Feeding Behavioural Studies with Freshwater Gammarus spp.: The Importance of a Standardised Methodology.

Freshwater Gammarids are common leaf-shredding detritivores, and they usually feed on naturally conditioned organic material, in other words leaf litter that is characterised by an increased palatability, due to the action and presence of microorganisms (Chaumot et al. 2015; Cummins 1974: Maltby et al. 2002). Gammarus spp. are biologically omnivorous organisms, so they are involved in shredding leaf litter and are also prone to cannibalism, predation behaviour (Kelly et al. 2002) and coprophagy when juveniles (McCahon and Pascoe 1988). Gammarus spp. is a keystone species (Woodward et al. 2008), and it plays an important role in the decomposition of organic matter (Alonso et al. 2009; Bundschuh et al. 2013) and is also a noteworthy prey for fish and birds (Andrén and Eriksson Wiklund 2013; Blarer and Burkhardt-Holm 2016). Gammarids are considered to be fairly sensitive to different contaminants (Ashauer et al. 2010; Bloor et al. 2005; Felten et al. 2008a; Lahive et al. 2015; Kunz et al. 2010); in fact Amphipods have been reported to be one of the most sensitive orders to metals and organic compounds (Wogram and Liess 2001), which makes them representative test organisms for ecotoxicological studies and valid sentinel species for assessing water quality status (Garcia-Galan et al. 2017).

Contaminant exposure effects in a changing climate: how multiple stressors can multiply exposure effects in the amphipod Hyalella azteca.

Global climate change (GCC) is likely to intensify the synergistic effects between altered physicochemical parameters [of changing habitats] and other anthropogenic threats, such as water pollution, posing increased risks to aquatic biodiversity. As such, it is critical to understand how organisms will respond to changes in water temperature and salinity in the presence of contaminants. We exposed the epibenthic amphipod Hyalella azteca to a 3 × 3 factorial treatment design of three temperatures and three salinities ranging from 12 to 18 °C and 0 to 8 parts per thousand (ppt), respectively, in combination with a low-level environmentally relevant concentration of the pyrethroid insecticide bifenthrin (1 ng/L). Effects on survival and swimming behavior were evaluated after 96 h exposure. Transcription of a select suite of genes was monitored at 24, 48, and 96 h using quantitative polymerase chain reaction (qPCR). Our results not only demonstrate that the changes in salinity and temperature result in negative effects to invertebrate survival, behavior, and gene response, but that the effects were significantly more pronounced in the presence of bifenthrin. This is particularly important since greater thermal fluctuations, changes in timing and extent of glacial melt, and changes in precipitation, could result in H. azteca experiencing lower temperatures at times that coincide with increased spraying of pyrethroids. These environmentally relevant exposures using the standard test species H. azteca provide essential information for understanding effects caused by GCC in conjunction with increasing pesticide use, further highlighting the need to incorporate GCC impacts into risk assessments of contaminants of concern.

Addressing the recovery of feeding rates in post-exposure feeding bioassays: Cyathura carinata as a case study.

Post-exposure bioassays are used in environmental assessment as a cost-effective tool, but the effects of organism's recovery after exposure to pollutant has not yet been addressed in detail. The recoveries of post-exposure feeding rates after being exposed to two sublethal concentrations of cadmium during two different exposure periods (48h and 96h) were evaluated under laboratory conditions using the estuarine isopod Cyathura carinata. Results showed that feeding depression was a stable endpoint up to 24h after cadmium exposure, which is useful for ecotoxicological bioassays.

Development of a Sublethal Chronic Toxicity Test for the Northern Trout Gudgeon, Mogurnda mogurnda, and Application to Uranium, Magnesium, and Manganese.

