Differential Legionella spp. survival between intracellular and extracellular forms in thermal spring environments

Legionella are commonly found in natural and man-made aquatic environments and are able to inhabit various species of protozoa. The relationship between the occurrence of Legionella spp. within protozoa and human legionellosis has been demonstrated; however, the proportions of intracellular and extracellular Legionella spp. in the aquatic environment were rarely reported. In this study, we developed a new method to differentiate intracellular and extracellular Legionella spp. in the aquatic environment. Water samples from three thermal spring recreational areas in southeastern Taiwan were collected and analyzed. For each water sample, concurrent measurements were performed for Legionella spp. and their free-living amoebae hosts. The overall detection rate was 32 % (16/50) for intracellular Legionella spp. and 12 % (6/50) for extracellular Legionella spp. The most prevalent host of Legionella spp. was Hartmannella vermiformis. The identified Legionella spp. differed substantially between intracellular and extracellular forms. The results showed that it may be necessary to differentiate intracellular and extracellular forms of Legionella spp.     

Aquatic arsenic: phytoremediation using floating macrophytes

Phytoremediation, a plant based green technology, has received increasing attention after the discovery of hyperaccumulating plants which are able to accumulate, translocate, and concentrate high amount of certain toxic elements in their above-ground/harvestable parts. Phytoremediation includes several processes namely, phytoextraction, phytodegradation, rhizofiltration, phytostabilization and phytovolatilization. Both terrestrial and aquatic plants have been tested to remediate contaminated soils and waters, respectively. A number of aquatic plant species have been investigated for the remediation of toxic contaminants such as As, Zn, Cd, Cu, Pb, Cr, Hg, etc. Arsenic, one of the deadly toxic elements, is widely distributed in the aquatic systems as a result of mineral dissolution from volcanic or sedimentary rocks as well as from the dilution of geothermal waters. In addition, the agricultural and industrial effluent discharges are also considered for arsenic contamination in natural waters. Some aquatic plants have been reported to accumulate high level of arsenic from contaminated water. Water hyacinth (Eichhornia crassipes), duckweeds (Lemna gibba, Lemna minor, Spirodela polyrhiza), water spinach (Ipomoea aquatica), water ferns (Azolla caroliniana, Azolla filiculoides, and Azolla pinnata), water cabbage (Pistia stratiotes), hydrilla (Hydrilla verticillata) and watercress (Lepidium sativum) have been studied to investigate their arsenic uptake ability and mechanisms, and to evaluate their potential in phytoremediation technology. It has been suggested that the aquatic macrophytes would be potential for arsenic phytoremediation, and this paper reviews up to date knowledge on arsenic phytoremediation by common aquatic macrophytes.     

Microbial and physicochemical parameters associated with Legionella contamination in hot water recirculation systems

Hot water recirculation systems (HWRS) in hotels and nursing homes, which are common in countries such as Spain, have been related to outbreaks of legionellosis. To establish the relationships of microbial and physicochemical parameters, especially protozoa, with the occurrence of Legionella in HWRS, 231 samples from hotels and nursing homes were analysed for Legionella, protozoa, heterotrophic plate counts (HPC) at 22 and 37 °C, Pseudomonas, metals, temperature and others. Legionella pneumophila was the dominant species isolated, and 22 % were sg. 1. The sampling method became particularly important in order to define which factors were involved on the occurrence of Legionella. Results showed that the bacteria and the accompanying microbiota were more abundant in the first flush water whose temperature was lower. The bacteria occurred in those samples with high HPC and were inversely correlated with high temperatures. Multivariate regression showed that a concentration above 1?×?10⁵?CFU/100 mL of HPC at 37 °C, Fe above 0.095 ppm and the presence of protozoa increased significantly the risk of Legionella colonization, while univariant regression showed that the presence of Cu above 0.76 ppm and temperature above 55 °C diminished it. Therefore, to reduce the risk associated with Legionella occurrence in HWRS these parameters should be taken into consideration.     

