Understanding pharmaceutical exposure and the potential for effects in marine biota: A survey of bonefish (Albula vulpes) across the Caribbean Basin.

Most research on pharmaceutical presence in the environment to date has focused on smaller scale assessments of freshwater and riverine systems, relying mainly on assays of water samples, while studies in marine ecosystems and of exposed biota are sparse. This study investigated the pharmaceutical burden in bonefish (Albula vulpes), an important recreational and artisanal fishery, to quantify pharmaceutical exposure throughout the Caribbean Basin. We sampled 74 bonefish from five regions, and analyzed them for 102 pharmaceuticals. We assessed the influence of sampling region on the number of pharmaceuticals, pharmaceutical assemblage, and risk of pharmacological effects. To evaluate the risk of pharmacological effects at the scale of the individual, we proposed a metric based on the human therapeutic plasma concentration (HTPC), comparing measured concentrations to a threshold of 1/3 the HTPC for each pharmaceutical. Every bonefish had at least one pharmaceutical, with an average of 4.9 and a maximum of 16 pharmaceuticals in one individual. At least one pharmaceutical was detected in exceedance of the 1/3 HTPC threshold in 39% of bonefish, with an average of 0.6 and a maximum of 11 pharmaceuticals exceeding in a Key West individual. The number of pharmaceuticals (49 detected in total) differed across regions, but the risk of pharmacological effects did not (23 pharmaceuticals exceeded the 1/3 HTPC threshold). The most common pharmaceuticals were venlafaxine (43 bonefish), atenolol (36), naloxone (27), codeine (27), and trimethoprim (24). Findings suggest that pharmaceutical detections and concentration may be independent, emphasizing the need to monitor risk to biota regardless of exposure diversity, and to focus on risk quantified at the individual level. This study supports the widespread presence of pharmaceuticals in marine systems and shows the utility of applying the HTPC to assess the potential for pharmacological effects, and thus quantify impact of exposure at large spatial scales.

Water temperature affects the biotransformation and accumulation of a psychoactive pharmaceutical and its metabolite in aquatic organisms.

Pharmaceutically active compounds (PhACs) have been shown to accumulate in aquatic and riparian food-webs. Yet, our understanding of how temperature, a key environmental factor in nature, affects uptake, biotransformation, and the subsequent accumulation of PhACs in aquatic organisms is limited. In this study, we tested to what extent bioconcentration of an anxiolytic drugs (temazepam and oxazepam) is affected by two temperature regimes (10 and 20 °C) and how the temperature affects the temazepam biotransformation and subsequent accumulation of its metabolite (oxazepam) in aquatic organisms. We used European perch (Perca fluviatilis) and dragonfly larvae (Sympetrum sp.), which represent predator and prey species of high ecological relevance in food chains of boreal and temperate aquatic ecosystems. Experimental organisms were exposed to target pharmaceuticals at a range of concentrations (0.2-6 µg L-1) to study concentration dependent differences in bioconcentration and biotransformation. We found that the bioconcentration of temazepam in perch was significantly reduced at higher temperatures. Also, temperature had a strong effect on temazepam biotransformation in the fish, with the production and subsequent accumulation of its metabolite (oxazepam) being two-fold higher at 20 °C compared to 10 °C. In contrast, we found no temperature dependency for temazepam bioconcentration in dragonfly larvae and no detectable biotransformation of the parent compound that would result in measurable concentrations of oxazepam in this organism. Our results highlight that while organisms may share the same aquatic ecosystem, their exposure to PhACs may change differently across temperature gradients in the environment.

Bioconcentration and behavioral effects of four benzodiazepines and their environmentally relevant mixture in wild fish.

