Neurochemical effects of benzodiazepine and morphine on freshwater mussels

The purpose of this study was to examine the neurochemical effects of morphine, diazepam, a common benzodiazepine, and an effluent concentrate on the endemic freshwater mussel Elliptio complanata. Mussels were exposed to the drugs and to the solid-phase concentrate of a municipal effluent and left to stand at 15 °C for 48 h. Neurochemical effects were determined by monitoring changes in dopamine, serotonin, glutamate and γ-aminobutyric acid (GABA) levels in the visceral mass (containing the nerve ganglia) of mussels. The activities of acetylcholinesterase (AChE), dopamine and serotonin-dependent adenylyl cyclase (ADC) were also determined in the mussels. Oxidative stress was determined by tracking changes in lipid peroxidation (LPO) in the mitochondrial and post-mitochondrial fractions. The results revealed that the drugs and the effluent extract were biologically active in mussels. Morphine reduced serotonin and increased dopamine in mussel tissues while reducing AChE activity and increasing GABA levels. This suggests the induction of a relaxation state in mussels. Diazepam also reduced serotonin levels but produced no change in dopamine levels. However, dopamine-sensitive ADC activity was readily activated, indicating the potential effect on opiate signaling. Diazepam increased glutamate levels slightly, but AChE remained stable. The increase in both dopamine ADC activity and glutamate concentrations was also associated with greater oxidative stress on the mitochondrial and post-mitochondrial fractions in cells. A comparison of the global response pattern of these drugs with those of the effluent extract revealed only a relative proximity to morphine. In conclusion, the data warrant more studies on the analysis of opiates and benzodiazepines in municipal effluents to better address the potential environmental hazard of these neuroactive drug classes to aquatic organisms.     

Context-dependent effects of freshwater mussels on stream benthic communities

1. We asked whether unionid mussels influence the distribution and abundance of co‐occurring benthic algae and invertebrates. In a yearlong field enclosure experiment in a south‐central U.S. river, we examined the effects of living mussels versus sham mussels (shells filled with sand) on periphyton and invertebrates in both the surrounding sediment and on mussel shells. We also examined differences between two common unionid species, Actinonaias ligamentina (Lamarck 1819) and Amblema plicata (Say 1817). 2. Organic matter concentrations and invertebrate densities in the sediment surrounding mussels were significantly higher in treatments with live mussels than treatments with sham mussels or sediment alone. Organic matter was significantly higher in the sediment surrounding Actinonaias than that surrounding Amblema. Actinonaias was more active than Amblema and may have increased benthic organic matter through bioturbation. 3. Living mussels increased the abundance of periphyton on shells and the abundance and richness of invertebrates on shells, whereas effects of sham mussels were similar to sediment alone. Differences in the amount of periphyton growing on the shells of the two mussel species reflected differences in mussel activity and shell morphology. 4. Differences between living and sham mussel treatments indicate that biological activities of mussels provide ecosystem services to the benthic community beyond the physical habitat provided by shells alone. In treatments containing live mussels we found significant correlations between organic matter and chlorophyll a concentrations in the sediment, organic matter concentrations and invertebrate abundance in the sediment and the amount of chlorophyll a on the sediment and invertebrate abundance. There were no significant correlations among these response variables in control treatments. Thus, in addition to providing biogenic structure as habitat, mussels likely facilitate benthic invertebrates by altering the availability of resources (algae and organic matter) through nutrient excretion and biodeposition. 5. Effects of mussels on sediment and shell periphyton concentrations, organic matter concentrations and invertebrate abundance, varied seasonally, and were strongest in late summer during periods of low water volume, low flow, and high water temperature. 6. Our study demonstrates that freshwater mussels can strongly influence the co‐occurring benthic community, but that effects of mussels are context‐dependent and may vary among species.     

