The last 50 years of organic contamination of a highly anthropized tributary of the Po River (Italy).

We examined the temporal profiles of many organic micropollutants analysed in a sediment core sampled from a highly anthropized tributary of the Po River, the Lambro River. Analysed for extractable organic halogens (EOX), total petroleum hydrocarbons (C10-C40TPH), polycyclic aromatic hydrocarbons (PAHs), common legacy pollutants (DDTs, PCBs), halogenated flame retardants (PBDEs, DBDPE, TBBPA-bis, TCBPA, TBBPA, HBCDs), organotins (TBT, TPhT), antimicrobials (TCS, TCC), fragrances (AHTN, HHCB) and phthalates (DMP, DEP, DnBP, BBP, DEHP, DnOP), the dated sediment core revealed the historical record of 50 years of chemical contamination discharged into the Lambro and thereby the Po River. In this regard, the peak levels of PCBs and DDTs found in Lambro sediments were also identified in other sediment cores collected from the Po River prodelta in the Adriatic Sea, thus hundreds of kilometres downstream (Combi et al., 2020). The highest risk to aquatic organisms was associated with decades of high levels of C10-C40 TPH, PBDEs, PCBs, PAHs, DDTs, EOX, TCC, AHTN and DEHP, which in different periods of the contamination history, showed exceedances of guideline/threshold values. C10-C40 TPH and TCC, for example, were very high in the 1960s, whereas PCBs, DDTs, and PBDEs, peaked from the 1980s onward. The corresponding sums of PEC quotients ranged between 0.48 and 28.63, with a mean value (±SD) for the entire recording period of 10.62 ± 9.83. Environmental legislations and improved wastewater treatments were the main drivers of the recent downward trends observed for most of the chemicals investigated. Floods in turn resulted in macroscopic yet temporary improvements in the chemical quality of the tributary, conveying contaminated sediments into the Po River.

Vertical profiles of trace elements in a sediment core from the Lambro River (northern Italy): Historical trends and pollutant transport to the Adriatic Sea.

River sediments generally act as a sink for trace elements but, when resuspended, they contribute to long-term downstream transport of contamination, which may finally reach the marine environment. This study analyzed these processes in a complex aquatic system that includes a contaminated tributary, the Lambro River (Northern Italy) and its recipient and main Italian watercourse, the Po River, with the prodelta in the Adriatic Sea. The study was conducted from a historical perspective which, covering the last 50 years, examined the main driving events such as the inputs of contaminants, the construction of WWTPs and the evolution of environmental legislation. The time trend of trace element contamination was analyzed in a sediment core collected in the Lambro River and dated 1962-2011. The highest enrichments were found for Hg, Zn, Cu, Pb and Cd, which showed similar trends, with EF maxima in the '60s-'90s (172, 56, 40, 28 and 21, respectively), following industrial and urban development, and a general decreasing pattern after the late '90s. Only in the 2000s the ecological risk associated with metal contamination showed mean PEC Quotients stably below 1. The results of a literature survey on sedimentary trace elements in the Po River and the prodelta for the last 50 years were then compared to the Lambro sediment core. A significant contribution to Cu, Zn, Pb, Hg and Cd contamination was proved to derive from Lambro sediment transport. In the prodelta, increasing Ni and Cr concentrations were also evidenced, likely as a result of enhanced soil erosion in the Po basin. This study highlights the key role of WWTPs, of lower-impact industrial processes and of environmental legislation in reducing contaminant inputs. It also emphasizes the active contribution of riverine sediment-bound contamination to long-distance marine sediment quality.

Embryo/larval toxicity and transcriptional effects in zebrafish (Danio rerio) exposed to endocrine active riverbed sediments.

Sediment toxicity plays a fundamental role in the health of inland fish communities; however, the assessment of the hazard potential of contaminated sediments is not a common objective in environmental diagnostics or remediation. This study examined the potential of transcriptional endpoints investigated in zebrafish (Danio rerio) exposed to riverbed sediments in ecotoxicity testing. Embryo-larval 10-day tests were conducted on sediment samples collected from five sites (one upstream and four downstream of the city of Milan) along a polluted tributary of the Po River, the Lambro River. Sediment chemistry showed a progressive downstream deterioration in river quality, so that the final sampling site showed up to eight times higher concentrations of, for example, triclosan, galaxolide, PAH, PCB, BPA, Ni, and Pb, compared with the uppermost site. The embryo/larval tests showed widespread toxicity although the middle river sections evidenced worse effects, as evidenced by delayed embryo development, hatching rate, larval survival, and growth. At the mRNA transcript level, the genes encoding biotransformation enzymes (cyp1a, gst, ugt) showed increasing upregulations after exposure to sediment from further downstream sites. The genes involved in antioxidant responses (sod, gpx) suggested that more critical conditions may be present at downstream sites, but even upstream of Milan there seemed to be some level of oxidative stress. Indirect evidences of potential apoptotic activity (bcl2/bax < 1) in turn suggested the possibility of genotoxic effects. The genes encoding for estrogen receptors (erα, erß1, erß2) showed exposure to (xeno)estrogens with a progressive increase after exposure to sediments from downstream sites, paralleled by a corresponding downregulation of the ar gene, likely related to antiandrogenic compounds. Multiple levels of thyroid disruption were also evident particularly in downstream zebrafish, as for thyroid growth (nkx2.1), hormone synthesis and transport (tg, ttr, d2), and signal transduction (trα, trß). The inhibition of the igf2 gene reasonably reflected larval growth inhibitions. Although none of the sediment chemicals could singly explain fish responses, principal component analysis suggested a good correlation between gene transcripts and the overall trend of contamination. Thus, the combined impacts from known and unknown covarying chemicals were proposed as the most probable explanation of fish responses. In summary, transcriptional endpoints applied to zebrafish embryo/larval test can provide sensitive, comprehensive, and timeliness information which may greatly enable the assessment of the hazard potential of sediments to fish, complementing morphological endpoints and being potentially predictive of longer studies.

Phosphorus content in a deep river sediment core as a tracer of long-term (1962-2011) anthropogenic impacts: A lesson from the Milan metropolitan area.

Reconstructions of past fluvial contamination through the analysis of deep sediment cores are rarely reported in literature. We examined the phosphorus fractions in a deep (2.6 m) sediment core of the Lambro River downstream of the highly anthropized Milan metropolitan area and upstream of the Po river the main Italian watercourse. The core covered the period 1962-2011. Total phosphorus concentrations resulted typical of a strongly impacted environment (4788 mg P kg DW-1 on average) with the highest concentrations related to the 1960s (7639 mg P kg DW-1) reflecting the period of maximum demographic growth. Afterwards, phosphorus concentrations decreased thanks to the infrastructural and legislative initiatives carried out in the 1980s and the 1990s to reduce the impact of urban point sources. Subsequently, total phosphorus concentrations stabilized on values around 3000 mg P kg DW-1 and did not diminish further, even after the second phase of infrastructural interventions carried out in the second half of the 2000s. This was related to the increasing relative impact of the combined sewer overflows in the sewage system and to the strong phosphorus enrichment of the basin. Most of the phosphorus was in inorganic forms (86% of the total) that have been identified as the final target of the domestic effluent inputs. The contribution of organic phosphorus was lower but constant over the period 1962-2011. It likely originated from the agricultural areas located south of the city of Milan. In conclusion, this study underlines how past interventions have been effective in reducing urban point sources but it also highlights the current difficulties related to the growing importance of other sources influenced by the surface runoff (i.e., combined sewer overflows and agriculture). The study also emphasizes a general phosphorus enrichment of the Lambro River basin and its impact on the Po River and the Adriatic Sea.

