Titanium (IV) oxide anatase nanoparticles as vectors for diclofenac: assessing the antioxidative responses to single and combined exposures in the aquatic macrophyte Egeria densa.

Titanium dioxide, frequently used in commonplace products, is now regularly detected in aquatic environments. Understanding its toxic effects on native biota is essential. However, combined toxicity with commonly occurring pollutants, such as the pharmaceutical diclofenac, may provide more insight into environmental situations. Therefore, the present study aimed to evaluate the effects of titanium dioxide and diclofenac, individually and combined, on the macrophyte Egeria densa. Diclofenac uptake and removal by the macrophyte were assessed. Diclofenac and titanium dioxide were mixed prior to exposure to allow binding, which was assessed. Toxicity of the individual compounds and the combination was evaluated by assaying enzymes as bioindicators of biotransformation and the antioxidative system. Cytosolic glutathione S-transferase and glutathione reductase activities were increased by diclofenac, titanium dioxide, and the combination. Both enzymes' activities were more significantly elevated by diclofenac and the combination than nanoparticles alone. Microsomal glutathione S-transferase was unaffected by diclofenac exposure but inhibited with titanium dioxide and the mixture. Diclofenac elicited the most significant response. Based on the data, the cytosolic enzymes effectively prevented damage.

Self-contamination from clothing in microplastics research.

Self-contamination should not be underestimated when quantifying microplastics (MPs) in environmental matrices. Standardised and validated methodologies for MP sampling, extraction, and analysis are lacking. The various applications of plastics in our society have made them ubiquitous, even in clothing, rendering MP self-contamination inevitable. In the present study, we sampled lake sediment, snow, and ice, purposefully wearing red overalls composed of cotton; fibres from which we could quantify using Fourier-Transform Infrared Spectroscopy (FTIR), serving as an indication of possible self-contamination from clothes. The suitability of cotton as a representation of MP contamination was also evaluated. For all detected fibres, 25 ± 1%, 20 ± 7%, and 8 ± 6% for snow, ice, and sediment, respectively, originated from sampling attire. These findings demonstrate that self-contamination can play a significant role when quantifying MP pollution, highlighting that sampling conducted to date might have overestimated the presence of MP or even contaminated MP-free samples.

Vegetables cultivated with exposure to pure and naturally occurring ß-N-methylamino-L-alanine (BMAA) via irrigation.

Bioaccumulation and biomagnification of ß-N-methylamino-L-alanine (BMAA), a potent neurotoxin, has been demonstrated in various food webs. It is alarming as this intensification of BMAA will result in exposure to higher concentrations from a direct cyanobacterial source. As more food items are being identified as a source of BMAA and with the large variations in BMAA content, the aim of the present study was to evaluate BMAA uptake by, and accumulation in, two commonly consumed vegetables, Lactuca sativa and Allium fistulosum. Plants exposed to pure BMAA in controlled laboratory experiments, as well as vegetables naturally irrigated with water containing a BMAA producing cyanobacterial bloom were evaluated during growth and ripening. In the laboratory exposures, free BMAA was detected in both the edible ripe parts of L. sativa and A. fistulosum after 60 days of exposure to a total of 4.5 µg BMAA. However, in the bloom exposure samples no BMAA could be detected in the ripe vegetables of A. fistulosum, Cucurbita pepo, or Brassica rapa chinensis. The study emphasises the need to further screen items for BMAA to understand the human exposure risk as well as the difference between BMAA uptake patterns with free BMAA and that contained in cyanobacterial cells.

Desmodesmus subspicatus co-cultured with microcystin producing (PCC 7806) and the non-producing (PCC 7005) strains of Microcystis aeruginosa.

Although microcystins (MCs) are the most commonly studied cyanotoxins, their significance to the producing organisms remains unclear. MCs are known as endotoxins, but they can be found in the surrounding environment due to cell lysis, designated as extracellular MCs. In the present study, the interactions between MC producing and the non-producing strains of Microcystis aeruginosa, PCC 7806 and PCC 7005, respectively, and a green alga, Desmodesmus subspicatus, were studied to better understand the probable ecological importance of MCs at the collapse phase of cyanobacterial blooms. We applied a dialysis co-cultivation system where M. aeruginosa was grown inside dialysis tubing for one month. Then, D. subspicatus was added to the culture system on the outside of the membrane. Consequently, the growth of D. subspicatus and MC contents were measured over a 14-day co-exposure period. The results showed that Microcystis negatively affected the green alga as the growth of D. subspicatus was significantly inhibited in co-cultivation with both the MC-producing and -deficient strains. However, the inhibitory effect of the MC-producing strain was greater and observed earlier compared to the MC-deficient strain. Thus, MCs might be considered as an assistant factor that, in combination with other secondary metabolites of Microcystis, reinforce the ability to outcompete co-existing species.

Phytoremediation: green technology for the removal of mixed contaminants of a water supply reservoir.

The Iraí Reservoir, a water supply in Brazil, is constantly impacted by anthropogenic activities such as waste inputs from agriculture, hospitals and urbanization, resulting toxic cyanobacterial blooms causing economic, social and environmental problems. This study assessed the concentration of some common contaminants of the Iraí Reservoir, namely paracetamol, diclofenac and microcystin-LR and tested whether a laboratory scale Green Liver System® would serve as a suitable technology to remove these contaminants. Further, the study investigated whether the pollutants caused adverse effects to the macrophytes using catalase as a biomarker for oxidative stress and investigated whether biotransformation (glutathione S-transferase) was a main route for detoxification. Egeria densa, Ceratophyllum demersum and Myriophyllum aquaticum were exposed to a mixture of the three contaminants for 14 days in a concentration range similar to those detected in the reservoir. The plants removed 93% of diclofenac and 100% of MC-LR after 14 days. Paracetamol could not be detected. Catalase and glutathione S-transferase enzyme activities remained unaltered after the 14-day exposure, indicating that the mixture did not cause oxidative stress. The study showed that the aquatic macrophytes used are suitable tools to apply in a Green Liver System® for the remediation of mixed pollutants.

Uptake and biotransformation of pure commercial microcystin-LR versus microcystin-LR from a natural cyanobacterial bloom extract in the aquatic fungus Mucor hiemalis.

OBJECTIVES: To evaluate the remediation efficiency of Mucor hiemalis by comparing media elimination, uptake, and biotransformation of microcystin-LR with exposure to pure toxin versus a crude bloom extract. RESULTS: With exposure to the extract, the elimination rate of microcystin-LR from the media, which was 0.28 ng MC-LR l-1 h-1, was significantly higher compared to that achieved with exposure to the pure toxin (0.16 ng MC-LR l-1 h-1) after 24 h. However, intracellular breakdown of microcystin-LR was significantly lower in the extract exposed pellets compared to the pure toxin treated fungal pellets over time. This coincided with reduced intracellular glutathione S-transferase activity with crude extract exposure which could be responsible for the detection of only the glutathione conjugate of microcystin-LR. CONCLUSION: This paper signifies the importance of using laboratory exposure scenarios which resemble conditions in nature to fully understand and evaluate remediation efficiency. There is merit in using M. hiemalis for mycoremediation of cyanotoxins in surface waters.

Pharmacological and pharmacokinetic properties of lanthipeptides undergoing clinical studies.

The intrinsic qualities of lanthipeptides for their use as therapeutic drugs present several challenges because of their properties, which include stability, solubility and bioavailability, which, under physiological conditions, are very low. Researches have encouraged clinical evaluation of a few compounds, such as mutacin 1140, microbisporicin, actagardine and duramycin, with pharmacokinetic profiles showing rapid distribution and elimination rates, good bioavailability and fecal excretion, as well as high protein binding. Local and parenteral administration are currently suitable to minimize environmental influences on lanthipeptides and ensure efficient activity. Nevertheless, valuable improvements on pharmacodynamic and pharmacokinetic properties may also permit systemic applications via enteral routes. Understanding how rational modifications influence the desired pharmacological and pharmacokinetic properties of these biomolecules would help to answer some specific questions about their susceptibility to environmental changes, mechanism of action and how to engineer other peptides of the same group to improve their clinical relevance.

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.

Uptake of the cyanobacterial neurotoxin, anatoxin-a, and alterations in oxidative stress in the submerged aquatic plant Ceratophyllum demersum.

