Eco-friendly management of harmful cyanobacterial blooms in eutrophic lakes through vertical flow multi-soil-layering technology.

Eutrophication has led to the widespread occurrence of cyanobacterial blooms. Toxic cyanobacterial blooms with high concentrations of microcystins (MCs) have been identified in the Lalla Takerkoust reservoir in Morocco. The objective of this study was to evaluate the efficiency of the Multi-Soil-Layering (MSL) ecotechnology in removing natural cyanobacterial blooms from the lake. Two MSL pilots were used in rectangular glass tanks (60 × 10 × 70 cm). They consisted of permeable layers (PLs) made of pozzolan and a soil mixture layer (SML) containing local soil, ferrous metal, charcoal and sawdust. The main difference between the two systems was the type of local soil used: sandy soil for MSL1 and clayey soil for MSL2. Both MSL pilots effectively reduced cyanobacterial cell concentrations in the treated water to very low levels (0.09 and 0.001 cells/mL). MSL1 showed a gradual improvement in MC removal from 52 % to 99 %, while MSL2 started higher at 90 % but dropped to 54% before reaching 86%. Both MSL systems significantly reduced organic matter levels (97.2 % for MSL1 and 95.8 % for MSL2). Both MSLs were shown to be effective in removing cyanobacteria, MCs, and organic matter with comparable performance.

Unlocking the potential of bacterioplankton-mediated microcystin degradation and removal: A bibliometric analysis of sustainable water treatment strategies.

Microcystins (MCs) constitute a significant threat to human and environmental health, urging the development of effective removal methods for these toxins. In this review, we explore the potential of MC-degrading bacteria as a solution for the removal of MCs from water. The review insights into the mechanisms of action employed by these bacteria, elucidating their ability to degrade and thus remove MCs. After, the review points out the influence of the structural conformation of MCs on their removal, particularly their stability at different water depths within different water bodies. Then, we review the crucial role played by the production of MCs in ensuring the survival and safeguarding of the enzymatic activities of Microcystis cells. This justifies the need for developing effective and sustainable methods for removing MCs from aquatic ecosystems, given their critical ecological function and potential toxicity to humans and animals. Thereafter, challenges and limitations associated with using MC-degrading bacteria in water treatment are discussed, emphasizing the need for further research to optimize the selection of bacterial strains used for MCs biodegradation. The interaction of MCs-degrading bacteria with sediment particles is also crucial for their toxin removal potential and its efficiency. By presenting critical information, this review is a valuable resource for researchers, policymakers, and stakeholders involved in developing sustainable and practical approaches to remove MCs. Our review highlights the potential of various applications of MC-degrading bacteria, including multi-soil-layering (MSL) technologies. It emphasizes the need for ongoing research to optimize the utilization of MC-degrading bacteria in water treatment, ultimately ensuring the safety and quality of water sources. Moreover, this review highlights the value of bibliometric analyses in revealing research gaps and trends, providing detailed insights for further investigations. Specifically, we discuss the importance of employing advanced genomics, especially combining various OMICS approaches to identify and optimize the potential of MCs-degrading bacteria.

Microcystin influence on soil-plant microbiota: Unraveling microbiota modulations and assembly processes in the rhizosphere of Vicia faba.

Microcystins (MCs) are frequently detected in cyanobacterial bloom-impacted waterbodies and introduced into agroecosystems via irrigation water. They are widely known as phytotoxic cyanotoxins, which impair the growth and physiological functions of crop plants. However, their impact on the plant-associated microbiota is scarcely tackled and poorly understood. Therefore, we aimed to investigate the effect of MCs on microbiota-inhabiting bulk soil (BS), root adhering soil (RAS), and root tissue (RT) of Vicia faba when exposed to 100 µg L-1 MCs in a greenhouse pot experiment. Under MC exposure, the structure, co-occurrence network, and assembly processes of the bacterial microbiota were modulated with the greatest impact on RT-inhabiting bacteria, followed by BS and, to a lesser extent, RAS. The analyses revealed a significant decrease in the abundances of several Actinobacteriota-related taxa within the RT microbiota, including the most abundant and known genus of Streptomyces. Furthermore, MCs significantly increased the abundance of methylotrophic bacteria (Methylobacillus, Methylotenera) and other Proteobacteria-affiliated genera (e.g., Paucibacter), which are supposed to degrade MCs. The co-occurrence network of the bacterial community in the presence of MCs was less complex than the control network. In MC-exposed RT, the turnover in community composition was more strongly driven by deterministic processes, as proven by the beta-nearest taxon index. Whereas in MC-treated BS and RAS, both deterministic and stochastic processes can influence community assembly to some extent, with a relative dominance of deterministic processes. Altogether, these results suggest that MCs may reshape the structure of the microbiota in the soil-plant system by reducing bacterial taxa with potential phytobeneficial traits and increasing other taxa with the potential capacity to degrade MCs.