Many international guidance documents for deriving water quality guideline values recommend the use of chronic toxicity data. For the tropical fish northern trout gudgeon, Mogurnda mogurnda, 96-h acute and 28-d chronic toxicity tests have been developed, but both tests have drawbacks. The 96-h toxicity test is acute and has a lethal endpoint; hence it is not a preferred method for guideline value derivation. The 28-d method has a sublethal (growth) endpoint, but is highly resource intensive and is high risk in terms of not meeting quality control criteria. The present study aimed to determine the feasibility of a 7-d larval growth toxicity test as an alternative to the 96-h survival and 28-d growth tests. Once the method was successfully developed, derived toxicity estimates for uranium, magnesium, and manganese were compared with those for other endpoints and tests lengths within the literature. As a final validation of the 7-d method, the sensitivity of the 7-d growth endpoint was compared with those of 14-, 21-, and 28-d exposures. Fish growth rate, based on length, over 7 d was significantly more sensitive compared with existing acute toxicity endpoints for magnesium and manganese, and was similarly sensitive to existing chronic toxicity endpoints for uranium. For uranium, the sensitivity of the growth endpoint over the 4 exposure periods was similar, suggesting that 7 d as an exposure duration is sufficient to provide an indication of longer term chronic growth effects. The sensitivity of the 7-d method, across the 3 metals tested, highlights the benefit of utilizing the highly reliable short-term 7-d chronic toxicity test method in future toxicity testing using M. mogurnda. Environ Toxicol Chem 2021;40:1596-1605. © 2021 Commonwealth of Australia. Environmental Toxicology and Chemistry © 2021 SETAC.

Assessing estuarine quality: A cost-effective in situ assay with amphipods.

In situ assays based on feeding depression can be powerful ecotoxicological tools that can link physiological organism-level responses to population and/or community-level effects. Amphipods are traditional target species for toxicity tests due to their high sensitivity to contaminants, availability in the field and ease of handling. However, cost-effective in situ assays based on feeding depression are not yet available for amphipods that inhabit estuarine ecosystems. The aim of this work was to assess a short-term in situ assay based on postexposure feeding rates on easily quantifiable food items with an estuarine amphipod. Experiments were carried out under laboratory conditions using juvenile Echinogammarus marinus as the target individual. When 60 Artemia franciscana nauplii (as prey) were provided per individual for a period of 30 min in dark conditions, feeding rates could be easily quantified. As an endpoint, postexposure feeding inhibition in E. marinus was more sensitive to cadmium contamination than mortality. Assay calibration under field conditions demonstrated the relevance of sediment particle size in explaining individual feeding rates in uncontaminated water bodies. An evaluation of the 48-h in situ bioassay based on postexposure feeding rates indicated that it is able to discriminate between unpolluted and polluted estuarine sites. Using the harmonized protocol described here, the in situ postexposure feeding assay with E. marinus was found to be a potentially useful, cost-effective tool for assessing estuarine sediment and water quality.

Establishing mussel behavior as a biomarker in ecotoxicology.

Most freshwater mussel species of the Unionoida are endangered, presenting a conservation issue as they are keystone species providing essential services for aquatic ecosystems. As filter feeders with limited mobility, mussels are highly susceptible to water pollution. Despite their exposure risk, mussels are underrepresented in standard ecotoxicological methods. This study aimed to demonstrate that mussel behavioral response to a chemical stressor is a suitable biomarker for the advancement of ecotoxicology methods that aids mussel conservation. Modern software and Hall sensor technology enabled mussel filtration behavior to be monitored real-time at very high resolution. With this technology, we present our method using Anodonta anatina and record their response to de-icing salt pollution. The experiment involved an environmentally relevant 'pulse-exposure' design simulating three subsequent inflow events. Three sublethal endpoints were investigated, Filtration Activity, Transition Frequency (number of changes from opened to closed, or vice versa) and Avoidance Behavior. The mussels presented a high variation in filtration behavior, behaving asynchronously. At environmentally relevant de-icing salt exposure scenarios, A. anatina behavior patterns were significantly affected. Treated mussels' Filtration Activity decreased during periods of very high and long de-icing salt exposure (p<0.001), however, increased during short de-icing salt exposure. Treated mussels' Transition Frequency increased during periods of very high and long de-icing salt exposure (p<0.001), which mirrored the Avoidance Behavior endpoint observed only by mussels under chemical stress. Characteristics of Avoidance Behavior were tighter shell closures with repeated and irregular shell movements which was significantly different to their undisturbed resting behavior (p<0.001). Additionally, we found that mussels were sensitive to a chemical stressor even when the mussel's valves were closed. Due to the effects of de-icing salt pollution on freshwater mussel behavior, we suggest better management practices for de-icing salt use be implemented. Our experimental method demonstrated that, with the application of current technologies, mussel behavioral response to a chemical stressor can be measured. The tested sublethal endpoints are suitable for mussel ecotoxicology studies. Avoidance Behavior proved to be a potentially suitable endpoint for calculating mussel behavior effect concentration. Therefore we recommend adult mussel behavior as a suitable biomarker for future ecotoxicological research. This method could be applied to other bivalve species and for physical and environmental stressors, such as particulate matter and temperature.