Gene expression and genotoxicity in Manila clam (Ruditapes philippinarum) modulated by sediment contamination and lagoon dynamics in the Po river delta.

The lagoons of the Po River delta are potentially exposed to complex mixtures of contaminants, nevertheless, there is a substantial lack of information about the biological effects of these contaminants in the Po delta lagoons. These environments are highly dynamic and the interactions between chemical and environmental stressors could prevent the proper identification of biological effects and their causes. In this study, we aimed to disentangle such interactions focusing on Manila clams, previously exposed to six lagoons of the Po delta, adopting three complementary tools: a) the detailed description via modelling techniques of lagoon dynamics for salinity and water temperature; b) the response sensitivity of a number of target genes (ahr, cyp4, ρ-gst, σ-gst, hsp22, hsp70, hsp90, ikb, dbh, ach, cat, Mn-sod, Cu/Zn-sod, cyp-a, flp, grx, TrxP) investigated in clam digestive glands by Real Time PCR; and c) the relevance of DNA adducts determined in clams as markers of exposure to genotoxic chemicals. The lagoons showed specific dynamics, and two of them (Marinetta and Canarin) could induce osmotic stress. A group of genes (ahr, cyp4, Mn-sod, σ-gst, hsp-22, cyp-a, TrxP) seemed to be associated with overall lagoon characteristics as may be described by salinity and its variations. Lagoon modelling and a second group of genes (hsp70, hsp90, cat, ikb, ach, grx, Cu/Zn-sod) also suggested that moderate increases of river discharge may imply worse exposure conditions. Oxidative stress seemed to be associated with such events but it was slightly evident also under normal exposure conditions. DNA adduct formation was mainly associated with overwhelmed antioxidant defences (e.g. low Cu/Zn-sod) or seemingly with their lack of response in due time. In Po delta lagoons, Manila clam can be affected by chemical and environmental factors which can contribute to induce oxidative stress, DNA adduct formation and, ultimately, to affect clam condition and health.

Hepatic gene expression profiles of a non-model cyprinid (Barbus plebejus) chronically exposed to river sediments.

In this study, we characterized the gene expression responses of the Padanian barbel (Barbus plebejus), a native benthivorous cyprinid with a very compromised presence within the fish community of the River Po. Barbel juveniles were exposed in the laboratory to two river sediments reflecting an upstream/downstream gradient of increasing contamination and collected from one of the most anthropized tributaries of the River Po. After 7months of exposure, hepatic transcriptional changes that were diagnostic of sediment exposure were assessed. We investigated a set of 24 genes involved in xenobiotic biotransformation (cyp1a, gstα, ugt), antioxidant defense (gpx, sod, cat, hsp70), trace metal exposure (mt-I, mt-II), DNA repair (xpa, xpc), apoptosis (bax, casp3), growth (igf2), and steroid (erα, erß1, erß2, ar, vtg) and thyroid (dio1, dio2, trα, trß, nis) hormone signaling pathways. In a consistent overall picture, the results showed that long-term sediment exposure mainly increased the levels of mRNAs encoding proteins involved in xenobiotic metabolism, oxidative stress defense, repair of DNA damage and activation of the apoptotic process. Transcript up-regulation of three receptor genes (erß2, ar, trß), likely representing compensatory responses to antagonistic/toxic effects, was also observed, confirming the exposure to disruptors of the reproductive and thyroidal axes. In contrast to expectations, a few genes showed no response (e.g., casp3) or even downregulation (vtg), further suggesting that the timing of exposure/assessment, potential compensatory effects or post-transcriptional modifications interact to modify the gene expression profiles, particularly during exposure to mixtures of contaminants.

Medium- and Long-Term Effects of Estrogenic Contaminants on the Middle River Po Fish Community as Reconstructed from a Sediment Core.

Recent studies showed that endocrine active compounds (EDs) capable to induce fish gonadal histopathologies, plasma vitellogenin and thyroid disruption, are transported by the River Lambro to the River Po, potentially affecting the fish community of the main Italian river. To assess whether fish relative abundance, composition and health were impaired by the River Lambro, a 3-year survey was undertaken in the main river. Results showed that the tributary supports in the River Po a denser fish community (+43 %), with a higher total biomass (+35 %). The survey also showed niche- and sensitivity-dependent effects, so that three benthopelagic species (bleak, topmouth gudgeon, and bitterling) were, for example, more abundant downstream from the tributary (up to 3.4×), but their sizes were significantly smaller. The present fish community was then compared with that described 30 years before in the same area of the Po River. This comparison highlighted that some fish species have disappeared and many have severely declined. To better evaluate this contrast, a sediment core of the Lambro tributary was analysed for the time trends of natural estrogens (E1, E2, E3), bisphenol A and alkylphenols. The results showed that during the last 50 years the River Lambro has been exposed to high estrogenic activities (16.1 ± 9.3 ng E2 equivalents/g), which inevitably affected also the River Po. In addition, at the time of the previous survey, six species of the main river had skewed sex ratios toward all-female populations, providing evidence that EDs and particularly (xeno)estrogens were already affecting the long-term viability of fish populations. Estrogens thus can be ascribed among the causal factors of fish qualitative and quantitative decline of the River Po, although long-term effects have been likely mitigated by nonconfinement of fish populations and nutrient enrichment.

Endocrine-disrupting chemicals in coastal lagoons of the Po River delta: sediment contamination, bioaccumulation and effects on Manila clams.

The large estuary that the River Po forms at its confluence into the Adriatic Sea comprises a multitude of transitional environments, including coastal lagoons. This complex system receives the nutrients transported by the River Po but also its load of chemical contaminants, which may pose a substantial (eco)toxicological risk. Despite the high ecological and economic importance of these vulnerable environments, there is a substantial lack of information on this risk. In light of the recent amendments of the European Water Framework Directive (2013/39/EU), the present study investigated the sediment contamination of six coastal lagoons of the Po delta and its effects on Manila clams (Ruditapes philippinarum), exposed in situ for 3 months. Sediment contamination and clam bioaccumulation of a wide range of chemicals, i.e. trace metals (Cd, Cr, Ni, Hg, Pb, As), polybrominated diphenyl ethers (PBDEs), alkylphenols (APs), organochlorine compounds (PCBs, DDTs), polycyclic aromatic hydrocarbons (PAHs) and organotins (TPhT, TBT), suggested a southward increase related to the riverine transports. Where the River Po influence was more direct, the concentrations of contaminants were higher, with nonylphenol and BDE-209 exceeding sediment quality guidelines. Biometric indicators suggested the influence of contamination on organism health; an inverse relationship between PBDEs in sediments and clam condition index has been found, as well as different biota-sediment accumulation factors (BSAFs) in the lagoons.

Exposing native cyprinid (Barbus plebejus) juveniles to river sediments leads to gonadal alterations, genotoxic effects and thyroid disruption.

Juveniles (50 days post hatch) of a native cyprinid fish (Barbus plebejus) were exposed for 7 months to sediments from the River Lambro, a polluted tributary impairing the quality of the River Po for tens of kilometers from their confluence. Sediments were collected upstream of the city of Milan and downstream at the closure of the drainage basin of the River Lambro. Chemical analyses revealed the presence of a complex mixture of bioavailable endocrine-active chemicals, with higher exposure levels in the downstream section of the tributary. Mainly characterized by brominated flame retardants, alkylphenols, polychlorinated biphenyls, and minor co-occurring personal care products and natural hormones, the sediment contamination induced reproductive disorders, as well as other forms of endocrine disruption and toxicity. In particular, exposed male barbel exhibited higher biliary PAH-like metabolites, overexpression of the cyp1a gene, vitellogenin production in all specimens, the presence of oocytes (up to 22% intersex), degenerative alterations in their testis, liver fat vacuolization, a marked depression of total thyroxine (T4) and triiodothyronine (T3) plasma levels, and genotoxic damages determined as hepatic DNA adducts. These results clearly demonstrate that Lambro sediments alone are responsible for recognizable changes in the structure and function of the reproductive and, in general, the endocrine system of a native fish species. In the real environment, exposure to waterborne and food-web sources of chemicals are responsible for additional toxic loads, and the present findings thus provide evidence for a causal role of this tributary in the severe decline observed in barbel in recent decades and raise concern that the fish community of the River Po is exposed to endocrine-mediated health effects along tens of kilometres of its course.

