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.

Seaweed Bioactive Compounds against Pathogens and Microalgae: Potential Uses on Pharmacology and Harmful Algae Bloom Control.

Cyanobacteria are found globally due to their adaptation to various environments. The occurrence of cyanobacterial blooms is not a new phenomenon. The bloom-forming and toxin-producing species have been a persistent nuisance all over the world over the last decades. Evidence suggests that this trend might be attributed to a complex interplay of direct and indirect anthropogenic influences. To control cyanobacterial blooms, various strategies, including physical, chemical, and biological methods have been proposed. Nevertheless, the use of those strategies is usually not effective. The isolation of natural compounds from many aquatic and terrestrial plants and seaweeds has become an alternative approach for controlling harmful algae in aquatic systems. Seaweeds have received attention from scientists because of their bioactive compounds with antibacterial, antifungal, anti-microalgae, and antioxidant properties. The undesirable effects of cyanobacteria proliferations and potential control methods are here reviewed, focusing on the use of potent bioactive compounds, isolated from seaweeds, against microalgae and cyanobacteria growth.

Microcystin Contamination in Irrigation Water and Health Risk.

Microcystins (MCs), natural hepatotoxic compounds produced by cyanobacteria, pose significant risks to water quality, ecosystem stability, and the well-being of animals, plants, and humans when present in elevated concentrations. The escalating contamination of irrigation water with MCs presents a growing threat to terrestrial plants. The customary practice of irrigating crops from local water sources, including lakes and ponds hosting cyanobacterial blooms, serves as a primary conduit for transferring these toxins. Due to their high chemical stability and low molecular weight, MCs have the potential to accumulate in various parts of plants, thereby increasing health hazards for consumers of agricultural products, which serve as the foundation of the Earth's food chain. MCs can bioaccumulate, migrate, potentially biodegrade, and pose health hazards to humans within terrestrial food systems. This study highlights that MCs from irrigation water reservoirs can bioaccumulate and come into contact with plants, transferring into the food chain. Additionally, it investigates the natural mechanisms that organisms employ for conjugation and the microbial processes involved in MC degradation. To gain a comprehensive understanding of the role of MCs in the terrestrial food chain and to elucidate the specific health risks associated with consuming crops irrigated with water contaminated with these toxins, further research is necessary.

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.

Monitoring of toxic cyanobacterial blooms in Lalla Takerkoust reservoir by satellite imagery and microcystin transfer to surrounding farms.

Cyanobacterial harmful algal blooms (CyanoHABs) threaten public health and freshwater ecosystems worldwide. In this study, our main goal was to explore the dynamics of cyanobacterial blooms and how microcystins (MCs) move from the Lalla Takerkoust reservoir to the nearby farms. We used Landsat imagery, molecular analysis, collecting and analyzing physicochemical data, and assessing toxins using HPLC. Our investigation identified two cyanobacterial species responsible for the blooms: Microcystis sp. and Synechococcus sp. Our Microcystis strain produced three MC variants (MC-RR, MC-YR, and MC-LR), with MC-RR exhibiting the highest concentrations in dissolved and intracellular toxins. In contrast, our Synechococcus strain did not produce any detectable toxins. To validate our Normalized Difference Vegetation Index (NDVI) results, we utilized limnological data, including algal cell counts, and quantified MCs in freeze-dried Microcystis bloom samples collected from the reservoir. Our study revealed patterns and trends in cyanobacterial proliferation in the reservoir over 30 years and presented a historical map of the area of cyanobacterial infestation using the NDVI method. The study found that MC-LR accumulates near the water surface due to the buoyancy of Microcystis. The maximum concentration of MC-LR in the reservoir water was 160 µg L-1. In contrast, 4 km downstream of the reservoir, the concentration decreased by a factor of 5.39 to 29.63 µgL-1, indicating a decrease in MC-LR concentration with increasing distance from the bloom source. Similarly, the MC-YR concentration decreased by a factor of 2.98 for the same distance. Interestingly, the MC distribution varied with depth, with MC-LR dominating at the water surface and MC-YR at the reservoir outlet at a water depth of 10 m. Our findings highlight the impact of nutrient concentrations, environmental factors, and transfer processes on bloom dynamics and MC distribution. We emphasize the need for effective management strategies to minimize toxin transfer and ensure public health and safety.

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.

Phytotoxic effects of a natural bloom extract containing microcystins on Lycopersicon esculentum.

The cyanobacterial toxins microcystins (MC) are known to affect many processes in plants. Their presence in the water used for irrigation may have considerable impact on the survivorship, growth and development of plants. In this study, a crude extract of a toxic cyanobacterial bloom from "Lalla Takerkoust" reservoir (Morocco) was used to study the effects of extract containing MC on tomato plants (Lycopersicon esculentum). Five MC variants: MC-LR, MC-FR, MC-LY, MC-(H4)-YR and DMC-LR were identified by HPLC in the cyanobacterial extract. Exposure of the seeds to the crude extract (containing 22.24 µg MC mL(-1)) caused a reduction of germination up to 85%. Experiments showed that 30 days exposure of plant to the cyanobacterial extract containing MC caused inhibition of L. esculentum growth and productivity, as well as harmful effects on photosystem II activity, measured by Fv/Fm fluorescence. An accumulation of nutrients Na(+), K(+) and Ca(2+) was also registered. The activity of peroxidase and phenolic content indicated that the extract caused an oxidative stress. The tissue necrosis of leaves was also a consequence of MC exposure indicating a disorder in the exposed plant metabolism.

Cyanobacterial extracts containing microcystins affect the growth, nodulation process and nitrogen uptake of faba bean (Vicia faba L., Fabaceae).

The use of irrigation water containing cyanobacterial toxins may generate a negative impact in both yield and quality of agricultural crops causing significant economic losses. We evaluated the effects of microcystins (MC) on the growth, nodulation process and nitrogen uptake of a Faba bean cultivar (Vicia faba L., Fabaceae), particularly the effect of MC on rhizobia-V. faba symbiosis. Three rhizobial strains (RhOF4, RhOF6 and RhOF21), isolated from nodules of local V. faba were tested. The exposure of rhizobia to MC showed that the toxins had a negative effect on the rhizobial growth especially at the highest concentrations of 50 and 100 µg/l. The germination of faba bean seeds was also affected by cyanotoxins. We registered germination rates of 75 and 68.75% at the toxin levels of 50 and 100 µg/l as compared to the control (100%). The obtained results also showed there was a negative effect of MC on plants shoot, root (dry weight) and total number of nodules per plant. Cyanotoxins exposure induced a significant effect on nitrogen assimilation by faba bean seedlings inoculated with selected rhizobial strains RhOF6 and RhOF21, while the effect was not significant on beans seedling inoculated with RhOF4. This behavior of tolerant rhizobia-legumes symbioses may constitute a very important pathway to increase soil fertility and quality and can represent a friendly biotechnological way to remediate cyanotoxins contamination in agriculture.

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).

Allelopatic effects of cyanobacteria extracts containing microcystins on Medicago sativa-Rhizobia symbiosis.

The eutrophication of water leads to massive blooms of cyanobacteria potentially producers of highly toxic substances: cyanotoxins, especially microcystins (MC). The contamination of water used for irrigation by these toxins, can cause several adverse effects on plants and microorganisms. In this work, we report the phytotoxic effects of microcystins on the development of symbiosis between the leguminous plant Medicago sativa (Alfalfa) and rhizobia strains. The exposure of rhizobial strains to three different concentrations 0.01, 0.05 and 0.1 µg MC ml(-1) led to decrease on the bacteria growth. The strains of rhizobia Rh L1, Rh L2, Rh L3 and Rh L4 reduced their growth to, respectively, 20.85%, 20.80%, 33.19% and 25.65%. The chronic exposure of alfalfa seeds and seedlings to different MC concentrations affects the whole stages of plant development. The germination process has also been disrupted with an inhibition, which reaches 68.34% for a 22.24 µg MC ml(-1). Further, seedlings growth and photosynthetic process were also disrupted. The toxins reduced significantly the roots length and nodule formation and leads to an oxidative stress. Thus, the MCs contained in lake water and used for irrigation affect the development of symbiosis between M. sativa and Rhizobia.

