Physiological and toxicological response of Microcystis aeruginosa BCCUSP232 exposed to Salvinia auriculata extracts.

Microcystis aeruginosa is one of the most predominant freshwater bloom-forming cyanobacterium found globally which is capable of producing toxic secondary metabolites including microcystins that might intoxicate animals and humans when contaminated water or food is ingested. Salvinia auriculata Aubl is one of the plants that might possess bioactive compounds capable of controlling growth and reproduction of M. aeruginosa. The present study aimed to determine the presence of bioactive compounds in S. auriculata extracts and determine alterations occurred in growth and reproduction of M. aeruginosa when exposed to these plant extracts. In addition, this investigation aimed to examine the influence of S. auriculata on antioxidant enzymes detected in M. aeruginosa. The results obtained demonstrated that the aqueous and ethanolic extracts of S. auriculata presented potential for control of cyanobacteria populations, exhibiting algicidal action on M. aeruginosa as well as interfering in antioxidant enzymes activities and parameters associated with oxidative stress. Phytochemical analyses demonstrated the presence of polyphenols and flavonoids content in both extracts. In addition, application of S. auriculata extracts did not produce cytogenotoxicity and/or mutagenicity utilizing Allium cepa test. Therefore, further studies are needed in order to identify and characterize the compounds responsible for these effects on M. aeruginosa and provide information regarding the possible application of S. auriculata in the treatment of drinking water.

Unveiling the link between Raphidiopsis raciborskii blooms and saxitoxin levels: Evaluating water quality in tropical reservoirs, Brazil.

The proliferation of Raphidiopsis raciborskii blooms has sparked concerns regarding potential human exposure to heightened saxitoxins (STXs) levels. Thus, comprehending how environmental elements drive the proliferation of this STXs-producing species can aid in predicting human exposure risks. This study aimed to explore the link between cyanobacteria R. raciborskii, STXs cyanotoxins, and environmental factors in 37 public supply reservoirs in the tropical region and assess potential health hazards these toxins pose in the reservoir waters. A Structural Equation Model was used to assess the impact of environmental factors (water volume and physical and chemical variables) on R. raciborskii biomass and STXs levels. Furthermore, the potential risk of STXs exposure from consuming untreated reservoir water was evaluated. Lastly, the cumulative distribution function (CDF) of STXs across the reservoirs was computed. Our findings revealed a correlation between R. raciborskii biomass and STXs concentrations. Total phosphorus emerged as a critical environmental factor positively influencing species biomass and indirectly affecting STXs levels. pH significantly influenced STXs concentrations, indicating different factors influencing R. raciborskii biomass and STXs. Significantly, for the first time, the risk of STXs exposure was gauged using the risk quotient (HQ) for untreated water consumption from public supply reservoirs in Brazil's semi-arid region. Although the exposure risks were generally low to moderate, the CDF underscored the risk of chronic exposure due to low toxin concentrations in over 90% of samples. These outcomes emphasize the potential expansion of R. raciborskii in tropical settings due to increased phosphorus, amplifying waterborne STXs levels and associated intoxication risks. Thus, this study reinforces the importance of nutrient control, particularly phosphorus regulation, as a mitigation strategy against R. raciborskii blooms and reducing STXs intoxication hazards.

Susceptibility of phytoplankton to the increasing presence of active pharmaceutical ingredients (APIs) in the aquatic environment: A review.

Human and veterinary pharmaceuticals either in the form of un-metabolized, incompletely metabolized, and metabolized drugs are increasingly present in aquatic ecosystems. These active pharmaceutical ingredients from pharmaceutical industries, hospitals, agricultural, and domestic discharges find their way into water systems - where they adversely affect non-target organisms like phytoplankton. Different aspects of phytoplankton life; ranging from growth, reproduction, morphology, physiology, biochemical composition, oxidative response, proteomics, and transcriptomics are altered by pharmaceuticals. This review discusses the currently available information on the susceptibility of phytoplankton to the ever-increasing presence of pharmaceutical products in the aquatic environment by focusing on the effect of APIs on the physiology, metabolome, and proteome profiles of phytoplankton. We also highlight gaps in literature concerning the salient underlining biochemical interactions between phytoplankton communities and pharmaceuticals that require an in-depth investigation. This is all in a bid to understand the imminent dangers of the contamination of water bodies with pharmaceutical products and how this process unfavorably affects aquatic food webs.

