Can short-term data accurately model long-term environmental exposures? Investigating the multigenerational adaptation potential of Daphnia magna to environmental concentrations of organic ultraviolet filters.

Organic ultraviolet filters (UVFs) are contaminants of concern, ubiquitously found in many aquatic environments due to their use in personal care products to protect against ultraviolet radiation. Research regarding the toxicity of UVFs such as avobenzone, octocrylene and oxybenzone indicate that these chemicals may pose a threat to invertebrate species; however, minimal long-term studies have been conducted to determine how these UVFs may affect continuously exposed populations. The present study modeled the effects of a 5-generation exposure of Daphnia magna to these UVFs at environmental concentrations. Avobenzone and octocrylene resulted in minor, transient decreases in reproduction and wet mass. Oxybenzone exposure resulted in > 40% mortality, 46% decreased reproduction, and 4-fold greater reproductive failure over the F0 and F1 generations; however, normal function was largely regained by the F2 generation. These results indicate that Daphnia are able to acclimate over long-term exposures to concentrations of 6.59 µg/L avobenzone, ∼0.6 µg/L octocrylene or 16.5 µg/L oxybenzone. This suggests that short-term studies indicating high toxicity may not accurately represent long-term outcomes in wild populations, adding additional complexity to risk assessment practices at a time when many regions are considering or implementing UVF bans in order to protect these most sensitive invertebrate species.

Fever integrates antimicrobial defences, inflammation control, and tissue repair in a cold-blooded vertebrate.

Multiple lines of evidence support the value of moderate fever to host survival, but the mechanisms involved remain unclear. This is difficult to establish in warm-blooded animal models, given the strict programmes controlling core body temperature and the physiological stress that results from their disruption. Thus, we took advantage of a cold-blooded teleost fish that offered natural kinetics for the induction and regulation of fever and a broad range of tolerated temperatures. A custom swim chamber, coupled to high-fidelity quantitative positional tracking, showed remarkable consistency in fish behaviours and defined the febrile window. Animals exerting fever engaged pyrogenic cytokine gene programmes in the central nervous system, increased efficiency of leukocyte recruitment into the immune challenge site, and markedly improved pathogen clearance in vivo, even when an infecting bacterium grew better at higher temperatures. Contrary to earlier speculations for global upregulation of immunity, we identified selectivity in the protective immune mechanisms activated through fever. Fever then inhibited inflammation and markedly improved wound repair. Artificial mechanical hyperthermia, often used as a model of fever, recapitulated some but not all benefits achieved through natural host-driven dynamic thermoregulation. Together, our results define fever as an integrative host response that regulates induction and resolution of acute inflammation, and demonstrate that this integrative strategy emerged prior to endothermy during evolution.

Freshwater early life growth influences partial migration in populations of Dolly Varden (Salvelinus malma malma).

Populations of northern Dolly Varden (Salvelinus malma malma) exhibit partial seaward migration, yet little is known about this phenomenon in Dolly Varden populations. Our study analyzed data from three different Dolly Varden populations in the western Canadian Arctic in order to determine if: (1) differences in size-at-first seaward migration exist between fish that migrate at early and late ages among populations inhabiting different river systems, and (2) annual growth influences anadromous or resident life history choice. Otolith strontium analysis and back-calculation were used to determine age- and size-at-first seaward migration, respectively. Differences in age- and size-at-first seaward migration were determined across river system and migration age. Back-calculated fish lengths were compared using a mixed effect model to determine how early growth influences migratory tactics (early or late aged smolt, or resident). Our results indicate that fish exhibiting faster early growth migrated in earlier years and at smaller sizes than slower growing fish, however size- and age-at first seaward migration varied by river system. Faster growing Dolly Varden tended to become either residents or early smolts, while slower growth was associated with smolting later in life. This is contrary to life history theory where the fastest growing fish in a population should mature as a resident. Our results indicate factors other than growth may be influencing life history 'decisions' in Dolly Varden. Future work on growth efficiencies and metabolic rates is needed to assess how they affect migratory behaviours.

A burning issue: The effect of organic ultraviolet filter exposure on the behaviour and physiology of Daphnia magna.