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.

Phytobeneficial bacteria improve saline stress tolerance in Vicia faba and modulate microbial interaction network.

Increased global warming, caused by climate change and human activities, will seriously hinder plant development, such as increasing salt concentrations in soils, which will limit water availability for plants. To ensure optimal plant growth under such changing conditions, microorganisms that improve plant growth and health must be integrated into agricultural practices. In the present work, we examined the fate of Vicia faba microbiota structure and interaction network upon inoculation with plant-nodulating rhizobia (Rhizobium leguminosarum RhOF125) and non-nodulating strains (Paenibacillus mucilaginosus BLA7 and Ensifer meliloti RhOL1) in the presence (or absence) of saline stress. Inoculated strains significantly improved plant tolerance to saline stress, suggesting either a direct or indirect effect on the plant response to such stress. To determine the structure of microbiota associated with V. faba, samples of the root-adhering soil (RAS), and the root tissues (RT) of seedlings inoculated (or not) with equal population size of RhOF125, BLA7 and RhOL1 strains and grown in the presence (or absence) of salt, were used to profile the microbial composition by 16S rRNA gene sequencing. The inoculation did not show a significant impact on the composition of the RT microbiota or RAS microbiota. The saline stress shifted the RAS microbiota composition, which correlated with a decrease in Enterobacteriaceae and an increase in Sphingobacterium, Chryseobacterium, Stenotrophomonas, Agrobacterium and Sinorhizobium. When the microbiota of roots and RAS are considered together, the interaction networks for each treatment are quite different and display different key populations involved in community assembly. These findings indicate that upon seed inoculation, community interaction networks rather than their composition may contribute to helping plants to better tolerate environmental stresses. The way microbial populations interfere with each other can have an impact on their functions and thus on their ability to express the genes required to help plants tolerate stresses.

Seaweed Essential Oils as a New Source of Bioactive Compounds for Cyanobacteria Growth Control: Innovative Ecological Biocontrol Approach.

The application of natural compounds extracted from seaweeds is a promising eco-friendly alternative solution for harmful algae control in aquatic ecosystems. In the present study, the anti-cyanobacterial activity of three Moroccan marine macroalgae essential oils (EOs) was tested and evaluated on unicellular Microcystis aeruginosa cyanobacterium. Additionally, the possible anti-cyanobacterial response mechanisms were investigated by analyzing the antioxidant enzyme activities of M. aeruginosa cells. The results of EOs GC-MS analyses revealed a complex chemical composition, allowing the identification of 91 constituents. Palmitic acid, palmitoleic acid, and eicosapentaenoic acid were the most predominant compounds in Cystoseira tamariscifolia, Sargassum muticum, and Ulva lactuca EOs, respectively. The highest anti-cyanobacterial activity was recorded for Cystoseira tamariscifolia EO (ZI = 46.33 mm, MIC = 7.81 µg mL-1, and MBC = 15.62 µg mL-1). The growth, chlorophyll-a and protein content of the tested cyanobacteria were significantly reduced by C. tamariscifolia EO at both used concentrations (inhibition rate >67% during the 6 days test period in liquid media). Furthermore, oxidative stress caused by C. tamariscifolia EO on cyanobacterium cells showed an increase of the activities of superoxide dismutase (SOD) and catalase (CAT), and malondialdehyde (MDA) concentration was significantly elevated after 2 days of exposure. Overall, these experimental findings can open a promising new natural pathway based on the use of seaweed essential oils to the fight against potent toxic harmful cyanobacterial blooms (HCBs).

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.

Potential control of toxic cyanobacteria blooms with Moroccan seaweed extracts.

Marine macroalgae are a promising source of diverse bioactive compounds with applications in the biocontrol of harmful cyanobacteria blooms (cyanoHABs). In this work, we evaluated the potential algicidal activities of 14 species of seaweed collected from the coast of Souiria Laqdima, Morocco. Methanol extracts were screened in solid and liquid medium against the growth of the toxic cyanobacteria Microcystis aeruginosa and the microalgae Chlorella sp. used as food supplement. The results in solid medium revealed that the algicidal activity was limited to M. aeruginosa with the extract of Bornetia secundiflora showing the highest growth inhibition activity against Microcystis (27.33 ± 0.33 mm), whereas the extracts of Laminaria digitata, Halopytis incurvus, Ulva lactuca, and Sargasum muticum showed no inhibition. In liquid medium, the results indicated that all methanolic extracts of different macroalgae tested have a significant inhibitory effect on M. aeruginosa compared with that of the negative control. The maximum inhibition rates of M. aeruginosa were produced by the extracts of Bifurcaria tuberculata, Codium elongatum, and B. secundiflora. Moreover, the extracts of B. secundiflora recorded the maximum inhibition rate of Chlorella sp. Overall, the results highlight the potential of the extracts from macroalgae to control toxic cyanobacteria species.

Microcystin-tolerant Rhizobium protects plants and improves nitrogen assimilation in Vicia faba irrigated with microcystin-containing waters.

Irrigation of crops with microcystins (MCs)-containing waters-due to cyanobacterial blooms-affects plant productivity and could be a way for these potent toxins entering the food chain. This study was performed to establish whether MC-tolerant rhizobia could benefit growth, nodulation, and nitrogen metabolism of faba bean plants irrigated with MC-containing waters. For that, three different rhizobial strains-with different sensitivity toward MCs-were used: RhOF96 (most MC-sensitive strain), RhOF125 (most MC-tolerant strain), or Vicz1.1 (reference strain). As a control, plants grown without rhizobia and fertilized by NH4NO3 were included in the study. MC exposure decreased roots (30-37 %) and shoots (up to 15 %) dry weights in un-inoculated plants, whereas inoculation with rhizobia protects plants toward the toxic effects of MCs. Nodulation and nitrogen content were significantly impaired by MCs, with the exception of plants inoculated with the most tolerant strain RhOF125. In order to deep into the effect of inoculation on nitrogen metabolism, the nitrogen assimilatory enzymes (glutamine synthetase (GS) and glutamate synthase (GOGAT)) were investigated: Fertilized plants showed decreased levels (15-30 %) of these enzymes, both in shoots and roots. By contrast, inoculated plants retained the levels of these enzymes in shoots and roots, as well as the levels of NADH-GOGAT activity in nodules. We conclude that the microcystin-tolerant Rhizobium protects faba bean plants and improves nitrogen assimilation when grown in the presence of MCs.

Microbiote shift in the Medicago sativa rhizosphere in response to cyanotoxins extract exposure.

The bloom-containing water bodies may have an impact due to cyanotoxins production on other microorganisms and aquatic plants. Where such water is being used for crops irrigation, the presence of cyanotoxins may also have a toxic impact on terrestrial plants and their rhizosphere microbiota. For that purpose, PCR-based 454 pyrosequencing was applied to phylogenetically characterize the bacterial community of Medicago sativa rhizosphere in response to cyanotoxins extract. This analysis revealed a wide diversity at species level, which decreased from unplanted soil to root tissues indicating that only some populations were able to compete for nutrients and niches in this selective habitat. Gemmatimonas, Actinobacteria, Deltaproteobacteria and Opitutae mainly inhabited the bulk soil, whereas, the root-adhering soil and the root tissues were inhabited by Gammaproteobacteria and Alphaproteobacteria. The proportion of these populations fluctuated in response to cyanotoxins extract exposure. Betaproteobacteria proportion increased in the three studied compartments, whereas Gammaproteobacteria proportion decreased except in the bulk soil. This study revealed the potential toxicity of cyanotoxins extract towards Actinobacteria, Gemmatimonas, Deltaproteobacteria, and Gammaproteobacteria, however Clostridia, Opitutae and bacteria related with Betaproteobacteria, were stimulated denoting their tolerance. Altogether, these data indicate that crop irrigation using cyanotoxins containing water might alter the rhizosphere functioning.