The prevalence of cyanobacterial blooms in fresh water bodies worldwide has become a serious environmental problem. The blooms can increase the occurrence of cyanobacterial neurotoxin, anatoxin-a, and this toxin can interact with aquatic plants and other pivotal components of aquatic ecosystems. Despite this, several questions regarding the uptake of the toxin by aquatic plants and its association with toxic effects still remain. This study investigated the uptake of anatoxin-a in relation to alterations in oxidative stress, estimated by changes in lipid peroxidation and tocopherol contents (alpha-, beta-, gamma-, and delta-tocopherol), in the submerged aquatic plant, Ceratophylum demersum, at environmentally relevant concentrations. Exposure to five different concentrations of anatoxin-a (0.005, 0.05, 0.5, 5 and 50µgl(-1)) for 24h increased concentrations in C. demersum in a dose-dependent manner. All four forms of tocopherols were elevated at low concentrations of anatoxin-a (0.005. 0.05. 0.5 and 5µgl(-1)). However, a decline in the four tocopherol forms along with a high level of lipid peroxidation was observed at 50µgl(-1) exposure dose. During 336-h exposure to 15µgl(-1) anatoxin-a, rapid toxin uptake during the first 24h and subsequent steady accumulation of the toxin were observed. The four tocopherol forms increased in response to anatoxin-a uptake, attaining their maximum levels together with a significant increase of lipid peroxidation after 12 or 24h. After 24-h exposure, the four tocopherol forms decreased gradually without recovery. The results clearly indicate that anatoxin-a uptake can cause a disturbance of the oxidative stress in the aquatic plant, and depending on the concentration and exposure duration, oxidative damage occurs.

Uptake of a cyanotoxin, ß-N-methylamino-L-alanine, by wheat (Triticum aestivum).

In order to study the uptake of the cyanobacterial neurotoxin ß-N-methylamino-l-alanine (BMAA) into the crop plant Triticum aestivum during germination and primary growth imbibed grains and 7-day-old seedlings were irrigated with 100 and 1000µg l(-1) BMAA for 4 days and 100µg l(-1) BMAA for 28 days. Content of derivatized free and protein-associated BMAA in seedlings, root and shoot tissue, respectively, were analyzed by LC-MS/MS. Free BMAA was only detected in seedlings exposed to 1000µg l(-1) BMAA, whereas protein-associated BMAA was found at both exposure concentrations. Irrigation with 100µgl(-1) BMAA led to an uptake of the neurotoxin into roots and shoots and to immediate protein-association. In roots, protein-associated BMAA was detectable after 5 days with peaking amounts after 14 days. Longer exposure did not cause further accumulation in roots. In contrast, protein-associated BMAA was detected in shoot samples after only 1 day. In shoots the highest amounts of protein-associated BMAA were found after 28 days. In turn, in both plant compartments free BMAA was below the measurable concentration.

The fate of the cyanobacterial toxin ß-N-methylamino-L-alanine in freshwater mussels.

The cyanobacterial neurotoxin, ß-N-methylamino-l-alanine (BMAA) has been suggested as a causative agent for certain neurodegenerative diseases. This cyanotoxin bioaccumulates in an array of aquatic organisms, in which it occurs as both a free amino acid and in a protein-associated form. This study was intended to investigate the environmental fate of BMAA by examining the metabolism of isotopically labeled BMAA in four freshwater mussel species. All species showed substantial uptake of BMAA from the culture media. Data showed no significant evidence for BMAA catabolism in any of the animals but did suggest metabolism via the reversible covalent modification of BMAA in freshwater mussels, a process that appears to be variable in different species.

Accumulation of microcystin congeners in different aquatic plants and crops--a case study from lake Amatitlán, Guatemala.

Microcystins (MCs) fate in natural environments can lead to its transfer into aquatic organisms, e.g. aquatic plants. Moreover, lakes in several countries sustain agriculture activities posing a serious health threat for the public. The case of Lake Amatitlán in Guatemala, was addressed to better understand MCs accumulation of four aquatic plants (Polygonum portoricensis, Eichhornia crassipes, Typha sp. and Hydrilla verticillata) coexisting with Microcystis aeruginosa blooms. These findings were further corroborated with an uptake/accumulation laboratory study. Finally crop products (Solanum lycopersicum and Capsicum annuum) irrigated with lake water were also evaluated for MCs. The obtained results suggest that Lake Amatitlán is highly contaminated with MCs (intra- and extracellular 1931 and 90 µg/L, respectively). In fruits of S. lycopersicum and C. annuum a concentration of 1.16 and 1.03 µg/kg dry weight (DW), respectively could be detected. All four aquatic plants showed a high MCs uptake capacity based on obtained bioconcentration factors (BCF) 165, 18, 16 and 11, respectively. These results were further corroborated in a laboratory study with 30 percent of total MCs taken up by H. verticillata within just 7 days. Additionally it was evidenced that all plants accumulated more MC-LR than other MCs congeners. Monitoring of crop products irrigated with lake water needs further consideration.

ß-N-Methylamino-L-alanine exposure alters defense against oxidative stress in aquatic plants Lomariopsis lineata, Fontinalis antipyretica, Riccia fluitans and Taxiphyllum barbieri.