Seed treatments containing neonicotinoids and fungicides reduce aquatic insect richness and abundance in midwestern USA–managed floodplain wetlands

Agrochemicals including neonicotinoid insecticides and fungicides are frequently applied as seed treatments on corn, soybeans, and other common row crops. Crops grown from pesticide-treated seed are often directly planted in managed floodplain wetlands and used as a soil disturbance or food resource for wildlife. We quantified invertebrate communities within mid-latitude floodplain wetlands and assessed their response to use of pesticide-treated seeds within the floodplain. We collected and tested aqueous and sediment samples for pesticides in addition to sampling aquatic invertebrates from 22 paired wetlands. Samples were collected twice in 2016 (spring [pre-water level drawdown] and autumn [post-water level flood-up]) followed by a third sampling period (spring 2017). Meanwhile, during the summer of 2016, a portion of study wetlands were planted with either pesticide-treated or untreated corn seed. Neonicotinoid toxic equivalencies (NI-EQs) for sediment (X??=?0.58 μg/kg), water (X??=?0.02 μg/L), and sediment fungicide concentrations (X??=?0.10 μg/kg) were used to assess potential effects on wetland invertebrates. An overall decrease in aquatic insect richness and abundance was associated with greater NI-EQs in wetland water and sediments, as well as with sediment fungicide concentration. Post-treatment, treated wetlands displayed a decrease in insect taxa-richness and abundance before recovering by the spring of 2017. Information on timing and magnitude of aquatic insect declines will be useful when considering the use of seed treatments for wildlife management. More broadly, this study brings attention to how agriculture is used in wetland management and conservation planning.

     

Eukaryotic diversity in premise drinking water using 18S rDNA sequencing: implications for health risks

The goal of this study was to characterize microbial eukaryotes over a 12-month period to provide insight into the occurrence of potential bacterial predators and hosts in premise plumbing. Nearly 6,300 partial 18S rRNA gene sequences from 24 hot (36.9–39.0 °C) and cold (6.8–29.1 °C) drinking water samples were analyzed and classified into major eukaryotic groups. Each major group, consisting of free-living amoebae (FLA)/protozoa, algae, copepods, dinoflagellates, fungi, nematodes, and unique uncultured eukaryotic sequences, showed limited diversity dominated by a few distinct populations, which may be characteristic of oligotrophic environments. Changes in the relative abundance of predators such as nematodes, copepods, and FLA appear to be related to temperature and seasonal changes in water quality. Sequences nearly identical to FLA such as Hartmannella vermiformis, Echinamoeba thermarmum, Pseudoparamoeba pagei, Protacanthamoeba bohemica, Platyamoeba sp., and Vannella sp. were obtained. In addition to FLA, various copepods, rotifers, and nematodes have been reported to internalize viral and bacterial pathogens within drinking water systems thus potentially serving as transport hosts; implications of which are discussed further. Increasing the knowledge of eukaryotic occurrence and their relationship with potential pathogens should aid in assessing microbial risk associated with various eukaryotic organisms in drinking water.     

Invertebrates control metal/metalloid sequestration and the quality of DOC/DON released during litter decay in slightly acidic environments

Plant litter and organic sediments are a main sink for metals and metalloids in aquatic ecosystems. The effect of invertebrate shredder (a key species in litter decay) on metal/metalloid fixation by organic matter is described only under alkaline water conditions whereas for slightly acidic waters nothing can be found. Furthermore, less is known about the effect of invertebrate shredders on the quality of dissolved organic carbon (DOC) and nitrogen (DON) released during litter decay. We conducted an experiment to investigate the impact of invertebrate shredder (Gammarus pulex) on metal/metalloid fixation/remobilization and on the quality of DOC/DON released under slightly acidic water conditions. During decomposition of leaf litter, invertebrate shredder facilitated significantly the emergence of smaller particle sizes of organic matter. The capacity of metal fixation was significantly higher in smaller particles (POM 2,000?C63???m) compared to original leaf litter and litter residues. Thus, G. pulex enhanced metal fixation by organic partition of sediments by increasing the amount of smaller particle of organic matter in aquatic ecosystems. In contrast, the capacity of metal/metalloid fixation in the smallest fraction of POM (<63???m) was lower compared with leaf residues in treatment without invertebrates. Remobilization of metals and metalloids was very low for all measured elements. A significant effect of invertebrates on quantitative formation of DOC/DON was confirmed. The quality of released DOC/DON, which may affect metal/metalloid remobilization, was also significantly affected by invertebrate shredders (e.g., more carboxylates). Hence, invertebrate shredder enhanced significantly the fixation of metals/metalloids into POM in slightly acidic environments.     