We studied the adverse effects of four benzodiazepines frequently measured in European surface waters. We evaluated bioaccumulation potential of oxazepam, bromazepam, temazepam, and clobazam in freshwater fish species - perch (Perca fluviatilis) and we conducted a series of behavioral trials to assess their potential to alter boldness, activity, and social behavior. All selected endpoints were studied individually for each target benzodiazepine and as a mixture of all tested compounds to assess possible combinatory effects. We used a three-dimensional automated tracking system to quantify the fish behavior. The four compounds bioconcentrated differently in fish muscle (temazepam > clobazam > oxazepam > bromazepam) at high exposure (9.1, 6.9, 5.7, 8.1 µg L-1, respectively) and low exposure (0.5, 0.5, 0.3, 0.4 µg L-1, respectively) concentrations. A significant amount of oxazepam was also measured in fish exposed to temazepam, most likely because of the metabolic transformation of temazepam within the fish. Bromazepam, temazepam, and clobazam significantly affected fish behavior at high concentration, while no statistically significant changes were registered for oxazepam. The studied benzodiazepines affected behavior in combination, because the mixture treatment significantly changed several important behavioral traits even at low concentration, while no single compound exposure had such an effect at that dose. Based on our results, we conclude that effects of pharmaceuticals on aquatic environments could be underestimated if risk assessments only rely on the evaluation of single compounds. More studies focused on the combinatory effects of environmentally relevant mixtures of pharmaceuticals are necessary to fill the gaps in this knowledge.

Interrelationship of cartilage composition and chondrocyte mechanics after a partial meniscectomy in the rabbit knee joint - Experimental and numerical analysis.

Site-specific and depth-dependent properties of cartilage were implemented within a finite element (FE) model to determine if compositional or structural changes in the tissue could explain site-specific alterations of chondrocyte deformations due to cartilage loading in rabbit knee joints 3 days after a partial meniscectomy (PM). Depth-dependent proteoglycan (PG) content, collagen content and collagen orientation in the cartilage extracellular matrix (ECM), and PG content in the pericellular matrix (PCM) were assessed with microscopic and spectroscopic methods. Patellar, femoral groove and samples from both the lateral and medial compartments of the femoral condyle and tibial plateau were extracted from healthy controls and from the partial meniscectomy group. For both groups and each knee joint site, axisymmetric FE models with measured properties were generated. Experimental cartilage loading was applied in the simulations and chondrocyte volumes were compared to the experimental values. ECM and PCM PG loss occurred within the superficial cartilage layer in the PM group at all locations, except in the lateral tibial plateau. Collagen content and orientation were not significantly altered due to the PM. The FE simulations predicted similar chondrocyte volume changes and group differences as obtained experimentally. Loss of PCM fixed charge density (FCD) decreased cell volume loss, as observed in the medial femur and medial tibia, whereas loss of ECM FCD increased cell volume loss, as seen in the patella, femoral groove and lateral femur. The model outcome, cell volume change, was also sensitive to applied tissue geometry, collagen fibril orientation and loading conditions.

Fish on steroids: Temperature-dependent effects of 17ß-trenbolone on predator escape, boldness, and exploratory behaviors.

Hormonal growth promoters (HGPs), widely used in beef cattle production globally, make their way into the environment as agricultural effluent-with potential impacts on aquatic ecosystems. One HPG of particular concern is 17ß-trenbolone, which is persistent in freshwater habitats and can affect the development, morphology and reproductive behaviors of aquatic organisms. Despite this, few studies have investigated impacts of 17ß-trenbolone on non-reproductive behaviors linked to growth and survival, like boldness and predator avoidance. None consider the interaction between 17ß-trenbolone and other environmental stressors, such as temperature, although environmental challenges confronting animals in the wild seldom, if ever, occur in isolation. Accordingly, this study aimed to test the interactive effects of trenbolone and temperature on organismal behavior. To do this, eastern mosquitofish (Gambusia holbrooki) were subjected to an environmentally-relevant concentration of 17ß-trenbolone (average measured concentration 3.0 ±â€¯0.2 ng/L) or freshwater (i.e. control) for 21 days under one of two temperatures (20 and 30 °C), after which the predator escape, boldness and exploration behavior of fish were tested. Predator escape behavior was assayed by subjecting fish to a simulated predator strike, while boldness and exploration were assessed in a separate maze experiment. We found that trenbolone exposure increased boldness behavior. Interestingly, some behavioral effects of trenbolone depended on temperature, sex, or both. Specifically, significant effects of trenbolone on male predator escape behavior were only noted at 30 °C, with males becoming less reactive to the simulated threat. Further, in the maze experiment, trenbolone-exposed fish explored the maze faster than control fish, but only at 20 °C. We conclude that field detected concentrations of 17ß-trenbolone can impact ecologically important behaviors of fish, and such effects can be temperature dependent. Such findings underscore the importance of considering the potentially interactive effects of other environmental stressors when investigating behavioral effects of environmental contaminants.

