Maranda Esterhuizen-Londt

Rise of toxic cyanobacterial blooms in temperate freshwater lakes: causes, correlations and possible countermeasures

ABSTRACT The progressive degradation of surface freshwater quality due to the mass proliferation of toxic cyanobacterial blooms is of growing global concern. The occurrence of cyanobacterial blooms is not a new phenomenon. However, a global increase in the frequency, duration, and distribution of toxic cyanobacterial blooms could be observed in the past decades. Evidence suggests that this trend might be attributed to a complex interplay of direct and indirect anthropogenic influences. The underlying causes and interrelations for this development have not been fully clarified. Nonetheless, all evidence points to the fact that mitigation of toxic cyanobacterial blooms will be a key challenge of the twenty-first century. This review addresses the underlying causes for the increased incidence of toxic cyanobacteria in temperate freshwater lakes and attempts to reveal possible reciprocities between bloom promoting factors. Selected approaches for the prevention of toxic cyanobacterial blooms as well as the mitigation of their potential negative impacts on humans will be presented.

LC-MS/MS method development for quantitative analysis of acetaminophen uptake by the aquatic fungus Mucor hiemalis.

Acetaminophen is a pharmaceutical, frequently found in surface water as a contaminant. Bioremediation, in particular, mycoremediation of acetaminophen is a method to remove this compound from waters. Owing to the lack of quantitative analytical method for acetaminophen in aquatic organisms, the present study aimed to develop a method for the determination of acetaminophen using LC-MS/MS in the aquatic fungus Mucor hiemalis. The method was then applied to evaluate the uptake of acetaminophen by M. hiemalis, cultured in pellet morphology. The method was robust, sensitive and reproducible with a lower limit of quantification of 5 pg acetaminophen on column. It was found that M. hiemalis internalize the pharmaceutical, and bioaccumulate it with time. Therefore, M. hiemalis was deemed a suitable candidate for further studies to elucidate its pharmaceutical tolerance and the longevity in mycoremediation applications.

Antioxidative stress responses in the floating macrophyte Lemna minor L. with cylindrospermopsin exposure.

Cylindrospermopsin toxicity and oxidative stress have been examined in aquatic animals, however, only a few studies with aquatic plants have been conducted focusing on the potential for bioaccumulation of cylindrospermopsin. The oxidative stress effects caused by cylindrospermopsin on macrophytes have not yet been specifically studied. The oxidative stress response of Lemna minor L. with exposure to cylindrospermopsin, was therefore tested in this study. The hydrogen peroxide concentration together with the activities of the antioxidant enzymes (catalase, peroxidase, glutathione reductase and glutathione S-transferase) were determined after 24h (hours) of exposure to varying concentrations (0.025, 0.25, 2.5 and 25μg/L) of cylindrospermopsin. Responses with longer exposure periods (48, 96, 168h) were tested only with exposure to 2.5 and 25μg/L cylindrospermopsin. Additionally, the content of the carotenoids was determined as a possible non-enzymatic antioxidant defence mechanism against cylindrospermopsin. The levels of hydrogen peroxide increased after 24h even at the lowest cylindrospermopsin exposure concentrations. Catalase showed the most representative antioxidant response observed after 24h and maintained its activity throughout the experiment. Catalase activity corresponded with the contents of hydrogen peroxide at 2.5 and 25μg/L cylindrospermopsin. The data suggest that glutathione S-transferase, glutathione reductase and the carotenoid content act together with catalase but are more sensitive to higher concentrations of cylindrospermopsin and after a longer exposure period (168h). The results indicate that cylindrospermopsin promotes oxidative stress in L. minor at concentrations of 2.5 and 25μg/L. However, L. minor has sufficient defence mechanisms in place against this cyanobacterial toxin. Even though L. minor exhibits the potential to managing and control cylindrospermopsin contamination in aquatic systems, further studies in tolerance limits to cylindrospermopsin, uptake and experiments with prolonged exposure periods of more than 7 days are required.