Tracing endocrine disrupting chemicals in a coastal lagoon (Sacca di Goro, Italy): sediment contamination and bioaccumulation in Manila clams.

The Water Framework Directive, recently amended with new priority substances (2013/39/EU), is meant to regulate the health status of European aquatic environments, including transitional waters. Despite the ecological and economic importance of transitional water bodies and, in particular, of coastal lagoons, a relevant example of this type of environments, little is known about their contamination by priority substances, particularly by endocrine disrupting chemicals (EDCs). In this study, a wide array of priority substances, all with recognised disrupting properties, was investigated in the Sacca di Goro Lagoon (Adriatic Sea, Italy), which receives freshwater from the Po River after draining the most urbanised and industrialised Italian regions. Flame retardants, alkylphenols, bisphenol A, natural and synthetic steroids, personal care products and legacy pollutants were investigated both in sediments and in the clam Ruditapes philippinarum collected from three sites in the lagoon. Sediments showed that most of the chemicals analysed could reach the lagoon ecosystem but their concentrations were below existing quality guidelines. Clams essentially reflected this condition although some concern was raised by polybrominated diphenyl ethers (PBDEs): the limit for the sum of six congeners set for biota in the European Directive (2013/39/EU) to protect human health was exceeded 4-5 times. No significant biota-sediment accumulation factors (BSAFs) were calculated. Nonylphenol, tonalide, PBDE, polychlorinated biphenyls and bisphenol A were the most abundant chemicals in clam tissues.

Decabromodiphenyl ether (BDE-209) enters the food web of the River Po and is metabolically debrominated in resident cyprinid fishes.

Decabromodiphenyl ether (BDE-209), the primary constituent of a widely used flame retardant formulation, is present at relatively high levels in sediments and macroinvertebrates of the River Po. Since it was demonstrated that BDE-209 can be biotransformed to smaller and more toxic polybrominated dipheyl ethers (PBDEs), the main objective of this study was to assess whether the large quantities of BDE-209 present in the River Po are bioavailable to the higher levels of the food web and are biotransformed in feral fishes. To this aim, 23 cyprinids, mainly common carp, were analysed for the hepatic contents of PBDEs. Contrary to sediments and invertebrates of the same area, no fish sample contained detectable levels of BDE-209. All fishes contained typical PBDE representatives, e.g. BDE-47, BDE-99, BDE-100, BDE-153 and BDE-154, but more importantly they contained three congeners, i.e. BDE-179, BDE-188 and BDE-202, which are not present in any technical formulations and are known products of BDE-209 debromination in fish. The age of carps had no effects on the bioaccumulation of PBDEs. Conversely, the contents of PCBs, which also were determined in the same fish samples, showed a positive correlation with age. Both groups of chemicals displayed a tendency to a higher contamination in male fish. This study shows that BDE-209 enters the food web of the River Po contributing to the load of lower brominated PBDEs and thus to the load of chemical stressors threatening the aquatic life of the major Italian watercourse.

Endocrine modulation, inhibition of ovarian development and hepatic alterations in rainbow trout exposed to polluted river water.

Under laboratory conditions, female rainbow trout were exposed to graded concentrations of water from the River Lambro, a polluted tributary of the River Po, and to the effluent of a large wastewater treatment plant which flows into the River Lambro. In field exposures, trout were held in cages in the River Po upstream and downstream from the confluence of the River Lambro. After 10-day (laboratory) and 30-day (laboratory and field) exposures, trout were examined for several chemical, biochemical and histological endpoints. The results indicated that exposure to complex mixtures of chemicals, including estrogen receptor agonists, aryl-hydrocarbon receptor agonists, and probably antiandrogens, had occurred. Exposure altered the plasma levels of 17ß-estradiol and testosterone, and some treatments also enhanced the activity of hepatic ethoxyresorufin O-deethylase. Gonadal histology showed varying levels of degenerative processes characterised by oocyte atresia, haemorrhages, melano-macrophage centres (MMCs), and oogonia proliferation. Liver histology showed less severe effects.

Polybrominated diphenyl ethers (PBDEs) in gammarids, caddisflies, and bed sediments of the lowland River Po.

Polybrominated diphenyl ethers (PBDEs) were investigated in sediments and invertebrates (gammarids and caddisflies) collected in the River Po, upstream and downstream from a polluted tributary. Besides a diffuse contamination by penta-BDE technical mixture, the river sediments identified the tributary as an important source to the main river of decabromodiphenyl ether (BDE-209), which peaked to 64 microg/g (OC) in the downstream stretch. At 10 km downstream from the tributary, a higher bioavailability was evident than at 22 km, and small gammarids accumulated at two/three times the levels of PBDEs found in large gammarids. The congener profiles of sediments and invertebrates were dominated by BDE-47, BDE-99 and BDE-209.

Estrogenicity profile and estrogenic compounds determined in river sediments by chemical analysis, ELISA and yeast assays.

An effects-directed strategy was applied to bed sediments of a polluted tributary in order to isolate and identify the major estrogenic chemicals it discharges into the River Po, the principal Italian watercourse. Sediment extract was concentrated by solid phase extraction and then fractioned into 10 fractions by reversed phase high performance liquid chromatography (RP-HPLC). Estrogenic activity of whole extract and fractions were determined using a recombinant yeast assay containing the human estrogen receptor (YES). The 10 fractions and whole extract were analysed for target compounds, e.g. estrone (E1), 17beta-estradiol (E2), estriol (E3), 4-nonylphenol (NP), 4-tert-octylphenol (t-OP), bisphenol A (BPA), using both liquid chromatography-tandem mass spectrometry (LC-MS/MS) and non-competitive enzyme-linked immunosorbent assays (ELISA). The YES assay determined high estrogenic activity in whole sediment (15.6 ng/g EE2 equivalents), and positive results for fractions nr 1, 2, 6, 7 and 8. E1, E3 and NP were the main estrogenic chemicals, however, other unidentified compounds contributed to sediment estrogenicity, particularly for polar fractions nr 1 and 2. A GC-MS screening performed in scan mode identified other potential contributors such as phthalates (DBP, BBP), and OP isomers. A next sampling campaign extended to other tributaries and receiving stretches of the River Po confirmed E1, E3 and NP as major estrogenic chemicals potentially threatening other sites of the main river. In general, target compound ELISAs have been shown to be suitable tools for a rapid screening of wide areas or large numbers of environmental samples for estrogenic risk. The potential for interferences suggests however to use cautiously the concentration values obtained from some of the immunoassays.

Polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in 0+ juvenile cyprinids and sediments of the Po River.

PBDE and PCB content has been determined in 0+ bleak (Cyprinus alburnus), nase (Chondrostoma soetta), gudgeon (Cyprinus gobio), chub (Cyprinus cephalus), and barbel (Barbus sp.) as well as in bed sediments sampled from the River Po upstream and downstream of the confluence of a tributary draining a highly industrialized and urbanized subbasin. Both groups of chemicals were present at higher levels in fish and sediments downstream from the confluence. In addition, whole-body concentrations of PBDEs and PCBs were different among species despite the young specimen age. The fact that PBDEs and PCBs were higher in benthivorous versus planktivorous fish, as well as in carnivorous versus herbivorous species, suggests that feeding behavior is a major controlling factor that may help differentiate the accumulation levels of 0+ juveniles. Of the five species, the pelagic/planktivorous cyprinid bleak (C. alburnus) showed the lowest concentrations (111 ng PBDE/g lipid weight [l.w.], 2016 ng PCB/g l.w.), whereas the benthic dweller and feeder barbel (Barbus sp.) had the highest concentrations of both groups of chemicals (259 ng PBDE/g l.w., 4785 ng PCB/g l.w.). The rank order of species contamination was essentially stable upstream and downstream from the tributary, and the congener contribution of PBDEs was also similar. In general, BDE-47 was the dominant congener, followed by BDE-100, -154, -153, and -28. BDE-209 dominated the PBDE congener profiles of sediments but was not found in any fish sample. Conversely, an unidentified hexa-BDE congener, which was not detected in sediments, was found in all fish species. The levels of PBDEs and PCBs determined in adult goby (Padogobius martensii), a small demersal predator also examined in the same river stretches, provided additional useful insights with which to interpret results.

Assessment of the environmental significance of heavy metal pollution in surficial sediments of the River Po.

The magnitude and ecological relevance of metal pollution of the middle Po river deriving from the River Lambro tributary was investigated by applying different (complementary) sediment quality assessment approaches: (1) comparisons of concentrations with regional reference data, and (2) comparisons with consensus-based sediment quality guidelines (SQGs), as well as by investigations of the partitioning patterns of target heavy metals (Cd, Cu, Ni, Pb, Zn). Total metal concentrations in the surficial sediments revealed significant pollution inputs on the whole river stretch investigated, with a distinct peak at the inlet of the River Lambro. Based on the geoaccumulation index of target heavy metals, the middle reach of River Po has to be considered as moderately polluted with Cd (1

Androgenic and antiandrogenic activities in water and sediment samples from the river Lambro, Italy, detected by yeast androgen screen and chemical analyses.

The river Lambro is the most polluted tributary of the river Po in North Italy and was chosen as a representative water course discharging industrialized areas. Water and sediment samples of the river Lambro were investigated regarding the presence of endocrine disrupting compounds. A combined procedure was used consisting of solid-phase extraction and HPLC based fractionation of samples, of screening for (anti)androgenic activity using the yeast androgen screen (YAS) and of chemical analysis using HPLC-MS/MS and GC-MS. Androgenic and antiandrogenic activities were found in specific fractions of the water and sediment while the total extracts showed antiandrogenic activity only. The chemical analysis of the fractions and total extracts with antiandrogenic activities revealed the presence of compounds with suspected antiandrogenic potency such as bisphenol A, iprodione, nonylphenol, p,p'-DDE and tert-octylphenol but other unknown compounds contributed mainly to the observed antiandrogenic activities. The antiandrogenic load of the river Lambro ranged between 1.34 and 17.1 microM flutamide-equivalents and may pose a risk to aquatic environments. Future screenings for EDC in the environment that have the potential to interfere with reproduction of aquatic organisms should be extended to different modes of actions including (anti)androgenic ones.

The accumulation levels of PAHs, PCBs and DDTs are related in an inverse way to the size of a benthic amphipod (Echinogammarus stammeri Karaman) in the River Po.