Ultraviolet (UV) filters are compounds utilized in many manufacturing processes and personal care products such as sunscreen to protect against UV-radiation. These highly lipophilic compounds are emerging contaminants of concern in aquatic environments due to their previously observed potential to bioaccumulate and exert toxic effects in marine ecosystems. Currently, research into the toxic effects of UV filter contamination of freshwater ecosystems is lacking, thus the present study sought to model the effects of acute and chronic developmental exposures to UV filters avobenzone, oxybenzone and octocrylene as well as a mixture of these substances in the freshwater invertebrate, Daphnia magna, at environmentally realistic concentrations. Median 48-hour effect and lethal concentrations were determined to be in the low mg/L range, with the exception of octocrylene causing 50% immobilization near environmental concentrations. 48-hour acute developmental exposures proved to behaviourally impair daphnid phototactic response; however, recovery was observed following a 19-day post-exposure period. Although no physiological disruptions were detected in acutely exposed daphnids, delayed mortality was observed up to seven days post-exposure at 200 µg/L of avobenzone and octocrylene. 21-day chronic exposure to 7.5 µg/L octocrylene yielded complete mortality within 7 days, while sublethal chronic exposure to avobenzone increased Daphnia reproductive output and decreased metabolic rate. 2 µg/L oxybenzone induced a 25% increase in metabolic rate of adult daphnids, and otherwise caused no toxic effects at this dose. These data indicate that UV filters can exert toxic effects in freshwater invertebrates, therefore further study is required. It is clear that the most well-studied UV filter, oxybenzone, may not be the most toxic to Daphnia, as both avobenzone and octocrylene induced behavioural and physiological disruption at environmentally realistic concentrations.

Early-life exposure to weathered, unweathered and dispersed oil has persisting effects on ecologically relevant behaviors in sheepshead minnow.

The Deepwater Horizon oil spill released 3.19 million barrels of crude oil into the Gulf of Mexico, making it the largest oil spill in U.S. history. Weathering and the application of dispersants can alter the solubility of compounds within crude oil, thus modifying the acute toxicity of the crude oil to aquatic life. The primary aim of our study was to determine the lasting impact of early-life stage sheepshead minnow (Cyprinodon variegatus variegatus) exposure to weathered, unweathered and dispersed crude oil on prey capture, male aggression, novel object interaction and global DNA methylation. Embryos were exposed from 1 to 10 dpf to water accommodations of crude oil and were raised to adulthood in artificial seawater. Our results suggest exposure to crude oil did not result in lasting impairment of complex behavioral responses of male sheepshead minnow. Exposure to dispersed weathered oil, however, decreased border dwelling in response to a novel object (i.e. decreased anxiety). Principal component analysis revealed that exposure to weathered oil had no overarching effect, but that unweathered crude oil increased variability in exploratory behaviors but decreased variability in anxiety-associated behaviors. Further work is needed to understand the effects of oil exposure on fish behavior and the potential ecological impact of subtle behavioral changes in fishes.

Field-collected crude oil, weathered oil and dispersants differentially affect the early life stages of freshwater and saltwater fishes.

The Deepwater Horizon (DWH) oil spill was the biggest in US history and released 3.19 million barrels of light crude oil into the Gulf of Mexico. In this study, we compared the toxicity of water accommodated fractions (WAFs) of naturally weathered crude oils, source oil, and source oil with dispersant mixtures and their effects on developing sheepshead minnow and zebrafish. Although a freshwater fish, zebrafish has been used as a model for marine oil spills owing to the molecular and genetic tools available and their amenability to lab care. Our study not only aimed to determine the effect of crude oil on early life stages of these two fish species, but also aimed to determine whether dissolved crude oil constituents were similar in fresh and saltwater, and if freshwater fish might be a suitable model to study marine spills. Weathering and dispersant had similar effects on WAF composition in both fresh and saltwater, except that the saltwater source oil + dispersant WAF had markedly higher PAH levels than the freshwater equivalent. WAF exposure differentially affected survival, as the LC50 values in %WAF for the zebrafish and sheepshead minnow exposures were 44.9% WAF (95% confidence interval (C.I.) 42.1-47.9) and 16.8% WAF (95% C.I. 13.7-20.5); respectively. Exposure increased heart rate of zebrafish embryos, whereas in sheepshead, source oil exposure had the opposite effect. WAF exposure altered mRNA expression of biotransformation makers, vitellogenin and neurodevelopment genes in both species. Muscle deformations were only found in oil-exposed zebrafish. This is one of the most comprehensive studies to date on crude oil toxicity, and highlights the species-specific differences in cardiotoxicity, estrogenic effects, biotransformation enzyme induction and potential neurotoxicity of crude oil exposure.

Assessment of raw and ozonated oil sands process-affected water exposure in developing zebrafish: Associating morphological changes with gene expression.

With the ever-increasing amounts of oil sands process-affected water (OSPW) accumulating from Canada's oil sands operations, its eventual release must be considered. As OSPW has been found to be both acutely and chronically toxic to aquatic organisms, remediation processes must be developed to lower its toxicity. Ozone treatment is currently being studied as a tool to facilitate the removal of organic constituents associated with toxicity. Biomarkers (e.g. gene expression) are commonly used when studying the effects of environmental contaminants, however, they are not always indicative of adverse effects at the whole organism level. In this study, we assessed the effects of OSPW exposure on developing zebrafish by linking gene expression to relevant cellular and whole organism level endpoints. We also investigated whether or not ozone treatment decreased biomarkers and any associated toxicity observed from OSPW exposure. The concentrations of classical naphthenic acids in the raw and ozonated OSPW used in this study were 16.9 mg/L and 0.6 mg/L, respectively. Ozone treatment reduced the total amount of naphthenic acids (NAs) in the OSPW sample by 92%. We found that exposure to both raw and ozonated OSPW had no effect on the survival of zebrafish embryos. The expression levels of biotransformation genes CYP1A and CYP1B were induced by raw OSPW exposure, with CYP1B being more highly expressed than CYP1A. In contrast, ozonated OSPW exposure did not increase the expression of CYP1A and only slightly induced CYP1B. A decrease in cardiac development and function genes (NKX2.5 and APT2a2a) was not associates with large changes in heart rate, arrhythmia or heart size. We did not find any indications of craniofacial abnormalities or of increased occurrence of apoptotic cells. Overall, our study found that OSPW was not overtly toxic to zebrafish embryos.

Growth and recovery of zebrafish embryos after developmental exposure to raw and ozonated oil sands process-affected water.

Due to the increasing volume of oil sands process-affect water (OSPW) and its toxicity to aquatic organisms, it is important to fully understand its effects and study remediation processes that will enable its release to the environment. Ozone treatment is currently being considered as a tool to expedite remediation, as it is known to degrade toxic organic compounds present in OSPW. In this study, we aimed to measure the effects of OSPW exposure on the growth, development and recovery of zebrafish (Danio rerio) embryos. We also used ozone-treated OSPW to determine whether ozonation negated any effects of raw OSPW exposure. As biomarkers of exposure, we assessed the expression of genes involved in neurodevelopment (ngn1, neuroD), estrogenicity (vtg), oxidative stress (sod1), and biotransformation (cyp1a, cyp1b). Our study found that exposure to both raw and ozonated OSPW did not impair growth of zebrafish embryos, however, otoliths of exposed embryos were smaller than those of control embryos. The expression levels of both cyp1a and cyp1b were induced by raw OSPW exposure. However, after the exposure period, expression levels of these genes returned to control levels within two days of residence in clean water. We found no changes in the expression levels of ngn1, neuroD and vtg genes with exposure to treated or untreated OSPW. Overall, our study found that raw OSPW exposure did not have many negative effects on zebrafish embryos and embryos appeared to recover relatively quickly after exposure ended. Furthermore, ozone treatment decreased the induction of cyp1a and cyp1b.

Sensory and Behavioral Responses of a Model Fish to Oil Sands Process-Affected Water with and without Treatment.