Derivation of aquatic predicted no-effect concentration (PNEC) for 2,4-dichlorophenol: comparing native species data with non-native species data

2,4-Dichlorophenol (2,4-DCP) is known as an important chemical intermediate and an environmental endocrine disruptor. There is no paper dealing with the predicted no-effect concentration (PNEC) of 2,4-DCP, mainly due to shortage of chronic and site-specific toxicity data. In the present study, toxicity data was obtained from the tests using six Chinese native aquatic species. The HC₅ (hazardous concentration for 5% of species) was derived based on the constructed species sensitivity distribution (SSD), which was compared with that derived from literature toxicity data of non-native species. For invertebrates, the survival no-observed effect concentrations (NOECs) were 0.05 and 1.00 mg L⁻¹ for Macrobrachium superbum and Corbicula fluminea, respectively. NOECs based on fishes’ growth were 0.10, 0.20 and 0.40 mg L⁻¹ for Mylopharyngodon piceus, Plagiognathops microlepis and Erythroculter ilishaeformis, respectively. For aquatic plant Soirodela polyrhiza, NOEC based on concentration of chlorophyll was 1.00 mg L⁻¹. A final PNEC calculated using the SSD approach with a 50% certainty based on different taxa ranged between 0.008 and 0.045 mg L⁻¹. There is no significant difference between HC₅ derived from native and that from non-native taxa.     

Phytoremediation Potential of Aquatic Macrophyte, <Emphasis Type="Italic">Azolla</Emphasis>

Aquatic macrophytes play an important role in the structural and functional aspects of aquatic ecosystems by altering water movement regimes, providing shelter to fish and aquatic invertebrates, serving as a food source, and altering water quality by regulating oxygen balance, nutrient cycles, and accumulating heavy metals. The ability to hyperaccumulate heavy metals makes them interesting research candidates, especially for the treatment of industrial effluents and sewage waste water. The use of aquatic macrophytes, such as Azolla with hyper accumulating ability is known to be an environmentally friendly option to restore polluted aquatic resources. The present review highlights the phytoaccumulation potential of macrophytes with emphasis on utilization of Azolla as a promising candidate for phytoremediation. The impact of uptake of heavy metals on morphology and metabolic processes of Azolla has also been discussed for a better understanding and utilization of this symbiotic association in the field of phytoremediation.     

Toxic effects of Hydrilla verticillata exposed to toluene, ethylbenzene and xylene and safety assessment for protecting aquatic macrophytes

Little information is available about the toxicity of toluene, ethylbenzene and xylene acting on macrophytes, and their toxicity data are rarely used in regulation and criteria decisions. The results extended the knowledge on toxic effects of toluene, ethylbenzene and xylene on aquatic plants. The responses of Hydrilla verticillata to these pollutants were investigated. Chlorophyll levels, lipid peroxidation, and antioxidant enzymes (superoxide dismutase and guaiacol peroxidase) showed diverse responses at different concentrations of toluene, ethylbenzene and xylene. The linear regression analyses were performed respectively, suggesting the concentrations of toluene, ethylbenzene and xylene expected to protect aquatic macrophytes were 7.30 mg L⁻¹, 1.15 mg L⁻¹ and 2.36 mg L⁻¹, respectively. This study emphasized that aquatic plants are also sensitive to organic pollutants as fishes and zooplanktons, indicating that macrophytes could be helpful in predicting the toxicity of these pollutants and should be considered in regulation and criteria decisions for aquatic environment protection.     