Tracing the fate of microcystins from irrigation water to food chains: Studies with Fragaria vulgaris and Meriones shawi.

Microcystins (MCs) are cyanobacterial toxins that can negatively impact human and animal health. This study investigated the bioaccumulation, transfer, depuration, and health risks of MCs in strawberry plants (Fragaria vulgaris) and Meriones shawi animals. The plants were irrigated with 1, 5, 10, and 20 µg/L MCs for 60 days (bioaccumulation phase) and then with clean water for 30 days (depuration phase). The harvested plants (roots and leaves) were then prepared in an aliquot form and used as feed for Meriones shawi. Liquid chromatography-mass spectrometry (LC/MS/MS) was used to measure MC concentrations in plant and animal tissues. The bioaccumulation of MCs was found to be highest in the roots, followed by leaves, fruits, liver, stomach, and fecal matter. The bioaccumulation factor (BAF) was highest in perlite (8.48), followed by roots (5.01), leaves (1.55), stomach (0.87), and fecal matter (1.18), indicating that the parts with high bioaccumulation factor had high translocation of MCs. The transfer of MCs to animal organs was low, and the daily toxin intake of adult consumers of strawberry fruit irrigated with 1, 5, 10, and 20 µg/L MC did not exceed the WHO-recommended limit of 0.04 µg MC-LR/Kg of bw/day. However, fruits from plants irrigated with 10 and 20 µg/L may pose a moderate health risk to children (25 Kg bw), and Meriones' consumption of leaves may pose a significant health risk. After the depuration phase, MC concentration in perlite, roots, leaves, and fruits decreased, indicating that depuration reduced the danger of MC transmission and bioaccumulation. The study also found that glutathione reductase and glutathione S-transferase activity were essential in the depuration of MCs in the tested plants. The findings suggest that legislation regulating the quality of irrigation water in terms of MC concentrations is necessary to prevent detrimental consequences to crops and human exposure.

Health risk assessment of lake water contaminated with microcystins for fruit crop irrigation and farm animal drinking.

The health risks linked to the consumption of microcystin-accumulating crops have been increasing worldwide in toxic cyanobloom-occurring regions. The bioaccumulation of microcystins (MCs) in agricultural produce at environmentally realistic concentrations is poorly investigated. In this field study, we assessed the health risks of MCs in raw water used for irrigating fruit crops (bioaccumulation) and watering farm animals in the Lalla Takerkoust agricultural region (Marrakesh, Morocco). Thus, MCs were extracted from water and fruit samples and quantified by enzyme-linked immunosorbent assay in order to calculate the health risk indicators. MCs posed a high health-risk level to poultry and horses, with estimated daily intakes (EDI) being 14- and 19-fold higher than the recommended limits (3.1 and 2.3 µg MC-LR L-1), respectively. Furthermore, pomegranate posed the same level of risk, with EDI being 22- and 53-fold higher than the limit dose (0.04 µg MC-LR kg-1) for adults and children, respectively. There was an urgent need for guidelines regarding water use and management in MC-polluted areas, besides the setup of nature-based tools for toxin removal from raw water used in farming practices. Moreover, MCs could contaminate the human food chain, which implies further investigations of their potential accumulation in livestock- and poultry-based food.

Bacterioplankton Associated with Toxic Cyanobacteria Promote Pisum sativum (Pea) Growth and Nutritional Value through Positive Interactions.

Research on Plant Growth-Promoting Bacteria (PGPB) has focused much more on rhizospheric bacteria. However, PGPB associated with toxic cyanobacterial bloom (TCB) could enter the rhizosphere through irrigation water, helping plants such as Pisum sativum L. (pea) overcome oxidative stress induced by microcystin (MC) and improve plant growth and nutritional value. This study aimed to isolate bacteria associated with toxic cyanobacteria, test PGPB properties, and inoculate them as a consortium to pea seedlings irrigated with MC to investigate their role in plant protection as well as in improving growth and nutritional value. Two bacterioplankton isolates and one rhizosphere isolate were isolated and purified on a mineral salt medium supplemented with 1000 µg/L MC and identified via their 16S rRNA gene. The mixed strains were inoculated to pea seedlings in pots irrigated with 0, 50, and 100 µg/L MC. We measured the morphological and physiological parameters of pea plants at maturity and evaluated the efficiency of the plant's enzymatic and non-enzymatic antioxidant responses to assess the role and contribution of PGPB. Both bacterioplankton isolates were identified as Starkeya sp., and the rhizobacterium was identified as Brevundimonas aurantiaca. MC addition significantly (p < 0.05) reduced all the growth parameters of the pea, i.e., total chlorophyll content, leaf quantum yield, stomatal conductance, carotenoids, and polyphenol contents, in an MC concentration-dependent manner, while bacterial presence positively affected all the measured parameters. In the MC treatment, the levels of the pea's antioxidant traits, including SOD, CAT, POD, PPO, GST, and ascorbic acid, were increased in the sterile pots. In contrast, these levels were reduced with double and triple PGPB addition. Additionally, nutritional values such as sugars, proteins, and minerals (Ca and K) in pea fruits were reduced under MC exposure but increased with PGPB addition. Overall, in the presence of MC, PGPB seem to positively interact with pea plants and thus may constitute a natural alternative for soil fertilization when irrigated with cyanotoxin-contaminated water, increasing the yield and nutritional value of crops.

Effects of Irrigation with Microcystin-Containing Water on Growth, Physiology, and Antioxidant Defense in Strawberry Fragaria vulgaris under Hydroponic Culture.

Over the last years, the use of artificial lakes and ponds to irrigate agricultural crops has been intensified and cultivation methods have been diversified. Hydroponics is a type of hydroculture which usually involves growing plants in an inert substrate, by using nutrient-enriched water to support plant growth. However, irrigating plants in hydroponic-based culture must be accompanied by monitoring the quality of irrigation water. The human health risks involved are mainly related to the proliferation of microcystin-producing cyanobacteria that contaminate water used for irrigation purposes. Strawberry (Fragaria vulgaris L.) is a widely cultivated plant of an increased economically importance worldwide. Its fruits provide essential elements for human nutrition; therefore, the study of its sensitivity to microcystins (MCs) is of paramount importance. The objective of this study was to evaluate the effects of MCs in irrigation water on the growth, physiology, and antioxidant defense system in F. vulgaris. In this study, strawberry seedlings at the three-leaf stage were grown in pots containing perlite under controlled conditions. Plants were exposed to a crude extract of Microcystis aeruginosa bloom at different concentrations of MCs (1, 5, 10, and 20 µg/L) for 60 days of exposure. The results showed that the highest concentrations of 10 and 20 µg/L induced a decrease in growth parameters. They resulted in root/shoot length decrease as well as number of leaves, roots/leaves dry and fresh weight. Furthermore, MCs reduced chlorophyll/carotenoid content, stomatal conductance, fluorescence, and total protein content of strawberry plants. At the same time, a significant increase in Malondialdehyde (MDA) (an indicator of lipid peroxidation), polyphenol, and sugar content were recorded in strawberry plants exposed to MCs at 5, 10, and 20 µg/L compared with the control. Additionally, superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), Polyphenoloxydase (PPO), and ascorbate peroxidase (APX) activities significantly increased in plants under MCs exposure. The oxidative stress was higher in plants exposed to 10 and 20 µg/L of MCs from the second harvest (after 60 days of exposure) compared to those from the first harvest (after 30 days). Overall, the results obtained in this study indicate an increasingly negative effect of MCs on strawberry plants grown in hydroponics even at concentrations (10 and 20 µg/L). This effect is more damaging on the roots after exposure (60 days).

Role of Rhizospheric Microbiota as a Bioremediation Tool for the Protection of Soil-Plant Systems from Microcystins Phytotoxicity and Mitigating Toxin-Related Health Risk.

Frequent toxic cyanoblooms in eutrophic freshwaters produce various cyanotoxins such as the monocyclic heptapeptides microcystins (MCs), known as deleterious compounds to plant growth and human health. Recently, MCs are a recurrent worldwide sanitary problem in irrigation waters and farmland soils due to their transfer and accumulation in the edible tissues of vegetable produce. In such cases, studies about the persistence and removal of MCs in soil are scarce and not fully investigated. In this study, we carried out a greenhouse trial on two crop species: faba bean (Vicia faba var. Alfia 321) and common wheat (Triticum aestivum var. Achtar) that were grown in sterile (microorganism-free soil) and non-sterile (microorganism-rich soil) soils and subjected to MC-induced stress at 100 µg equivalent MC-LR L-1. The experimentation aimed to assess the prominent role of native rhizospheric microbiota in mitigating the phytotoxic impact of MCs on plant growth and reducing their accumulation in both soils and plant tissues. Moreover, we attempted to evaluate the health risk related to the consumption of MC-polluted plants for humans and cattle by determining the estimated daily intake (EDI) and health risk quotient (RQ) of MCs in these plants. Biodegradation was liable to be the main removal pathway of the toxin in the soil; and therefore, bulk soil (unplanted soil), as well as rhizospheric soil (planted soil), were used in this experiment to evaluate the accumulation of MCs in the presence and absence of microorganisms (sterile and non-sterile soils). The data obtained in this study showed that MCs had no significant effects on growth indicators of faba bean and common wheat plants in non-sterile soil as compared to the control group. In contrast, plants grown in sterile soil showed a significant decrease in growth parameters as compared to the control. These results suggest that MCs were highly bioavailable to the plants, resulting in severe growth impairments in the absence of native rhizospheric microbiota. Likewise, MCs were more accumulated in sterile soil and more bioconcentrated in root and shoot tissues of plants grown within when compared to non-sterile soil. Thereby, the EDI of MCs in plants grown in sterile soil was more beyond the tolerable daily intake recommended for both humans and cattle. The risk level was more pronounced in plants from the sterile soil than those from the non-sterile one. These findings suggest that microbial activity, eventually MC-biodegradation, is a crucial bioremediation tool to remove and prevent MCs from entering the agricultural food chain.

Impacts of Microcystins on Morphological and Physiological Parameters of Agricultural Plants: A Review.

Cyanobacteria are a group of photosynthetic prokaryotes that pose a great concern in the aquatic environments related to contamination and poisoning of wild life and humans. Some species of cyanobacteria produce potent toxins such as microcystins (MCs), which are extremely aggressive to several organisms, including animals and humans. In order to protect human health and prevent human exposure to this type of organisms and toxins, regulatory limits for MCs in drinking water have been established in most countries. In this regard, the World Health Organization (WHO) proposed 1 µg MCs/L as the highest acceptable concentration in drinking water. However, regulatory limits were not defined in waters used in other applications/activities, constituting a potential threat to the environment and to human health. Indeed, water contaminated with MCs or other cyanotoxins is recurrently used in agriculture and for crop and food production. Several deleterious effects of MCs including a decrease in growth, tissue necrosis, inhibition of photosynthesis and metabolic changes have been reported in plants leading to the impairment of crop productivity and economic loss. Studies have also revealed significant accumulation of MCs in edible tissues and plant organs, which raise concerns related to food safety. This work aims to systematize and analyze the information generated by previous scientific studies, namely on the phytotoxicity and the impact of MCs especially on growth, photosynthesis and productivity of agricultural plants. Morphological and physiological parameters of agronomic interest are overviewed in detail in this work, with the aim to evaluate the putative impact of MCs under field conditions. Finally, concentration-dependent effects are highlighted, as these can assist in future guidelines for irrigation waters and establish regulatory limits for MCs.

Protective Role of Native Rhizospheric Soil Microbiota Against the Exposure to Microcystins Introduced into Soil-Plant System via Contaminated Irrigation Water and Health Risk Assessment.

Microcystins (MCs) produced in eutrophic waters may decrease crop yield, enter food chains and threaten human and animal health. The main objective of this research was to highlight the role of rhizospheric soil microbiota to protect faba bean plants from MCs toxicity after chronic exposure. Faba bean seedlings were grown in pots containing agricultural soil, during 1 month under natural environmental conditions of Marrakech city in Morocco (March-April 2018) and exposed to cyanobacterial extracts containing up to 2.5 mg·L-1 of total MCs. Three independent exposure experiments were performed (a) agricultural soil was maintained intact "exposure experiment 1"; (b) agricultural soil was sterilized "exposure experiment 2"; (c) agricultural soil was sterilized and inoculated with the rhizobia strain Rhizobium leguminosarum RhOF34 "exposure experiment 3". Overall, data showed evidence of an increased sensitivity of faba bean plants, grown in sterilized soil, to MCs in comparison to those grown in intact and inoculated soils. The study revealed the growth inhibition of plant shoots in both exposure experiments 2 and 3 when treated with 2.5 mg·L-1 of MCs. The results also showed that the estimated daily intake (EDI) of MCs, in sterilized soil, exceeded 2.18 and 1.16 times the reference concentrations (0.04 and 0.45 µg of microcysin-leucine arginine (MC-LR). Kg-1 DW) established for humans and cattle respectively, which raises concerns about human food chain contamination.