Four different aquatic plants, the Pteridophyte Lomariopsis lineata and the Bryophytes Fontinalis antipyretica, Riccia fluitans and Taxiphyllum barbieri, were tested for their capacity to absorb the neurotoxin ß-N-Methylamino-L-alanine (BMAA) from water and thus their possible applicability in a "Green Liver System". After exposure to 10 and 100 µg L(-1) BMAA for 1, 3, 7 and 14 days exposure concentration of medium and tissue were analyzed by LC-MS/MS. The amount removed by the plants within only 1 day was equal to the biological degradation of 14 days. Comparing the "BMAA-removal" capacity of the 4 tested aquatic plants R. fluitans, L. lineata and T. barbieri turned out to be most effective in cleaning the water from this cyanobacterial toxin by up to 97% within 14 days. Activity of the antioxidant enzymes peroxidase (POD) and catalase (CAT), as well as biotransformation enzyme glutathione S-transferase (GST) was compared between exposed and control plants to determine possible harmful effects induced by BMAA. Whereas the Bryophytes displayed increased POD activity and subsequent adaptation when exposed to the lower concentration, as well as partly inhibited antioxidant response at the higher applied BMAA concentration, the Pteridophyte L. lineata reacted with increased POD activity during the whole experiment and increased GST activity after longer exposure for 14 days. To give a recommendation of the suitability of an aquatic plant to be used for sustainable phytoremediation of contaminated water, testing of removal capacity of specific contaminants as well as studying general physiological parameters giving hint on survivability in such environments has to be combined.

Phytoremediation of diclofenac using the Green Liver System: Macrophyte screening to system optimization.

Green Liver Systems employ the ability of macrophytes to take up, detoxify (biotransform), and bioaccumulate pollutants; however, these systems require optimization to target specific pollutants. In the present study, the aim was to test the applicability of the Green Liver System for diclofenac remediation considering the effects of selected variables. As a starting point, 42 macrophyte life forms were evaluated for diclofenac uptake. With the three best performing macrophytes, the system efficiency was evaluated at two diclofenac concentrations, one environmentally relevant and that other significantly higher (10 µg/L and 150 µg/L) and in two system sizes (60 L and 1000 L) as well as at three flow rates (3, 7, and 15 L/min). The effect of single species and combinations on removal efficiency was also considered. The highest internalization percentage was recorded in Ceratophyllum spp., Myriophyllum spp., and Egeria densa. Phytoremediation efficiency with species combinations was far superior to utilizing only a single macrophyte type. Furthermore, the results indicate that the flow rate significantly affected the removal efficiency of the pharmaceutical tested, with the highest remediation efficiency obtained with the highest flow rate. System size did not significantly affect phytoremediation; however, increase diclofenac concentration reduced the systems performance significantly. When planning the setup of a Green Liver System for wastewater remediation, basic knowledge about the water, i.e., pollutant types and flow, must be utilized during planning to optimize remediation. Various macrophytes show diverse uptake efficiencies for different contaminants and should be selected based on the pollutant composition of the wastewater.

In vivo oxidative stress responses of the freshwater basket clam Corbicula javanicus to microplastic fibres and particles.

Microplastics have been detected in several aquatic organisms, especially bivalves such as clams, oysters, and mussels. To understand the ecotoxicological implication of microplastic accumulation in biota, it is crucial to investigate effects at the physiological level to identify knowledge gaps regarding the threat posed to the environment and assist decision-makers to set the necessary priorities. Typically, xenobiotics elicit an overproduction of reactive oxygen species in organisms, resulting in oxidative stress and cellular damage when not combated by the antioxidative system. Therefore, the present study aimed to establish the impacts of microplastic particles and fibres on the freshwater basket clam Corbicula javanicus. We measured the oxidative stress responses following microplastic exposure as the specific activities of the antioxidative enzymes glutathione S-transferase and catalase. When exposed to polyester fibres from the fleece jackets, the enzyme activities increased in the clams, while the enzyme activities decreased with high-density polyethylene microplastic fragments from bottle caps. All the exposures showed that the adverse effects on the antioxidative response system were elicited, indicating the negative ecotoxicological implications of microplastic pollution.

Ecotoxicological implications of leachates from concrete demolition debris on oligochaetes: survival and oxidative stress status.