If oil sands process-affected water (OSPW) is to be returned to the environment, a desire is that it not adversely affect aquatic life. We investigated whether a relevant model fish (rainbow trout, Oncorhynchus mykiss) could detect OSPW using its olfactory sense (smell) and whether exposure to it would result in behavioral changes. We also investigated whether ozonation of OSPW, which lowers the concentration of organic compounds attributed with toxicity (naphthenic acids), would ameliorate any observed adverse effects. We found that OSPW, regardless of ozonation, evoked olfactory tissue responses similar to those expected of natural odorants, suggesting that fish could smell OSPW. In 30 min OSPW exposures, olfactory responses to a food odorant and a pheromone were reduced to a similar degree by OSPW, again regardless of ozonation. However, olfactory responses returned within minutes of exposure cessation. In contrast, in longer (7 d) exposures, olfactory responses remained impaired, but not in fish that had received ozone-treated OSPW. In the behavioral assay, fish avoided an introduced plume of OSPW, and this response was not affected by ozonation. Taken together, our data suggest that fish smell OSPW, that they may use this sense to mount an avoidance response, and that, if they cannot avoid it, their sensory responses may be impaired, unless the OSPW has received some remediation.

Mallards (Anas platyrhynchos) and wastewater ponds, Part I: Mallard ducks overwintering at a northern wastewater treatment pond.

In northern urban areas, wastewater treatment ponds (WWTPs) may provide a thermal refuge during winter (~10°C) that is used by normally migratory mallards (Anas platyrhynchos). On the ponds, mallards may experience stress due to crowding, or through the ingestion of WWTP water, be exposed to a diverse array of synthetic chemicals, which may have adverse health effects. Photographic sampling was used to assess mallard sex ratios and behavioural patterns throughout the late winter on wastewater ponds in Edmonton, Canada. The WWTP mallard population was large (>1000 birds), but temporally variable and consistently male-dominated. Locomotion and dabbling were the primary behaviors observed; aggression was rarely observed, which suggests crowding stress was low or absent. Mallard abundance tended to be higher at lower air temperatures, suggesting that WWTP ponds acted as a thermal refuge. Stable isotope analysis of carbon and nitrogen from duck feces and potential food sources indicated that mallards were not feeding at the site, or on invertebrates or select waste grain from offsite. Rather, ducks either consumed an undetermined food source or were feeding very little. Taken together, the data suggest that winter use of northern WWTP ponds may serve as an alternative to migration, whether this strategy benefits or harms mallards likely depends on winter severity, and not on WWTP pond characteristics or water quality.

Comparison of Diluted Bitumen (Dilbit) and Conventional Crude Oil Toxicity to Developing Zebrafish.

To facilitate pipeline transport of bitumen, it is diluted with natural gas condensate, and the resulting mixture, "dilbit", differs greatly in chemical composition to conventional crude oil. Despite the risk of accidental dilbit release, the effects of dilbit on aquatic animals are largely unknown. In this study, we compared the toxicity of water accommodated fractions (WAFs) of dilbit and two conventional crude oils, medium sour composite and mixed sweet blend, to developing zebrafish. Mortality and pericardial edema was lowest in dilbit WAF-exposed embryonic zebrafish but yolk sac edema was similar in all exposures. Shelter-seeking behavior was decreased by dilbit and conventional crude WAF exposures, and continuous swimming behavior was affected by all tested WAF exposures. Regardless of WAF type, monoaromatic hydrocarbon content (largely made up of benzene, toluene, ethylbenzene, and xylene (BTEX)) was a more accurate predictor of lethality and pericardial edema than polycyclic aromatic hydrocarbon (PAH) content. Our results suggest that the toxicity of dilbit to a model fish is less than or similar to that of conventional crudes.

Chemical avoidance responses of fishes.