The effect of oxytetracycline on physiological and enzymatic defense responses in aquatic plant species Egeria densa, Azolla caroliniana, and Taxiphyllum barbieri

ABSTRACT Oxytetracycline is an antibiotic widely employed in aquaculture to control and treat bacterial diseases of fish. Due to ineffective wastewater treatment, antibiotic residues from fish ponds are directly released into surface and groundwater affecting the environment. The Green Liver System® is a sustainable and cost-effective water treatment based on the ability of aquatic plants to biotransformation xenobiotics. To expand the application range of this system, Egeria densa, Azolla caroliniana, and Taxiphyllum barbieri were tested in response to oxytetracycline exposure. The aquatic plants were exposed to 0.5, 2.0, 5.0, and 25 μg/L oxytetracycline (n = 4) for 24 h in order to analyze the physiological responses (photosynthetic pigment contents and hydrogen peroxide formation -- H2O2), the biotransformation process (activity of glutathione S-transferase), and the antioxidant defense (glutathione reductase; peroxidase; and catalase) responses. There were no statistical differences in the photosynthetic pigment contents and H2O2 level between the treatment and control groups in the three aquatic plant species. It can be concluded that oxytetracycline does not provoke physiological damage to the tested species within 24 h of exposure. Nevertheless, the antioxidant defense mechanism was activated. Peroxidase and catalase were the most prominent antioxidant enzymes in the three plant species, whereas glutathione S-transferase activity was significantly increased in A. caroliniana and T. barbieri.

Oxidative stress responses in the animal model, Daphnia pulex exposed to a natural bloom extract versus artificial cyanotoxin mixtures

In the natural environment, Daphnia spp. are constantly exposed to a complex matrix of biomolecules, especially during cyanobacterial bloom events. When cyanobacterial cells decay, not only are toxic secondary metabolites known as cyanotoxins released, but also multiple other secondary metabolites, some of which act as enzyme inhibitors. The present study examined the effects of such a natural toxin matrix (crude extract from a bloom) versus artificial toxin mixtures in terms of oxidative stress in Daphnia pulex. The results indicate that there is no significant effect on the survival of D. pulex. However, exposure to the bloom extract resulted in increased lipid peroxidation over a shorter exposure period and reduced antioxidative enzyme activities when compared to the artificial mixtures. The daphnids also needed a longer recovery time to reduce the increased cellular hydrogen peroxide concentration associated with the exposure to the crude extract than with the artificial mixtures. The results indicate a significant difference between the bloom crude extract and the two synthetic mixtures for all stress markers tested, indicating enhanced toxicity of the bloom extract.

Development and validation of an in‐house quantitative analysis method for cylindrospermopsin using hydrophilic interaction liquid chromatography–tandem mass spectrometry: Quantification demonstrated in 4 aquatic organisms

The cyanobacterial toxin cylindrospermopsin (CYN) is of great concern in aquatic environments because of its incidence, multiple toxicity endpoints, and, therefore, the severity of health implications. It may bioaccumulate in aquatic food webs, resulting in high exposure concentrations to higher-order trophic levels, particularly humans. Because of accumulation at primary levels resulting from exposure to trace amounts of toxin, a sensitive analytical technique with proven aquatic applications is required. In the present study, a hydrophilic interaction liquid chromatographic-tandem mass spectrometric method with a lower limit of detection of 200 fg on column (signal-to-noise ratio = 3, n = 9) and a lower limit of quantification of 1 pg on column (signal-to-noise ratio = 11, n = 9) with demonstrated application in 4 aquatic organisms is described. The analytical method was optimized and validated with a linear range (r(2) = 0.999) from 0.1 ng mL(-1) to 100 ng mL(-1) CYN. Mean recovery of the extraction method was 98 ± 2%. Application of the method was demonstrated by quantifying CYN uptake in Scenedesmus subspicatus (green algae), Egeria densa (Brazilian waterweed), Daphnia magna (water flea), and Lumbriculus variegatus (blackworm) after 24 h of static exposure to 50 μg L(-1) CYN. Uptake ranged from 0.05% to 0.11% of the nominal CYN exposure amount. This constitutes a sensitive and reproducible method for extraction and quantification of unconjugated CYN with demonstrated application in 4 aquatic organisms, which can be used in further aquatic toxicological investigations.

