Environmental health assessment of warming coastal ecosystems in the tropics – Application of integrative physiological indices Carolina Madeira a,b, ⁎, Vanessa Mendonça a , Miguel C. Leal a,c , Augusto A.V. Flores d , Henrique N. Cabral a , Mário S. Diniz b , Catarina Vinagre a a MARE - Marine and Environmental Sciences Centre, Faculdade de Ciências da Universidade de Lisboa, Campo Grande, 1749-016, Lisboa, Portugal b UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal c Department of Fish Ecology & Evolution, Centre for Ecology, Evolution and Biogeochemistry, Swiss Federal Institute of Aquatic Science and Technology (Eawag), Seestrasse 79, 6047 Kastanienbaum, Switzerland d Cebimar - Centro de Biologia Marinha, Universidade De São Paulo, Rod. Manoel Hipólito do Rego, Km 131.5, São Sebastião, SP, Brazil HIGHLIGHTS • Chronic exposure to elevated tempera- ture tested in tropical intertidal reef an- imals • Markers of molecular damage, ROS scavenging and body condition assessed with indices • Markers at molecular and cellular level showed deleterious effects in fitness. • Markers at tissue and whole body level showed positive or no effects in fitness. • Indices are efficient holistic methods to monitor thermal change in natural populations. GRAPHICAL ABSTRACT abstract article info Article history: Received 27 April 2018 Received in revised form 11 June 2018 Accepted 12 June 2018 Available online xxxx Editor: Daniel Wunderlin According to climate science, ocean warming is one of the current and future greatest threats to coastal ecosys- tems. Projection scenarios for the end of this century show that tropical intertidal ecosystems are particularly at risk. In this study we optimized and tested a holistic method for bio-monitoring present and projected thermal pressure in such ecosystems, in order to assess organism vulnerability to ocean warming. Several species repre- sentative of different animal groups (fish, crustaceans and gastropods) were collected from the field and sub- jected to an experimental trial for 28 days, testing two temperatures: control (present seawater summer temperature) and elevated temperature (+3 °C, projected seawater temperature anomaly for 2100). Muscle samples were collected weekly to quantify several biomarkers of: i) macromolecular damage (protein unfolding and denaturation, and lipid peroxidation), ii) reactive oxygen species (ROS) scavengers (antioxidant enzymes), and iii) body condition (energy reserves and body mass). These biomarkers were combined in integrated bio- marker response (IBR) indices, either in three separate stress response categories (as previously defined) or in a unique combined analysis of overall physiological performance. Both approaches suggest that temperature af- fected IBRs, with increasing temperatures significantly impairing the overall health of individuals. Biomarkers of lower levels of biological organization indicated deleterious effects of temperature, whereas biomarkers of higher levels suggested maintenance of performance after chronic exposure. Overall indices combining the estimates of biomarkers across levels of biological organization are essential to predict the vulnerability of species, or Keywords: Tropical rocky reef Ocean warming Physiological performance Stress biomarkers IBRs Environmental health assessment Science of the Total Environment 643 (2018) 28–39 ⁎ Corresponding author at: MARE - Marine and Environmental Sciences Centre, Faculdade de Ciências da Universidade de Lisboa, Campo Grande, 1749-016, Lisboa, Portugal. E-mail address: scmadeira@fc.ul.pt (C. Madeira). https://doi.org/10.1016/j.scitotenv.2018.06.152 0048-9697/© 2018 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv