | - Catherine E Wazniak
- Maryland Department of Natural Resources, Resource Assessment Service, Annapolis, MD 21401, USA
| | - Julia S Lankton
- U.S. Geological Survey, National Wildlife Health Center, Madison, WI 53711, USA
| | - Peter C McGowan
- U.S. Fish and Wildlife Service, Chesapeake Bay Field Office, Annapolis, MD 21401, USA
| | - Serguei V Drovetski
- U.S. Geological Survey, Eastern Ecological Science Center at the Patuxent Research Refuge, Beltsville, MD 20705, USA
| | - Todd A Egerton
- Virginia Department of Health, Division of Shellfish Safety and Waterborne Hazards, Norfolk, VA 23510, USA
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8
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Breidenbach JD, French BW, Gordon TT, Kleinhenz AL, Khalaf FK, Willey JC, Hammersley JR, Mark Wooten R, Crawford EL, Modyanov NN, Malhotra D, Teeguarden JG, Haller ST, Kennedy DJ. Microcystin-LR aerosol induces inflammatory responses in healthy human primary airway epithelium. ENVIRONMENT INTERNATIONAL 2022; 169:107531. [PMID: 36137425 DOI: 10.1016/j.envint.2022.107531] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/17/2022] [Revised: 08/24/2022] [Accepted: 09/16/2022] [Indexed: 06/16/2023]
Abstract
Harmful algal blooms plague bodies of freshwater globally. These blooms are often composed of outgrowths of cyanobacteria capable of producing the heptapeptide Microcystin-LR (MC-LR) which is a well-known hepatotoxin. Recently, MC-LR has been detected in aerosols generated from lake water. However, the risk for human health effects due to MC-LR inhalation exposure have not been extensively investigated. In this study, we exposed a fully differentiated 3D human airway epithelium derived from 14 healthy donors to MC-LR-containing aerosol once a day for 3 days. Concentrations of MC-LR ranged from 100 pM to 1 µM. Although there were little to no detrimental alterations in measures of the airway epithelial function (i.e. cell survival, tissue integrity, mucociliary clearance, or cilia beating frequency), a distinct shift in the transcriptional activity was found. Genes related to inflammation were found to be upregulated such as C-C motif chemokine 5 (CCL5; log2FC = 0.57, p = 0.03) and C-C chemokine receptor type 7 (CCR7; log2FC = 0.84, p = 0.03). Functionally, conditioned media from MC-LR exposed airway epithelium was also found to have significant chemo-attractive properties for primary human neutrophils. Additionally, increases were found in the concentration of secreted chemokine proteins in the conditioned media such as CCL1 (log2FC = 5.07, p = 0.0001) and CCL5 (log2FC = 1.02, p = 0.046). These results suggest that MC-LR exposure to the human airway epithelium is capable of inducing an inflammatory response that may potentiate acute or chronic disease.
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Affiliation(s)
| | - Benjamin W French
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Tamiya T Gordon
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Andrew L Kleinhenz
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Fatimah K Khalaf
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA; College of Pharmacy, University of Alkafeel, Najaf, Iraq
| | - James C Willey
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | | | - R Mark Wooten
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Erin L Crawford
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Nikolai N Modyanov
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Deepak Malhotra
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - Justin G Teeguarden
- Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, USA; Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, USA
| | - Steven T Haller
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA
| | - David J Kennedy
- College of Medicine and Life Sciences, University of Toledo, Toledo, OH, USA.
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9
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Hill D, Lang J, McCord J, Strynar M, Rosal C, Schmid J, Le TT, Chernoff N. Variability of Microcystin-LR Standards Available from Seven Commercial Vendors. Toxins (Basel) 2022; 14:toxins14100705. [PMID: 36287973 PMCID: PMC9611723 DOI: 10.3390/toxins14100705] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/02/2022] [Revised: 10/07/2022] [Accepted: 10/10/2022] [Indexed: 12/05/2022] Open
Abstract
Microcystins (MCs) are a large group of heptapeptide cyanobacterial toxins commonly produced in harmful algal blooms (HABs) and associated with adverse health effects in wildlife, livestock, pets, and humans. MC chemical standards are extracted from cyanobacteria biomass rather than produced synthetically and are used in water assessment methods and toxicological studies. MC standards are generally supplied in less than 1 mg quantities, and verification of the mass can only be accomplished by analytical chemistry methods using a certified reference of the specific MC for comparison. Analytical quantification of MCs in environmental samples and toxicology studies using accurate doses of test chemicals administered to experimental animals rely on the availability and accuracy of chemical standards. To check the accuracy and purity of available standards, seven individual microcystin-LR (MCLR) standards were purchased from separate commercial vendors and analyzed to determine the actual mass supplied and identify the presence of potential contaminants. To determine the effect of varying toxin mass in toxicological studies, each MCLR standard was administered to CD-1 mice in doses based on mass purchased, by a single 40 µg/kg intraperitoneal injection. The measured mass purchased varied from the vendor label mass by more than 35% for two of the seven MCLR standards. Contaminants, including trifluoroacetic acid (TFA), were identified in four of the seven samples. Comparative in vivo hepatotoxicity between vendor samples closely reflected the actual amount of MCLR present in each standard and demonstrated the toxicological impact of varying cyanotoxin mass.
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Affiliation(s)
- Donna Hill
- Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Research Triangle Park, NC 27711, USA
- Correspondence:
| | - Johnsie Lang
- Arcadis (United States), 4204 Technology Dr, Durham, NC 27704, USA
| | - James McCord
- Center for Environmental Measurement and Modeling, United States Environmental Protection Agency, Research Triangle Park, NC 27711, USA
| | - Mark Strynar
- Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Research Triangle Park, NC 27711, USA
| | | | - Judith Schmid
- Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Research Triangle Park, NC 27711, USA
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10
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LC-MS/MS Validation and Quantification of Cyanotoxins in Algal Food Supplements from the Belgium Market and Their Molecular Origins. Toxins (Basel) 2022; 14:toxins14080513. [PMID: 36006175 PMCID: PMC9415669 DOI: 10.3390/toxins14080513] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2022] [Revised: 07/19/2022] [Accepted: 07/22/2022] [Indexed: 02/04/2023] Open
Abstract
Food supplements are gaining popularity worldwide. However, harmful natural compounds can contaminate these products. In the case of algae-based products, the presence of toxin-producing cyanobacteria may cause health risks. However, data about the prevalence of algal food supplements on the Belgian market and possible contaminations with cyanotoxins are scarce. Therefore, we optimized and validated a method based on Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry to quantify eight microcystin congeners and nodularin in algal food supplements. Our analytical method was successfully validated and applied on 35 food supplement samples. Nine out of these samples contained microcystin congeners, of which three exceeded 1 µg g−1, a previously proposed guideline value. Additionally, the mcyE gene was amplified and sequenced in ten products to identify the taxon responsible for the toxin production. For seven out of these ten samples, the mcyE gene could be amplified and associated to Microcystis sp. EFSA and posology consumption data for algal-based food supplements were both combined with our toxin prevalence data to establish different toxin exposure scenarios to assess health risks and propose new guideline values.
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11
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Mrdjen I, Lee J, Weghorst CM, Knobloch TJ. Impact of Cyanotoxin Ingestion on Liver Cancer Development Using an At-Risk Two-Staged Model of Mouse Hepatocarcinogenesis. Toxins (Basel) 2022; 14:toxins14070484. [PMID: 35878222 PMCID: PMC9320861 DOI: 10.3390/toxins14070484] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/07/2022] [Revised: 07/05/2022] [Accepted: 07/06/2022] [Indexed: 12/04/2022] Open
Abstract
Exposure to cyanobacterial hepatotoxins has been linked to the promotion and increased incidence of liver cancer in pre-clinical and epidemiologic studies. The family of hepatotoxins, microcystins (MCs), are produced by over 40 cyanobacterial species found in harmful algal blooms (HABs) worldwide, with MC-LR being the most common and potent MC congener. In the current study, we hypothesized that the low-dose chronic ingestion of Microcystis cyanotoxins via drinking water would promote liver carcinogenesis in pre-initiated mice. Four groups of C3H/HeJ mice received one intraperitoneal (i.p.) injection of diethylnitrosamine (DEN) at 4 weeks of age. Three weeks later, the mice were administered ad libitum drinking water containing one of the following: (1) reverse osmosis, deionized water; (2) water containing 500 mg/L phenobarbital (PB500); (3) water with purified MC-LR (10 µg/L) added; or (4) water containing lysed Microcystis aeruginosa (lysate; 10 µg/L total MCs). The exposure concentrations were based on environmentally relevant concentrations and previously established Ohio EPA recreational water MC guidelines. Throughout the 30-week exposure, mouse weights, food consumption, and water consumption were not significantly impacted by toxin ingestion. We found no significant differences in the number of gross and histopathologic liver lesion counts across the treatment groups, but we did note that the PB500 group developed lesion densities too numerous to count. Additionally, the proportion of lesions classified as hepatocellular carcinomas in the MC-LR group (44.5%; p < 0.05) and lysate group (55%; p < 0.01) was significantly higher compared to the control group (14.9%). Over the course of the study, the mice ingesting the lysate also had a significantly lower survival probability (64.4%; p < 0.001) compared to water (96.8%), PB500 (95.0%), and MC-LR (95.7%) exposures. Using cyanotoxin levels at common recreational water concentration levels, we demonstrate the cancer-promoting effects of a single cyanotoxin MC congener (MC-LR). Furthermore, we show enhanced hepatocarcinogenesis and significant mortality associated with combinatorial exposure to the multiple MCs and bioactive compounds present in lysed cyanobacterial cells—a scenario representative of the ingestion exposure route, such as HAB-contaminated water and food.
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Affiliation(s)
- Igor Mrdjen
- College of Public Health, Environmental Health Sciences, The Ohio State University, Columbus, OH 43210, USA; (I.M.); (J.L.); (C.M.W.)
| | - Jiyoung Lee
- College of Public Health, Environmental Health Sciences, The Ohio State University, Columbus, OH 43210, USA; (I.M.); (J.L.); (C.M.W.)
- Department of Food Science and Technology, The Ohio State University, Columbus, OH 43210, USA
| | - Christopher M. Weghorst
- College of Public Health, Environmental Health Sciences, The Ohio State University, Columbus, OH 43210, USA; (I.M.); (J.L.); (C.M.W.)
| | - Thomas J. Knobloch
- College of Public Health, Environmental Health Sciences, The Ohio State University, Columbus, OH 43210, USA; (I.M.); (J.L.); (C.M.W.)
- Correspondence:
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12
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Baliu-Rodriguez D, Peraino NJ, Premathilaka SH, Birbeck JA, Baliu-Rodriguez T, Westrick JA, Isailovic D. Identification of Novel Microcystins Using High-Resolution MS and MS n with Python Code. ENVIRONMENTAL SCIENCE & TECHNOLOGY 2022; 56:1652-1663. [PMID: 35018784 DOI: 10.1021/acs.est.1c04296] [Citation(s) in RCA: 49] [Impact Index Per Article: 16.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/14/2023]
Abstract
Cyanotoxins called microcystins (MCs) are highly toxic and can be present in drinking water sources. Determining the structure of MCs is paramount because of its effect on toxicity. Though over 300 MC congeners have been discovered, many remain unidentified. Herein, a method is described for the putative identification of MCs using liquid chromatography (LC) coupled with high-resolution (HR) Orbitrap mass spectrometry (MS) and a new bottom-up sequencing strategy. Maumee River water samples were collected during a harmful algal bloom and analyzed by LC-MS with simultaneous HRMS and MS/MS. Unidentified ions with characteristic MC fragments (135 and 213 m/z) were recognized as possible novel MC congeners. An innovative workflow was developed for the putative identification of these ions. Python code was written to generate the potential structures of unidentified MCs and to assign ions after the fragmentation for structural confirmation. The workflow enabled the putative identification of eight previously reported MCs for which standards are not available and two newly discovered congeners, MC-HarR and MC-E(OMe)R.
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Affiliation(s)
- David Baliu-Rodriguez
- Department of Chemistry and Biochemistry, University of Toledo, Toledo, Ohio 43606, United States
| | - Nicholas J Peraino
- Department of Chemistry, Wayne State University, Detroit, Michigan 48202, United States
| | - Sanduni H Premathilaka
- Department of Chemistry and Biochemistry, University of Toledo, Toledo, Ohio 43606, United States
| | - Johnna A Birbeck
- Department of Chemistry, Wayne State University, Detroit, Michigan 48202, United States
| | | | - Judy A Westrick
- Department of Chemistry, Wayne State University, Detroit, Michigan 48202, United States
| | - Dragan Isailovic
- Department of Chemistry and Biochemistry, University of Toledo, Toledo, Ohio 43606, United States
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13
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A Summer of Cyanobacterial Blooms in Belgian Waterbodies: Microcystin Quantification and Molecular Characterizations. Toxins (Basel) 2022; 14:toxins14010061. [PMID: 35051038 PMCID: PMC8780180 DOI: 10.3390/toxins14010061] [Citation(s) in RCA: 16] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/19/2021] [Revised: 01/08/2022] [Accepted: 01/12/2022] [Indexed: 12/04/2022] Open
Abstract
In the context of increasing occurrences of toxic cyanobacterial blooms worldwide, their monitoring in Belgium is currently performed by regional environmental agencies (in two of three regions) using different protocols and is restricted to some selected recreational ponds and lakes. Therefore, a global assessment based on the comparison of existing datasets is not possible. For this study, 79 water samples from a monitoring of five lakes in Wallonia and occasional blooms in Flanders and Brussels, including a canal, were analyzed. A Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) method allowed to detect and quantify eight microcystin congeners. The mcyE gene was detected using PCR, while dominant cyanobacterial species were identified using 16S RNA amplification and direct sequencing. The cyanobacterial diversity for two water samples was characterized with amplicon sequencing. Microcystins were detected above limit of quantification (LOQ) in 68 water samples, and the World Health Organization (WHO) recommended guideline value for microcystins in recreational water (24 µg L−1) was surpassed in 18 samples. The microcystin concentrations ranged from 0.11 µg L−1 to 2798.81 µg L−1 total microcystin. For 45 samples, the dominance of the genera Microcystis sp., Dolichospermum sp., Aphanizomenon sp., Cyanobium/Synechococcus sp., Planktothrix sp., Romeria sp., Cyanodictyon sp., and Phormidium sp. was shown. Moreover, the mcyE gene was detected in 75.71% of all the water samples.
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14
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da Silva CG, Duque MD, Freire Nordi CS, Viana-Niero C. New insights into toxicity of microcystins produced by cyanobacteria using in silico ADMET prediction. Toxicon 2021; 204:64-71. [PMID: 34742780 DOI: 10.1016/j.toxicon.2021.11.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2021] [Revised: 10/28/2021] [Accepted: 11/01/2021] [Indexed: 11/16/2022]
Abstract
In silico methodologies can be used in the discovery of new drugs for measuring toxicity, predicting effects of substances not yet analyzed by in vivo methodologies. The ADMET Predictor® software (absorption, distribution, metabolism, elimination, and toxicity [ADMET]) was used in this work to predict toxic effects of microcystin variants MC-LR, MC-YR, MC-RR, and MC-HarHar. In the case of rodents, predictive results for all analyzed variants indicated carcinogenic potential. The predictive model of respiratory sensitivity in this group differentiated microcystins into 2 categories: sensitizer (MC-LR and -YR) and non-sensitizer (MC-HarHar and -RR). Predictive results for humans indicated that MC-LR and -RR are phospholipidosis inducers; on the other hand, MC-LR showed the highest predictive value of permeability in rabbit cornea and probability of crossing lipoprotein barriers (MC-LR>-YR>-HarHar>-RR). Considering bioavailable fractions, microcystins are more likely to cause biological effects in rats than humans, showing significant differences between models. The results of ADMET predictions add valuable information on microcystin toxicity, especially in the case of variants not yet studied experimentally.
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Affiliation(s)
- Cristiane Gonçalves da Silva
- Departamento de Microbiologia, Imunologia e Parasitologia [Department of Microbiology, Immunology and Parasitology], Universidade Federal de São Paulo [Federal University of São Paulo], Rua Botucatu, 862, São Paulo, SP, Zip Code: 04023-901, Brazil; Departamento de Ciências Farmacêuticas [Department of Pharmaceutical Sciences], Universidade Federal de São Paulo, Rua São Nicolau, 210, Diadema, SP, Zip Code: 09913-030, Brazil
| | - Marcelo Dutra Duque
- Departamento de Ciências Farmacêuticas [Department of Pharmaceutical Sciences], Universidade Federal de São Paulo, Rua São Nicolau, 210, Diadema, SP, Zip Code: 09913-030, Brazil
| | - Cristina Souza Freire Nordi
- Departamento de Ciências Ambientais [Department of Environmental Sciences], Universidade Federal de São Paulo, Rua Prof. Artur Riedel, 275, Diadema, SP, Zip Code: 09972-270, Brazil.
| | - Cristina Viana-Niero
- Departamento de Microbiologia, Imunologia e Parasitologia [Department of Microbiology, Immunology and Parasitology], Universidade Federal de São Paulo [Federal University of São Paulo], Rua Botucatu, 862, São Paulo, SP, Zip Code: 04023-901, Brazil
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15
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Ait Abderrahim L, Taibi K, Boussaid M, Al-Shara B, Ait Abderrahim N, Ait Abderrahim S. Allium sativum mitigates oxidative damages induced by Microcystin-LR in heart and liver tissues of mice. Toxicon 2021; 200:30-37. [PMID: 34217748 DOI: 10.1016/j.toxicon.2021.06.018] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/22/2021] [Revised: 06/27/2021] [Accepted: 06/30/2021] [Indexed: 10/20/2022]
Abstract
Microcystins (MCs) are hepatotoxic cyanotoxins implicated in several incidents of human and animal toxicity. Microcystin-(Lysine, Arginine) or MC-LR is the most toxic and encountered variant of MCs where oxidative stress plays a key role in its toxicity. This study investigated the oxidative damages induced in the liver and heart of Balb/C mice by an intraperitoneal injected acute dose of MC-LR. Thereafter, the potential protective effect of garlic (Allium sativum) extract supplementation against such damages was assessed through the evaluation of oxidative stress and cytotoxicity markers. Lipid peroxidation (LPO), carbonyl content (CC), glutathione content (GSH), alkaline phosphatase activity (ALP), lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH) activities were measured. Results showed important oxidative damages in hepatic and cardiac cells of mice injected with the toxin. However, these damages have been significantly reduced in mice supplemented with garlic extract. Thus, this study demonstrated for the first time the effective use of garlic as an antioxidant agent against oxidative damages induced by MC-LR. As well, this study supports the use of garlic as a potential remedy against pathologies related to toxic agents.
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Affiliation(s)
- Leila Ait Abderrahim
- Faculty of Life and Natural Sciences, Ibn Khaldoun University, Karman Campus, 14000, Tiaret, Algeria.
| | - Khaled Taibi
- Faculty of Life and Natural Sciences, Ibn Khaldoun University, Karman Campus, 14000, Tiaret, Algeria
| | - Mohamed Boussaid
- Faculty of Life and Natural Sciences, Ibn Khaldoun University, Karman Campus, 14000, Tiaret, Algeria
| | - Baker Al-Shara
- Institute of Biological Sciences, Faculty of Science, University Malaya, Kuala Lumpur, 50603, Malaysia
| | | | - Sabrina Ait Abderrahim
- Faculty of Life and Natural Sciences, Ibn Khaldoun University, Karman Campus, 14000, Tiaret, Algeria
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16
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Mills MC, Evans MV, Lee S, Knobloch T, Weghorst C, Lee J. Acute cyanotoxin poisoning reveals a marginal effect on mouse gut microbiome composition but indicates metabolic shifts related to liver and gut inflammation. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 2021; 215:112126. [PMID: 33721663 DOI: 10.1016/j.ecoenv.2021.112126] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/21/2020] [Revised: 02/26/2021] [Accepted: 03/01/2021] [Indexed: 06/12/2023]
Abstract
Freshwater harmful algal blooms (HABs) are a major environmental health problem worldwide. HABs are caused by a predominance of cyanobacteria, some of which produce potent toxins. The most ubiquitous cyanotoxin is microcystin (MC) and the congener MC-LR is the most studied due to its toxicity. Short-term exposure to toxins can cause gut microbiome disturbances, but this has not been well described with MC-LR exposure. This study investigated the gut microbial communities of mice from a prior study, which identified significant liver toxicity from ingestion of MC-LR daily for 8 days. CD-1 mice were divided into three dosage groups: control, low exposure (sub-lethal MC-LR concentration), and high exposure (near-lethal MC-LR concentration). Fecal samples were analyzed using 16S rRNA sequencing. Results revealed that at population level, there were no significant shifts in bacterial diversity or the microbial community structure over the exposure period. However, there were significant differences between male and female mice. Predictive functional gene analysis indicated that several metabolic pathways were significantly different in the high dose group before exposure and following 7 doses of MC-LR, as well as between the control and high dose groups on Day 8. Significant differentially abundant taxa were also identified contributing to these pathways. Several pathways, including superpathway of N-acetylneuraminate degradation, were related to liver and gut inflammation. The outcome of this study suggests a need for in-depth investigation of metabolic activity and other functions in the gut in future studies, as well as potential consideration of the role of sex in MC-LR toxicity.
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Affiliation(s)
- Molly C Mills
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States; Environmental Sciences Graduate Program, The Ohio State University, Columbus, OH, United States
| | - Morgan V Evans
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States; College of Veterinary Medicine, Department of Veterinary Preventative Medicine, The Ohio State University, Columbus, OH, United States
| | - Seungjun Lee
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States
| | - Thomas Knobloch
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States
| | - Christopher Weghorst
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States
| | - Jiyoung Lee
- College of Public Health Division of Environmental Health Sciences, The Ohio State University, Columbus, OH, United States; Environmental Sciences Graduate Program, The Ohio State University, Columbus, OH, United States; Department of Food Science & Technology, The Ohio State University, Columbus, OH, United States.
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17
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Shi L, Du X, Liu H, Chen X, Ma Y, Wang R, Tian Z, Zhang S, Guo H, Zhang H. Update on the adverse effects of microcystins on the liver. ENVIRONMENTAL RESEARCH 2021; 195:110890. [PMID: 33617868 DOI: 10.1016/j.envres.2021.110890] [Citation(s) in RCA: 63] [Impact Index Per Article: 15.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/30/2020] [Revised: 02/10/2021] [Accepted: 02/11/2021] [Indexed: 06/12/2023]
Abstract
Microcystins (MCs) are the most common cyanobacteria toxins in eutrophic water, which have strong hepatotoxicity. In the past decade, epidemiological and toxicological studies on liver damage caused by MCs have proliferated, and new mechanisms of hepatotoxicity induced by MCs have also been discovered and confirmed. However, there has not been a comprehensive and systematic review of these new findings. Therefore, this paper summarizes the latest advances in studies on the hepatotoxicity of MCs to reveal the effects and mechanisms of hepatotoxicity induced by MCs. Current epidemiological studies have confirmed that symptoms or signs of liver damage appear after human exposure to MCs, and a long time of exposure can even lead to liver cancer. Toxicological studies have shown that MCs can affect the expression of oncogenes by activating cell proliferation pathways such as MAPK and Akt, thereby promoting the occurrence and development of cancer. The latest evidence shows that epigenetic modifications may play an important role in MCs-induced liver cancer. MCs can cause damage to the liver by inducing hepatocyte death, mainly manifested as apoptosis and necrosis. The imbalance of liver metabolic homeostasis may be involved in hepatotoxicity induced by MCs. In addition, the combined toxicity of MCs and other toxins are also discussed in this article. This detailed information will be a valuable reference for further exploring of MCs-induced hepatotoxicity.
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Affiliation(s)
- Linjia Shi
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Xingde Du
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Haohao Liu
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Xinghai Chen
- Department of Chemistry and Biochemistry, St Mary's University, San Antonio, TX, USA
| | - Ya Ma
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Rui Wang
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Zhihui Tian
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Shiyu Zhang
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China
| | - Hongxiang Guo
- College of Life Sciences, Henan Agricultural University, Zhengzhou, Henan, China
| | - Huizhen Zhang
- College of Public Health, Zhengzhou University, Zhengzhou, Henan, China.
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18
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Xiao W, Zhong Q, Sun F, Wang W, Zhao Z, Gu K. Differences in Abnormal Water Metabolism between SD Rats and KM Mice Intoxicated by Microcystin-RR. INTERNATIONAL JOURNAL OF ENVIRONMENTAL RESEARCH AND PUBLIC HEALTH 2021; 18:1900. [PMID: 33669356 PMCID: PMC7920292 DOI: 10.3390/ijerph18041900] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/15/2020] [Revised: 02/09/2021] [Accepted: 02/10/2021] [Indexed: 01/17/2023]
Abstract
The effects of microcystin-RR (MC-RR) on water metabolism were studied on Sprague-Dawley (SD) rats and KunMing (KM) mice. In the single dose toxicity test, polydipsia, polyuria, hematuria and proteinuria were found in group of rats receiving a MC-RR dose of 574.7 μg/kg, and could be relieved by dexamethasone (DXM). Gradient damage was observed in kidney and liver in rats with gradient MC-RR doses of 574.7, 287.3, and 143.7 μg/kg. No significant water metabolic changes or kidney injuries were observed in mice treated with MC-RR doses of 210.0, 105.0, and 52.5 μg/kg. In the continuous exposure test, in which mice were administrated with 140.0, 70.0, and 35.0 μg/kg MC-RR for 28 days, mice in the 140.0 μg/kg group presented increasing polydipsia, polyuria, and liver damage. However, no anatomic or histological changes, including related serological and urinary indices, were found in the kidney. In summary, abnormal water metabolism can be induced by MC-RR in rats through kidney injury in single dose exposure; the kidney of SD rats is more sensitive to MC-RR than that of KM mouse; and polydipsia and polyuria in mice exposed to MC-RR for 28 days occurred but could not be attributed to kidney damage.
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Affiliation(s)
| | | | | | | | | | - Kangding Gu
- MOE Key Lab of Environment and Health, Institute of Environmental Medicine, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; (W.X.); (Q.Z.); (F.S.); (W.W.); (Z.Z.)
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19
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Chernoff N, Hill D, Lang J, Schmid J, Farthing A, Huang H. Dose-Response Study of Microcystin Congeners MCLA, MCLR, MCLY, MCRR, and MCYR Administered Orally to Mice. Toxins (Basel) 2021; 13:86. [PMID: 33498948 PMCID: PMC7911753 DOI: 10.3390/toxins13020086] [Citation(s) in RCA: 18] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2020] [Revised: 01/18/2021] [Accepted: 01/19/2021] [Indexed: 01/25/2023] Open
Abstract
Microcystins are common freshwater cyanobacterial toxins that affect liver function. The toxicities of five microcystin congeners (microcystin-LA (MCLA), MCLR, MCLY, MCRR, and MCYR) commonly observed in harmful algal blooms (HABs) were evaluated in BALB/c mice after a single oral administration of doses ranging from those that were no observed adverse effect levels (NOAELs) to lowest observed adverse effect levels (LOAELs). Animals were monitored for changes in behavior and appearance, and euthanized 24 h after dosing. Test endpoints included clinical changes, necropsy observations, and serum indicators of hepatic toxicity and general homeostasis. Doses were 0.5-7 mg/kg MCLA, 0.5-11 mg/kg MCLR, 1-7 mg/kg MCLY, 7-22 mg/kg MCRR, and 3-11 mg/kg MCYR. MCLA at 3 mg/kg elevated liver/body weight ratio and liver score, ALT, AST, and GLDH, indicating hepatic toxicity, reduced serum glucose and highly elevated total serum bilirubin. MCLR and MCLY induced similar effects with LOAELs of 5 mg/kg, although a greater extent and severity of effects were observed in MCLR animals. MCRR exposure at 22 mg/kg was associated with reduced serum glucose. MCYR induced scattered liver effects at 7 mg/kg and reduced serum glucose levels at 5 mg/kg. The results indicate significant differences in congener-induced toxicity after microcystin exposure.
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Affiliation(s)
- Neil Chernoff
- Center for Public Health and Environmental Assessment, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA; (D.H.); (J.S.)
| | - Donna Hill
- Center for Public Health and Environmental Assessment, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA; (D.H.); (J.S.)
| | - Johnsie Lang
- Oak Ridge Institute for Science and Education, Oak Ridge, TN 37831, USA; (J.L.); (A.F.); (H.H.)
| | - Judith Schmid
- Center for Public Health and Environmental Assessment, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA; (D.H.); (J.S.)
| | - Amy Farthing
- Oak Ridge Institute for Science and Education, Oak Ridge, TN 37831, USA; (J.L.); (A.F.); (H.H.)
| | - Hwa Huang
- Oak Ridge Institute for Science and Education, Oak Ridge, TN 37831, USA; (J.L.); (A.F.); (H.H.)
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Guilin Z, Pengyu Z, Wei L, Fengqi H, Chen F, Yu Y, Yue H, Yuting T. Reduction of gut microbial diversity and short chain fatty acids in BALB/c mice exposure to microcystin-LR. ECOTOXICOLOGY (LONDON, ENGLAND) 2020; 29:1347-1357. [PMID: 32772242 DOI: 10.1007/s10646-020-02254-9] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Accepted: 07/09/2020] [Indexed: 06/11/2023]
Abstract
Gut microbiota has been shown to play critical roles in host health. The present study was to determine the toxicological effects of microcystin-LR (MCLR) on gut microbial community and metabolites using 16S rDNA sequencing and gas chromatography-mass spectrometry (GC-MS). MCLR was administered to BALB/c mice by gavage for eight weeks. Results of the microbial alpha-diversity (Sobs, Chao1, ACE and Shannon indexes) decreased in MCLR-treated group versus controls. Phylum Candidatus Saccharibacteria decreased significantly in MCLR-treated group versus controls. Correspondingly, more than thirties genera in relative abundance decreased, especially short chain fatty acid (SCFA)-producing bacteria (e.g., Alistipes and Ruminococcus). These results indicated that the gut microbial community structure was remarkably changed by MCLR. Furthermore, concentrations of SCFAs were significantly decreased after MCLR exposure (P < 0.01), where butyrate decreased as high as 4.9-fold. Consequently, sub-chronic exposure to MCLR could not only alter the microbial composition but metabolites. This study offered novel insights into the toxic mechanism of MCs from gut microbiota, and facilitated further clarification of risks to human health from MCs exposure.
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Affiliation(s)
- Zhang Guilin
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Zhang Pengyu
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Li Wei
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China.
- Wuhan Haidian Foreign Language Shi Yan School, Wuhan, 430220, China.
| | - Hao Fengqi
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Fangmei Chen
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Yang Yu
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Hou Yue
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
| | - Tao Yuting
- Changchun University of Science and Technology, School of Life Science and Technology, Changchun, 130022, China
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Toxin Degradation by Rumen Microorganisms: A Review. Toxins (Basel) 2020; 12:toxins12100664. [PMID: 33092236 PMCID: PMC7590051 DOI: 10.3390/toxins12100664] [Citation(s) in RCA: 50] [Impact Index Per Article: 10.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2020] [Revised: 10/07/2020] [Accepted: 10/13/2020] [Indexed: 12/21/2022] Open
Abstract
Animal feeds may contain exogenous compounds that can induce toxicity when ruminants ingest them. These toxins are secondary metabolites originating from various sources including plants, bacteria, algae and fungi. Animal feed toxins are responsible for various animal poisonings which negatively impact the livestock industry. Poisoning is more frequently reported in newly exposed, naïve ruminants while ‘experienced’ ruminants are observed to better tolerate toxin-contaminated feed. Ruminants can possess detoxification ability through rumen microorganisms with the rumen microbiome able to adapt to utilise toxic secondary metabolites. The ability of rumen microorganisms to metabolise these toxins has been used as a basis for the development of preventative probiotics to confer resistance against the poisoning to naïve ruminants. In this review, detoxification of various toxins, which include plant toxins, cyanobacteria toxins and plant-associated fungal mycotoxins, by rumen microorganisms is discussed. The review will include clinical studies of the animal poisoning caused by these toxins, the toxin mechanism of action, toxin degradation by rumen microorganisms, reported and hypothesised detoxification mechanisms and identified toxin metabolites with their toxicity compared to their parent toxin. This review highlights the commercial potential of rumen inoculum derived probiotics as viable means of improving ruminant health and production.
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Sedan D, Malaissi L, Vaccarini CA, Ventosi E, Laguens M, Rosso L, Giannuzzi L, Andrinolo D. [D-Leu 1]MC-LR Has Lower PP1 Inhibitory Capability and Greater Toxic Potency than MC-LR in Animal and Plant Tissues. Toxins (Basel) 2020; 12:toxins12100632. [PMID: 33019556 PMCID: PMC7600089 DOI: 10.3390/toxins12100632] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2020] [Revised: 09/15/2020] [Accepted: 09/21/2020] [Indexed: 01/04/2023] Open
Abstract
Two microcystins, MC-LR and [D-Leu1]MC-LR, present in La Plata Basin blooms, are differentiated by substitution of D-Alanine for D-Leucine at position 1. Our objective was to evaluate acute toxicity of [D-Leu1]MC-LR and MC-LR in mice (N:NIH Swiss) and beans (Phaseolus vulgaris). We observed variations in [D-Leu1]MC-LR lethal doses with respect to those reported for MC-LR (100 μg/kg), with an increased liver/body weight ratio and intrahepatic hemorrhages in mice exposed to 50–200 μg [D-Leu1]MC-LR/kg and slight steatosis after a single 25 μg [D-Leu1]MC-LR/kg i.p. dose. Our study in the plant model showed alterations in germination, development, morphology and TBARs levels after a single contact with the toxins during imbibition (3.5 and 15 µg/mL), those treated with [D-Leu1]MC-LR being more affected than those treated with the same concentration of MC-LR. Protein phosphatase 1 (PP1) IC50 values were 40.6 nM and 5.3 nM for [D-Leu1]MC-LR and MC-LR, respectively. However, the total phosphatase activity test in root homogenate showed 60% inhibition for [D-Leu1]MC-LR and 12% for MC-LR. In mouse liver homogenate, 50% inhibition was observed for [D-Leu1]MC-LR and 40% for MC-LR. Our findings indicate the need for further research into [D-Leu1]MC-LR toxicity since together with oxidative stress, the possible inhibition of other phosphatases could explain the differences detected in the potency of the two toxins.
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Affiliation(s)
- Daniela Sedan
- Center for Environmental Research (CIM), National Council for Scientific and Technical Research (CONICET), National University of La Plata (UNLP), La Plata 1900, Argentina
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Luciano Malaissi
- Center for Environmental Research (CIM), National Council for Scientific and Technical Research (CONICET), National University of La Plata (UNLP), La Plata 1900, Argentina
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Cristian Adrián Vaccarini
- Center for Environmental Research (CIM), National Council for Scientific and Technical Research (CONICET), National University of La Plata (UNLP), La Plata 1900, Argentina
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Ezequiel Ventosi
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Martín Laguens
- Pathology B Cathedra, School of Medical Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Lorena Rosso
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
| | - Leda Giannuzzi
- Research Center in Food Cryotechnology (CIDCA), National Council for Scientific and Technical Research (CONICET), La Plata 1900, Argentina
| | - Darío Andrinolo
- Center for Environmental Research (CIM), National Council for Scientific and Technical Research (CONICET), National University of La Plata (UNLP), La Plata 1900, Argentina
- Area of Toxicology, School of Exact Sciences, National University of La Plata (UNLP), La Plata 1900, Argentina
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Kong X, Ma J, Le-Clech P, Wang Z, Tang CY, Waite TD. Management of concentrate and waste streams for membrane-based algal separation in water treatment: A review. WATER RESEARCH 2020; 183:115969. [PMID: 32721703 DOI: 10.1016/j.watres.2020.115969] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/22/2020] [Revised: 05/09/2020] [Accepted: 05/21/2020] [Indexed: 06/11/2023]
Abstract
Frequent occurrence of harmful algal blooms (HABs) and red tides in freshwater and seawater poses serious threats to water treatment and drives the application of membrane-based technologies in algal separation. Despite the high removal efficiency of algal cells and their metabolites (e.g. organic matter and toxins) by membranes, the generation of concentrate and waste streams presents a major challenge. In this paper, we review the scenarios under which membrane-based processes are integrated with algal separation, with particular attention given to (i) drinking water production and desalination at low algal concentrations and (ii) cyanobacteria-laden water treatment/desalination. The concentrate and waste streams from backwashing and membrane cleaning in each scenario are characterised with this information facilitating a better understanding of the transport of algal cells and metabolites in membrane processes. Current strategies and gaps in managing concentrate and waste streams are identified with guidance and perspectives for future studies discussed in an Eisenhower framework.
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Affiliation(s)
- Xiangtong Kong
- UNSW Water Research Centre, School of Civil and Environmental Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia.
| | - Jinxing Ma
- UNSW Water Research Centre, School of Civil and Environmental Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia.
| | - Pierre Le-Clech
- UNESCO Centre for Membrane Science and Technology, School of Chemical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia.
| | - Zhiwei Wang
- State Key Laboratory of Pollution Control and Resource Reuse, School of Environmental Science and Engineering, Tongji University, Shanghai, 200092, China; Shanghai Institute of Pollution Control and Ecological Security, Shanghai, 200092, China.
| | - Chuyang Y Tang
- Department of Civil Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong, China.
| | - T David Waite
- UNSW Water Research Centre, School of Civil and Environmental Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia; Shanghai Institute of Pollution Control and Ecological Security, Shanghai, 200092, China.
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The Comparative Toxicity of 10 Microcystin Congeners Administered Orally to Mice: Clinical Effects and Organ Toxicity. Toxins (Basel) 2020; 12:toxins12060403. [PMID: 32570788 PMCID: PMC7354475 DOI: 10.3390/toxins12060403] [Citation(s) in RCA: 45] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2020] [Revised: 06/16/2020] [Accepted: 06/16/2020] [Indexed: 12/16/2022] Open
Abstract
Microcystins (MCs) are common cyanobacterial toxins that occur in freshwaters worldwide. Only two of the >200 MC variants have been tested for potential toxicity after oral exposure. This paper reports on the toxicity of 10 different MC congeners identified in algal blooms, microcystin-LR (MCLR), MCLA, MCLF, MCLW, MCLY, MCRR, [Asp3]MCRR, [Asp3,Dhb7]MCRR, MCWR, and MCYR after single administrations to BALB/c mice. In a preliminary MCLR dose–response study of 3 to 9 mg/kg doses, ≥5 mg/kg induced clinical changes, increased serum levels of ALT, AST, and GLDH, liver congestion, increased liver/body weight ratios, and reduced serum glucose and total protein. Based on the extent of these effects, the 10 congeners were administered as single 7 mg/kg oral doses and toxicity evaluated. The greatest toxicity was observed with MCLA and MCLR including a high percentage of moribundity. In addition to eliciting effects similar to those listed above for MCLR, MCLA also induced serum alterations indicative of jaundice. MCLY, and MCYR induced changes like those noted with MCLR, but to lesser extents. MCLW and MCLF exhibited some serum and morphological changes associated with hepatic toxicity, while there were few indications of toxicity after exposures to MCRR, [Asp3]MCRR, [Asp3,Dhb7]MCRR, or MCWR. These data illustrate a wide spectrum of hepatic effects and different potencies of these MC congeners.
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Díez-Quijada L, Medrano-Padial C, Llana-Ruiz-Cabello M, Cătunescu GM, Moyano R, Risalde MA, Cameán AM, Jos Á. Cylindrospermopsin-Microcystin-LR Combinations May Induce Genotoxic and Histopathological Damage in Rats. Toxins (Basel) 2020; 12:E348. [PMID: 32466519 PMCID: PMC7354441 DOI: 10.3390/toxins12060348] [Citation(s) in RCA: 21] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/08/2020] [Revised: 05/21/2020] [Accepted: 05/23/2020] [Indexed: 12/28/2022] Open
Abstract
Cylindrospermopsin (CYN) and microcystins (MC) are cyanotoxins that can occur simultaneously in contaminated water and food. CYN/MC-LR mixtures previously investigated in vitro showed an induction of micronucleus (MN) formation only in the presence of the metabolic fraction S9. When this is the case, the European Food Safety Authority recommends a follow up to in vivo testing. Thus, rats were orally exposed to 7.5 + 75, 23.7 + 237, and 75 + 750 μg CYN/MC-LR/kg body weight (b.w.). The MN test in bone marrow was performed, and the standard and modified comet assays were carried out to measure DNA strand breaks or oxidative DNA damage in stomach, liver, and blood cells. The results revealed an increase in MN formation in bone marrow, at all the assayed doses. However, no DNA strand breaks nor oxidative DNA damage were induced, as shown in the comet assays. The histopathological study indicated alterations only in the highest dose group. Liver was the target organ showing fatty degeneration and necrotic hepatocytes in centrilobular areas, as well as a light mononuclear inflammatory periportal infiltrate. Additionally, the stomach had flaking epithelium and mild necrosis of epithelial cells. Therefore, the combined exposure to cyanotoxins may induce genotoxic and histopathological damage in vivo.
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Affiliation(s)
- Leticia Díez-Quijada
- Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González n2, 41012 Sevilla, Spain; (L.D.-Q.); (C.M.-P.); (M.L.-R.-C.); (Á.J.)
| | - Concepción Medrano-Padial
- Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González n2, 41012 Sevilla, Spain; (L.D.-Q.); (C.M.-P.); (M.L.-R.-C.); (Á.J.)
| | - María Llana-Ruiz-Cabello
- Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González n2, 41012 Sevilla, Spain; (L.D.-Q.); (C.M.-P.); (M.L.-R.-C.); (Á.J.)
| | - Giorgiana M. Cătunescu
- University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Calea Mănăștur 3-5, 400372 Cluj-Napoca, Romania;
| | - Rosario Moyano
- Department of Pharmacology, Toxicology and Legal and Forensic Medicine, Faculty of Veterinary Medicine, University of Córdoba, Campus de Rabanales, 14014 Córdoba, Spain;
| | - Maria A. Risalde
- Animal Pathology Department. Faculty of Veterinary Medicine, University of Córdoba, Campus Universitario de Rabanales s/n, 14014 Cordoba, Spain;
- Instituto Maimonides de Investigación Biomédica de Córdoba (IMIBIC)-Hospital Universitario Reina Sofía de Córdoba-Universidad de Córdoba, Avenida Menendez Pidal s/n, 14006 Cordoba, Spain
| | - Ana M. Cameán
- Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González n2, 41012 Sevilla, Spain; (L.D.-Q.); (C.M.-P.); (M.L.-R.-C.); (Á.J.)
| | - Ángeles Jos
- Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González n2, 41012 Sevilla, Spain; (L.D.-Q.); (C.M.-P.); (M.L.-R.-C.); (Á.J.)
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Liu J, Huang Y, Cai F, Dang Y, Liu C, Wang J. MicroRNA-181a regulates endoplasmic reticulum stress in offspring of mice following prenatal microcystin-LR exposure. CHEMOSPHERE 2020; 240:124905. [PMID: 31563103 DOI: 10.1016/j.chemosphere.2019.124905] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/17/2019] [Revised: 09/17/2019] [Accepted: 09/17/2019] [Indexed: 06/10/2023]
Abstract
Microcystin-LR (MCLR) was commonly regarded as a potent hepatotoxin and has been reported to cause neurotoxicity. This study was aimed to investigate how maternal MCLR exposure during pregnancy alters behavioral responses in offspring mice and the possible molecular mechanism involved in this procedure. Three doses of MCLR solutions (0, 3 or 15 μg/kg body weight) were administered subcutaneously to pregnant C57bl/6 from gestation day (GD) 6-19. Our results showed that MCLR prenatal exposure led to the impairment of learning and memory function in offspring on postnatal days (PND) 35, accompanied by endoplasmic reticulum (ER) stress and neuronal apoptosis in hippocampal CA1 regions of mice. Sixteen miRNAs in hippocampus of pups on PND 35 were significantly affected by MCLR exposure with the markedly decreased transcription of miR-181a-5p. We then found that miR-181a-5p was down-regulated, accompanied by activation of ER stress after prenatal exposure to MCLR using qPCR analysis. Furthermore, glucose-regulated protein, 78kDa/binding immunoglobulin protein (Grp78/BIP), a major ER chaperone and signaling regulator, was identified as a target of miR-181a-5p. Our study showed that miR-181a could lead to a decrease in the mRNA expression and protein levels of Grp78 by directly binding to its 3'-untranslated region (3'-UTR) in primary hippocampal neurons. Our findings indicate that the up-regulation of Grp78 mediated by inhibition of miR-181a-5p is a possible mechanism resulting in ER stress and cognitive impairment in pups following prenatal MCLR exposure.