The combined effect of clethodim (herbicide) and nitrogen variation on allelopathic interactions between Microcystis aeruginosa and Raphidiopsis raciborskii.

The large-scale use of herbicides deteriorates water quality and threatens aquatic biodiversity. Unfortunately, there are few studies on the ecological effects of herbicides on toxin-producing strains of cyanobacteria under changing nutrient conditions. The objective of the present study was to investigate the effects of the herbicide clethodim and nitrogen variation on the allelopathic interactions and toxin production of Microcystis aeruginosa BCCUSP232 and Raphidiopsis raciborskii (formerly known as Cylindrospermopsis raciborskii) ITEPA1. M. aeruginosa had increased cell density when exposed to the clethodim (H +) (23.55 mg/L), whereas the highest cell density of R. raciborskii was observed in the treatment with clethodim plus limited nitrogen. Also, the cell-free exudate of R. raciborskii significantly stimulated the growth of M. aeruginosa on day 3 of the experiment. The concentration of chlorophyll-a in M. aeruginosa cultures generally increased in all the treatments, while in R. raciborskii cultures, the opposite occurred. Total microcystins (MCs) content of M. aeruginosa in the mixed cultures was 68% higher in nitrogen-enriched conditions than the control. A similar increase in MC content occurred in M. aeruginosa unialgal culture treated with R. raciborskii exudate. Total saxitoxin concentration was 81% higher in mixed cultures of R. raciborskii simultaneously exposed to high nitrogen and clethodim. Similarly, unialgal cultures of R. raciborskii exposed to either high nitrogen or clethodim had higher saxitoxins concentrations than the control. The intracellular H2O2 content of M. aeruginosa cultures decreased, whereas, in R. raciborskii cultures, it increased during exposure to high nitrogen and clethodim. Only R. raciborskii had a significant variation in peroxidase activity. The activities of glutathione S-transferase of both strains were higher in the presence of clethodim. These results revealed that nitrogen enrichment and the presence of clethodim might lead to the excessive proliferation of M. aeruginosa and R. raciborskii and increased production of cyanotoxins in aquatic environments.

Clethodim (herbicide) alters the growth and toxins content of Microcystis aeruginosa and Raphidiopsis raciborskii.

Increased agricultural intensification goes with the widespread use of herbicides that adversely affect aquatic biodiversity. The effects of herbicides on toxin-producing cyanobacteria have been poorly studied. The present study aimed to investigate the toxicological and physiological effects of the herbicide clethodim on Raphidiopsis raciborskii (a.k.a. Cylindrospermopsis raciborskii) ITEPA1 and Microcystis aeruginosa BCCUSP232. On day four of the experiment, the exposure to 25 mg/L clethodim resulted in the highest cell density of R. raciborskii. Similarly, exposure to the 1, 5, 20, and 50 mg/L clethodim treatments resulted in the highest cell densities of M. aeruginosa on day 4 of the experiment. Medium effect concentrations (EC50) after 96 h of exposure of both strains to clethodim were 192.98 mg/L and 168.73 mg/L for R. raciborskii and M. aeruginosa, respectively. The presence of clethodim significantly increased the total microcystin content of M. aeruginosa compared to the control cultures. At 400 mg/L, total saxitoxins content of R. raciborskii was 27% higher than that of the control cultures on day 4. In contrast, cultures exposed to 100 mg/L clethodim had the lowest saxitoxins levels per cell quota. There was an increase in the levels of intracellular hydrogen peroxide in both species during exposure to clethodim, which was followed by significant changes (p < 0.05) in the activity of antioxidant enzymes such as peroxidase and superoxide dismutase. These results revealed that the presence of low levels of clethodim in the aquatic environment might lead to the excessive proliferation of cyanobacteria and alteration of their cyanotoxins content.

Effect of flavonoids isolated from Tridax procumbens on the growth and toxin production of Microcystis aeruginos.

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.

Potential human health risk assessment of cylindrospermopsin accumulation and depuration in lettuce and arugula.