The effects of zinc oxide nanoparticles on the metallome in freshwater mussels

The use of zinc oxide nanoparticles (nanoZnO) as sunscreens has raised concerns about their safety and release in the aquatic environment through swimming activities and within municipally treated wastewaters. This study's purpose was to examine the effects of nanoZnO on the elemental composition (metallome) in exposed freshwater mussels, Elliptio complanata. Mussels were exposed for 21 days to an environmentally realistic (low) concentration (2 μg/L) of nanoZnO and zinc chloride. The mussels were also exposed to a physically and chemically treated municipal effluent (ME), both alone and in the presence of both forms of Zn. The metallome profile was characterized by the following 15 elements in gills, digestive gland and gonad tissues: Ag, Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, V and Zn. The levels of metallothioneins (MT) and lipid peroxidation (LPO) in the digestive gland were also measured as biomarkers of toxic effects. The data revealed that exposure to nanoZnO increased the total levels of Zn, MT and LPO in the digestive gland. Discriminate function analysis revealed that the digestive gland responded the most to exposure to either nanoZnO or Zn^2 +. For nanoZnO, the observed changes in Al, As and Mo in the digestive gland offered the best discrimination from dissolved Zn^2 +. Co-exposure of nanoZnO with the ME changed the metallome profile closer to dissolved Zn^2 +, suggesting a common interaction site within the ME. This was observed in changes in Ni, Cu, Se and Zn in the digestive gland of exposed mussels. Canonical analysis of essential and non-essential elements revealed that exposure to nanoZnO increased the relationships between LPO and the sum of essential elements in the digestive gland. Conversely, exposure to dissolved Zn^2 + and the ME decreased the relationship between the sum of non-essential elements and LPO and MT. In conclusion, the use of a “metallomic” approach was used to discriminate changes following exposure to nanoZnO and dissolved Zn in freshwater mussels and provided insights into the interaction of forms of Zn in ME towards mussels.     

Direct and transgenerational effects of an experimental heatwave on early life stages in a freshwater snail

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The effects of endangered freshwater pearl mussels on channel morphology and flow in a low-gradient sandy river

Hydrobiologia - Conservation of ecosystem engineers, which modulates the surrounding habitat by causing physical state changes in biotic or abiotic materials, is important for maintaining the...     

Development of a flow-through system to create occluding thrombus

Occlusive thrombosis accounts for many heart attacks and strokes. These acute events are difficult to catch in patients and animal test methods may be misleading because anti-thrombotic therapeutics often do not cross-react with different species. This paper presents a new flow-through system that leads to rapid occlusive thrombosis in arterial flow conditions. Whole porcine blood is perfused through a tubular test section. The growing thrombus is visualized in real time from early platelet attachment, through accumulation, to occlusion. The progression of flow rate reduction provides a clear distinguishing parameter between thrombus formation and embolization. Thrombus growth rate is a linear function of very high shear rate beyond 40,000 s(-1). The histology of the thrombus reveals predominantly platelet accumulation and growth as a rough surface with tendrils. This flow-through system may be useful for the economic testing of new anti-thrombosis therapies.     

Adaptation of a freshwater anammox population to high salinity wastewater

For the successful application of anaerobic ammonium oxidation (anammox) in wastewater practice it is important to know how to seed new anammox reactors with biomass from existing reactors. In this study, a new high salinity anammox reactor was inoculated with biomass from a freshwater system. The changes in activity and population shifts were monitored. It was shown that freshwater anammox bacteria could adapt to salt concentrations as high as 30 gl(-1) provided the salt concentration was gradually increased. Higher salt concentrations reversibly inhibited anammox bacteria. The nitrogen removal efficiency and maximum anammox activity of the salt adapted sludge was very similar to the reference freshwater sludge. Fluorescence in situ hybridization analysis revealed that the freshwater anammox species Candidatus "Kuenenia stuttgartiensis" was the dominant in both salt adapted sludge and freshwater sludge. These results show that gradual adaptation may be the key to successful seeding of anammox bioreactors.     

Assessment of nutritional subsidies to freshwater mussels using a multiple natural abundance isotope approach

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Biodiversity on the brink: an assessment of conservation strategies for North American freshwater mussels

Fish and zooplankton populations of nine Ethiopian freshwater lakes were quantitatively sampled along a North–South gradient. Differences in altitude and latitude resulted in a temperature gradient from North to South. We tested three hypotheses: (1) the degree of zooplanktivory decreases with water temperature, i.e. from North to South; (2) the degree of zooplanktivory increases with the abundance of large-bodied zooplankton; and (3) the pattern of zooplanktivory in eutrophic Ethiopian water bodies differs from other tropical and temperate water bodies. Proportions of zooplanktivory in the fish communities did not show a geographical trend, but mainly depended on fish species, zooplankton density and the availability of large-bodied cladocerans. The degree of zooplanktivory in eutrophic Ethiopian water bodies differs from other eutrophic water bodies, both temperate and tropical. In Ethiopia, the degree of zooplanktivory can be both low and high, in contrast with other tropical water bodies where zooplanktivory is generally low and with temperate eutrophic water bodies where it is generally high. As a result, predation pressure on zooplankton by fish varies dramatically amongst Ethiopian water bodies.     

