This comprehensive field survey on indigenous European chub (Squalius cephalus L.) presents, for the first time, site-specific variability of trace metal concentrations in the gut content, gastrointestinal tissue and two gastrointestinal sub-cellular fractions, operationally defined as metal-sensitive fraction (S50, which was isolated at 50,000 × g and contains total water soluble proteins), and metal detoxified fraction (heat-treated S50 (HT S50), which contains heat-stable proteins like metallothioneins). At five sampling sites along the Sava River in Croatia 1 to 5-year-old chub were collected in the post-spawning period (September) in order to estimate if metal concentrations in fish intestine are related to their levels in the gut content or fish age. Concentrations of essential metals (Zn, Fe, Cu, Mn) and non-essential Cd decrease in the gut content as follows: Fe > Mn > Zn > Cu > Cd, while in the gastrointestinal tissue: Zn > Fe > Cu ≥ Mn > Cd. Observed difference in metal abundance between the gut content and gastrointestinal tissue points to the selective metal absorption in fish intestine. Relationship among metal concentrations in the gastrointestinal tissue and two sub-cellular fractions (S50/HT S50) is significant for all analysed metals, with Spearman correlation coefficients (r) at p < 0.01 for Zn 0.84/0.73, Cu 0.73/0.73, Fe 0.62/0.58, Mn 0.81/0.78, Cd 0.81/0.82. Site-specific differences point to the age-related increase of gastrointestinal Cu, Mn and Cd towards the downstream sites, while significant correlation between metal concentrations in the gut content and fish age exists only for Mn. In the sub-cellular gastrointestinal fractions, site-specific differences were not recorded on total water-soluble protein and metallothionein concentrations, which might be ascribed to the constitutional level.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Andres, S., Ribeyre, F., Tourencq, J. N., & Boudou, A. (2000). Interspecific comparison of cadmium and zinc contamination in the organs of four fish species along a polymetallic pollution gradient (Lot River, France). Science of the Total Environment, 248(1), 11–25.
Berntssen, M. H. G., Hylland, K., Bonga, S. E. W., & Maage, A. (1999). Toxic levels of dietary copper in Atlantic salmon (Salmo salar L.) parr. Aquatic Toxicology, 46(2), 87–99.
Berntssen, M. H. G., Aspholm, O. O., Hylland, K., Bonga, S. E. W., & Lundebye, A.-K. (2001). Tissue metallothionein, apoptosis and cell proliferation responses in Atlantic salmon (Salmo salar L.) parr fed elevated dietary cadmium. Comparative Biochemistry and Physiology. C, 128(3), 299–310.
Bury, N. R., Walker, P. A., & Glover, C. N. (2003). Nutritive metal uptake in teleost fish. Journal of Experimental Biology, 206, 11–23.
Campbell, P. G. C., Clearwater, S. J., Brown, P. B., Fisher, N. S., Hogstrand, C., Lopez, G. R., Mayer, L. M., & Meyer, J. S. (2005). Digestive physiology, chemistry and nutrition. In J. S. Meyer, W. J. Adams, K. V. Brix, S. N. Luoma, D. R. Mount, W. A. Stubblefield, & C. M. Wood (Eds.), Toxicity of dietborne metals to aquatic organisms (pp. 13–57). Brussels: Society of environmental toxicology and chemistry (SETAC).
Chowdhury, M. J., Baldisserotto, B., & Wood, C. M. (2005). Tissue-specific cadmium and metallothionein levels in rainbow trout chronically acclimated to waterborne or dietary cadmium. Archives of Environmental Contamination and Toxicology, 48(3), 381–390.
Clearwater, S. J., Baskin, S. J., Wood, C. M., & McDonald, D. G. (2000). Gastrointestinal uptake and distribution of copper in rainbow trout. Journal of Experimental Biology, 203(16), 2455–2466.
Creighton, N., & Twining, J. (2010). Bioaccumulation from food and water of cadmium, selenium and zinc in an estuarine fish, Ambassis jacksoniensis. Marine Pollution Bulletin, 60, 1815–1821.