Harmful Cyanobacterial Blooms (HCBs): innovative green bioremediation process based on anti-cyanobacteria bioactive natural products.

Over the last decades, Harmful Cyanobacterial Blooms (HCBs) represent one of the most conspicuous hazards to human health in freshwater ecosystems, due to the uses of the water for drinking, recreation and aquaculture. Cyanobacteria are one of the main biological components in freshwater ecosystems and they may proliferate in nutrients rich ecosystems causing severe impacts at different levels. Therefore, several methods have been applied to control cyanobacterial proliferation, including physical, chemical and biological strategies. However, the application of those methods is generally not very efficient. Research on an eco-friendly alternative leading to the isolation of new bioactive compounds with strong impacts against harmful cyanobacteria is a need in the field of water environment protection. Thus, this paper aims to give an overview of harmful cyanobacterial blooms and reviews the state of the art of studying the activities of biological compounds obtained from plants, seaweeds and microorganisms in the cyanobacterial bloom control.

Moroccan actinobacteria with promising activity against toxic cyanobacteria Microcystis aeruginosa.

In recent decades, harmful cyanobacterial blooms (HCBs) have become a severe hazard for human health mainly in drinking water resources and are responsible for serious ecological disturbances in freshwater ecosystems. The present study aims to explore the potential of actinobacteria isolated from sediment samples collected from Moroccan salt river to control HCBs mainly through Microcystis aeruginosa lysis. In order to investigate the possible anti-cyanobacterial response mechanisms, the antioxidant enzyme activities of M. aeruginosa cells were analysed. Anti-cyanobacterial activity was tested using the agar cylinder method against the toxic cyanobacteria Microcystis aeruginosa. Amongst the twenty-three isolates tested, only one showed promising anti-cyanobacterial activities with inhibition zone (ZI) equal to 22.00 mm, minimum inhibitory concentration (MIC) equal to 19.53 mg/L and minimum bactericidal concentration MBC equal to 39.06 mg/L. Phylogenetic analysis of the near-complete 16S rRNA gene sequence indicated that the strain DS1R1 belongs to the genus Streptomyces and has the highest similarity (100%) to Streptomyces sp. Indeed, M. aeruginosa growth, chlorophyll-a and protein content were significantly reduced by Streptomyces sp. DS1R1 extract. Superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) were significantly elevated after treatment with Streptomyces sp. DS1R1 extract. These experimental findings provided insights in the development of a new eco-friendly procedure based on the use of actinobacteria for toxic cyanobacterial bloom bio-control.

Mode of action and fate of microcystins in the complex soil-plant ecosystems.

Over the last decades, global warming has increasingly stimulated the expansion of cyanobacterial blooms in freshwater ecosystems worldwide, in which toxic cyanobacteria produce various congeners of cyanotoxins, mainly dominated by microcystins (MCs). MCs introduced into agricultural soils have deleterious effects on the germination, growth and development of plants and their associated microbiota, leading to remarkable yield losses. Phytotoxicity of MCs may refer to the inhibition of phosphatases activity, generating deleterious reactive oxygen species, altering gene functioning and phytohormones translocation within the plant. It is also known that MCs can pass through the root membrane barrier, translocate within plant tissues and accumulate into different organs, including edible ones. Also, MCs impact the microbial activity in soil via altering plant-bacterial symbioses and decreasing bacterial growth rate of rhizospheric microbiota. Moreover, MCs can persist in agricultural soils through adsorption to clay-humic acid particles and results in a long-term contact with the plant-microflora complex. However, their bioavailability to plants and half-life in soil seem to be influenced by biodegradation process and soil physicochemical properties. This review reports the latest and most relevant information regarding MCs-phytotoxicity and impact on soil microbiota, the persistence in soil, the degradation by native microflora and the bioaccumulation within plant tissues.