Behaviour and physiology are linked in the responses of freshwater mussels to drought

1. Droughts have become prevalent in the south‐eastern U.S.A. and are predicted to become more common in the future. Drought conditions have particularly strong negative effects on sessile aquatic organisms with limited dispersal ability. This study explored the linkages between physiological tolerance, behavioural response and survival of three species of freshwater mussels subjected to drought. 2. To assess physiological tolerance, we measured survival under desiccation at 25, 35 and 45 °C in the laboratory. To assess behavioural responses, we tracked horizontal and vertical movement of mussels in a drying stream reach. 3. Uniomerus tetralasmus showed the greatest desiccation tolerance, Lampsilis straminea was intermediate, and Pyganodon grandis had the lowest tolerance at all temperatures. 4. In the drying stream reach, U. tetralasmus showed little horizontal movement and quickly became stranded. The other two species tracked the receding water. It was not until the pool had been reduced to c. 10% of its original size that ≥50% of P. grandis and L. straminea became stranded. 5. Uniomerus tetralasmus and L. straminea burrowed in response to becoming stranded; however, both species burrowed only shallowly (3–4 cm), presumably because burrowing to greater depths did not convey increasing thermal refuge. Pyganodon grandis rarely burrowed. No P. grandis survived the 15‐week drought, while 45% of L. straminea and 77% of U. tetralasmus survived. 6. Three strategies emerged for freshwater mussels to cope with drought conditions: tracking (intolerant), track then burrow (semi‐tolerant) and burrowing (tolerant). Results suggest that drought poses the greatest threat to intolerant trackers, while tolerant burrowers are the most resistant to drought conditions.     

Handling effects on survival and growth of plain pocketbook Lampsilis cardium (Rafinesque, 1820) freshwater mussels

Hydrobiologia - Handling freshwater mussels may have negative impacts on their survival, growth, or reproduction, and this may affect our interpretation of how mussels respond to research and...     

Biofilm formation on brass coupons exposed to a cooling system of an oil refinery

Brass coupons (70% Cu 30% Zn) were exposed to a cooling freshwater system of an oil refinery, in order to investigate susceptibility of the metal to biofilm formation. The coupons were fixed on bypasses at points which allowed the circulation of makeup, cooling and return water. The number of aerobic, anaerobic and sulfate-reducing bacteria was determined in both the planktonic and the sessile phases. Maximum bacterial concentrations were detected in the cooling water, corresponding to 2.1 ± 0.1 × 10⁶ CFU ml⁻¹ (planktonic phase) and 1.3 ± 0.2 × 10⁵ CFU cm⁻² (sessile phase) for aerobic bacteria and to 3.2 ± 0.3 × 10⁵ cells ml⁻¹ (planktonic phase) and 6.2 ± 0.7 × 10⁵ cells cm⁻² (sessile phase) for anaerobic bacteria. Sulfate-reducing bacteria (SRB) were observed only in the planktonic phase, being found in greater numbers in the return water. Scanning electron microscopy (SEM) analysis indicated that biofilm formation occurred at the three monitored sites and showed a diversity in cell morphology. Nonetheless, no evidence of corrosion was observed on the brass coupons during the experimental period. Received 22 May 1997/ Accepted in revised form 19 September 1997     

Effects of a freshwater canal discharge on polychaete assemblages inhabiting an exposed sandy beach in Uruguay

This paper analyzes the spatial and temporal effects of a freshwater discharge (Andreoni Canal) on polychaete assemblages and the corresponding habitat in a sandy beach of Uruguay. Bimonthly, we examined 17 environmental variables plus polychaete abundance, richness, and diversity of three sites: ‘Andreoni’ (highly disturbed) at the canal mouth, ‘Coronilla’ (moderately disturbed), 1 km far away, and ‘Barra del Chuy’ (less disturbed), 13 km north from Andreoni Canal. The highly disturbed site showed significantly lower values of salinity, beach width, swash width, slope, chlorophyll a, and significantly higher values of sediment organic matter. Polychaete abundance and diversity significantly decreased towards the canal and were positively correlated with spatial variations in salinity, which was the most important factor structuring polychaete assemblages. The results highlighted the negative effects of this artificial freshwater discharge on the sandy beach environment and stress the usefulness of polychaetes as effective and reliable ecological indicators for these ecosystems.     