Dallinger, R., & Kautzky, H. (1985). The importance of contaminated food for the uptake of heavy metals by rainbow trout (Salmo gairdneri): a field study. Oecologia, 67, 82–89.
Dragun, Z., & Raspor, B. (2005). Direct determination of Cd in NaCl containing metallothionein fractions of different red mullet tissues by graphite furnace atomic absorption spectrometry. Journal of Analytical Atomic Spectrometry, 20(10), 1121–1123.
Dragun, Z., & Raspor, B. (2008). Copper determination by ETAAS in fish tissue cytosols with minimal sample pre-treatment. Atomic Spectroscopy, 29(3), 107–113.
Dragun, Z., Raspor, B., & Podrug, M. (2007). The influence of the season and the biotic factors on the cytosolic metal concentrations in the gills of the European chub (Leuciscus cephalus L.). Chemosphere, 69(6), 911–919.
Dragun, Z., Raspor, B., & Roje, V. (2008). The labile metal concentrations in Sava River water assessed by diffusive gradients in thin films. Chemical Speciation and Bioavailability, 20, 33–46.
Dragun, Z., Podrug, M., & Raspor, B. (2009). The assessment of natural causes of metallothionein variability in the gills of European chub (Squalius cephalus L.). Comparative Biochemistry and Physiology. C, 150, 209–217.
Dragun, Z., Roje, V., Mikac, N., & Raspor, B. (2009). Preliminary assessment of total dissolved trace metal concentrations in Sava River water. Environmental Monitoring and Assessment, 159(1), 99–110.
Erk, M., Ivanković, D., Raspor, B., & Pavičić, J. (2002). Evaluation of different purification procedures for the electrochemical quantification of mussel metallothioneins. Talanta, 57, 1211–1218.
European Parliament and the Council of the European Union (EPC EU). (2008). Directive 2008/105/EC of the European Parliament and of the Council on environmental quality standards in the field of water policy, amending and subsequently repealing Council Directives 82/176/EEC, 83/513/EEC, 84/156/EEC, 84/491/EEC, 86/280/EEC, and amending Directive 2000/60/EC of the European Parliament and of the Council. Official Journal of the European Union, 348, 84–97.
Farkas, A., Salánki, J., & Specziár, A. (2003). Age- and size-specific patterns of heavy metals in the organs of freshwater fish Abramis brama L. populating a low-contaminated site. Water Research, 37, 959–964.
Filipović Marijić, V., & Raspor, B. (2006). Age and tissue dependent metallothionein and cytosolic metal distribution in a native Mediterranean fish, Mullus barbatus, from the Eastern Adriatic Sea. Comparative Biochemistry and Physiology. C, 143, 382–387.
Filipović Marijić, V., & Raspor, B. (2007a). Metal exposure assessment in native fish, Mullus barbatus L., from the Eastern Adriatic Sea. Toxicology Letters, 168(3), 292–301.
Filipović Marijić, V., & Raspor, B. (2007b). Metallothinein in intestine of red mullet, Mullus barbatus as a biomarker of copper exposure in the coastal marine areas. Marine Pollution Bulletin, 54(7), 935–940.
Filipović Marijić, V., & Raspor, B. (2010). The impact of fish spawning on metal and protein levels in gastrointestinal cytosol of indigenous European chub. Comparative Biochemistry and Physiology. C, 152, 133–138.
Giguère, A., Campbell, P. G. C., Hare, L., McDonald, D. G., & Rasmussen, J. B. (2004). Influence of lake chemistry and fish age on cadmium, copper, and zinc concentrations in various organs of indigenous yellow perch (Perca flavescens). Canadian Journal of Fisheries and Aquatic Sciences, 61(9), 1702–1716.
Habeković, D., Aničić, I., & Safner, R. (1993). Growth dynamics of the chub fish in the River Sava [Dinamika rasta klena u rijeci Savi, in Croatian]. Ribarstvo, 48(3), 79–88.
Handy, R. D., Sims, D. W., Giles, A., Campbell, H. A., & Musonda, M. M. (1999). Metabolic trade-off between locomotion and detoxification for maintenance of blood chemistry and growth parameters by rainbow trout (Oncorhynchus mykiss) during chronic dietary exposure to copper. Aquatic Toxicology, 47, 23–41.