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Affiliation(s)
- Jue Liu
- Department of Pharmacy, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430014, China
| | - Yangyang Huang
- Fisheries College, Huazhong Agricultural University, Wuhan, 430070, China
| | - Fei Cai
- Hubei Province Key Laboratory on Cardiovascular, Cerebrovascular, and Metabolic Disorders, Hubei University of Science and Technology, Xianning, 437100, Hubei, China; Department of Pharmacology, Hubei University of Science and Technology, Xianning, 437100, China
| | - Yao Dang
- Fisheries College, Huazhong Agricultural University, Wuhan, 430070, China
| | - Chunsheng Liu
- Fisheries College, Huazhong Agricultural University, Wuhan, 430070, China
| | - Jianghua Wang
- Fisheries College, Huazhong Agricultural University, Wuhan, 430070, China.
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Protective effects of melatonin and N-acetyl cysteine against oxidative stress induced by microcystin-LR on cardiac muscle tissue. Toxicon 2019; 169:38-44. [PMID: 31465783 DOI: 10.1016/j.toxicon.2019.08.005] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/18/2019] [Revised: 08/19/2019] [Accepted: 08/21/2019] [Indexed: 12/18/2022]
Abstract
Microcystin Leucine-Arginine (MC-LR) is a toxin produced by the cyanobacteria Microcystis aeruginosa. It is the most encountered and toxic type of cyanotoxins. Oxidative stress was shown to play a role in the pathogenesis of microcystin LR by the induction of intracellular reactive oxygen species (ROS) formation that oxidize and damage cellular macromolecules. In the present study we examined the effect of acute MC-LR dose on the cardiac muscle of BALB/c mice. Afterwards, melatonin and N-acetyl cysteine (NAC) were assayed and evaluated as potential protective and antioxidant agents against damages generated by MC-LR. For this purpose, thirty mice were assigned into six groups of five mice each. The effect of MC-LR was first compared to the control group supplied with distilled water, then compared to the other groups supplied with melatonin and NAC. The experiment lasted 10 days after which animals were euthanized. Biomarkers of toxicity such as alkaline phosphatase activity, lipid peroxidation, protein carbonyl content, reduced glutathione content, serum lactate dehydrogenase and serum sorbitol dehydrogenase were assayed. Results showed that toxin treated mice have experienced significant oxidative damage in their myocardial tissue as revealed by noticeable levels of oxidative stress biomarkers and by the reduction in alkaline phosphatase activity. Whereas, melatonin and NAC treated mice manifested lesser oxidative damages. Our findings suggest a potential therapeutic use of melatonin and N-acetyl cysteine as antioxidant protective agents against oxidative damage induced by MC-LR.
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Svirčev Z, Lalić D, Bojadžija Savić G, Tokodi N, Drobac Backović D, Chen L, Meriluoto J, Codd GA. Global geographical and historical overview of cyanotoxin distribution and cyanobacterial poisonings. Arch Toxicol 2019; 93:2429-2481. [DOI: 10.1007/s00204-019-02524-4] [Citation(s) in RCA: 220] [Impact Index Per Article: 36.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/26/2019] [Accepted: 07/18/2019] [Indexed: 10/26/2022]
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Mecina GF, Chia MA, Cordeiro-Araújo MK, Bittencourt-Oliveira MDC, Varela RM, Torres A, González Molinillo JM, Macías FA, da Silva RMG. Effect of flavonoids isolated from Tridax procumbens on the growth and toxin production of Microcystis aeruginos. AQUATIC TOXICOLOGY (AMSTERDAM, NETHERLANDS) 2019; 211:81-91. [PMID: 30954847 DOI: 10.1016/j.aquatox.2019.03.011] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/24/2019] [Revised: 03/14/2019] [Accepted: 03/14/2019] [Indexed: 06/09/2023]
Abstract
The excessive proliferation of toxin producing cyanobacteria constitutes a significant health risk to the environment and humans. This is due to the contamination of potable water and accumulation of cyanotoxins in plant and animal tissues. As a means of controlling bloom forming cyanobacteria, secondary metabolites with pro-oxidative activities from plants are used to treat water bodies contaminated with cyanobacterial blooms and their associated toxins. The objective of the present study was to evaluate the mechanism of action of extract, fractions and isolated flavonoids of Tridax procumbens L. on Microcystis aeruginosa (Kützing) Kützing. by monitoring changes in growth, oxidative stress, antioxidant response, and cyanatoxin microcystins (MCs) production. The extract, fraction 3 and the isolated flavonoids significantly reduced the cell density of the cyanobacterium. Furthermore, the extract and fraction 3 increased the production of reactive oxygen species, induced lipid peroxidation, and altered antioxidant enzyme activities of M. aeruginosa. The total MCs content of the cyanobacterium was negatively affected by the presence of the extract, fractions and isolated flavonoids. The present study show that T. procumbens has secondary metabolites that are capable of interfering with the physiology and microcystins production of M. aeruginosa. These characteristics are promising for the control of this noxious cyanobacterium in aquatic ecosystems.
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Affiliation(s)
- Gustavo Franciscatti Mecina
- São Paulo State University (UNESP), School of Sciences, Humanities and Languages Assis, Department of Biotechnology, Laboratory of Herbal Medicine and Natural Products, Avenida Dom Antônio 2100, CEP: 19806-900, Assis, São Paulo, Brazil; São Paulo State University (UNESP), Institute of Chemistry, Rua Prof. Francisco Degni 55, CEP: 14800-060, Araraquara, São Paulo, Brazil
| | - Mathias Ahii Chia
- Department of Botany, Ahmadu Bello University, Zaria, 810001 Nigeria
| | - Micheline Kézia Cordeiro-Araújo
- Departamento de Ciências Biológicas, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, São Paulo, Brazil
| | - Maria do Carmo Bittencourt-Oliveira
- Departamento de Ciências Biológicas, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, São Paulo, Brazil
| | - Rosa Maria Varela
- Allelopathy Group, Department of Organic Chemistry, Institute of Biomolecules (INBIO), Campus de Excelencia Internacional 6 (ceiA3), School of Science, University of Cadiz, C/República Saharaui 7, 11510, Puerto Real, Cadiz, Spain
| | - Ascensión Torres
- Allelopathy Group, Department of Organic Chemistry, Institute of Biomolecules (INBIO), Campus de Excelencia Internacional 6 (ceiA3), School of Science, University of Cadiz, C/República Saharaui 7, 11510, Puerto Real, Cadiz, Spain
| | - José María González Molinillo
- Allelopathy Group, Department of Organic Chemistry, Institute of Biomolecules (INBIO), Campus de Excelencia Internacional 6 (ceiA3), School of Science, University of Cadiz, C/República Saharaui 7, 11510, Puerto Real, Cadiz, Spain
| | - Francisco Antonio Macías
- Allelopathy Group, Department of Organic Chemistry, Institute of Biomolecules (INBIO), Campus de Excelencia Internacional 6 (ceiA3), School of Science, University of Cadiz, C/República Saharaui 7, 11510, Puerto Real, Cadiz, Spain
| | - Regildo Márcio Gonçalves da Silva
- São Paulo State University (UNESP), School of Sciences, Humanities and Languages Assis, Department of Biotechnology, Laboratory of Herbal Medicine and Natural Products, Avenida Dom Antônio 2100, CEP: 19806-900, Assis, São Paulo, Brazil; São Paulo State University (UNESP), Institute of Chemistry, Rua Prof. Francisco Degni 55, CEP: 14800-060, Araraquara, São Paulo, Brazil.
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Impact of Microcystin-LR on Liver Function Varies by Dose and Sex in Mice. Toxins (Basel) 2018; 10:toxins10110435. [PMID: 30373283 PMCID: PMC6266648 DOI: 10.3390/toxins10110435] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2018] [Revised: 10/22/2018] [Accepted: 10/26/2018] [Indexed: 12/18/2022] Open
Abstract
Microcystin (MC) exposure is an increasing concern because more geographical locations are covered with cyanobacterial blooms as eutrophication and bloom-favoring environmental factors become more prevalent worldwide. Acute MC exposure has been linked to gastrointestinal distress, liver toxicity, and death in extreme circumstances. The goal of this study was to provide an accurate and comprehensive description of MC-LRs impacts on liver pathology, clinical chemistry, and gap junction intercellular communication (GJIC) in CD-1 male and female mice. Mice were exposed to 0, 3000, and 5000/4000 µg/kg/day MC-LR, daily for 7 days, and were necropsied on Day 8. Blood samples for clinical chemistry analysis were processed to serum, while liver sections were fixed for histopathology or evaluated for GJIC using fluorescent cut-load dye. Results show a dose-dependent relationship with MC-LR exposure and hepatocellular hypertrophy, degradation, and necrosis. Clinical chemistry parameters alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, and cholesterol increased significantly in MC-LR exposed mice. Clinical chemistry parameter analysis showed significantly increased susceptibility to MC-LR in females compared to males. Changes in GJIC were not noted, but localization of hepatotoxicity near the central veins and midlobular areas was seen. Future toxicity studies involving MCs should consider response differences across sexes, differing MC congeners, and combinatorial exposures involving other cyanotoxins.
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Scoglio S. Microcystins in water and in microalgae: Do microcystins as microalgae contaminants warrant the current public alarm? Toxicol Rep 2018; 5:785-792. [PMID: 30105209 PMCID: PMC6086208 DOI: 10.1016/j.toxrep.2018.07.002] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/24/2018] [Revised: 07/08/2018] [Accepted: 07/24/2018] [Indexed: 12/31/2022] Open
Abstract
Microcystins have been the subject of increasingly alarming popular and scientific articles, which have taken as their unquestionable foundation the provisional Guideline of 1 μg/L established by the WHO Panel on microcystins levels in water, and mechanically translated by the Oregon government as 1 μg/g of Klamath Aphanizomenon flos aquae microalgae. This article underlines the significant limitations and ultimately scientific untenability of the WHO Guideline on microcystins in water, for being based on testing methodologies which may lead to a significant overestimation of the toxicity of microcystins. I propose criteria for the realization of new experimental studies on the toxicity of microcystins, based on the essential understanding that drinking water is contaminated by whole cyanobacterial microalgae rather than purified microcystins, while it is important to differentiate between water and cyanobacterial supplements. It is indeed a mistake to automatically apply standards that are proper for water to cyanobacterial supplements, as they have different concentrations of the antioxidant substances that inactivate or significantly reduce the toxicity of microcystins, a fact that also require that each cyanobacterial supplement be tested individually and through realistic testing methodologies.
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Chen L, Giesy JP, Xie P. The dose makes the poison. THE SCIENCE OF THE TOTAL ENVIRONMENT 2018; 621:649-653. [PMID: 29197283 DOI: 10.1016/j.scitotenv.2017.11.218] [Citation(s) in RCA: 41] [Impact Index Per Article: 5.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/21/2017] [Revised: 11/18/2017] [Accepted: 11/19/2017] [Indexed: 06/07/2023]
Abstract
Some microcystins (MCs) might cause hepatotoxicity in animals and humans. MC-LR is also a tumor promoter and a suspect carcinogen. In 2010, the International Agency for Research on Cancer (IARC) classified MC-LR as a possible human carcinogen (Group 2B). Recently, an article entitled "Long-term, low-dose exposure to microcystin toxin does not increase the risk of liver tumor development or growth in mice" was published in Hepatology Research by Meaghan Labine and Gerald Y. Minuk. However, the experimental design was flawed and the conclusion is misleading. 1μg/L MC-LR in drinking water is the provisional guideline value established by the World Health Organization (WHO) for humans in 1998, based on a tolerable daily intake (TDI) of 0.04μg/kg body mass (BM). Assuming the mice drink 1.5mL/10g BM of water per day, the exposure dose would be 0.15μg/kg/d BM, about 270-fold less than 40μg/kg/d, the no-observed-adverse-effect level (NOAEL). Thus, the dose of MC-LR was too small and "unlikely to result in liver tumor development or enhance existing tumor growth", even with a long-term (28weeks) exposure. Presumably, they didn't consider inter-species variations between mice and humans, including toxicokinetics and toxicodynamics. Ranges of "low-dose" MCs for animals and humans should be defined. Also, the authors misunderstood or misrepresented several previous studies. Before drawing final conclusions on the carcinogenicity of MCs, further well-designed experiments are warranted.
