Maria Gonzalez-Rey

Effects of non-steroidal anti-inflammatory drug (NSAID) diclofenac exposure in mussel Mytilus galloprovincialis.

In recent years, research studies have increasingly focused on assessing the occurrence of active pharmaceutical ingredients (APIs) in ecosystems. However, much remains unknown concerning the potential effects on APIs on non-target organisms due to the complexity of the mode of action, reactivity and bioconcentration potential for each specific drug. The non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF) is one of the most frequently detected APIs in surface waters worldwide and has recently been included in the list of priority substances under the European Commission. In this study, mussels (Mytilus galloprovincialis) were exposed to an environmentally relevant nominal concentration of DCF (250 ng L(-1)) over 15 days. The responses of several biomarkers were assessed in the mussel tissues: condition index (CI); superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and phase II glutathione-S-transferase (GST) activities, lipid peroxidation levels (LPO) associated with oxidative stress, acetylcholinesterase (AChE) activity related to neurotoxic effects and vitellogenin-like proteins linked to endocrine disruption. This study demonstrated significant induction of SOD and GR activities in the gills in addition to high CAT activity and LPO levels in the digestive gland. Phase II GST remained unaltered in both tissues, while the up-regulation of the AChE activity was directly related to the vitellogenin-like protein levels in exposed females, indicating an alteration in the estrogenic activity, rather than a breakdown in cholinergic neurotransmission function. This study confirmed that DCF at a concentration often observed in surface water induces tissue-specific biomarker responses. Finally, this study also revealed the importance of a multi-biomarker approach when assessing the potentially deleterious effects in a species that may be vulnerable to the continuously discharge of APIs into the ecosystems; this approach provides crucial new information regarding the unknown effects of DCF.

Does selective serotonin reuptake inhibitor (SSRI) fluoxetine affects mussel Mytilus galloprovincialis

Fluoxetine (FLX) the active pharmaceutical ingredient (API) in Prozac(®) is a widely prescribed psychoactive drug which ubiquitous occurrence in the aquatic environment is associated to a poor removal rate in waste-water treatment plant (WWTP) systems. This API acts as a selective serotonin reuptake inhibitor (SSRI) frequently reported to cause disrupting effects in non-target species. The objective of this study includes a multibiomarker response evaluation on mussel Mytilus galloprovincialis during two weeks exposure to 75 ng L(-1) FLX assessing antioxidant enzymes activities--superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); lipid peroxidation (LPO), acetylcholinesterase (AChE) neurotoxic response and endocrine disruption through alkali-labile phosphates (ALP) indirect measurement of vitellogenin-like proteins. Results show transient tissue-specific enzymatic responses and damage affecting mostly mussel gills. However, the clear ALP levels inhibition throughout time in both sex-differentiated gonads gives evidence to FLX reinforced action as an endocrine disruptor rather than an oxidative or neurotoxic inducer.

Effects of active pharmaceutical ingredients mixtures in mussel Mytilus galloprovincialis.

Active pharmaceutical ingredients (APIs) are emergent environmental contaminants widely detected in surface waters as result of incomplete waste water treatment plant (WWTP) removal processes and improper disposal. The assessment of potential effects of APIs on non-target organisms is still scarce since besides presenting multiple chemical structures, properties and modes of action, these compounds occur as complex mixtures. This study comprises a 15-day exposure of mussels Mytilus galloprovincialis to mixtures (at environmentally relevant nominal concentrations) of non-steroidal inflammatory drugs ibuprofen (IBU) and diclofenac (DCF) (250 ng L(-1) each) and selective serotonin reuptake inhibitor (SSRI) fluoxetine (FLX) (75 ng L(-1)) (MIX 1) along with the addition of classical pro-oxidant copper (Cu) (5 μg L(-1)) (MIX 2). The goals included the assessment of oxidative stress, neurotoxic and endocrine effects on this sentinel species applying both a multibiomarker and gene expression (here and later gene expression is taken as synonym to gene transcription, although it is acknowledged that it is also affected by, e.g. translation, and mRNA and protein stability) analysis approaches. The results revealed a swifter antioxidant response in digestive glands than in gills induced by MIX 1, nevertheless the presence of Cu in MIX 2 promoted a higher lipid peroxidation (LPO) induction. Neither mixture altered acetylcholinesterase (AChE) activity, while both triggered the formation of vitellogenin-like proteins in females confirming the xenoestrogenic effect of mixtures. All these results varied with respect to those obtained in previous single exposure essays. Moreover, RT-PCR analysis revealed a catalase (CAT) and CYP4Y1 gene expression down- and upregulation, respectively, with no significant changes in mRNA levels of genes encoding superoxide dismutase (SOD) and glutathione-S-transferase (GST). Finally, this study highlights variable tissue and time-specific biomarker responses and gene expression alterations, which along with several interactions between each mixture component on each biomarker confirm the susceptibility of mussels to API mixtures.