Exposure to an anti-androgenic herbicide negatively impacts reproductive physiology and fertility in Xenopus tropicalis.

Amphibians are threatened on a global scale and pollutants may be contributing to population declines, but how chemicals impact on their reproduction is poorly understood. We conducted a life cycle analysis to investigate the impacts of early life exposure to two anti-androgens (exposure until completion of metamorphosis;stage 66): flutamide, (50 µg/L)/linuron (9 and 45 µg/L)) on sexual development and breeding competence in Xenopus tropicalis. Our analyses included: mRNA levels of dmrt1, cyp17, amh, cyp19, foxl2 and ar (tadpoles/metamorphs), gonadal histomorphology (metamorphs/adults), mRNA levels of ar/gr (adult male brain/gonad/forelimb), testosterone/corticosterone levels (adult males), secondary sexual characteristics (forelimb width/nuptial pad: adult males) and breeding competence (amplexus/fertility: adult males). Compared to controls, feminised sex ratios and increased number of spermatogonia (adults) were observed after exposure to flutamide and the lower linuron concentration. Exposure to the lower linuron concentration also resulted in demasculinisation of secondary sexual characteristics and reduced male fertility. Flutamide exposure resulted in masculinisation of the nuptial pad and elevated mRNA levels of dmrt1, cyp17, amh and foxl2 in brains (metamorphs). Testosterone levels were higher in all treatment groups, however, overall few effects were observed in response to the higher linuron concentration. Our findings advance understanding of reproductive biology of X. tropicalis and illustrate negative effects of linuron on reproductive processes at a concentration measured in freshwater environments.

Size-dependent trapping behavior and optical emission study of NaYF4 nanorods in optical fiber tip tweezers.

Trapping of NaYF4:Er/Yb/Gd nanorods using an original optical fiber-tip tweezers is reported. Depending on their length, nanorods are reproducibly trapped in single or dual fiber tip configurations. Short rods of 600 nm length are trapped with two fiber tips facing each other. In contrary, long rods (1.9 µm) can be stably trapped at the apex of one single fiber tip and at a second stable trapping position 5 µm away from the tip. The up-conversion emission of trapped long nanorods is studied as a function of the position on the nanorod and in three orthogonal directions. The experimental results are discussed using numerical simulations based on exact Maxwell Stress Tensor approach.

No evidence of increased growth or mortality in fish exposed to oxazepam in semi-natural ecosystems.

An increasing number of short-term laboratory studies on fish reports behavioral effects from exposure to aquatic contaminants or raised carbon dioxide levels affecting the GABAA receptor. However, how such GABAergic behavioral modifications (GBMs) impact populations in more complex natural systems is not known. In this study, we induced GBMs in European perch (Perca fluviatilis) via exposure to a GABA agonist (oxazepam) and followed the effects on growth and survival over one summer (70days) in replicated pond ecosystems. We hypothesized that anticipated GBMs, expressed as anti-anxiety like behaviors (higher activity and boldness levels), that increase feeding rates in laboratory assays, would; i) increase growth and ii) increase mortality from predation. To test our hypotheses, 480 PIT tagged perch of known individual weights, and 12 predators (northern pike, Esox lucius) were evenly distributed in 12 ponds; six control (no oxazepam) and six spiked (15.5±4µgl-1 oxazepam [mean±1S.E.]) ponds. Contrary to our hypotheses, even though perch grew on average 16% more when exposed to oxazepam, we found no significant difference between exposed and control fish in growth (exposed: 3.9±1.2g, control: 2.9±1g [mean±1S.E.], respectively) or mortality (exposed: 26.5±1.8individuals pond-1, control: 24.5±2.6individuals pond-1, respectively). In addition, we show that reduced prey capture efficiency in exposed pike may explain the lack of significant differences in predation. Hence, our results suggest that GBMs, which in laboratory studies impact fish behavior, and subsequently also feeding rates, do not seem to generate strong effects on growth and predation-risk in more complex and resource limited natural environments.

Early in situ changes in chondrocyte biomechanical responses due to a partial meniscectomy in the lateral compartment of the mature rabbit knee joint.

We determined the biomechanical responses of chondrocytes to indentation at specific locations within the superficial zone of cartilage (i.e. patellar, femoral groove, femoral condylar and tibial plateau sites) taken from female New Zealand white rabbits three days after a partial meniscectomy in the lateral compartment of a knee joint. Confocal laser scanning microscopy combined with a custom indentation system was utilized to image chondrocyte responses at sites taken from ten contralateral and experimental knee joints. Cell volume, height, width and depth changes, global, local axial and transverse strains and Young׳s moduli were determined. Histological assessment was performed and proteoglycan content from the superficial zone of each site was determined. Relative to contralateral group cells, patellar, femoral groove and lateral femoral condyle cells in the experimental group underwent greater volume decreases (p < 0.05), due to smaller lateral expansions (with greater decreases in cell height only for the lateral femoral condyle cells; p < 0.05) whereas medial femoral and medial tibial plateau cells underwent smaller volume decreases (p < 0.05), due to less deformation in cell height (p < 0.05). Proteoglycan content was reduced in the patellar (p > 0.05), femoral groove, medial femoral condyle and medial tibial plateau experimental sites (p < 0.05). The findings suggest: (i) cell biomechanical responses to cartilage loading in the rabbit knee joint can become altered as early as 3 days after a partial meniscectomy, (ii) are site-specific, and (iii) occur before alterations in tissue mechanics or changes detectable with histology.

Site-specific cell-tissue interactions in rabbit knee joint articular cartilage.

Relationships between cartilage structure and superficial in situ chondrocyte deformations were investigated from 6 different knee joint locations (n=10 knees). Depth dependent cartilage structure and composition were quantified with microscopic/microspectroscopic methods. Medial tibial cartilages had the lowest superficial collagen content, highest collagen orientation angle, and highest proteoglycan content in the pericellular matrix relative to that in the extracellular matrix, coupled with the largest chondrocyte deformations. In contrast, femoral groove and lateral tibial cartilages had the highest superficial collagen contents, lowest collagen orientation angles, and low normalized proteoglycan contents in the pericellular matrix, coupled with the smallest chondrocyte deformations. To study cell-tissue interactions further, observations (n=57) from all locations were pooled and a multivariable linear regression was performed. Cell width deformations (R2=0.57) correlated with collagen orientation angle (standardized regression coefficient ß=0.398) and collagen content (ß=-0.402). Cell height deformations (R2=0.52) also correlated with collagen orientation (ß=-0.248) and collagen content (ß=0.455). Cell volume change upon cartilage compression (R2=0.41) correlated with collagen content (ß=0.435) and proteoglycan content (ß=0.279). In conclusion, higher collagen and proteoglycan contents combined with lower collagen orientation angle in the extracellular matrix were related to reductions in superficial chondrocyte deformations. Also, a steep gradient of proteoglycan content from the extracellular to the pericellular matrix was associated with increased cell deformation, particularly in the medial tibial plateau cartilage.

Bioaccumulation of five pharmaceuticals at multiple trophic levels in an aquatic food web - Insights from a field experiment.