HRN EN 14011: (2005). Fish sampling by electric power [Uzorkovanje riba električnom strujom, in Croatian].
Kamunde, C., Clayton, C., & Wood, C. M. (2002). Waterborne vs. dietary copper uptake in rainbow trout and the effects of previous waterborne copper exposure. American Journal of Physiology - Regulatory, Integrative and Comparative Physiology, 283(1), R69–R78.
Krča, S., Žaja, R., Čalić, V., Terzić, S., Grubešić, M. S., Ahel, M., & Smital, T. (2007). Hepatic biomarker responses to organic contaminants in feral chub (Leuciscus cephalus)—laboratory characterization and field study in the Sava River, Croatia. Environmental Toxicology and Chemistry, 26(12), 2620–33.
Langston, W. J., Chesman, B. S., Burt, G. R., Pope, N. D., & McEvoy, J. (2002). Metallothionein in liver of eels Anguilla anguilla from the Thames Estuary: an indicator of environmental quality? Marine Environmental Research, 53, 263–293.
Linde, A. R., Sánchez-Galán, S., Klein, D., García-Vázquez, E., & Summer, K. H. (1999). Metallothionein and heavy metals in brown trout (Salmo trutta) and European eel (Anguilla anguilla): a comparative study. Ecotoxicology and Environmental Safety, 44, 168–173.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin Phenol reagent. Journal of Biological Chemistry, 193, 265–275.
Mason, A. Z., & Jenkins, K. D. (1995). Metal detoxification in aquatic organisms. In A. Tessier & D. R. Turner (Eds.), Metal Speciation and Bioavailability in Aquatic Systems (pp. 479–608). Chichester: John Wiley and Sons.
Miramand, P., Lafaurie, M., Fowler, S. W., Lemaire, P., Guary, J. C., & Bentley, D. (1991). Reproductive cycle and heavy metals in the organs of red mullet, Mullus barbatus (L), from the northwestern Mediterranean. Science of the Total Environment, 103, 47–56.
Ojo, A. A., & Wood, C. M. (2007). In vitro analysis of the bioavailability of six metals via the gastro-intestinal tract of the rainbow trout (Oncorhynchus mykiss). Aquatic Toxicology, 83, 10–23.
Oliveira, M., Ahmad, I., Maria, V. L., Serafim, A., Bebianno, M. J., Pacheco, M., & Santos, M. A. (2010). Hepatic metallothionein concentrations in the golden grey mullet (Liza aurata)—relationship with environmental metal concentrations in a metal-contaminated coastal system in Portugal. Marine Environmental Research, 69, 227–233.
Olsvik, P. A., Gundersen, P., Andersen, R. A., & Zachariassen, K. E. (2001). Metal accumulation and metallothionein in brown trout, Salmo trutta, from two Norwegian rivers differently contaminated with Cd, Cu and Zn. Comparative Biochemistry and Physiology. C, 128(2), 189–201.
Podrug, M., & Raspor, B. (2009). Seasonal variation of the metal (Zn, Fe, Mn) and metallothionein concentrations in the liver cytosol of the European chub (Squalius cephalus L.). Environmental Monitoring and Assessment, 157, 1–10.
Podrug, M., Raspor, B., Erk, M., & Dragun, Z. (2009). Protein and metal concentrations in two fractions of hepatic cytosol of the European chub (Squalius cephalus L.). Chemosphere, 75, 843–849.
Rainbow, P. S., Luoma, S. N., & Wang, W.-X. (2011). Trophically available metal—a variable feast. Environmental Pollution, 159(10), 2347–2349.
Rashed, M. N. (2001a). Cadmium and lead levels in fish (Tilapia nilotica) tissues as biological indicator for lake water pollution. Environmental Monitoring and Assessment, 68(1), 78–89.
Rashed, M. N. (2001b). Monitoring of environmental heavy metals in fish from Nasser Lake. Environment International, 27, 27–33.