Physiological and antioxidant responses of Medicago sativa-rhizobia symbiosis to cyanobacterial toxins (Microcystins) exposure.

Toxic cyanobacteria in freshwaters can induce potent harmful effects on growth and development of plants irrigated with contaminated water. In this study, the effect of cyanobacteria extract containing Microcystins (MC) on Medicago sativa-rhizobia symbiosis was investigated in order to explore plants response through biomass production, photosynthetic pigment and antioxidant enzymes analysis: Peroxidase (POD), Polyphenoloxidase (PPO) and Catalase (CAT). Alfalfa plants were inoculated with two endosymbiotic rhizobial strains: RhOL1 (MC less sensitive strain) and RhOL3 (MC more sensitive strain), to evaluate the rhizobial contribution on the plant response cultured under cyanobacterial toxins stress. The two rhizobia strains were identified as Ensifer meliloti by sequence analysis of their rrs and atpD genes. The chronic exposure to MC extract showed shoot, root and nodules dry weight decrease, in both symbiosis cultures. The rate of decline in plants inoculated with RhOL3 was higher than that in symbiosis with RhOL1 mainly at 20 µg L(-1) of MC. Cyanotoxins also reduced photosynthetic pigment content and generated an oxidative stress observed at cellular level. POD, PPO and CAT activities were significantly increased in leaves, roots and nodules of alfalfa plants exposed to MC. These enzyme activities were higher in plants inoculated with RhOL3 especially when alfalfa plants were exposed to 20 µg L(-1) of MC. The present paper reports new scientific finding related to the behavior of rhizobia-M. sativa associations to MC (Microcystins) for later recommendation concerning the possible use of these symbiosis face to crops exposure to MC contaminated water irrigation.

Physiological and biochemical defense reactions of Vicia faba L.-Rhizobium symbiosis face to chronic exposure to cyanobacterial bloom extract containing microcystins.

The presence of cyanotoxins, mainly microcystins (MCs), in surface freshwater represents a serious health risk to aquatic organisms living in the water body, as well as terrestrial animals and plants that are in contact with contaminated water. Consequently, the use of MCs contaminated water for irrigation represents a hazard for cultivated plants and could induce severe economical losses due to crops' yield reduction. The experimental approach undertaken in this work was exposing Vicia faba seedlings (inoculated with a Rhizobium strain resistant to MCs), to water supplemented with cyanobacterial crude extract containing total microcystins at a concentration of 50 and 100 µg/L (environmental relevant concentrations of MCs dissolved in the raw irrigation water from Lalla Takerkoust Lake-Marrakesh region). After chronic MCs exposure (2 months), biological and physiological parameters (plant growth, nitrogen uptake, mineral assimilation, and oxidative defense mechanisms) were evaluated. The results obtained showed evidence that chronic exposure to cyanobacterial bloom extract containing MCs strongly affected the physiological and biological plants activities; reduction of dry matter, photosynthetic activity, nodule number, and nitrogen assimilation. At the same time, an increase of oxidative stress was observed, as deduced from a significant increase of the activities of peroxidase, catalase, polyphenoloxidase, and phenylalanine ammonia lyase in leaves, roots, and nodules of faba bean plants exposed to cyanotoxins, especially at 100 µg/L of MCs. This experimentation constitutes a simulation of the situation related to cyanotoxins chronic exposure of seedlings-plants via the contaminated irrigation water. For this reason, once should take into consideration the possibility of contamination of agricultural crops and the quality of irrigation water should be by the way monitored for cyanotoxins biohazard.