Pharmaceuticals derived from manufacturing and human consumption contaminate surface waters worldwide. To what extent such pharmaceutical contamination accumulates and disperses over time in different compartments of aquatic food webs is not well known. In this study we assess to what extent five pharmaceuticals (diphenhydramine, oxazepam, trimethoprim, diclofenac, and hydroxyzine) are taken up by fish (European perch) and four aquatic invertebrate taxa (damselfly larvae, mayfly larvae, waterlouse, and ramshorn snail), by tracing their bioconcentrations over several months in a semi-natural large-scale (pond) system. The results suggest both significant differences among drugs in their capacity to bioaccumulate and differences among species in uptake. While no support for in situ uptake of diclofenac and trimethoprim was found, oxazepam, diphenhydramine, and hydroxyzine were detected in all analyzed species. Here, the highest bioaccumulation factor (tissue:water ratio) was found for hydroxyzine. In the food web, the highest concentrations were found in the benthic species ramshorn snail and waterlouse, indicating that bottom-living organism at lower trophic positions are the prime receivers of the pharmaceuticals. In general, concentrations in the biota decreased over time in response to decreasing water concentrations. However, two interesting exceptions to this trend were noted. First, mayfly larvae (primarily grazers) showed peak concentrations (a fourfold increase) of oxazepam, diphenhydramine, and hydroxyzine about 30days after initial addition of pharmaceuticals. Second, perch (top-predator) showed an increase in concentrations of oxazepam throughout the study period. Our results show that drugs can remain bioavailable for aquatic organism for long time periods (weeks to months) and even re-enter the food web at a later time. As such, for an understanding of accumulation and dispersion of pharmaceuticals in aquatic food webs, detailed ecological knowledge is required.

Site-dependent biomechanical responses of chondrocytes in the rabbit knee joint.

Biomechanical responses of chondrocytes were determined in specific locations within the superficial zone of patellar, femoral groove, femoral condyle and tibial plateau cartilages obtained from female New Zealand White rabbits. A confocal laser scanning microscope combined with a custom indentation system was utilized for experimentation. Changes in cell volumes and dimensions (i.e. cell height, width and depth) due to loading, global, local axial and transverse strains were determined for each site. Tissue composition and structure was analysed at each indentation site with digital densitometry, polarized light microscopy and Fourier transform infrared imaging spectroscopy. Patellar cells underwent greater volume decreases (compared to femoral groove cells; p<0.05) primarily due to greater decreases in cell height (p<0.05), consistent with greater levels of both global and local axial strains (p<0.05). Lateral condyle cells underwent greater volume decreases (compared to lateral plateau cells; p<0.05) primarily due to greater decreases in cell height, consistent with greater levels of tissue strains (p<0.05). Medial condyle cells underwent smaller volume decreases (compared to medial plateau cells; p<0.05) primarily due to elevated cell expansions in the depth direction, which was consistent with greater levels of minor transverse strains (p<0.05). Site-dependent differences in collagen orientation angles agreed conceptually with the observed cell dimensional changes. Chondrocyte biomechanical responses were highly site-dependent and corresponded primarily with the orientation of the collagen fibrils. The observed differences were thought to be due to the different biomechanical loading conditions at each site.

Long-Term Persistence of an Anxiolytic Drug (Oxazepam) in a Large Freshwater Lake.

Production and human consumption of pharmaceuticals result in contamination of surface waters worldwide. Little is known about the long-term (i.e., over decades) fate of pharmaceuticals in aquatic systems. Here, we show that the most prescribed anxiolytic in Sweden (oxazepam) persists in its therapeutic form for several decades after being deposited in a large freshwater lake. By comparing sediment cores collected in 1995 and 2013, we demonstrate that oxazepam inputs from the early 1970s remained in the sediments until sampling in 2013, despite in situ degradation processes and sediment diagenesis. In laboratory and pond experiments, we further reveal that therapeutic forms of oxazepam can persist over several months in cold (5 °C) lake water free from UV light. We conclude that oxazepam can persist in lakes over a time scale much longer than previously realized and that levels can build up in lakes due to both a legacy of past inputs and a growing urban population.

Alterations in subchondral bone plate, trabecular bone and articular cartilage properties of rabbit femoral condyles at 4 weeks after anterior cruciate ligament transection.