Still challenging: the ecological function of the cyanobacterial toxin microcystin – What we know so far

Abstract Microcystins (MCs) are the most commonly studied cyanotoxins. While these past studies have mainly focused on the toxicity of MCs, the evolutionary history of life has shown that toxicity can be considered as an assigned role to MCs. Nowadays, there is a growing interest in understanding the importance of cyanotoxins in any of the physiological processes or beyond at the ecological level. This review evaluates variously proposed intracellular and extracellular functions of MCs and how they benefit the producing cyanobacterium. However, the strain-specific and divergent laboratory and field results obtained to date have made it difficult to generalize.

Physiological responses of Cladophora glomerata to cyanotoxins: a potential new phytoremediation species for the Green Liver Systems

Due to increased pollution of potable water sources as a consequence of eutrophication and anthropogenic xenobiotics, sustainable water purification is an essential concern. Therefore, the Green Liver System, a natural, economic and sustainable water purification system employing the biotransformation capabilities of aquatic plants, was developed. To expand the capacities and applications of this system, new aquatic plants are continually evaluated for their potential to remediate various aquatic pollutants. In this study, the potential of Cladophora glomerata to internalize cyanotoxins, microcystins (MCs) and anatoxin-a, and consequently its ability to cope with the subsequent oxidative stress associated with toxin-uptake were investigated. C. glomerata was able to take up all three of the tested MC congeners as well as anatoxin-a, similarly to previous toxin internalizations reported for aquatic plants such as Ceratophyllum demersum, Myriophyllum spicatum and Hydrilla versiculata. The antioxidative stress defense of C. glomerata proved to efficiently endure the toxin-uptake with no adverse effects. Subsequently, the uptake potential of C. glomerata was investigated at lab-scale by exposure to the three MC congeners and anatoxin-a collectively. After a period of seven days, 95–97% of the MCs and 100% of anatoxin-a were removed from the exposure media. C. glomerata therefore, is a suitable candidate to be incorporated in future Green Liver Systems.

Toxicity and Toxin Composition of Microcystis aeruginosa from Wangsong Reservoir

ObjectiveThe increasing world population, resulting in increased anthropogenic water pollution, is negatively impacting the limited available water resources. In South Korea, this similarly affects the water quality of reservoirs. As water is a basic necessity for life, water quality monitoring is essential but typically does not include toxicity testing. However, as toxic bloom event frequencies are increasing, this previously neglected aspect becomes pertinent. Therefore, in the present study, the toxin composition and toxicity of a Microcystis aeruginosa strain isolated from a persistent bloom in lake Wangsong, South Korea, was investigated.MethodsA combination of bioassays and chemical analysis was used for this purpose. The bioassay species included terrestrial and aquatic plants, an alga, a rotifer, a tubificid annelid, and crustaceans, representing various trophic levels.ResultsThe strain was found to produce microcystin-LR, -RR, and YR, as well as β-N-methylamino-L-alanine. The bioassays indicated that the primary producers were less sensitive to the crude extract.ConclusionThe presence or absence of a visible cyanobacterial bloom is also not an indication of the toxins that may be present in the afflicted waters, and thus does not predict exposure risk. Similarly, the presence and absence of toxins and mixtures thereof does not indicate the ecological effect. Therefore, it would be advantages to include toxicity testing into routine water testing regimes to better understand the impact of harmful algal blooms.

Fate of Enrofloxacin in Lake Sediment: Biodegradation, Transformation Product Identification, and Ecotoxicological Implications

ABSTRACT Various pharmaceutical drugs are being detected in different environmental compartments such as surface waters, groundwater, and sediment; a major concern since they are biologically active substances which can interfere with biological systems affecting the native biota. Among these drugs, antimicrobials are especially worrisome mainly due to the development of bacterial resistance. The aims of this study were to investigate if enrofloxacin, an emergent antibiotic pollutant, could be biodegraded in lake sediment, identify its break down products and to determine if these products have antimicrobial properties or are toxic. Three biodegradation products were identified and the antibiotic susceptibility assay proved that the products formed did not display antibiotic effects. Ecotoxicity testing with green algae suggested that the degradation products do not cause adverse effects statistically. However, it is suggested that further investigations are needed to identify the mechanism of degradation and the microbes involved.