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Affiliation(s)
- Liang Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; University of Chinese Academy of Sciences, Beijing 100049, China.
| | - John P Giesy
- Department of Veterinary Biomedical Sciences and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan S7N5B3, Canada.
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining 810016, China.
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Nielsen DS, Shepherd NE, Xu W, Lucke AJ, Stoermer MJ, Fairlie DP. Orally Absorbed Cyclic Peptides. Chem Rev 2017; 117:8094-8128. [PMID: 28541045 DOI: 10.1021/acs.chemrev.6b00838] [Citation(s) in RCA: 294] [Impact Index Per Article: 36.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022]
Abstract
Peptides and proteins are not orally bioavailable in mammals, although a few peptides are intestinally absorbed in small amounts. Polypeptides are generally too large and polar to passively diffuse through lipid membranes, while most known active transport mechanisms facilitate cell uptake of only very small peptides. Systematic evaluations of peptides with molecular weights above 500 Da are needed to identify parameters that influence oral bioavailability. Here we describe 125 cyclic peptides containing four to thirty-seven amino acids that are orally absorbed by mammals. Cyclization minimizes degradation in the gut, blood, and tissues by removing cleavable N- and C-termini and by shielding components from metabolic enzymes. Cyclization also folds peptides into bioactive conformations that determine exposure of polar atoms to solvation by water and lipids and therefore can influence oral bioavailability. Key chemical properties thought to influence oral absorption and bioavailability are analyzed, including molecular weight, octanol-water partitioning, hydrogen bond donors/acceptors, rotatable bonds, and polar surface area. The cyclic peptides violated to different degrees all of the limits traditionally considered to be important for oral bioavailability of drug-like small molecules, although fewer hydrogen bond donors and reduced flexibility generally favored oral absorption.
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Affiliation(s)
- Daniel S Nielsen
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
| | - Nicholas E Shepherd
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
| | - Weijun Xu
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
| | - Andrew J Lucke
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
| | - Martin J Stoermer
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
| | - David P Fairlie
- Division of Chemistry and Structural Biology, and ‡Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Institute for Molecular Bioscience, The University of Queensland , Brisbane, QLD 4072, Australia
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Buratti FM, Manganelli M, Vichi S, Stefanelli M, Scardala S, Testai E, Funari E. Cyanotoxins: producing organisms, occurrence, toxicity, mechanism of action and human health toxicological risk evaluation. Arch Toxicol 2017; 91:1049-1130. [DOI: 10.1007/s00204-016-1913-6] [Citation(s) in RCA: 385] [Impact Index Per Article: 48.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2016] [Accepted: 12/13/2016] [Indexed: 12/11/2022]
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Affiliation(s)
- Junguo Ma
- College of Life Science, Henan Normal University, Xinxiang, China
| | - Xiaoyu Li
- College of Life Science, Henan Normal University, Xinxiang, China
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Microcystin-LR induced liver injury in mice and in primary human hepatocytes is caused by oncotic necrosis. Toxicon 2016; 125:99-109. [PMID: 27889601 DOI: 10.1016/j.toxicon.2016.11.254] [Citation(s) in RCA: 28] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/21/2016] [Revised: 11/16/2016] [Accepted: 11/22/2016] [Indexed: 11/22/2022]
Abstract
Microcystins are a group of toxins produced by freshwater cyanobacteria. Uptake of microcystin-leucine arginine (MC-LR) by organic anion transporting polypeptide 1B2 in hepatocytes results in inhibition of protein phosphatase 1A and 2A, and subsequent cell death. Studies performed in primary rat hepatocytes demonstrate prototypical apoptosis after MC-LR exposure; however, no study has directly tested whether apoptosis is critically involved in vivo in the mouse, or in human hepatocytes. MC-LR (120 μg/kg) was administered to C57BL/6J mice and cell death was evaluated by alanine aminotransferase (ALT) release, caspase-3 activity in the liver, and histology. Mice exposed to MC-LR had increases in plasma ALT values, and hemorrhage in the liver, but no increase in capase-3 activity in the liver. Pre-treatment with the pan-caspase inhibitor z-VAD-fmk failed to protect against cell death measured by ALT, glutathione depletion, or hemorrhage. Administration of MC-LR to primary human hepatocytes resulted in significant toxicity at concentrations between 5 nM and 1 μM. There were no elevated caspase-3 activities and pretreatment with z-VAD-fmk failed to protect against cell death in human hepatocytes. MC-LR treated human hepatocytes stained positive for propidium iodide, indicating membrane instability, a marker of necrosis. Of note, both increases in PI positive cells, and increases in lactate dehydrogenase release, occurred before the onset of complete actin filament collapse. In conclusion, apoptosis does not contribute to MC-LR-induced cell death in the in vivo mouse model or in primary human hepatocytes in vitro. Thus, targeting necrotic cell death mechanisms will be critical for preventing microcystin-induced liver injury.
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Jung JM, Lee J, Kim J, Kim KH, Kim HW, Jeon YJ, Kwon EE. Enhanced thermal destruction of toxic microalgal biomass by using CO2. THE SCIENCE OF THE TOTAL ENVIRONMENT 2016; 566-567:575-583. [PMID: 27236623 DOI: 10.1016/j.scitotenv.2016.05.161] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/26/2016] [Revised: 05/16/2016] [Accepted: 05/22/2016] [Indexed: 06/05/2023]
Abstract
This work confirmed that dominant microalgal strain in the eutrophic site (the Han River in Korea) was Microcystis aeruginosa (M. aeruginosa) secreting toxins. Collected and dried microalgal biomass had an offensive odor due to microalgal lipid, of which the content reached up to 2±0.2wt.% of microalgal biomass (dry basis). This study has validated that the offensive odor is attributed to the C3-6 range of volatile fatty acids (VFAs), which was experimentally identified by the non-catalytic transformation of triglycerides (TGs) and free fatty acids (FFAs) in microalgal biomass into fatty acid methyl esters (FAMEs). In particular, this study mechanistically investigated the influence of CO2 in the thermal destruction (i.e., pyrolysis) of hazardous microalgal biomass in order to achieve dual purposes (i.e., thermal disposal of hazardous microalgal biomass and energy recovery). The influence of CO2 in pyrolysis of microalgal biomass was identified as 1) the enhanced thermal cracking behaviors of volatile organic compounds (VOCs) from the thermal degradation of microalgal biomass and 2) the direct gas phase reaction between CO2 and VOCs. These identified influences of CO2 in pyrolysis of microalgal biomass significantly enhanced the generation of CO: the enhanced generation of CO in the presence of CO2 was 590% at 660°C, 1260% at 690°C, and 3200% at 720°C. In addition, two identified influences of CO2 (i.e., enhanced thermal cracking and direct gas phase reaction) occurred simultaneously and independently. The identified gas phase reaction in the presence of CO2 was only initiated at temperatures higher than 500°C, which was different from the Boudouard reaction. Lastly, the experimental work justified that exploiting CO2 as a reaction medium and/or chemical feedstock will provide new technical approaches for controlling syngas ratio and in-situ air pollutant control without using catalysts.
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Affiliation(s)
- Jong-Min Jung
- Department of Environment and Energy, Sejong University, Seoul 05006, Republic of Korea
| | - Jechan Lee
- Department of Environment and Energy, Sejong University, Seoul 05006, Republic of Korea
| | - Jieun Kim
- Department of Environment and Energy, Sejong University, Seoul 05006, Republic of Korea
| | - Ki-Hyun Kim
- Department of Civil and Environmental Engineering, Hanyang University, Seoul 04763, Republic of Korea
| | - Hyung-Wook Kim
- Department of Biological Science and Technology, Sejong University, Seoul 05006, Republic of Korea
| | - Young Jae Jeon
- Department of Microbiology, Pukyong National University, Busan 48513, Republic of Korea.
| | - Eilhann E Kwon
- Department of Environment and Energy, Sejong University, Seoul 05006, Republic of Korea.
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Cordeiro-Araújo MK, Chia MA, Arruda-Neto JDDT, Tornisielo VL, Vilca FZ, Bittencourt-Oliveira MDC. Microcystin-LR bioaccumulation and depuration kinetics in lettuce and arugula: Human health risk assessment. THE SCIENCE OF THE TOTAL ENVIRONMENT 2016; 566-567:1379-1386. [PMID: 27267723 DOI: 10.1016/j.scitotenv.2016.05.204] [Citation(s) in RCA: 48] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/03/2016] [Revised: 05/23/2016] [Accepted: 05/29/2016] [Indexed: 06/06/2023]
Abstract
Microcystin-LR (MC-LR) is one of the most toxic and common microcystins (MCs) variant found in aquatic ecosystems. Little is known about the possibility of recovering microcystins contaminated agricultural crops. The objectives of this study were to determine the bioaccumulation and depuration kinetics of MC-LR in leaf tissues of lettuce and arugula, and estimate the total daily intake (ToDI) of MC-LR via contaminated vegetables by humans. Arugula and lettuce were irrigated with contaminated water having 5 and 10μgL(-1) of MC-LR for 7days (bioaccumulation), and subsequently, with uncontaminated water for 7days (depuration). Quantification of MC-LR was performed by LC-MS/MS. The one-compartment biokinetic model was employed for MC-LR bioaccumulation and depuration data analysis. MC-LR was only accumulated in lettuce. After 7days of irrigation with uncontaminated water, over 25% of accumulated MC-LR was still retained in leaf tissues of plants treated with 10μgL(-1) MC-LR. Total daily toxin intake by adult consumers (60kg-bw) exceeded the 0.04μgMC-LRkg(-1) limit recommended by WHO. Bioaccumulation was found to be linearly proportional to the exposure concentration of the toxin, increasing over time; and estimated to become saturated after 30days of uninterrupted exposure. On the other hand, MC-LR depuration was less efficient at higher exposure concentrations. This is because biokinetic half-life calculations gave 2.9 and 3.7days for 5 and 10μgL(-1) MC-LR treatments, which means 29-37days are required to eliminate the toxin. For the first time, our results demonstrated the possibility of MC-LR decontamination of lettuce plants.