The hydrosphere is a repository for all of our waste and mistakes, be they sewage, garbage, process-affected waters, runoff, and gases. For fish living in environments receiving undesirable inputs, moving away seems an obvious way to avoid harm. While this should occur, there are numerous examples where it will not. The inability to avoid harmful environments may lead to sensory impairments that in turn limit the ability to avoid other dangers or locate benefits. For avoidance to occur, the danger must first be perceived, which may not happen if the fish is 'blinded' in some capacity. Second, the danger must be recognized for what it is, which may also not happen if the fish is cognitively confused or impaired. Third, it is possible that the fish may not be able to leave the area, or worse, learns to prefer a toxic environment. Concerning generating regulations around avoidance, there are two possibilities: that an avoidance threshold be used to set guidelines for effluent release with the intention of driving fishes away; the second is to set a contaminant concentration that would not affect the avoidance or attraction responses to other cues. With the complexities of the modern world in which we release diverse pollutants, from light to municipal effluents full of 1000s of chemicals, to the diversity present in ecosystems, it is impossible to have avoidance data on every stimulus-species combination. Nevertheless, we may be able to use existing avoidance response data to predict the likelihood of avoidance of untested stimuli. Where we cannot, this review includes a framework that can be used to direct new research. This review is intended to collate existing avoidance response data, provide a framework for making decisions in the absence of data, and suggest studies that would facilitate the prediction of risk to fish health in environments receiving intentional and unintentional human-based chemical inputs.

Sublethal toxicity of chlorpyrifos to salmonid olfaction after hypersaline acclimation.

Salmonid habitats can be impacted by several environmental factors, such as salinization, which can also affect salmonid tolerance to anthropogenic stressors, such as pesticides. Previous studies have shown that hypersaline acclimation enhances the acute toxicity of certain organophosphate and carbamate pesticides to euryhaline fish; however, sublethal impacts have been far less studied. The current study aims to determine how hypersaline acclimation and exposure to the organophosphate chlorpyrifos (CPF) impact salmonid olfaction. Combined acclimation and exposure to CPF was shown to impact rainbow trout olfaction at the molecular, physiological, and behavioral levels. Concurrent exposure to hypersalinity and 0.5µg/L CPF upregulated four genes (chloride intracellular channel 4, G protein zgc:101761, calcium calmodulin dependent protein kinase II delta, and adrenergic alpha 2C receptor) that inhibit olfactory signal transduction. At the physiological level, hypersalinity and chlorpyrifos caused a decrease in sensory response to the amino acid l-serine and the bile salt taurocholic acid. Combined acclimation and exposure also negatively impacted behavior and reduced the avoidance of a predator cue (l-serine). Thus, acclimation to hypersaline conditions and exposure to environmentally relevant concentrations of chlorpyrifos caused an inhibition of olfactory signal transduction leading to a decreased response to odorants and impairment of olfactory mediated behaviors.

UV and hydrogen peroxide treatment restores changes in innate immunity caused by exposure of fish to reuse water.

The purpose of this study was to assess the innate immunity of goldfish exposed to reuse water, and UV/H2O2-treated reuse water, using a real-time flow-through exposure system. The reuse water generated by ultrafiltration of finished wastewater from the municipal wastewater treatment plant was analyzed for the presence of a panel of 20 herbicides/fungicides and 46 pharmaceuticals and personal care products (PPCP). There was a seasonal variation in the profile and concentrations of xenobiotics in reuse water with lowest levels occurring in the summer. The innate immunity parameters assessed were cytokine (IFNγ, IL-1ß, IL-10, TNFα2), and cytokine receptor (TNFR1, TNFR2, IFNGR1, IFNGR2) gene expression, and phagocytosis of kidney leukocyte subpopulations. Assessment of innate immunity parameters was done after acute (7 days) and sub chronic (30 and 60 days) exposure to reuse water, UV/H2O2-treated reuse water, and activated carbon-treated reuse water (ACT; control), during spring, summer and fall of 2012. Temporal (acute versus sub chronic) as well as seasonal differences in innate immunity of fish exposed to reuse water were observed. The acute exposure of fish to reuse water caused significant down-regulation in cytokine gene expression in different organs of fish (kidney, spleen, liver) and phagocytic ability of different kidney leukocyte subpopulations. The immune gene expression and phagocytosis of kidney leukocytes of fish returned to ACT control levels after sub chronic exposure suggesting that fish have habituated to the reuse water exposure. The changes in gene expression after acute exposure were related to variations in the profile of xenobiotics in reuse water during different seasons. The efficiency of xenobiotic removal using UV/H2O2 ranged between 1.6 and 100% indicating that treatment of reuse water using high dose UV/H2O2 was only partially effective in removing the xenobiotics, as assessed by both chemical analyses and measurement of innate immune responsiveness of the fish. Furthermore, exposure of fish to reuse water and UV/H2O2-treated reuse water generated in the spring and fall caused greater changes in innate immunity after acute exposure, compared to fish exposed to ACT reuse water, indicating that the remediation of reuse water, should be considered in order to protect aquatic and public health.