OBJECTIVE: To quantify early osteoarthritic-like changes in the structure and volume of subchondral bone plate and trabecular bone and properties of articular cartilage in a rabbit model of osteoarthritis (OA) induced by anterior cruciate ligament transection (ACLT). METHODS: Left knee joints from eight skeletally mature New Zealand white rabbits underwent ACLT surgery, while the contralateral (CTRL) right knee joints were left unoperated. Femoral condyles were harvested 4 weeks after ACLT. Micro-computed tomography imaging was applied to evaluate the structural properties of subchondral bone plate and trabecular bone. Additionally, biomechanical properties, structure and composition of articular cartilage were assessed. RESULTS: As a result of ACLT, significant thinning of the subchondral bone plate (P < 0.05) was accompanied by significantly reduced trabecular bone volume fraction and trabecular thickness in the medial femoral condyle compartment (P < 0.05), while no changes were observed in the lateral compartment. In both lateral and medial femoral condyles, the equilibrium modulus and superficial zone proteoglycan (PG) content were significantly lower in ACLT than CTRL joint cartilage (P < 0.05). Significant alterations in the collagen orientation angle extended substantially deeper into cartilage from the ACLT joints in the lateral femoral condyle relative to the medial condyle compartment (P < 0.05). CONCLUSIONS: In this model of early OA, significant changes in volume and microstructure of subchondral bone plate and trabecular bone were detected only in the femoral medial condyle, while alterations in articular cartilage properties were more severe in the lateral compartment. The former finding may be associated with reduced joint loading in the medial compartment due to ACLT, while the latter finding reflects early osteoarthritic changes in the lateral compartment.

In vitro glycation of articular cartilage alters the biomechanical response of chondrocytes in a depth-dependent manner.

OBJECTIVE: To determine if increasing cartilage cross-links through in vitro glycation of cartilage explants can alter the biomechanical response of chondrocytes to compressive deformation. METHOD: Bovine osteochondral explants were either incubated with cell culture solution supplemented with (n = 7) or without (n = 7) ribose for 42 h in order to induce glycation. Deformation-induced changes in cell volume, dimensions and local tissue strains were determined through confocal laser scanning microscopy (CLSM) and the use of a custom built micro-compression device. Osteochondral explants were also utilized to demonstrate changes in depth-wise tissue properties, biomechanical tissue properties and cross-links such as pentosidine (Pent), hydroxylysyl pyridinoline (HP) and lysyl pyridinoline (LP). RESULTS: The ribose treated osteochondral samples experienced reduced cell volume deformation in the upper tissue zone by ∼ 8% (P = 0.005), as compared the control samples, through restricting cell expansion. In the deeper tissue zone, cell volume deformation was increased by ∼ 12% (P < 0.001) via the transmission of mechanical signals further into the tissue depth. Biomechanical testing of the ribose treated osteochondral samples demonstrated an increase in the equilibrium and dynamic strain dependent moduli (P < 0.001 and P = 0.008, respectively). The biochemical analysis revealed an increase in Pent cross-links (P < 0.001). Depth-wise tissue property analyses revealed increased levels of carbohydrate content, greater levels of fixed charge density and an increased carbohydrate to protein ratio from 6 to 16%, 55-100% and 72-79% of the normalized tissue thickness (from the surface), respectively, in the ribose-treated group (P < 0.05). CONCLUSION: In vitro glycation alters the biomechanical response of chondrocytes in cartilage differently in upper and deeper zones, offering possible insights into how aging could alter cell deformation behavior in cartilage.

Effect of anaerobic digestion at 35, 55 and 60 °C on pharmaceuticals and organic contaminants.

The application of treated sewage sludge on farmland is a suggested method for recycling nutrients and reducing demand for commercial fertilizer. However, sludge needs to be safe from possible contaminants which can cause acute and long-term health and environmental problems. Residual pharmaceuticals and organic contaminants are mentioned as emerging threats since wastewater treatment plants are not designed to degrade these substances. The aim of this study was to screen and evaluate the presence, and reduction, of pharmaceuticals and polycyclic aromatic hydrocarbons (PAHs) during anaerobic digestion of mixed primary and waste-activated sludge at 35, 55 and 60 °C and during pasteurization at 70 °C. The study showed the difficulty of analysing pharmaceutical compounds in low concentrations in the sludge matrix. No general reduction of these compounds was seen during treatment, but for individual substances some reduction occurred. The PAHs were generally not reduced during digestion or pasteurization, but for three substances (indeno[1,2,3-cd]pyrene and dibenzo[a,h]anthracene (analysed together) and benzo[g,h,i]perylene) reduction (up to 60%) during digestion was seen. Digestion at 35 and 55 °C resulted in about the same order of reduction of the three individual PAHs, which was higher than for digestion at 60 °C.

