Speciation of lead, copper, zinc and antimony in water draining a shooting range—Time dependant metal accumulation and biomarker responses in brown trout (Salmo trutta L.) Lene Sørlie Heier a, ⁎, Ivar B. Lien b , Arnljot E. Strømseng b , Marita Ljønes b , Bjørn Olav Rosseland a , Knut-Erik Tollefsen c , Brit Salbu a a Norwegian University of Life Sciences (UMB), P.O. Box 5003, N-1432 Aas, Norway b Norwegian Defence Research Establishment (FFI), P.O. Box 23, N-2027 Kjeller, Norway c Norwegian Institute for Water Research (NIVA), Gaustadalleèn 21, N-0349 Oslo, Norway abstract article info Article history: Received 8 December 2008 Received in revised form 27 February 2009 Accepted 4 March 2009 Available online 31 March 2009 Keywords: Metals Speciation Brown trout Metal accumulation Biomarker response The speciation of Pb, Cu, Zn and Sb in a shooting range run-off stream were studied during a period of 23 days. In addition, metal accumulation in gills and liver, red blood cell ALA-D activity, hepatic metallothionine (Cd/Zn-MT) and oxidative stress index (GSSG/ tGSH levels) in brown trout (Salmo trutta L.) exposed to the stream were investigated. Fish, contained in cages, were exposed and sampled after 0, 2, 4, 7, 9, 11 and 23 days of exposure. Trace metals in the water were fractionated in situ according to size (nominal molecular mass) and charge properties. During the experimental period an episode with higher runoff occurred resulting in increased levels of metals in the stream. Pb and Cu were mainly found as high molecular mass species, while Zn and Sb were mostly present as low molecular mass species. Pb, Cu and Sb accumulated on gills, in addition to Al origination from natural sources in the catchment. Pb, Cu and Sb were also detected at elevated concentration in the liver. Blood glucose and plasma Na and Cl levels were significantly altered during the exposure period, and are attributed to elevated concentrations of Pb, Cu and Al. A significant suppression of ALA-D was detected after 11 days. Significant differences were detected in Cd/Zn-MT and oxidative stress (tGSH/GSSG) responses at Day 4. For Pb the results show a clear link between the HMM (high molecular mass) positively charged Pb species, followed by accumulation on gills and liver and a suppression in ALA-D. Thus, high flow episodes can remobilise metals from the catchment, inducing stress to aquatic organisms. © 2009 Elsevier B.V. All rights reserved. 1. Introduction Firing ranges are locations were substantial amounts of ammunition are deposited on the ground. The use of small arms ammunition causes significant deposits of lead (Pb), antimony (Sb), copper (Cu) and zinc (Zn). In the Norwegian small arms training areas, the deposition of metals in 2006 was estimated to be approximately 103, 73, 12 and 7 t of Pb, Cu, Zn and Sb, respectively (Reistad et al., 2008). Pb, Cu and Sb are considered to be of major concern, due to the amounts released and the potential negative effect on living organisms. Of the total Norwegian lead emissions in 2005 (total 240 t), ammunition was the major source (66%) followed by fishing equipment (24%), while for instance industrial deposits only constituted 3%. In addition, approximately 200 t of lead has been deposited as a result of atmospheric long-range transport from other countries in Europe. However, the contribution from long-range transport has been reduced since the late 1970s, and is today only 10% of the 1977 level (Norwegian Pollution Control Authority, 2008). The total Norwegian emissions of Cu (2005) was 1067 t, where fish farming net impregnation represented the greatest source (54%) followed by anti- fouling preparations (26%). Both Pb and Cu are on the Norwegian Pollution Control Authority priority list of chemicals to be reduced before 2010. Sb and Zn, however, are not considered as key pollutants. Information on Sb is also relatively scarce. Metals deposited on the soil can be present as particles or associated with soil components. Metals can be mobilized from solid phases due to particle weathering, changes in pH or redox conditions and complexa- tion with organics, and subsequently transported in the aquatic phase. During a 15-year surveillance program of aquatic metal pollution in military training areas, elevated levels of Pb, Cu, Zn and Sb were found in several areas (Rognerud and Forsvarsbygg, 2006). Maximum concen- trations of Pb, Cu and Zn have been detected in the range 50–80 μg/L, while Sb has been detected to a maximum of 20 μg/L (Strømseng, pers. comm.). High concentrations are mainly found in shooting ranges established on marshy ground (peat) where the corrosion rate of the projectiles are high due to acidic conditions and the mobility of the metals is high due to chelating humic ligands. Thus, runoff enriched in Science of the Total Environment 407 (2009) 4047–4055 ⁎ Corresponding author. P.O. Box 5003, Institute for Plant and Environmental Science (IPM), Norwegian University of Life Sciences, 1432 Aas, Norway. Tel.: +47 64965540; fax: +47 64966007. E-mail address: lene.sorlie.heier@umb.no (L.S. Heier). 0048-9697/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.scitotenv.2009.03.002 Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv