Organochlorine compounds in European catfish (Silurus glanis) living in river areas under the influence of a chlor-alkali plant (Ebro River basin).

European catfish, Silurus glanis, were used as sentinel organisms of the influence of recent and past discharges of organochlorine compounds (OCs) from a chlor-alkali plant located in the Ebro River. The fish concentrations of hexachlorobenzene (HCB), polychlorobiphenyls (PCBs) and DDTs were very high along the last 100 km of the river, including the irrigation channels, e.g. 1.2-27 ng/g wet weight of HCB, 6.3-100 ng/g ww of PCBs and 1-270 ng/g ww of total DDT compounds. These concentrations were much higher than those found upstream from the chlor-alkali discharge site, 0.2 ng/g ww for HCB, 5.6 ng/g ww for PCBs and 7.5 ng/g for DDT compounds. These concentrations were also standing out among those previously described in this fish species. The European catfish collected in sites under lower water flows, Ribarroja reservoir and irrigation channels, showed higher muscle lipid content, 1.09-7.2%, than those from sites of higher current intensities, river bed, 0.27%-0.67%. In these lower water current areas catfish exhibited OC ww concentrations that were correlated to % lipids. These differences suggest that normalization to lipid content is necessary for comparison of the OC accumulation in specimens from riverine systems living under different flow intensities. Accordingly, OC concentrations referred to lipid content showed more uniform downriver distribution which was consistent with a single focal point as main source of these compounds for the European catfish collected in the last 100 km of river stretch. This geographic distribution was also consistent with the uniform composition of PCB congeners in the studied European catfish. The distribution of DDT compounds was predominated by 4,4'-DDE which is common in most currently examined fish from aquatic environments. However, it included a high proportion of 4,4'-DDD and 2,4'-DDD which was consistent with the high contribution of benthic organisms from anoxic environments in the diet of these fish.

Methylmercury levels and bioaccumulation in the aquatic food web of a highly mercury-contaminated reservoir.

The low Ebro River basin (NE Spain) represents a particular case of chronic and long-term mercury pollution due to the presence of an industrial waste (up to 436 µg/g of Hg) coming from a chlor-alkali plant Albeit high total mercury (THg) levels have been previously described in several aquatic species from the surveyed area, methylmercury (MeHg) values in fish individuals have never been reported. Accordingly, in order to investigate bioaccumulation patterns at different levels of the aquatic food web of such polluted area, crayfish and various fish species, were analysed for THg and MeHg content. At the hot spot, THg mean values of crayfish muscle tissue and hepatopancreas were 10 and 15 times, respectively, greater than the local background level. Higher mean THg concentrations were detected in piscivorous (THg=0.848 ± 0.476 µg/g wet weight (ww); MeHg=0.672 ± 0.364 µg/g ww) than in non-piscivorous fish (THg=0.305 ± 0.163 µg/g ww; MeHg=0.278 ± 0.239 µg/g ww). Although these results indicated that THg in fish increased significantly with increasing trophic position, the percentage of the methylated form of Hg was not strongly influenced by differences in relative trophic position. This is an important finding, since the fraction of THg as MeHg in the top fish predator was unexpectedly lower than for other species of the aquatic food chain. Moreover, mean THg concentrations in piscivorous fish exceed the maximum level recommended for human consumption. From our findings, it is clear that for this specific polluted system, speciation becomes almost mandatory when risk assessment is based on MeHg, since single measurements of THg are inadequate and could lead to an over- or under-estimation of contamination levels.

Prevalence and sequence comparison of Phyllodistomum folium from zebra mussel and from freshwater fish in the Ebro River.

We utilised DNA analysis to detect the presence of the digenean Phyllodistomum folium in three cyprinid species, Scardinius erythrophthalmus, Cyprinus carpio and Rutilus rutilus. DNA sequencing of the region containing the genes ITS1-5.8S-ITS2 revealed 100% sequence identity between DNA from the sporocysts found in zebra mussels and DNA from adults located in the urinary system of 29 cyprinid fish. A second genetically different (variation=1.6%) sequence was observed in two samples from R. rutilus. In our opinion, the existence of a complex of species reported as P. folium is supported by recent genetic studies, including our own results. The overall prevalence of P. folium in mussels from the Ebro River was 4.67% in 2006, although during the summer months the rates frequently exceeded 10%.

Integrated biological and chemical analysis of organochlorine compound pollution and of its biological effects in a riverine system downstream the discharge point.

Pollution in riverine systems, along with its biological effects, may propagate downstream even at considerable distances. We analyzed the organochlorine compound (OC) pollution in a section of the low Ebro River (Northeast Spain) downstream a long-operating chlor-alkali plant. Maximal levels of OCs and of their associated dioxin-like biological activity occurred in residue samples from the plant, and persisted in river sediments some 40km downstream (Xerta site). Biological analysis at multiple organization levels in local carp (Cyprinus carpio, EROD, Cyp1A mRNA expression in the liver, hepatosomatic index, condition factor, and micronuclei index in peripheral blood) showed a similar pattern, with a maximal impact in Ascó, few kilometers downstream the plant, and a clear reduction at Xerta. This combination of chemical, molecular, cellular and physiological data allowed the precise assessment of the negative impact of the chlor-alkali plant on the quality of river sediments and on fish, and suggests that sediments may be a reservoir for toxic substances even in dynamic environments like rivers.

Physiological responses to mercury in feral carp populations inhabiting the low Ebro River (NE Spain), a historically contaminated site.

The low Ebro River course (Northeast Spain) is historically affected by mercury pollution due to a chlor-alkali plant operating at the town of Flix for more than a century. River sediments analysed during the last 10 years showed high mercury levels in the river section starting just downstream the factory and spanning some 90km, down to the river delta. The possible environmental impact was studied by a combination of field and laboratory studies. Mercury concentrations in liver, kidney and muscle of feral carp (Cyprinus carpio) sampled downstream Flix were one to two orders of magnitude higher than those from carps sampled upstream Flix. Elevated levels of mercury in these samples associated with significant increases on the concentration of reduced glutathione (GSH) in liver and on mRNA expression of two metallothionein genes, MT1 and MT2, in kidney and, partially, in scales, but not in liver. Conversely, no biochemical evidence for oxidative stress or DNA damage was found in these tissues. Non-contaminated carps subjected to intraperitoneal mercury injection resulted in a 20-fold increase of MT1 and MT2 mRNA levels in carp kidney, with minimal changes in liver levels. Our data suggests the coordinate increase of metallothionein mRNA in kidney and of GSH in liver constitutes an excellent marker of exposure to sub-toxic mercury levels in carps. This study also demonstrates that apparently healthy fish populations may exceed the mercury contamination acceptable for human consumption.