Computerized sperm motility analysis in toxicity bioassays: A new approach to pore water quality assessment $ Adele Fabbrocini a,n , Michele Di Stasio b , Raffaele D’Adamo a a Consiglio Nazionale delle Ricerche—Istituto di Scienze Marine, UOS Lesina, via Pola 4, 71010 Lesina (FG), Italy b Consiglio Nazionale delle Ricerche—Istituto di Scienze dell’Alimentazione, via Roma 52, 83100 Avellino, Italy article info Article history: Received 15 December 2009 Received in revised form 28 April 2010 Accepted 2 May 2010 Keywords: Toxicity bioassays Sea urchin Computerized sperm motility analysis Embryonic development Microtox s Pore water Mediterranean lagoon abstract The aim of this study was to test the sensitivity of computerized sperm motility analysis in the sea urchin Paracentrotus lividus as the endpoint in toxicity bioassays. The tested matrices were pore water samples collected in an agriculture-impacted Mediterranean lagoon, Lake Varano (Italy). Two standardized bioassays were also conducted as controls, the P. lividus spermiotoxicity test and the Vibrio fischeri (Microtox s ) test. VCL (curvilinear velocity), VSL (straight line velocity), VAP (average path velocity), and the percentage of rapid spermatozoa recorded by the Sperm Class Analyzer s system showed high sensitivity and discrimination ability, to a degree comparable with the larval development endpoint of the spermiotoxicity test. The test evaluated in this study requires small volumes of matrices, involves minimal sample manipulation, and can easily be extended to many other bioindicator species. It may therefore be considered a promising ‘‘quick response tool’’ following hazardous events that may adversely affect an aquatic ecosystem. & 2010 Elsevier Inc. All rights reserved. 1. Introduction Coastal ecosystems are currently subject to the impact of numerous human activities that lead to the input of a variety of pollutants of agricultural, urban, or industrial origin. This is particularly true of lagoon environments, which ‘‘act as inter- mediaries for material discharged from the main-land’’ (Cognetti and Maltagliati, 2000). These substances tend to be adsorbed by particulate matter and finally accumulate in the sediment (Bufflap and Allen, 1995; Nipper, 2000), which can thus be considered both the main sink and source of contaminants in marine coastal areas (Liß and Ahlf, 1997; Libralato et al., 2008). Sediment quality can be assessed using a variety of approaches. In recent years, ecotoxicological tests have been used more and more frequently and are now standardized in inter- laboratory comparisons (US EPA, 1994; ASTM, 1997). Toxicity bioassays give a rapid estimation of the bioavailable fraction of toxicants, even at low doses, and take account of the synergistic and/or antagonistic effects of all components interacting with the biota (McCauley et al., 2000; Kiss et al., 2003). The choice of organisms (biological indicators) for bioassays must take into account many factors, such as sensitivity and reliability, distribu- tion and environmental relevance, availability over the course of the year; furthermore, a toxicity test also requires a reproducible endpoint that can be accurately, predictably, and reliably measured (Chapman, 2002a). Due to their high sensitivity, the gametes and embryos of aquatic species are widely used in ecotoxicological bioassays (His et al., 1999; Nipper, 2000; Losso et al., 2007a; Beiras and Bellas, 2008) and the effects of many toxicants on sperm motility and capacity for fertilization have been investigated (Khan and Weis, 1987; Warnau et al., 1996; Rosety et al., 2003) as well the possible interactions of pollutants with the energy metabolism and the cell viability (Dreanno et al., 1999; Rurangwa et al., 2002; Flajˇ shans et al., 2004). Developments in the application of computerized sperm motility analyses to aquatic species have made it possible to put motility evaluation on a quantitative basis (Kime et al., 2001; Liu et al., 2007), and to describe the physical chemical and physiological factors controlling sperm motility in aquatic species (Cosson et al., 1995; Dreanno et al., 1999; Cosson, 2004). The use of computer-assisted sperm motility analysis provides a rapid, quanti- tative, and above all objective assessment of sperm quality, giving real-time information on sperm motility characteristics such as curvilinear, straight line, and angular velocity, which are not manually observable but have been shown to be positively correlated with capacity for fertilization (Linhart et al., 2000; Rurangwa et al., 2001; Au et al., 2002). In addition, predicting the Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/ecoenv Ecotoxicology and Environmental Safety 0147-6513/$ - see front matter & 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.ecoenv.2010.05.003 $ All the assays carried out with sea urchins were performed in accordance with national and institutional guidelines for the protection of wildlife animal welfare. n Corresponding author. Fax: + 39 0882 991352. E-mail addresses: adele.fabbrocini@fg.ismar.cnr.it (A. Fabbrocini), distasio@isa.cnr.it (M. Di Stasio), raffaele.dadamo@fg.ismar.cnr.it (R. D’Adamo). Ecotoxicology and Environmental Safety 73 (2010) 1588–1595