Ecotoxicologial evaluation of wastewater ozonation based on detritus-detritivore interactions

Advanced oxidation technologies such as ozonation have been proposed to improve removal efficiency of micropollutants during wastewater treatment. In a meta-analysis of peer-reviewed literature, we found no ecotoxicological effects of wastewater ozonation on invertebrates (n = 82), but significant adverse effects on bacteria (n = 24) and fish (n = 5). As information on functional endpoints or trophic interactions is lacking, we applied a bioassay relating to leaf litter decomposition to fill this gap. Leaf discs exposed to ozone-treated wastewater with a high (1.04 mg O₃ (mg DOC)⁻¹, n = 49) ozone concentration were significantly preferred by an aquatic detritivore, Gammarus fossarum, over discs conditioned in wastewater not treated with ozone. This effect might have been mediated by reduced bacterial and elevated fungal biomass, and appears to be the first demonstration of wastewater ozonation impacts on invertebrates and an associated ecosystem process. In accordance with the food-choice trials, chemical analyses revealed significantly decreased concentrations of organic micropollutants in wastewater treated with ozone at high concentrations. Thus, food-choice trials as applied here hold promise to assess environmental effects of advanced oxidation technologies in wastewater treatment and appear to be a valuable complement to the ecotoxicological toolbox in general.     

Long-term effects of heavy metals on aouatic plants

In long-term experiments lasting up to 73 days the effect of rather low levels of zinc, copper, lead and cadmium on the growth and metal uptake was studied by investigating four aquatic plant species: Elodea nuttallii, Callitriche plataycarpa, Spirodela polyrhiza and Lemma gibba. Except Elodea, which was already very sensitive to 5 μmol Cu 1^?1, no differentiation in growth or mortality could be detected depending on species or elements. There was a clear differentiation between the uptake levels of the heavy metals with regard to the plant species, resulting in a higher heavy metal content in de submerged species in comparison to the floating ones. For zinc, lead and cadmium, an equal ratio was detected between the concentration in the medium and in the plant tissue independent of the plant species. The involvement of roots in element absorption by aquatic plants and the possibility of using aquatic plants as indicators of heavy metal pollution in Dutch waters are discussed.     

Molecular docking: A potential tool to aid ecotoxicity testing in environmental risk assessment of pharmaceuticals

A cocktail of human pharmaceuticals pollute aquatic environments and there is considerable scientific uncertainty about the effects that this may have on aquatic organisms. Human drug target proteins can be highly conserved in non target species suggesting that similar modes of action (MoA) may occur. The aim of this work was to explore whether molecular docking offers a potential tool to predict the effects of pharmaceutical compounds on non target organisms. Three highly prescribed drugs, diclofenac, ibuprofen and levonorgestrel which regularly pollute freshwater environments were used as examples. Their primary drug targets are cyclooxygenase 2 (COX2) and progesterone receptor (PR). Molecular docking experiments were performed using these drugs and their primary drug target homologues for Danio rerio, Salmo salar, Oncorhynchus mykiss, Xenopus tropicalis, Xenopus laevis and Daphnia pulex. The results show that fish and frog COX2 enzymes are likely to bind diclofenac and ibuprofen in the same way as humans but that D. pulex would not. Binding will probably lead to inhibition of COX function and reduced prostaglandin production. Levonorgestrel was found to bind in the same binding pocket of the progesterone receptor in frogs and fish as the human form. This suggests implications for the fecundity of fish and frogs which are exposed to levonorgestrel. Chronic ecotoxicological effects of these drugs reported in the literature support these findings. Molecular docking may provide a valuable tool for ecotoxicity tests by guiding selection of test species and incorporating the MoA of drugs for relevant chronic test end points in environmental risk assessments.     

Whole effluent toxicity assessment at a wastewater treatment plant upgraded with a full-scale post-ozonation using aquatic key species

Ozonation as final wastewater (WW) polishing step, following conventional activated sludge treatment is increasingly implemented in sewage treatment for contaminant degradation to prevent surface water pollution. While the oxidative degradation of chemicals has been extensively investigated, the in vivo toxicological characteristics of ozonated whole effluents are rarely a matter of research.In the present study, whole effluents were toxicologically evaluated with an in vivo test battery before and after full-scale ozonation and subsequent sand filtration on site at a treatment plant. One aquatic plant (duckweed, Lemna minor) and five invertebrate species of different systematic groups (Lumbriculus variegatus, Chironomus riparius, Potamopyrgus antipodarum, Daphnia magna) were exposed to the effluents in a flow-through-designed test system with a test duration of 7-28 d.None of the considered toxicity endpoints correlated with the pollutant elimination. A tendency towards an increased toxicity after ozonation was apparent in three of the test systems showing [statistically] significant adverse effects in the L. variegatus toxicity test (decrease in reproduction and biomass). After sand filtration, adverse effects were reduced to a similar level like after conventional treatment. Solely the Daphnia reproduction test revealed beneficial effects after ozonation in combination with sand filtration.Results of the test battery indicate the formation of adverse oxidation products during WW ozonation. L. variegatus appeared to be the most sensitive of the five test species. Sand filtration effectively removes or detoxifies toxic oxidation products, as toxic effects were subsequently reduced to the level after conventional treatment.     

Lethal and sublethal effects of four essential oils on the egg parasitoids Trissolcus basalis

The essential oils from leaves of Schinus molle var. areira, Aloysia citriodora, Origanum vulgare and Thymus vulgaris have showed potential as phytoinsecticides against the green stink bug, Nezara viridula. In this work were evaluated their toxicological and behavioral effects on the parasitoid Trissolcus basalis, a biological control agent of this pest insect. Essential oils were obtained via hydrodestillation from fresh leaves. Insecticide activity in T. basalis females was evaluated in direct contact and fumigation bioassays. Behavioral effects were evaluated in olfactometer bioassays. To evaluate the residual toxicity, females of the parasitoids were exposed to oil residues; in these insects, the sublethal effects were evaluated (potential parasitism and survivorship of immature stages). The essential oils from O. vulgare and T. vulgaris proved to be highly selective when used as fumigant and did not change parasitoid behavior. After one week, the residues of these oils were harmless and did not show sublethal effects against T. basalis. According with these results, essential oils have potential applications for the integrated management of N. viridula.     

Aquatic ecotoxicity assessment of a new natural formicide

Background, aim and scope

Agrochemicals could reach aquatic ecosystems and damage ecosystem functionality. Natural formicide could be an alternative to use in comparison with the more toxic formicides available on the market. Thus, the objective of this study was to assess the ecotoxicity of the new natural formicide Macex? with a battery of classical aquatic ecotoxicity tests.

Material and methods

Bacteria (Aliivibrio fischeri), algae (Pseudokirchneriella subcapitata), hydra (Hydra attenuata), daphnids (Daphnia magna), and fish (Danio rerio) tests were performed in accordance with international standardized methodologies.

Results

In the range of formicide concentrations tested (0.03 to 2.0?g?L^?1) EC₅₀ values varied from 0.49 to >2.0?g?L^?1, with P. subcapitata being the most sensitive species and H. attenuata and D. rerio the most tolerant species to this product in aqueous solutions.

Conclusions

This new formicide preparation can be classed as a product of low toxicity compared to the aquatic ecotoxicity of the most common commercialized formicides.     

Nitrate causes deleterious effects on the behaviour and reproduction of the aquatic snail Potamopyrgus antipodarum (Hydrobiidae, Mollusca)

Nitrate (NO₃ ^?) is present in aquatic ecosystems as a natural component of the nitrogen cycle. However, in the last decades, several human activities are the causes of the rising amounts of organic matter and inorganic nitrogen nutrients in aquatic ecosystems, causing notable increase of nitrate above background natural levels. In spite of the toxicity of nitrate to aquatic animals, there are relatively few studies on the chronic toxicity of this compound to invertebrates. The aim of our study is to assess the effect of chronic (35 days) exposure to nitrate on the behaviour (velocity of movement) and reproduction (number of newborns) of the aquatic snail Potamopyrgus antipodarum. Four actual concentrations of nitrate were used (21.4, 44.9, 81.8 and 156.1 mg?N-NO₃ ^?/L). In each treatment, 12 animals were individually monitored for velocity (weekly) and newborn production (every 3–4 days). Velocity was recorded using quantitative video monitoring. Our results showed that nitrate did not cause mortality, but it reduced the velocity of movement (at 44.9, 81.8 and 156.1 mg?N-NO₃ ^?/L) and number of live newborns (in all tested concentrations). Reproductive impairment was caused at realistic nitrate concentrations which is relevant to the risk assessment of this compound. Our study contributes to the knowledge of the chronic effects of nitrate on the behaviour and reproduction of an aquatic snail.     

The relation between Acid Volatile Sulfides (AVS) and metal accumulation in aquatic invertebrates: Implications of feeding behavior and ecology

The present study evaluates the relationship between Acid Volatile Sulfides (AVS) and metal accumulation in invertebrates with different feeding behavior and ecological preferences. Natural sediments, pore water and surface water, together with benthic and epibenthic invertebrates were sampled at 28 Flemish lowland rivers. Different metals as well as metal binding sediment characteristics including AVS were measured and multiple regression was used to study their relationship with accumulated metals in the invertebrates taxa.Bioaccumulation in the benthic taxa was primarily influenced by total metal concentrations in the sediment. Regarding the epibenthic taxa metal accumulation was mostly explained by the more bioavailable metal fractions in both the sediment and the water. AVS concentrations were generally better correlated with metal accumulation in the epibenthic invertebrates, rather than with the benthic taxa. Our results indicated that the relation between AVS and metal accumulation in aquatic invertebrates is highly dependent on feeding behavior and ecology.     

Heavy metals and arsenic fixation into freshwater organic matter under Gammarus pulex L. influence

Organic sediments are a main sink for metal pollutants in aquatic systems. However, factors that make sediments a sink of metals and metalloids are still not clear. Consequently, we investigate the role of invertebrate shredders (Gammarus pulex L.) on quality of metal and arsenic fixation into organic partitions of sediment in the course of litter decay with laboratory microcosm experiments. During the decomposition of leaf litter, G. pulex significantly facilitated the development of small particles of organic matter. The capacity of metal fixation was significantly higher in smaller particles than leaf litter and litter residuals. Thus, G. pulex enhanced metal fixation into the organic partition of sediments by virtue of increasing the amount smaller particles in the aquatic system. Furthermore, invertebrates have a significant effect on formation of dissolved organic matter and remobilization of cobalt, molybdenum and cesium, but no significant effect on remobilization of all other measured elements.     

Ionic liquids as lubricants or lubrication additives: An ecotoxicity and biodegradability assessment

This paper reports on the (eco)toxicity and biodegradability of ionic liquids considered for application as lubricants or lubrication additives. Ammonium- and pyrrolidinium-based cations combined with methylsulphate, methylsulphonate and/or (CF₃SO₂)₂N⁻ anions were investigated in tests to determine their aquatic toxicity using water fleas Daphnia magna, green algae Selenastrum capricornutum and marine bacteria (Vibrio fischeri). Additional test systems with an isolated enzyme (acetylcholinesterase) and isolated leukaemia cells from rats (IPC-81) were used to assess the biological activity of the ionic liquids. These compounds generally exhibit low acute toxicity and biological activity. Their biodegradability was screened according to OECD test procedures 301 B and 301 F. For choline and methoxy-choline ionic liquids ready biodegradability was observed within 5 or 10 d, respectively. Some of the compounds selected have a considerable potential to contribute to the development of more sustainable products and processes.     

Evaluation and in situ assessment of photodegradation of polyaromatic hydrocarbons in semipermeable membrane devices deployed in ocean water

Semipermeable membrane devices (SPMDs) were deployed in water using four different methods: a typical SPMD cage with and without a mesh cover, a bowl chamber and without any protection. In addition to routinely used performance reference compounds (PRCs), perdeuterated dibenz[a,h]anthracene was added. Due to its high sampler to water partition coefficient no measurable clearance due to diffusion was expected during the deployment period, hence any observed loss could be attributed to photodegradation. The loss of PRCs was measured and SPMD-based water concentrations determined. Results showed that a typical SPMD deployment cage covered with mesh provided the best protection from photodegradation. Samplers which had undergone the highest photodegradation underestimated PAH water concentrations by up to a factor of 5 compared to the most protected SPMDs. This study demonstrates that the potential for photodegradation needs to be addressed when samplers are deployed in water of low turbidity.