Ls and methods Source data Previously published information obtained throughout a Mussel Watch monitoring carried out right after POS (April prilIntegrative biomarker indices for marine ecosystem wellness assessmentwere utilised to construct integrative biomarker indices (Marigomez et al. ; Orbea et al. ; Cajaraville et al. ; OrtizZarragoitia et al. ; Garmendia et al. a,b,c). Briefly,mussels,Mytilus galloprovincialis,have been collected along the coast of Galicia and Biscay Bay in localities in April,July and September and extended to localities in April,July and October and April (Fig In each locality,mussels ( cm shell length) had been collected and preprocessed right away soon after sampling,as detailed in earlier reports,and further on selected biomarkers,digestive gland histopathology and gamete improvement had been determined for every sample as summarized beneath. Biochemical biomarkers As detailed in preceding reports (Marigomez et al. ; Orbea et al. from which information have already been obtained,the digestive gland of mussels was dissected out in the field and instantly frozen in liquid nitrogen for biochemical analyses. AOX was determined spectrophotometrically. Because changes in AOX in response to POS followed a bellshaped profile,with induction at low and inhibition at high exposure levels,two components can be distinguished: exposure (AOXexp) and effect (AOXeff) elements (Garmendia et al. c): AOXexp eAOXiAOXo,and AOXeff e(AOXoAOXi)(AOXi); where “AOXi” is the AOX measured and “AOXo” may be the reference value at eachseason based on the available literature (Cancio et al. ; Garmendia et al Cytochemical biomarkers As detailed within the preceding paper (Garmendia et al. a) from which data have already been obtained for the present study,the digestive gland of 5 mussels was dissected out within the field immediately soon after sampling and processed to decide lysosomal responses by image analysis on cryotome sections. LP was calculated by subjective grading right after the histochemical demonstration of Nacetylhexosaminidase. VvL was determined by image evaluation following the histochemical demonstration of glucuronidase activity. The Epithelial Response Index (ERI) was calculated on the basis of the VvBAS values,according to the following formulae: ERI e(VvBASiget Hesperidin VvBASoVvBASo); exactly where “VvBASi” could be the VvBAS measured and “VvBASo” is definitely the reference worth,as outlined by the offered literature (Diez ; Marigomez et al. ; Garmendia et al. ,b). Theoretically,ERI values go as much as (high impact) with ERI B for the reference condition. Nevertheless,since hitherto the highest VvBAS identified are often under . lmlm,virtually,ERI might be normally under . Digestive gland tissue histopathology The data on the prevalence and intensity of individual inflammatory responses or parasitosis have been obtained within a preceding study (Garmendia et al. b),in which PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26698565 parasites and histopathological alterations were scored making use of either quantitative or semiquantitative scales. Intensity values of these inflammatory responses and parasitic infestations had been made use of to estimate their corresponding cumulative intensity (CIIR and CIPI,respectively),which may possibly present epizootiological indication of health impairment in mussel populations (i.e. enhanced activity from the systemic immune response or augmented susceptibility to disease; Garmendia et al. b): CIIR SPIRNHIR and CIPI SPPINHPI; exactly where NH would be the quantity of specimens presenting inflammatory responses (NHIR) or hosting parasites (NHPI),and SP would be the score corresponding to each and every inflammatory response (S.