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Affiliation(s)
- Micheline Kézia Cordeiro-Araújo
- Department of Biological Sciences, Luiz de Queiroz College of Agriculture, University of São Paulo, Av. Pádua Dias, 11, São Dimas, CEP 13418-900 Piracicaba, SP, Brazil; Botany Graduate Program, Rural and Federal University of Pernambuco, R. Dom Manoel de Medeiros, S/N, Dois Irmãos, CEP 52171-030 Recife, PE, Brazil
| | - Mathias Ahii Chia
- Department of Biological Sciences, Luiz de Queiroz College of Agriculture, University of São Paulo, Av. Pádua Dias, 11, São Dimas, CEP 13418-900 Piracicaba, SP, Brazil
| | - João Dias de Toledo Arruda-Neto
- Institute of Physics, University of São Paulo, Rua do Matão Travessa R Nr.187, CEP 05508-090, Cidade Universitária, SP, Brazil; FESP - São Paulo Engineering College, Av. Nove de Julho, 5520, CEP 01406-200 São Paulo, SP, Brazil
| | | | - Franz Zirena Vilca
- CENA, University of São Paulo, Av. Centenário, 303, CEP 13400-970 Piracicaba, SP, Brazil
| | - Maria do Carmo Bittencourt-Oliveira
- Department of Biological Sciences, Luiz de Queiroz College of Agriculture, University of São Paulo, Av. Pádua Dias, 11, São Dimas, CEP 13418-900 Piracicaba, SP, Brazil; Botany Graduate Program, Rural and Federal University of Pernambuco, R. Dom Manoel de Medeiros, S/N, Dois Irmãos, CEP 52171-030 Recife, PE, Brazil.
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Zhao Y, Shi K, Su X, Xie L, Yan Y. Microcystin-LR induces dysfunction of insulin secretion in rat insulinoma (INS-1) cells: Implications for diabetes mellitus. JOURNAL OF HAZARDOUS MATERIALS 2016; 314:11-21. [PMID: 27107231 DOI: 10.1016/j.jhazmat.2016.04.019] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/16/2015] [Revised: 03/26/2016] [Accepted: 04/09/2016] [Indexed: 06/05/2023]
Abstract
Microcystins (MCs) are the most frequent cyanobacterial toxins observed in freshwater systems. Accumulating evidence suggests that MCs pose a serious threat to public health. However, the contributions of the exposure of MCs to the occurrence of human diseases remain largely unknown. This study provides the evidence of the effects of MC-LR on pancreatic β-cell function through the exposure of rat insulinoma (INS-1) cells to 0, 10, 20, or 40μM MC-LR for 72h and explores the underlying molecular mechanisms. Our results demonstrate that exposure to MC-LR for 72h suppresses cell viability, disturbs glucose-stimulated insulin secretion (GSIS), and decreases the expression of insulin protein. Moreover, MC-LR disrupts the cell cycle distribution and increases cell apoptosis at 20 or 40μM for 72h, respectively, indicating that the β-cell mass would be decreased by MC-LR exposure. A transcriptomic analysis revealed several key genes (e.g., Pdx-1, Neurod1, and Abcc8) involved in insulin secretion are significantly differentially expressed in INS-1 cells in response to MC-LR exposure. In addition, several signal transduction pathways associated with diabetes (e.g., type 1 and 2 diabetes) were also identified compared with the control cells. We recommend that MC be considered as a new environmental factor that promotes diabetes development. The identified key genes or pathways may potentially contribute to the future therapies in the environmental contaminants induced β-cell damage.
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Affiliation(s)
- Yanyan Zhao
- State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, 73 East Beijing Road, Nanjing 210008, PR China.
| | - Kun Shi
- State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, 73 East Beijing Road, Nanjing 210008, PR China
| | - Xiaomei Su
- State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, 73 East Beijing Road, Nanjing 210008, PR China
| | - Liqiang Xie
- State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, 73 East Beijing Road, Nanjing 210008, PR China.
| | - Yunjun Yan
- Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, PR China
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Clark SP, Davis MA, Ryan TP, Searfoss GH, Hooser SB. Hepatic Gene Expression Changes in Mice Associated with Prolonged Sublethal Microcystin Exposure. Toxicol Pathol 2016; 35:594-605. [PMID: 17654400 DOI: 10.1080/01926230701383210] [Citation(s) in RCA: 45] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
Microcystin-LR (MCLR) is an acute hepatotoxicant and suspected carcinogen. Previous chronic studies have individually described hepatic morphologic changes, or alterations in the cytoskeleton, cell signaling or redox pathways. The objective of this study was to characterize chronic effects of MCLR in wild-type mice utilizing gene array analysis, morphology, and plasma chemistries. MCLR was given daily for up to 28 days. RNA from the 28-day study was hybridized onto mouse genechip arrays. RNA from 4 hours, 24 hours, 4 days, 1 day, and 28 days for selected genes was processed for quantitative-PCR. Increases in plasma hepatic enzyme activities and decreases in total protein, albumin and glucose concentrations were identified in MCLR-treated groups at 14 and 28 days. Histologically, marked hepatokaryomegaly was identified in the 14-day MCLR group with the addition of giant cells at 28 days. Major gene transcript changes were identified in the actin organization, cell cycle, apoptotic, cellular redox, cell signaling, albumin metabolism, and glucose homeostasis pathways, and the organic anion transport polypeptide system. Using toxicogenomics, we have identified key molecular pathways involved in chronic sublethal MCLR exposure in wild-type mice, genes participating in those critical pathways and related them to cellular and morphologic alterations seen in this and other studies.
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Affiliation(s)
- Shawn P Clark
- Department of Veterinary Pathobiology, Purdue University, Indiana 47907, USA.
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Nishizawa T, Neagari Y, Miura T, Asayama M, Murata K, Harada KI, Shirai M. Molecular Analysis of the Cyanobacterial Community in Gastric Contents of Egrets with Symptoms of Steatitis. Open Microbiol J 2015; 9:160-6. [PMID: 26668668 PMCID: PMC4676040 DOI: 10.2174/1874285801509010160] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/29/2014] [Revised: 05/03/2015] [Accepted: 08/01/2015] [Indexed: 11/22/2022] Open
Abstract
Many deaths of wild birds that have drunk water contaminated with hepatotoxic microcystin-producing cyanobacteria have been reported. A mass death of egrets and herons with steatitis were found at the agricultural reservoir occurring cyanobacterial waterblooms. This study aimed to verify a hypothesis that the egrets and herons which died in the reservoir drink microcystin-producing cyanobacteria and microcystin involves in the cause of death as well as the symptoms of steatitis. The cyanobacterial community in gastric contents of egrets and herons that died from steatitis was assessed using cyanobacterial 16S rRNA-based terminal-restriction fragment length polymorphism (T-RFLP) profiling and a cyanobacterial 16S rRNA-based clone library analysis. In addition, PCR amplification of the mcyB-C region and the mcyG gene, involved in microcystin biosynthesis, was examined. The cyanobacterial community in the gastric contents of two birds showed a simplistic composition. A comparison of cyanobacterial T-RFLP profiling and cloned sequences suggested that the genus Microcystis predominated in both samples of egrets died. Although we confirmed that two egrets which died in the reservoir have taken in cyanobacterial waterblooms containing the genus Microcystis, no mcy gene was detected in both samples according to the mcy gene-based PCR analysis. This study is the first to show the profiling and traceability of a cyanobacterial community in the gastric contents of wild birds by molecular analysis. Additionally, we consider causing symptoms of steatitis in the dead egrets.
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Affiliation(s)
| | - Yasuko Neagari
- Laboratory for Intellectual Fundamentals for Environmental Studies, National Institute for Environmental Studies, Ibaraki 305-8506, Japan
| | - Takamasa Miura
- College of Agriculture, Ibaraki University, Ibaraki 300-0393, Japan
| | - Munehiko Asayama
- College of Agriculture, Ibaraki University, Ibaraki 300-0393, Japan
| | - Koichi Murata
- College of Bioresource Sciences, Nihon University, Kanagawa 252-0880, Japan
| | - Ken-Ichi Harada
- Graduate School of Environmental and Human Science and Faculty of Pharmacy, Meijo University, Aichi 468-8503, Japan
| | - Makoto Shirai
- College of Agriculture, Ibaraki University, Ibaraki 300-0393, Japan
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Bautista AC, Moore CE, Lin Y, Cline MG, Benitah N, Puschner B. Hepatopathy following consumption of a commercially available blue-green algae dietary supplement in a dog. BMC Vet Res 2015; 11:136. [PMID: 26087767 PMCID: PMC4472392 DOI: 10.1186/s12917-015-0453-2] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/06/2015] [Accepted: 06/09/2015] [Indexed: 11/15/2022] Open
Abstract
Background Dietary supplement use in both human and animals to augment overall health continues to increase and represents a potential health risk due to the lack of safety regulations imposed on the manufacturers. Because there are no requirements for demonstrating safety and efficacy prior to marketing, dietary supplements may contain potentially toxic contaminants such as hepatotoxic microcystins produced by several species of blue-green algae. Case presentation An 11-year-old female spayed 8.95 kg Pug dog was initially presented for poor appetite, lethargy polyuria, polydipsia, and an inability to get comfortable. Markedly increased liver enzyme activities were detected with no corresponding abnormalities evident on abdominal ultrasound. A few days later the liver enzyme activities were persistently increased and the dog was coagulopathic indicating substantial liver dysfunction. The dog was hospitalized for further care consisting of oral S-adenosylmethionine, silybin, vitamin K, and ursodeoxycholic acid, as well as intravenous ampicillin sodium/sulbactam sodium, dolasetron, N-acetylcysteine, metoclopramide, and intravenous fluids. Improvement of the hepatopathy and the dog’s clinical status was noted over the next three days. Assessment of the dog’s diet revealed the use of a commercially available blue-green algae dietary supplement for three-and-a-half weeks prior to hospitalization. The supplement was submitted for toxicology testing and revealed the presence of hepatotoxic microcystins (MCs), MC-LR and MC-LA. Use of the supplement was discontinued and follow-up evaluation over the next few weeks revealed a complete resolution of the hepatopathy. Conclusions To the authors’ knowledge, this is the first case report of microcystin intoxication in a dog after using a commercially available blue-green algae dietary supplement. Veterinarians should recognize the potential harm that these supplements may cause and know that with intervention, recovery is possible. In addition, more prudent oversight of dietary supplement use is recommended for our companion animals to prevent adverse events/intoxications.
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Affiliation(s)
| | - Caroline E Moore
- Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA, USA.
| | - Yanping Lin
- Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA, USA.
| | | | - Noemi Benitah
- Red Bank Veterinary Hospital, Tinton Falls, NJ, USA.
| | - Birgit Puschner
- California Animal Health and Food Safety Laboratory System, Davis, CA, USA. .,Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA, USA.
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Lin J, Chen J, He J, Chen J, Yan Q, Zhou J, Xie P. Effects of microcystin-LR on bacterial and fungal functional genes profile in rat gut. Toxicon 2015; 96:50-6. [PMID: 25617596 DOI: 10.1016/j.toxicon.2015.01.011] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/26/2014] [Revised: 10/29/2014] [Accepted: 01/20/2015] [Indexed: 10/24/2022]
Abstract
The short-term exposure to microcystin-LR (MC-LR, one of the most common and toxic variants generated by toxigenic cyanobacteria) induced gut dysfunction such as generation of reactive oxygen species, cell erosion and deficient intestinal absorption of nutrients. However, till now, little is known about its impact on gut microbial community, which has been considered as necessary metabolic assistant and stresses resistant entities for the host. This study was designed to reveal the shift of microbial functional genes in the gut of rat orally gavaged with MC-LR. GeoChip detected a high diversity of bacterial and fungal genes involved in basic metabolic processes and stress resistance. The results showed that the composition of functional genes was significantly changed in rat gut after one week of exposure to MC-LR, and we found some relatively enriched genes that are involved in carbon degradation including chitin, starch and limonene metabolism, and these genes were mainly derived from fungal and bacterial pathogens. In addition, we found large amounts of significantly enriched genes relevant to degradation of the specific carbon compounds, aromatics. The dysbiosis of bacterial and fungal flora gave an implication of pathogens invasion. The enriched gene functions could be linked to acute gastroenteritis induced by MC-LR.