Raspor, B., Paić, M., & Erk, M. (2001). Analysis of metallothioneins by the modified Brdička procedure. Talanta, 55, 109–115.
Raspor, B., Dragun, Z., & Erk, M. (2005). Examining the suitability of mussel digestive gland to serve as a biomonitoring target organ. Archives of Industrial Hygiene and Toxicology, 56(2), 141–149.
Rotchell, J. M., Clarke, K. R., Newton, L. C., & Bird, D. J. (2001). Hepatic metallothionein as a biomarker for metal contamination: age effects and seasonal variation in European flounders (Pleuronectes flesus) from the Severn Estuary and Bristol Channel. Marine Environmental Research, 52, 151–171.
Saha, M., Sarkar, S. K., & Bhattacharya, B. (2006). Interspecific variation in heavy metal body concentrations in biota of Sunderban mangrove wetland, northeast India. Environment International, 32(2), 203–207.
Schlekat, C. E., Kidd, K. A., Adams, W. J., Baird, D. J., Farag, A. M., Maltby, L., & Stewart, A. R. (2005). Toxic effects of dietborne metals: field studies. In J. S. Meyer, W. J. Adams, K. V. Brix, S. N. Luoma, D. R. Mount, W. A. Stubblefield, & C. M. Wood (Eds.), Toxicity of dietborne metals to aquatic organisms (pp. 113–152). Brussels: Society of environmental toxicology and chemistry (SETAC).
Singha Roy, U., Chattopadhyay, B., Datta, S., & Kumar Mukhopadhyay, S. (2011). Metallothionein as a biomarker to assess the effects of pollution on Indian Major carp species from wastewater-fed fishponds of East Calcutta wetlands (a Ramsar Site). Environmental Research, Engineering and Management, 4(58), 10–17.
Standard Operation Procedure (1999). Preparation of S50-fraction from fish tissue (unapproved rev. 01). 1st Workshop in the frame of BEQUALM programme, 13-14 September 1999, NIVA, Oslo.
Staniskiene, B., Matusevicius, P., Budreckiene, R., & Skibniewska, K. A. (2006). Distribution of heavy metals in tissues of freshwater fish in Lithuania. Polish Journal of Environmental Studies, 15(4), 585–591.
Sun, L.-T., & Jeng, S.-S. (1998). Comparative zinc concentrations in tissues of common carp and other aquatic organisms. Zoological Studies, 37, 184–190.
Ünlü, E., Akba, O., Sevim, S., & Gümgüm, B. (1996). Heavy metal levels in Mullet, Liza abu (Heckel, 1843) (Mugilidae) from the Tigris River, Turkey. Fresenius Environmental Bulletin, 5, 107–112.
Van Campenhout, K., Bervoets, L., & Blust, R. (2003). Metallothionein concentrations in natural populations of gudgeon (Gobio gobio): relationship with metal concentrations in tissues and environment. Environmental Toxicology and Chemistry, 22(7), 1548–1555.
Watanabe, T., Kiron, V., & Satoh, S. (1997). Trace minerals in fish nutrition. Aquaculture, 151, 185–207.
Financial support by the Ministry of Science, Education and Sport of Republic Croatia (project no. 098-0982934-2721) is acknowledged. The authors are also grateful for the valuable help in the fieldwork to all EU FP6 SARIB Project participants and for the useful advice concerning metal determination on HR ICP-MS to Dr. Nevenka Mikac, Dr. Vibor Roje and Željka Fiket, B.Sc. and on AAS to Dr. Zrinka Dragun. We appreciate the assistance of the Meteorological and Hydrological Service Office for reporting the data on hydrological parameters of the Sava River water.
About this article
Cite this article
Filipović Marijić, V., Raspor, B. Site-Specific Gastrointestinal Metal Variability in Relation to the Gut Content and Fish Age of Indigenous European Chub from the Sava River. Water Air Soil Pollut 223, 4769–4783 (2012). https://doi.org/10.1007/s11270-012-1233-2
- Essential metals
- Total proteins
- Gastrointestinal tissue/sub-cellular fractions
- Freshwater fish