Advanced analytical mass spectrometric techniques and bioassays to characterize untreated and ozonated oil sands process-affected water.

Oil sands process-affected water (OSPW) is a toxic and poorly biodegradable mixture of sand, silt, heavy metals, and organics. In this study, qualitative and quantitative comparisons of naphthenic acids (NAs) were done using ultraperformance liquid chromatography time-of-flight mass spectrometry (UPLC TOF-MS), Fourier transform ion cyclotron resonance (FT-ICR) MS, and ion mobility spectrometry (IMS). The unique combination of these analyses allowed for the determination and correlation of NAs, oxidized NAs, and heteroatom (sulfur or nitrogen) NAs. Despite its lower resolution, UPLC-TOF MS was shown to offer a comparable level of reliability and precision as the high resolution FT-ICR MS. Additionally, the impacts of ozonation (35 mg/L utilized ozone dose) and subsequent NAs degradation on OSPW toxicity were assessed via a collection of organisms and toxicity end points using Vibrio fischeri (nonspecific), specific fish macrophage antimicrobial responses, and fish olfactory responses. Fish macrophages exposed to ozonated OSPW for 1 week showed higher production of reactive oxygen and nitrogen intermediates; however, after 12 weeks the responses were reduced significantly. Fish olfactory tests suggested that OSPW interfered with their perception of odorants. Current results indicate that the quantification of NAs species, using novel analytical methods, can be combined with various toxicity methods to assess the efficiency of OSPW treatment processes.

Acute exposure to a common suspended sediment affects the swimming performance and physiology of juvenile salmonids.

To study the effects of an acute exposure to turbidity generated by suspended sediment, we examined swimming performance (Ucrit) and related metabolic parameters in individual and groups of juvenile trout at three different concentrations of calcium carbonate. To investigate differences among strains or provenience, we compared one strain of rainbow trout (Oncorhynchus mykiss; RBT) and one strain of brown trout (Salmo trutta; BNT) from a common hatchery and one RBT strain from a separate hatchery. In general, trout swum individually or in groups exhibited a decrease in Ucrit as turbidity increased. Both RBT strains were more similar to each other and were impaired to a larger extent in swimming performance than BNT, which was less impacted. For groups, indicators of aerobic metabolism were elevated while those of anaerobic metabolism were depressed. Specifically, citrate synthase activities and glucose levels tended to be greater while plasma lactate and LDH activities were reduced. Lactate and LDH levels in individually swum trout under sediment exposure suggest a greater similarity of fish from the same provenience. We suggest that acute exposures to environmentally relevant turbidities generated by fine suspended sediment may cause a reduced Ucrit, and that these changes may be related to changes in the utilization of aerobic and anaerobic pathways.

Growth differentiation factor 6 as a putative risk factor in neuromuscular degeneration.