Antihistamines and aquatic insects: bioconcentration and impacts on behavior in damselfly larvae (Zygoptera).

Because aquatic insects use histamines as neurotransmitters, adverse impacts on aquatic insects living in aquatic environments that receive antihistamines with wastewater effluent are plausible. In this study, we exposed damselfly larvae to low concentrations of two commonly used antihistamines (Hydroxyzine and Fexofenadine, 360 ± 42 and 2,200 ± 43 ng l(-1), respectively), and recorded damselfly larvae behavior before and after exposure. Further, after the second set of behavioral assays was performed, we quantified bioconcentration of the antihistamines in the damselfly bodies. Our results showed significant changes in damselfly behavior following antihistamine exposure. After Hydroxyzine exposure, the damselfly larvae became less active, and they showed reduced fleeing response (i.e. increased boldness) after being exposed to Fexofenadine, the latter also being significantly different from the non-exposed (control) individuals. Further, we found high levels of bioconcentration in the damselflies; Hydroxyzine showed an average bioconcentration factor (BCF) of 2000. As such, our results indicate that low concentrations of antihistamines can have sub-lethal effects on aquatic insects manifested as behavioral changes, and that bioconcentration of these substances can be high. Therefore, the need to investigate the impact of emergent aquatic contaminants also on aquatic insects, and on behaviors that are of ecological importance, is further highlighted.

Dilute concentrations of a psychiatric drug alter behavior of fish from natural populations.

Environmental pollution by pharmaceuticals is increasingly recognized as a major threat to aquatic ecosystems worldwide. A variety of pharmaceuticals enter waterways by way of treated wastewater effluents and remain biochemically active in aquatic systems. Several ecotoxicological studies have been done, but generally, little is known about the ecological effects of pharmaceuticals. Here we show that a benzodiazepine anxiolytic drug (oxazepam) alters behavior and feeding rate of wild European perch (Perca fluviatilis) at concentrations encountered in effluent-influenced surface waters. Individuals exposed to water with dilute drug concentrations (1.8 micrograms liter(-1)) exhibited increased activity, reduced sociality, and higher feeding rate. As such, our results show that anxiolytic drugs in surface waters alter animal behaviors that are known to have ecological and evolutionary consequences.

Foreign body reaction to a retained surgical sponge (gossypiboma) mimicking an implant associated sarcoma in a dog after a tibial plateau levelling osteotomy.

A dog was presented with the complaint of an acute onset left pelvic limb lameness three years after a right tibial plateau levelling osteotomy had been performed. Radiographs taken at the time of presentation showed signs that were consistent with a diagnosis of an implant associated sarcoma. At revision surgery, a retained surgical sponge was identified, leading to a diagnosis of a gossypiboma. This is the first reported case of a gossypiboma as a complication of a tibial plateau levelling osteotomy surgery.

Prioritising pharmaceuticals for environmental risk assessment: Towards adequate and feasible first-tier selection.

The presence of pharmaceuticals in the aquatic environment, and the concerns for negative effects on aquatic organisms, has gained increasing attention over the last years. As ecotoxicity data are lacking for most active pharmaceutical ingredients (APIs), it is important to identify strategies to prioritise APIs for ecotoxicity testing and environmental monitoring. We have used nine previously proposed prioritisation schemes, both risk- and hazard-based, to rank 582 APIs. The similarities and differences in overall ranking results and input data were compared. Moreover, we analysed how well the methods ranked seven relatively well-studied APIs. It is concluded that the hazard-based methods were more successful in correctly ranking the well-studied APIs, but the fish plasma model, which includes human pharmacological data, also showed a high success rate. The results of the analyses show that the input data availability vary significantly; some data, such as logP, are available for most API while information about environmental concentrations and bioconcentration are still scarce. The results also suggest that the exposure estimates in risk-based methods need to be improved and that the inclusion of effect measures at first-tier prioritisation might underestimate risks. It is proposed that in order to develop an adequate prioritisation scheme, improved data on exposure such as degradation and sewage treatment removal and bioconcentration ability should be further considered. The use of ATC codes may also be useful for the development of a prioritisation scheme that includes the mode of action of pharmaceuticals and, to some extent, mixture effects.