DNA perseverance of microorganisms exposed to silica: an experimental study

The persistence of DNA from microorganisms exposed to various concentrations of SiO₂ (ranging from 0 to 3000 p.p.m.) was monitored over time. The impact of silica mineralization or silicification on the longevity of 16S rRNA and 16 s rDNA genes from whole cells of Bacillus subtilis and Escherichia coli K12 was quantified using real‐time polymerase chain reaction (RT‐PCR), and cells were visualized using optical microscopy. For B. subtilis, DNA longevity decreased in experiments with higher levels of SiO₂ (1000 and 3000 p.p.m.), in comparison to zero or low (100 p.p.m.) levels. For B. subtilis, cell viability was greatest in the absence of silica, and markedly decreased in the presence of any concentration of silica. Survival of Escherichia coli cells, on the other hand, was not sensitive to silica in the solution. All cells died at similar rates over the 180 days they were monitored, decreasing to about 1% survival. DNA longevity for E. coli did appear to be enhanced to some degree by the presence of 1000 p.p.m. silica, but higher or lower concentrations showed no increased longevity in comparison to the no‐silica control. Overall, findings of this study do not support the hypothesis that siliceous environments provide enhanced protection and preservation of DNA over time. However, results of this study do provide guidelines on the persistence of DNA that might be expected in modern silica‐rich environments, which may be an important factor for proper characterization of present‐day microbial communities.     

Linking flow and upper thermal limits of freshwater mussels to inform environmental flow benchmarks

1. Freshwater ecosystems are experiencing shifts in the natural range and variation of water temperatures due to anthropogenic activity, and these shifts can negatively affect survival, growth, and reproduction of aquatic species. Among the groups most affected are freshwater mussels of the family Unionidae. Knowledge of sublethal and lethal effects on mussels from changes in water temperature are largely unknown, especially for species from arid and semi-arid regions such as the south-western U.S.A. This limits the ability to assess, forecast, and adaptively manage this threat for those species and to understand how temperature influences population performance and community structure.
2. To determine the effects of elevated water temperature on mussels from the south-western U.S.A., we evaluated the upper thermal tolerances of adults of three species (Amblema plicata, Cyclonaias necki, and Fusconaia mitchelli) from the Guadalupe River. Mussels were acclimated to 27°C and then tested across a range of experimental temperatures (30–39°C) in standard acute (96-hr) and chronic (10-day) laboratory tests. The acute and chronic thresholds identified in thermal tolerance testing were then related to in situ water temperature and flows using a uniform continuous above-threshold analysis, which evaluates the duration and frequency of continuous events above a specified temperature threshold.
3. Median lethal temperature in 96-hr tests averaged 36.4°C and ranged from 33.7 to 37.5°C, while the chronic 10-day tests averaged 35.9°C and ranged from 32.4 to 37.5°C. Thermal tolerances of F. mitchelli were significantly lower than both A. plicata and C. necki, and the uniform continuous above-threshold analysis showed that temperature affecting 5% of the population thresholds were exceeded for F. mitchelli in the Guadalupe River at both acute (96-hr) and chronic (10-day) values (30.5 and 28.4°C, respectively).
4. Findings from this study indicate that freshwater mussels from the arid and semi-arid regions of the south-west U.S.A. are already at risk from rising environmental temperatures and altered hydrology. However, by incorporating laboratory thermal tolerance estimates with in situ temperature and discharge data, we provide a range of hydrologic thresholds to inform environmental flow recommendations and potentially mitigate thermal stress occurring during periods of low flow. In addition, this method can be readily adapted to other arid regions to guide flow recommendations or assess whether flow standards are sufficient to protect freshwater mussel populations during severe droughts and low flow periods.