Mutation of Glass bottom boat, the Drosophila homologue of the bone morphogenetic protein or growth/differentiation factor (BMP/GDF) family of genes in vertebrates, has been shown to disrupt development of neuromuscular junctions (NMJ). Here we tested whether this same conclusion can be broadened to vertebrate BMP/GDF genes. This analysis was also extended to consider whether such genes are required for NMJ maintenance in post-larval stages, as this would argue that BMP genes are viable candidates for analysis in progressive neuromuscular disease. Zebrafish mutants harboring homozygous null mutations in the BMP-family gene gdf6a were raised to adulthood and assessed for neuromuscular deficits. Fish lacking gdf6a exhibited decreased endurance (∼ 50%, p = 0.005) compared to wild type, and this deficit progressively worsened with age. These fish also presented with significantly disrupted NMJ morphology (p = 0.009), and a lower abundance of spinal motor neurons (∼ 50%, p<0.001) compared to wild type. Noting the similarity of these symptoms to those of Amyotrophic Lateral Sclerosis (ALS) model mice and fish, we asked if mutations in gdf6a would enhance the phenotypes observed in the latter, i.e. in zebrafish over-expressing mutant Superoxide Dismutase 1 (SOD1). Amongst younger adult fish only bigenic fish harboring both the SOD1 transgene and gdf6a mutations, but not siblings with other combinations of these gene modifications, displayed significantly reduced endurance (75%, p<0.05) and strength/power (75%, p<0.05), as well as disrupted NMJ morphology (p<0.001) compared to wild type siblings. Bigenic fish also had lower survival rates compared to other genotypes. Thus conclusions regarding a role for BMP ligands in effecting NMJ can be extended to vertebrates, supporting conservation of mechanisms relevant to neuromuscular degenerative diseases. These conclusions synergize with past findings to argue for further analysis of GDF6 and other BMP genes as modifier loci, potentially affecting susceptibility to ALS and perhaps a broader suite of neurodegenerative diseases.

Zebrafish (Danio rerio) as a model for the study of aging and exercise: physical ability and trainability decrease with age.

A rapidly aging global population has motivated the development and use of models for human aging. Studies on aging have shown parallels between zebrafish and humans at the internal organization level; however, few parallels have been studied at the whole-organism level. Furthermore, the effectiveness of exercise as a method to mitigate the effects of aging has not been studied in zebrafish. We investigated the effects of aging and intermittent exercise on swimming performance, kinematics and behavior. Young, middle-aged and old zebrafish (20-29, 36-48 and 60-71% of average lifespan, respectively) were exercised to exhaustion in endurance and sprint swimming tests once a week for four weeks. Both endurance and sprint performance decreased with increased age. Swimming performance improved with exercise training in young and middle-aged zebrafish, but not in old zebrafish. Tail-beat amplitude, which is akin to stride length in humans, increased for all age groups with training. Zebrafish turning frequency, which is an indicator of routine activity, decreased with age but showed no change with exercise. In sum, our results show that zebrafish exhibit a decline in whole-organism performance and trainability with age. These findings closely resemble the senescence-related declines in physical ability experienced by humans and mammalian aging models and therefore support the use of zebrafish as a model for human exercise and aging.

Inhibition of P-glycoprotein in the blood-brain barrier alters avermectin neurotoxicity and swimming performance in rainbow trout.

The importance of the blood brain barrier (BBB) and the contribution to its function by the efflux transporter P-glycoprotein (P-gp) in teleosts were examined using the P-gp substrates and central nervous system neurotoxins ivermectin (22,23-dihydroavermectin B1a+22,23-dihydroavermectin B1b) [IVM]) and emamectin benzoate (4″-deoxy-49″epimethylaminoavermectin B1 benzoate [EB]). Trout were injected intraperitoneally with 0.01-1.0 and 1-50mg/kg of IVM or EB, respectively either alone or in combination with cyclosporin A (CsA: a P-gp substrate) at 1mg/kg. IVM affected the swimming performance (critical swimming speed, burst swimming distance, and schooling) at significantly lower concentrations than EB. When fish were exposed to IVM or EB in the presence of CsA, alterations to swimming were increased, suggesting that competition for P-gp in the BBB by CsA increased IVM and EB penetration into the CNS and decreased swimming capabilities. The effect of co-administration of CsA on swimming-related toxicity was different between IVM and EB-treated fish; EB toxicity was increased to a greater extent than IVM toxicity. The greater chemosensitization effect of EB vs. IVM was examined using a P-gp competitive inhibition assay in isolated trout hepatocytes with rhodamine 123 as a substrate. At the cellular level, IVM was a more potent inhibitor of P-gp than EB, which allowed for a greater accumulation of R123 in hepatocytes. These results provide evidence for a role of P-gp in the BBB of fish, and suggest that this protein protects fish from environmental neurotoxins.