Occurrence of pharmaceutical compounds and pesticides in aquatic systems

This paper deals with the detection and quantification of APIs and other priority substances in the Arade River estuary (Portugal) providing the usefulness of Polar Organic Compound Integrative Samplers (POCIS). Thirteen APIs were detected whose variation was site and time dependent. Caffeine was at the highest concentration (804±209 ng/L) followed by theophylline (184±44 ng/L). Other APIs were analgesic, anticonvulsant, non-steroidal anti-inflammatory drugs, anti-lipidemic, anxiolytic and antidepressants. Twenty pesticides comprising atrazine, diuron, isoproturon, terbutryn and simazine included in the Water Framework Directive priority list were also site and time dependent. Carbendazim occurred at the highest concentration (45±18 ng/L at site 1) but atrazine, diuron, isoproturon and simazine levels were below the Environmental Quality Standards. Although the summer impact was unclear, the results highlighted POCIS suitability for profiling these contaminants. This is to our knowledge the first study concerning APIs and pesticides in this area.

Detoxification mechanisms in shrimp: Comparative approach between hydrothermal vent fields and estuarine environments

Hydrothermal vents are extreme deep-sea habitats that, due to their singular features, still intrigue scientific communities. Swift growth rates and profuse biomass of biological communities can be observed, despite of their inherently unstable physical-chemical and toxic conditions, indicating that organisms inhabiting this environment must be well adapted to these inhospitable conditions. The caridean shrimp, Chorocaris chacei, Mirocaris fortunata and Rimicaris exoculata, together with bathymodiolid mussels, dominate the vent fauna along the Mid-Atlantic Ridge (MAR). Crustacean species are widely used as biological indicators of environmental alterations, since they play a key ecological role as planktivorous grazers, epibenthic scavengers or as prey species. The biological consequences of the hydrothermal metal-rich environment in shrimp species are still largely unknown. Therefore, the aim of this study was the determination of the metal levels (Ag, Cd, Cu, Fe, Mn and Zn), metallothioneins (MT) and lipid peroxidation (LPO) in shrimp species collected in Rainbow, Lucky Strike and Menez-Gwen vent sites, in order to evaluate their different adaptation strategies toward metals when compared with two common coastal shrimp species (Palaemon elegans and Palaemonetes varians) from a fairly unpolluted estuarine system in south Portugal (Ria Formosa). Results show significant differences in metal concentrations, MT levels and lipid peroxidation between vent and coastal shrimp and also between shrimp species from the same site. This indicates that biochemical responses in both vent and coastal shrimp are affected not only by the environmental characteristics but also by inter-specific differences. Nevertheless, these responses apparently grant a successful adaptation for the survival in a metal-extreme environment.

Incidence of intersex in male clams Scrobicularia plana in the Guadiana Estuary (Portugal)

Endocrine disrupting chemicals (EDCs) known to be present in the marine environment, have great potential to influence reproductive status in aquatic organisms. A widespread incidence of intersex linked with EDCs exposure has been reported in molluscs and fish species. The species Scrobicularia plana is inherently gonochoristic; nonetheless, the feminisation of male clams was described in estuaries from the United Kingdom. Thus, this study reports the presence of oocytes in the testis of male clams of S. plana collected in the Guadiana River (Portugal). Samples were collected between June and September (2007), and gonad tissue analysed by light microscopy. Intersex with several degrees of intensity was detected during the first three months of sampling, in which the percentage of male clams affected varied between 5.9, 58.3 and 71.4%. Little is known about the causes of intersex in S. plana, being the urban, industrial and agricultural discharges possible major sources of induction. In the Guadiana River, the presence of several EDCs may be directly linked to the incidence of the intersex condition detected in Scrobicularia plana.