The accumulation of polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and DDTs were investigated in gammarids captured at three sites along the middle River Po; the first was located upstream and the other two were at increasing distances downstream of the confluence of a polluted tributary, the River Lambro. Using a GC-MS technique, the levels of PCBs, PAHs and DDTs were determined separately in large and small gammarids as well as in the fine fraction of sediment samples collected along the sites of capture. Results confirm the River Lambro as a source of these chemicals to the River Po, and show that bioaccumulation differences exist between small and large individuals, the former being more contaminated particularly by PCBs and DDTs. This is likely the result of several interacting factors such as contaminant bioavailability, gammarid-size effects on kinetic parameters and feeding selectivity. The bioaccumulation patterns of PCBs and DDTs, and their higher biota-sediment accumulation factors (BSAF), are consistent with chemical properties and suggest a dietary disequilibrium found only 10 km downstream from the tributary, and in smaller amphipods. Present results show that gammarids may represent an additional source of contaminants, particularly of chlorinated compounds, to the many organisms feeding on them, with a higher risk for those which prey selectively on smaller gammarids.

Quality assessment of bed sediments of the Po River (Italy).

Comprehensive and contemporary evaluations of physical, chemical and toxicological endpoints have been performed on bed sediments of the Po River, the major Italian watercourse. Two extensive sampling campaigns were conducted in summer and winter low-flow conditions. Composite sediment samples were collected from ten reaches of the main river: the first was located in the upper region (ambient control), and the others downstream of the confluences of nine principal tributaries. The two sampling programs were paralleled by contemporary investigations on the macroinvertebrate community. The particle-size composition along the Po River showed a relatively uniform distribution of fine sand, a progressive downstream decrease of coarse sands and a corresponding increase of fine materials. The levels of polychlorinated biphenyls (PCB), polycyclic aromatic hydrocarbons (PAH), extractable organo halides (EOX), Cd, Cr, Cu, Hg, Ni, Pb and Zn were determined in sediment fine particles (< 63 microm), and showed marked changes across the ten river reaches. Their longitudinal trends, as those of organic carbon and total nitrogen, were very similar and largely independent of the survey season. Sediment quality benchmarks were used to evaluate sediment chemistry, and, although the overall level of contamination was from moderate to low, the reaches located downstream of the tributaries Dora Riparia, Dora Baltea, Lambro and Oglio were considered to be at risk. Sediments were tested for toxicity on Oncorhynchus mykiss, Ceriodaphnia dubia, Raphidocelis subcapitata and Vibrio fischeri. The toxicity tests were conducted both with sediment extracts and whole samples. Sediment extracts showed toxic potentials that were consistent with the spatial distribution of contaminants. Whole-sediment toxicity showed moderate/low effects which also included false positives and negatives. Alterations of the macroinvertebrate community were found for many kilometers downstream of Dora Riparia, and with a seasonal dependence, also in other reaches of the Italian river. Principal component analysis (PCA) was used to describe the longitudinal and temporal changes of the Po River, and allowed the selection of the most useful and discriminating indicators.

Polychlorinated dibenzo-p-dioxins and dibenzofurans in River Po sediments.

The River Po is the main Italian river draining one of the most populated and industrialised regions in Italy. As part of a monitoring project to assess environmental quality in the River Po, we measured the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in bottom sediments collected along the whole course of the river, from the spring to the delta, downstream from the confluence of its main tributaries. The aim was to investigate the level of contamination in the main Italian river and the contribution of contaminant loads from the subbasins. Composite sediment samples were collected in summer and winter surveys in low-flow water conditions and analysed by HRGC-HRMS for PCDD and PCDF homologue groups and for the 2,3,7,8-substituted congeners. The spatial trend observed in the Po River for PCDD and PCDF concentrations varied depending on the load of contaminants received from the tributaries and the processes of sedimentation. The sum of PCDD and PCDF concentrations, and the toxic equivalent content, ranged from 121 to 814 and from 1.3 to 13 ng/kg dry weight sediment, respectively. These levels of contamination seem lower than in the sediments of rivers draining highly industrialised areas. The PCDD and PCDF homologue profiles in all the samples were very similar, suggesting a common source of this contamination in the River Po. Principal component analysis suggested that widespread sources, such as urban runoff and domestic wastewaters, are probably the main cause of these levels of PCDDs and PCDFs.