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Affiliation(s)
- Juan Lin
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China; Graduate University of Chinese Academy of Sciences, Beijing 10049, PR China
| | - Jun Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China
| | - Jun He
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China; Graduate University of Chinese Academy of Sciences, Beijing 10049, PR China
| | - Jing Chen
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China; Graduate University of Chinese Academy of Sciences, Beijing 10049, PR China
| | - Qingyun Yan
- Key Laboratory of Biodiversity and Conservation of Aquatic Organisms, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
| | - Jizhong Zhou
- Institute for Environmental Genomics, Department of Microbiology and Plant Biology, University of Oklahoma, Norman, OK 73019, USA
| | - Ping Xie
- Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, PR China.
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Ziková A, Kopp R. Impacts of microcystin, a cyanobacterial toxin, on laboratory rodents in vivo. ACTA UNIVERSITATIS AGRICULTURAE ET SILVICULTURAE MENDELIANAE BRUNENSIS 2014. [DOI: 10.11118/actaun200856050263] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
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Manubolu M, Madawala SRP, Dutta PC, Malmlöf K. In vitro biodegradation of cyanotoxins in the rumen fluid of cattle. BMC Vet Res 2014; 10:110. [PMID: 24885733 PMCID: PMC4018535 DOI: 10.1186/1746-6148-10-110] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2013] [Accepted: 05/01/2014] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND In countries around the Baltic Sea grazing ruminants have access to and drink, surface water from lakes, rivers and in several coastal regions. The water quality of these naturally occurring reservoirs affects performance and health of livestock. In the Baltic Sea both microcystin (MC) and nodularin (NOD) occurs as cyclic peptides and have hepatotoxic effects. Although cattle obviously have died after consuming contaminated water very little information is available as to how susceptible ruminants are to the toxins produced by cyanobacteria. The critical question as to whether the rumen microflora might constitute a protective shield is unresolved. For this reason our aim is to investigate a possible degradation rate of these toxins in rumen. RESULTS The ability of rumen microorganisms to degrade certain important cyanotoxins (MC-LR, YR, RR and NOD) was studied in vitro by incubating with rumen fluid at three different concentrations (0.05, 0.5 and 5 μg/mL) for 3 h. The degradation efficiencies were determined by LC-MS (ESI) positive mode. Degradation was observed in the following order MC-RR 36%, NOD 35%, MC-RR 25% and MC-LR 8.9% at lower concentrations within 3 h. However, average degradation was observed at concentration of 0.5 μg/mL. No degradation was observed in higher concentrations for entire 3 h. The present results reveal that the degradation was both dose and time dependent. CONCLUSIONS In conclusion the present results suggest that the rumen microbial flora may protect ruminants from being intoxicated by Cyanotoxins.
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Affiliation(s)
- Manjunath Manubolu
- Department of Anatomy, Physiology and Biochemistry, Faculty of Veterinary Medicine and Animal Science, SLU, Box 7011 750 07 Uppsala, Sweden.
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Effects of the amino acid constituents of microcystin variants on cytotoxicity to primary cultured rat hepatocytes. Toxins (Basel) 2013; 6:168-79. [PMID: 24380975 PMCID: PMC3920255 DOI: 10.3390/toxins6010168] [Citation(s) in RCA: 25] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2013] [Revised: 12/13/2013] [Accepted: 12/24/2013] [Indexed: 12/11/2022] Open
Abstract
Microcystins, which are cyclic heptapeptides produced by some cyanobacterial species from algal blooms, strongly inhibit serine/threonine protein phosphatase and are known as hepatotoxins. Microcystins have many structural variations, yet insufficient information is available on the differences in the cytotoxic potentials among the structural variants. In this study, the cytotoxicities of 16 microcystin variants at concentrations of 0.03–10 μg/mL to primary cultured rat hepatocytes were determined by measuring cellular ATP content, and subsequently determined by their 50% inhibitory concentration (IC50). Differences in the amino acid constituents were associated with differences in cytotoxic potential. [d-Asp3, Z-Dhb7] microcystin-LR exhibited the strongest cytotoxicity at IC50 of 0.053 μg/mL among the microcystin variants tested. Furthermore, [d-Asp3, Z-Dhb7] microcystin-HtyR was also highly cytotoxic. These results suggest that both d-Asp and Z-Dhb residues are important in determining the cytotoxic potential of microcystin variants.
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In vivo effects of microcystins and complex cyanobacterial biomass on rats (Rattus norvegicus var. alba): Changes in immunological and haematological parameters. Toxicon 2013; 73:1-8. [DOI: 10.1016/j.toxicon.2013.06.016] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/14/2012] [Revised: 06/21/2013] [Accepted: 06/26/2013] [Indexed: 01/31/2023]
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Vareli K, Jaeger W, Touka A, Frillingos S, Briasoulis E, Sainis I. Hepatotoxic seafood poisoning (HSP) due to microcystins: a threat from the ocean? Mar Drugs 2013; 11:2751-68. [PMID: 23921721 PMCID: PMC3766863 DOI: 10.3390/md11082751] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2013] [Revised: 07/15/2013] [Accepted: 07/16/2013] [Indexed: 01/11/2023] Open
Abstract
Cyanobacterial blooms are a major and growing problem for freshwater ecosystems worldwide that increasingly concerns public health, with an average of 60% of blooms known to be toxic. The most studied cyanobacterial toxins belong to a family of cyclic heptapeptide hepatotoxins, called microcystins. The microcystins are stable hydrophilic cyclic heptapeptides with a potential to cause cell damage following cellular uptake via organic anion-transporting proteins (OATP). Their intracellular biologic effects presumably involve inhibition of catalytic subunits of protein phosphatases (PP1 and PP2A) and glutathione depletion. The microcystins produced by cyanobacteria pose a serious problem to human health, if they contaminate drinking water or food. These toxins are collectively responsible for human fatalities, as well as continued and widespread poisoning of wild and domestic animals. Although intoxications of aquatic organisms by microcystins have been widely documented for freshwater ecosystems, such poisonings in marine environments have only occasionally been reported. Moreover, these poisonings have been attributed to freshwater cyanobacterial species invading seas of lower salinity (e.g., the Baltic) or to the discharge of freshwater microcystins into the ocean. However, recent data suggest that microcystins are also being produced in the oceans by a number of cosmopolitan marine species, so that Hepatotoxic Seafood Poisoning (HSP) is increasingly recognized as a major health risk that follows consumption of contaminated seafood.
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Affiliation(s)
- Katerina Vareli
- Department of Biological Applications and Technologies, University of Ioannina, 45110 Ioannina, Greece; E-Mail:
- Interscience Molecular Oncology Laboratory, Human Cancer Biobank Center, University of Ioannina, 45110 Ioannina, Greece; E-Mails: (A.T.); (E.B.)
| | - Walter Jaeger
- Department of Clinical Pharmacy and Diagnostics, University of Vienna, A-1090 Vienna, Austria; E-Mail:
| | - Anastasia Touka
- Interscience Molecular Oncology Laboratory, Human Cancer Biobank Center, University of Ioannina, 45110 Ioannina, Greece; E-Mails: (A.T.); (E.B.)
| | - Stathis Frillingos
- Laboratory of Biological Chemistry, School of Medicine, University of Ioannina, 45110 Ioannina, Greece; E-Mail:
| | - Evangelos Briasoulis
- Interscience Molecular Oncology Laboratory, Human Cancer Biobank Center, University of Ioannina, 45110 Ioannina, Greece; E-Mails: (A.T.); (E.B.)
| | - Ioannis Sainis
- Interscience Molecular Oncology Laboratory, Human Cancer Biobank Center, University of Ioannina, 45110 Ioannina, Greece; E-Mails: (A.T.); (E.B.)
- Author to whom correspondence should be addressed; E-Mail: ; Tel.: +30-26-510-075-57; Fax: +30-26-510-070-64
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Baker L, Sendall BC, Gasser RB, Menjivar T, Neilan BA, Jex AR. Rapid, multiplex-tandem PCR assay for automated detection and differentiation of toxigenic cyanobacterial blooms. Mol Cell Probes 2013; 27:208-14. [PMID: 23850895 DOI: 10.1016/j.mcp.2013.07.001] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2013] [Revised: 07/01/2013] [Accepted: 07/01/2013] [Indexed: 11/26/2022]
Abstract
Cyanobacterial blooms are a major water quality issue and potential public health risk in freshwater, marine and estuarine ecosystems globally, because of their potential to produce cyanotoxins. To date, a significant challenge in the effective management of cyanobacterial has been an inability of classical microscopy-based approaches to consistently and reliably detect and differentiate toxic from non-toxic blooms. The potential of cyanobacteria to produce toxins has been linked to the presence of specific biosynthetic gene clusters. Here, we describe the application of a robotic PCR-based assay for the semi-automated and simultaneous detection of toxin biosynthesis genes of each of the toxin classes characterized to date for cyanobacteria [i.e., microcystins (MCYs), nodularins (NODs), cylindrospermopsins (CYNs) and paralytic shellfish toxins (PSTs)/saxitoxins (SXTs)]. We demonstrated high sensitivity and specificity for each assay using well-characterized, cultured isolates, and establish its utility as a quantitative PCR using DNA, clone and cell-based dilution series. In addition, we used 206 field-collected samples and 100 known negative controls to compare the performance of each assay with conventional PCR and direct toxin detection. We report a diagnostic specificity of 100% and a sensitivity of ≥97.7% for each assay.
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Affiliation(s)
- Louise Baker
- Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria 3010, Australia
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Huang X, Zhang Y, Xiao W, Ye X, Zhong Q, Gu K. Comparison of response indices to toxic microcystin-LR in blood of mice. CHEMOSPHERE 2013; 92:563-569. [PMID: 23659964 DOI: 10.1016/j.chemosphere.2013.04.006] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/25/2012] [Revised: 03/30/2013] [Accepted: 04/01/2013] [Indexed: 06/02/2023]
Abstract
In order to investigate the response indices to toxic microcystin-LR (MC-LR) in blood of mice, concentrations of free and total MC-LR in blood and tissues, accompanied by serous parameters in series including some enzymatic activities, hematology and the function of leukocytes, were determined in mice exposed to the toxin ranging from 3.125 to 25.000 μg kg(-1)day(-1) by intraperitoneal injection for 7 days. On the 7th day, the ratios of mass of free MC-LR in serum to the mass of MC-LR in given dose were 3.843-4.555%, while the ratios of total MC-LR in liver were 34.465-38.567%. Comparing the overall experimental results, the three most sensitive indices are total MC-LR in the liver, the phagocytic index and reactive oxygen species (ROS) which have shown significant differences between the lowest dose group and the control group. Alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase had proportional correlations with the MC-LR exposure doses, and the hematology of the majority of blood cells and the volume of erythrocytes were also influenced by the toxin. The alterations of some cytokines and the ROS of leukocytes were observed. The results of the studies suggest that measurement of MC-LR in blood is powerful and clear evidence to indicate that subjects have been exposed to MC-LR and can be used to discriminate from other causes leading to hepatic lesions although it is not as sensitive as other indices that are usually as useful complements to reflect the liver function.
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Affiliation(s)
- Xianing Huang
- Institute of Environmental Medicine, MOE Key Lab of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, PR China
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