Microcystis are known for their potential ability to synthesize toxins, mainly microcystins (MCs). In order to evaluate the effects of temperature on chlorophyll a (Chl a), growth, physiological responses and toxin production of a native Microcystis aeruginosa, we exposed the cells to low (23°C) and high (29°C) temperature in addition to a 26°C control treatment. Exponential growth rate was significantly higher at 29°C compared to 23°C and control, reaching 0.43, 0.32 and 0.33 day −1 respectively. In addition, there was a delay of the start of exponential growth at 23°C. However, the intracellular concentration of Chl a decreased significantly due to temperature change. A significant increase in intracellular ROS was observed in coincidence with the activation of enzymatic antioxidant catalase (CAT) during the first two days of exposure to 23°and 29°C in comparison to the control experiment, decreasing thereafter to nearly initial values. in cells exposed to 29°C. The same trend was observed for all other MCs except for the least abundant MC-LR which showed a continuous increase during exposure time. Our results suggest a high ability of M. aeruginosa to perceive ROS and to rapidly initiate antioxidant defenses with a differential response on MC production.
The effects of prolonged exposure to microcystins (MCs) on health are not yet sufficiently understood and this type of poisoning is often undiagnosed. Even though chronic exposure has been linked with liver cancer and alterations have been described in liver damage marker enzymes in exposed populations, there are not profile parameters that indicate prolonged exposure to microcystins. The aim of this work is to determine, based on an animal model of prolonged exposure to successive i.p. doses of 25 μg MC-LR/kg body weight, several plasma parameters which could be useful as exposure biomarkers. Hemoglobin (Hb) and methemoglobin (MetHb) levels were determined on blood samples. We also studied plasma levels of hydroperoxides (ROOHs), α-tocopherol, glutathione and lipid profile as well as superoxide dismutase (SOD) and catalase (CAT) erythrocyte activities. In addition, the determination of MC-LR levels in liver, kidney, plasma, urine and feces of treated mice was carried out. We found that alteration in MetHb, ROOHs, glutathione, α-tocopherol levels, SOD activity and plasma lipid profile, correlates with those expected if the alteration derived from hepatic damage. The alterated plasma paramenters together with MC-LR determination could be used as biomarkers, helpful tools in screening and epidemiological studies.
RATIONALE: High-resolution mass spectrometry was applied to the study of a Microcystis aeruginosa strain previously reported as a [D-Leu 1 ]MC-LR producer. Detailed analysis revealed new microcystin (MC) variants produced from the strain, and seven of these were previously unreported variants. This work shows the importance of mass accuracy for the identification of unknown MCs. METHODS: The M. aeruginosa strain was isolated from a bloom sample collected from Argentina and acclimated to lab conditions. The MC variants in the strain were separated by UV/Vis detection-guided high-performance liquid chromatography, and their structures were unambiguous determined by tandem mass spectrometry (MS/MS). RESULTS: A simple strategy was developed for quickly locating the low-abundance MC precursors from complex samples. MS/MS anlysis revealed ten MC variants produced from the strain, of which seven have never been reported before. CONCLUSIONS: This work shows the interference of isobarics and isomers in the study of unknown MCs, and, therefore, high mass accuracy is important to avoid false assignments. Moreover, the peak list provided here (30-50 fragments unambiguously assigned for ten MCs) can be used as a reference for the discovery of MCs from environmental samples.
The aim of this study was to evaluate the effects of short-term (hours) exposure to solar UV radiation (UVR, 280-400 nm) on the physiology of Microcystis aeruginosa. Three solar radiation treatments were implemented: (i) PAR (PAR, 400-700 nm), (ii) TUVA (PAR + UVAR, 315-700 nm) and (iii) TUVR (PAR + UVAR + UVBR, 280-700 nm). Differential responses of antioxidant enzymes and the reactive oxygen species (ROS) production to UVR were observed. Antioxidant enzymes were more active at high UVR doses. However, different responses were observed depending on the exposure to UVAR or UVBR and the dose level. No effects were observed on the biomass, ROS production or increased activity of superoxide dismutase (SOD) and catalase (CAT) compared to the control when UVR + PAR doses were lower than 9875 kJ m. For intermediate doses, UVR + PAR doses between 9875 and 10 275 kJ m, oxidative stress increased while resistance was imparted through SOD and CAT in the cells exposed to UVAR. Despite the increased antioxidant activity, biomass decrease and photosynthesis inhibition were observed, but no effects were observed with added exposure to UVBR. At the highest doses (UVR + PAR higher than 10 275 kJ m), the solar UVR caused decreased photosynthesis and biomass with only activation of CAT by UVBR and SOD and CAT by UVAR. In addition, for such doses, a significant decrease of microcystins (MCs, measured as MC-LR equivalents) was observed as a consequence of UVAR. This study facilitates our understanding of the SOD and CAT protection according to UVAR and UVBR doses and cellular damage and reinforces the importance of UVR as an environmental stressor. In addition, our results support the hypothesized antioxidant function of MCs.
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