The cyanobacterial toxin cylindrospermopsin (CYN) has become a globally important secondary metabolite due to the negative effect it has on human and animal health. As a means of evaluating the risk of human exposure to CYN, the bioaccumulation and depuration of the toxin in lettuce (Lactuca sativa L.) and arugula (Eruca sativa Mill.) were investigated, after irrigation with contaminated water. The vegetables were irrigated for 7days with CYN (3, 5 and 10µg/L) contaminated water (bioaccumulation phase), and subsequently, irrigated for 7days with uncontaminated distilled water (depuration phase). In general, the bioaccumulation of CYN in both vegetables decreased with increasing exposure concentration. Bioconcentration factor (BCF) of CYN increased with the progression of the experiment at 3.0µg/L CYN, while the reverse occurred at 5 and 10µg/L CYN. In arugula, BCF increased at all CYN exposure concentrations throughout the study. The depuration of CYN decreased with increasing exposure concentration but was highest in the plants of both species with the highest bioaccumulation of CYN. Specifically, in plants previously irrigated with water contaminated with 3, 5 and 10µg/L CYN, the depuration of the toxin was 60.68, 27.67 and 18.52% for lettuce, and 47, 46.21 and 27.67% for arugula, respectively. Human health risks assessment revealed that the consumption of approximately 10 to 40g of vegetables per meal will expose children and adults to 1.00-6.00ng CYN/kg body mass for lettuce and 2.22-7.70ng CYN/kg body mass for arugula. The irrigation of lettuce and arugula with contaminated water containing low CYN concentrations constitutes a potential human exposure route.

Response of Microcystis aeruginosa BCCUSP 232 to barley (Hordeum vulgare L.) straw degradation extract and fractions.

The eutrophication of aquatic ecosystems is a serious environmental problem that leads to increased frequency of cyanobacterial blooms and concentrations of cyanotoxins. These changes in aquatic chemistry can negatively affect animal and human health. Environment-friendly methods are needed to control bloom forming cyanobacteria. We investigated the effect of Hordeum vulgare L. (barley) straw degradation extract and its fractions on the growth, oxidative stress, antioxidant enzyme activities, and microcystins content of Microcystis aeruginosa (Kützing) Kützing BCCUSP232. Exposure to the extract significantly (p<0.05) inhibited the growth of M. aeruginosa throughout the study, whereas only the highest concentration of fractions 1 and 2 significantly (p<0.05) reduced the growth of the cyanobacterium on day 10 of the experiment. The production of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzyme activities were significantly (p<0.05) altered by the extract and fractions 1 and 2. Phytochemical profiling of the extract and its fractions revealed that the barley straw degradation process yielded predominantly phenolic acids. These results demonstrate that barley straw extract and its fractions can efficiently interfere with the growth and development of M. aeruginosa under laboratory conditions.

Cylindrospermopsin induced changes in growth, toxin production and antioxidant response of Acutodesmus acuminatus and Microcystis aeruginosa under differing light and nitrogen conditions.

Growing evidence suggests that some bioactive metabolites (e.g. cyanotoxins) produced by cyanobacteria have allelopathic potential, due to their inhibitory or stimulatory effects on competing species. Although a number of studies have shown that the cyanotoxin cylindrospermopsin (CYN) has variable effects on phytoplankton species, the impact of changing physicochemical conditions on its allelopathic potential is yet to be investigated. We investigated the physiological response of Microcystis aeruginosa (Cyanobacteria) and Acutodesmus acuminatus (Chlorophyta) to CYN under varying nitrogen and light conditions. At 24h, higher microcystins content of M. aeruginosa was recorded under limited light in the presence of CYN, while at 120h the lower levels of the toxins were observed in the presence of CYN under optimum light. Total MCs concentration was significantly (p<0.05) lowered by CYN after 120h of exposure under limited and optimum nitrogen conditions. On the other hand, there were no significant (p>0.05) changes in total MCs concentrations after exposure to CYN under high nitrogen conditions. As expected, limited light and limited nitrogen conditions resulted in lower cell density of both species, while CYN only significantly (p<0.05) inhibited the growth of M. aeruginosa. Regardless of the light or nitrogen condition, the presence of CYN increased internal H2O2 content of both species, which resulted in significant (p<0.05) changes in antioxidant enzyme (catalase, peroxidase, superoxide dismutase and glutathione S-transferase) activities. The oxidative stress caused by CYN was higher under limited light and limited nitrogen. These results showed that M. aeruginosa and A. acuminatus have variable response to CYN under changing light and nitrogen conditions, and demonstrate that need to consider changes in physicochemical conditions during ecotoxicological and ecophysiological investigations.

Does anatoxin-a influence the physiology of Microcystis aeruginosa and Acutodesmus acuminatus under different light and nitrogen conditions?

Due to changing global climatic conditions, a lot of attention has been given to cyanobacteria and their bioactive secondary metabolites. These conditions are expected to increase the frequency of cyanobacterial blooms, and consequently, the concentrations of cyanotoxins in aquatic ecosystems. Unfortunately, there are very few studies that address the effects of cyanotoxins on the physiology of phytoplankton species under different environmental conditions. In the present study, we investigated the effect of the cyanotoxin anatoxin-a (ATX-A) on Microcystis aeruginosa (cyanobacteria) and Acutodesmus acuminatus (chlorophyta) under varying light and nitrogen conditions. Low light (LL) and nitrogen limitation (LN) resulted in significant cell density reduction of the two species, while the effect of ATX-A on M. aeruginosa was not significant. However, under normal (NN) and high nitrogen (HN) concentrations, exposure to ATX-A resulted in significantly (p < 0.05) lower cell density of A. acuminatus. Pigment content of M. aeruginosa significantly (p < 0.05) declined in the presence of ATX-A, regardless of the light condition. Under each light condition, exposure to ATX-A caused a reduction in total microcystin (MC) content of M. aeruginosa. The detected MC levels varied as a function of nitrogen and ATX-A concentrations. The production of reactive oxygen species (H2O2) and antioxidant enzyme activities of both species were significantly altered by ATX-A under different light and nitrogen conditions. Our results revealed that under different light and nitrogen conditions, the response of M. aeruginosa and A. acuminatus to ATX-A was variable, which demonstrated the need for different endpoints of environmental factors during ecotoxicological investigations.

Microcystin-LR bioaccumulation and depuration kinetics in lettuce and arugula: Human health risk assessment.

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.

Lettuce irrigated with contaminated water: Photosynthetic effects, antioxidative response and bioaccumulation of microcystin congeners.

The use of microcystins (MCs) contaminated water to irrigate crop plants represents a human health risk due to their bioaccumulation potential. In addition, MCs cause oxidative stress and negatively influence photosynthetic activities in plants. The present study was aimed at investigating the effect of MCs on photosynthetic parameters and antioxidative response of lettuce. Furthermore, the bioaccumulation factor (BAF) of total MCs, MC-LR and MC-RR in the vegetable after irrigation with contaminated water was determined. Lettuce crops were irrigated for 15 days with water containing cyanobacterial crude extracts (Microcystis aeruginosa) with MC-LR (0.0, 0.5, 2.0, 5.0 and 10.0 µg L(-1)), MC-RR (0.0, 0.15, 0.5, 1.5 and 3.0 µg L(-1)) and total MCs (0.0, 0.65, 2.5, 6.5 and 13.0 µg L(-1)). Increased net photosynthetic rate, stomatal conductance, leaf tissue transpiration and intercellular CO2 concentration were recorded in lettuce exposed to different MCs concentrations. Antioxidant response showed that glutathione S-transferase activity was down-regulated in the presence of MCs. On the other hand, superoxide dismutase, catalase and peroxidase activities were upregulated with increasing MCs concentrations. The bioaccumulation factor (BAF) of total MCs and MC-LR was highest at 6.50 and 5.00 µg L(-1), respectively, while for MC-RR, the highest BAF was recorded at 1.50 µg L(-1) concentration. The amount of total MCs, MC-LR and MC-RR bioacumulated in lettuce was highest at the highest exposure concentrations. However, at the lowest exposure concentration, there were no detectable levels of MC-LR, MC-RR and total MCs in lettuce. Thus, the bioaccumulation of MCs in lettuce varies according to the exposure concentration. In addition, the extent of physiological response of lettuce to the toxins relies on exposure concentrations.