Urbanization and population growth demand the construction of structures to facilitate the need for space, and old infrastructures must make space for new ones leading to demolition and concrete debris. In addition to demolition, aging and weather are factors leading to concrete deterioration and, thus, a new challenge as an environmental pollutant. Studies on how concrete debris and leachate affect biota in the environment are limited. The present study aimed to understand the effects of leachate from various sizes of concrete debris on the three oligochaete species Enchytraeus crypticus, Tubifex, and Lumbriculus variegatus. Acute toxicity testing was carried out to determine the adverse effects over time. The oligochaetes' survival was monitored as well as the activity of the biotransformation enzyme glutathione S-transferase and the antioxidative enzyme catalase as indicators of the oxidative stress status. Leachate from the smallest concrete particle size (<1 mm) was found to be the most toxic as it caused, on average, 6-fold increased oligochaete mortality compared to the larger pieces (2-5 cm) after 96 h of exposure, potentially due to the larger surface area facilitating the release of toxicants. Substrate buffered the toxic effect of the leachate with 42 ± 12% fewer mortalities and reduced adverse effects on the enzymes. Of the three oligochaetes, E. crypticus was the most resilient to the concrete leachate. The study is the first to investigate the effects of concrete leachate on oligochaetes.

Study of the antioxidant response of several bean variants to irrigation with water containing MC-LR and cyanobacterial crude extract.

Cyanobacterial toxins have adverse effects on both terrestrial and aquatic plants. Agricultural plants may come in contact with cyanobacterial toxins when surface waters are used for its irrigation. In the present study the effects of MC-LR and cyanobacterial crude extract on three variants of bean by irrigating the seedlings with water containing 5 µg L(-1) MC-LR were assessed. Study of the antioxidant enzymes activities glutathione-S-transferase (GST), peroxidases (POD) and glutathione reductase (GR) was performed in leaves and roots of the seedling exposed to the cyanotoxin compared to a control group and to a group exposed to cyanobacterial crude extract without MCs. Damage of oxidative status was observed in view of the fact that the antioxidant enzymes activities were altered. It is interesting to point out that each variant of bean was affected in a very different way, being GST the most altered enzyme and the B1 the most sensitive variant showing the greatest alterations in the majority of the studied enzymes.

Testing of Eluates from Waterproof Building Materials for Potential Environmental Effects Due to the Behavior of Enchytraeus albidus.

In order to determine the potential environmental impact of construction products, it is necessary to evaluate their influence on organisms exposed to them or their eluates under environmental conditions. The behavior of the white worm Enchytraeus albidus is a useful tool for assessing the potential environmental impact of construction products in contact with water and soil. This study investigates the environmental effects of eluates from two construction products, a reactive waterproofing product, and an injection resin, on the reproduction and avoidance behavior of E. albidus. The eluates were prepared according to existing guidelines. The soil used for the tests was moistened with the eluates of the construction products. The reproduction results of the worms were collected after six weeks of exposure. Offsprings were counted under the microscope and statistically analyzed. Results from the avoidance behavior were collected after 48 h of exposure, and results were compared with the reproduction results. The eluates from both construction products induced significant changes in the reproduction behavior of E. albidus. Undiluted or only slightly diluted eluates of the injection resin drastically reduced the reproduction of the worms, whereas the leaches of the reactive waterproofing product only had a minor effect. The avoidance results for the injection resin indicates that its presence in the habitat is clearly detrimental to the survival of E. albidus, while the avoidance results for the waterproofing resin showed an initial avoidance of the eluates, but no harmful effects were observed. The avoidance test is a way of rapid toxicity screening of environmental samples when time is a critical parameter to measure possible environmental effects. This study shows that ecotoxicological tests using Enchytraeids are a valuable and important tool for understanding the mode of action of eluates from construction products in the environment.

Reviewing Interspecies Interactions as a Driving Force Affecting the Community Structure in Lakes via Cyanotoxins.

The escalating occurrence of toxic cyanobacterial blooms worldwide is a matter of concern. Global warming and eutrophication play a major role in the regularity of cyanobacterial blooms, which has noticeably shifted towards the predomination of toxic populations. Therefore, understanding the effects of cyanobacterial toxins in aquatic ecosystems and their advantages to the producers are of growing interest. In this paper, the current literature is critically reviewed to provide further insights into the ecological contribution of cyanotoxins in the variation of the lake community diversity and structure through interspecies interplay. The most commonly detected and studied cyanobacterial toxins, namely the microcystins, anatoxins, saxitoxins, cylindrospermopsins and ß-N-methylamino-L-alanine, and their ecotoxicity on various trophic levels are discussed. This work addresses the environmental characterization of pure toxins, toxin-containing crude extracts and filtrates of single and mixed cultures in interspecies interactions by inducing different physiological and metabolic responses. More data on these interactions under natural conditions and laboratory-based studies using direct co-cultivation approaches will provide more substantial information on the consequences of cyanotoxins in the natural ecosystem. This review is beneficial for understanding cyanotoxin-mediated interspecies interactions, developing bloom mitigation technologies and robustly assessing the hazards posed by toxin-producing cyanobacteria to humans and other organisms.