     

A feasibility study of a Salix viminalis gravel hydroponic system to renovate primary settled wastewater

A Salix viminalis/gravel system based on hydroponics was developed for wastewater renovation in order to avoid the problems of soil damage and pollution associated with long-term application of wastewater to soil. For such a system to work the mineral elements applied must match closely the requirements of the tree species. To examine this the growth and nutrient uptake of S. viminalis in wastewater was compared with that in Long Ashton nutrient solution (1/4 strength). S. viminalis grew more slowly in wastewater than in Long Ashton solution, but exhibited no obvious deficiency or toxicity symptoms. Since industrial wastewaters often contain metals, the extent to which copper might inhibit wastewater treatment in this system was also examined. S. viminalis was grown in wastewater amended with 10 and 100 ppm copper. Trees were unaffected by wastewater with 10 ppm copper when compared to trees grown in wastewater alone. Wastewater containing with 100 ppm copper was too toxic for the trees to thrive and wastewater treatment was reduced. Treatment efficiencies for unamended wastewater were 57.7% for nitrogen, 90.6% for phosphorus and 24.9% for potassium. These efficiencies are much greater than those quoted for a Salix/soil system, and thus Salix/gravel systems may have potential for wastewater treatment in environmentally sensitive areas or situations.     

Effect of anthropogenic feeding regimes on activity rhythms of laboratory mussels exposed to natural light

Anthropogenic disturbance may affect animal behaviour and should generally be minimised. We examined how anthropogenic disturbance (24 h food deprivation) affected circadian rhythms in laboratory mussels Mytilus edulis exposed to natural light in the absence of tides. Repeated measures data were collected on mussel gape angle, exhalant pumping and valve adduction using a Hall sensor system over eight consecutive 24 h periods when exposed to two feeding conditions after 24 h food deprivation. Mussels (fed once per day at either midday or midnight) exposed to natural light showed a clear day–night rhythm with increased nocturnal activity: significantly greater gape angle, increased exhalant pumping and had significantly higher valve adduction rates. However, circadian rhythms were less clear directly after anthropogenic food deprivation, in terms of the circadian rhythm in gape angle becoming significantly more apparent over the following days. Unlike mussels fed at midnight, those fed at midday displayed no significant change in gape angle from the hour before to the hour after they were fed, i.e. mussels given food at midday reacted to this food less than mussels fed at midnight. We suggest that independent of feeding time, laboratory mussels exposed to natural light and free from anthropogenic disturbance increase feeding activity at night because their circadian rhythms are strongly influenced by light levels. This study emphasises that the behaviour of animals in the laboratory and in the wild can be altered by anthropogenic disturbances such as vibrations caused by experimental setups and artificial illumination at night.     

Toxicological effects of primary-treated urban wastewaters, before and after ozone treatment, on freshwater mussels (Elliptio complanata)

This study examined the toxic potential of a primary-treated municipal effluent, before and after ozonation, in freshwater mussels. Animals were exposed to various concentrations (0, 1, 3, 10 and 20% v/v) of a primary-treated effluent and also after a treatment with ozone at 10 mg/L in continuous flow-through mode for seven weeks. A suite of biomarkers was used to assess the potential toxic effects of various contaminants typically present in municipal wastewaters: heavy metal metabolism (metallothioneins and labile zinc), cytochrome P4501A1 and 3A4, glutathione S-transferase activities (biotransformation of organic compounds), lipid peroxidation and xanthine oxidoreductase (oxygen radical scavenging), DNA damage, mitochondrial electron transport activity at various temperatures and gonad lipid levels (cellular energy allocation) and aspartate transcarbamoylase and dihydrofolate reductase (gonad activity). On the one hand, some biomarkers, including metallothioneins, labile zinc, glutathione S-transferase, cytochrome P4503A4 activity, dehydrofolate reductase and aspartate transcarbamoylase, were readily decreased. In contrast, these biomarkers, cytochrome P4501A1, gill lipid peroxidation, DNA strand breaks in gills and digestive gland, mitochondrial electron transport at high and low temperatures (temperature-dependent activity) and total gonad lipids, were readily increased. In general, ozone treatment reduced adverse effects by either decreasing the intensity of the toxic responses or increasing the threshold concentration. For gill lipid peroxidation, however, intensity was greater at a higher threshold concentration. Ozone treatment eliminated the temperature sensitivity of the mitochondrial electron transport system, indicating a loss of interaction between temperature and urban pollution in terms of energy expenditure in mussels. Ozone treatment could significantly decrease either the toxic potency or intensity of urban pollutants at the expense of increased oxidative stress in gills of freshwater mussels.