Polycyclic aromatic hydrocarbons affect survival and development of common snapping turtle (Chelydra serpentina) embryos and hatchlings

Polycyclic aromatic hydrocarbons (PAHs) are toxic compounds found in the John Heinz National Wildlife Refuge in Philadelphia, Pennsylvania. We assessed the impact of PAHs and crude oil on snapping turtle development and behavior by exposing snapping turtle eggs from the Refuge and from three clean reference sites to individual PAHs or a crude oil mixture at stage 9 of embryonic development. Exposure to PAHs had a significant effect on survival rates in embryos from one clean reference site, but not in embryos from the other sites. There was a positive linear relationship between level of exposure to PAHs and severity of deformities in embryos collected from two of the clean reference sites. Neither righting response nor upper temperature tolerance (critical thermal maximum, CTM) of snapping turtle hatchlings with no or minor deformities was significantly affected by exposure to PAHs.     

Absorption of current use pesticides by snapping turtle (Chelydra serpentina) eggs in treated soil

Reptiles often breed within agricultural and urban environments that receive frequent pesticide use. Consequently, their eggs and thus developing embryos may be exposed to pesticides. Our objectives were to determine (i) if turtle eggs are capable of absorbing pesticides from treated soil, and (ii) if pesticide absorption rates can be predicted by their chemical and physical properties. Snapping turtle (Chelydra serpentina) eggs were incubated in soil that was treated with 10 pesticides (atrazine, simazine, metolachlor, azinphos-methyl, dimethoate, chlorpyrifos, carbaryl, endosulfan (I and II), captan, and chlorothalonil). There were two treatments, consisting of pesticides applied at application rate equivalents of 1.92 or 19.2 kg a.i/ha. Eggs were removed after one and eight days of exposure and analyzed for pesticides using gas chromatography coupled with a mass selective detector (GC-MSD) or high performance liquid chromatography (HPLC). Absorption of pesticides in eggs from soil increased with both magnitude and duration of exposure. Of the 10 pesticides, atrazine and metolachlor generally had the greatest absorption, while azinphos-methyl had the lowest. Chlorothalonil was below detection limits at both exposure rates. Our preliminary model suggests that pesticides having the highest absorption into eggs tended to have both low sorption to organic carbon or lipids, and high water solubility. For pesticides with high water solubility, high vapor pressure may also increase absorption. As our model is preliminary, confirmatory studies are needed to elucidate pesticide absorption in turtle eggs and the potential risk they may pose to embryonic development.     

Snapping turtles (Chelydra serpentina) as bioindicators in Canadian areas of concern in the Great Lakes Basin. II. Changes in hatching success and hatchling deformities in relation to persistent organic pollutants

Hatching success and deformities in snapping turtle hatchlings (Chelydra serpentina) were evaluated using eggs collected from 14 sites in the Canadian lower Great Lakes, including Areas of Concern (AOC), between 2001 and 2004. Eggs were analyzed for PCBs, PBDEs, and pesticides. Between 2002 and 2004, hatchling deformity rates were highest in two AOCs (18.3-28.3%) compared to the reference sites (5.3-11.3%). Hatching success was poorest in three AOCs (71.3-73.1%) compared to the reference sites (86.0-92.7%). Hatching success and deformity rates were generally poorer in 2001 compared to 2002-2004, irrespective of the study location and could be due to egg handling stress in 2001. Hatching success and deformities were generally worst from the Wheatley Harbour, St. Lawrence River (Cornwall), Detroit River, and Hamilton Harbour AOCs. Associations between contaminant burdens with embryonic development were sufficiently poor that the biological relevance is questionable. Stressors not measured may have contributed to development abnormalities.     

Total mercury concentrations in an industrialized catchment,the Thur River basin (north-eastern France): geochemical background level and contamination factors

River bottom sediments and soils were collected from the industrialized Thur River basin (north-eastern France) to assess mercury contamination. The regional geochemical background level of total mercury was evaluated to calculate mercury contamination factors (Fc) in soils and river bottom sediments. Our estimate of the mean background mercury levels in river sediments and soils, not affected by human activities, was 232 ng x g(-1) (range: 27-406 ng x g(-1)). Sediments contaminated by the effluent from a chlor-alkali plant yielded the highest contamination factors (Fc=1784). Contamination factors of surficial soils within 1 km of the industrial site range from 6.3 to 43.6. This contamination is attributed to diffuse atmospheric deposition from this local plant. However, even upstream from this industrial area elevated contamination factors were recorded for river bottom sediments (Fc=3.2 to 26.4) and for one alluvial soil profile (Fc=10). This is possibly due to past pollution resulting from waste water discharges. Mercury contamination in the different horizons of alluvial soils is not correlated with soil organic carbon content, but may be the result of occasional accidental pollution arising from the introduction of contaminated suspended particulate matter by the Thur River during periods of flooding.     

First evidence of polybrominated diphenyl ether (flame retardants) effects in feral barbel from the Ebro River basin (NE, Spain)

Polybrominated diphenyl ethers (PBDEs) are chemicals of environmental concern due to their lipophilic, persistent and bioaccumulable characteristics as well as for their potential endocrine disrupting role. Former studies carried out in a tributary of the Cinca river (Ebro basin, NE Spain) revealed high levels of PBDEs in fish due to the discharges of effluents rich in PBDEs coming from a nearby industrial park in Barbastro. In this study, several biomarkers of pollutants exposure were measured in barbel, Barbus graellsii, before (upstream) and after (downstream) the main industrial site (Barbastro) in the Vero river. The results evidenced an enhanced hepatic phase I and II metabolism (measured as reductases, glutathione S transferase and uridinediphospho-glucuronosyltransferase), and of the antioxidant enzyme glutathione peroxidase. Conversely, fishes collected from downstream reaches had their phase I CYP1A dependent ethoxyresorufin O-deethylase, antioxidant diaphorase and brain cholinesterase activities depleted. In addition, the histological study of the liver and kidney of these fish evidenced an increase of the number and size of macrophage aggregates in most individuals collected downstream. Bivariate correlated analyses showed that the above mentioned biomarkers were correlated to measured PBDE congeners, thus indicating that the observed biological effects were unlikely to be related to other environmental factors than PBDEs. Overall, the measured biochemical and histological markers provide new evidence that in field exposed fish, PBDEs levels were associated with high activities of phase I and II metabolic enzymes, oxidative stress in liver, neurotoxicity in brain and histopathological effects in both liver and kidney.     

PCBs and organochlorine pesticides in blubber biopsies from free-ranging St. Lawrence River Estuary beluga whales (Delphinapterus leucas), 1994-1998

For the first time, organochlorine (OC) contaminants were measured in blubber biopsies from free-ranging St. Lawrence River Estuary beluga whales (Delphinapterus leucas), to compare contaminant levels to those previously measured in dead stranded belugas. PCBs, DDTs, toxaphene and chlordane-related compounds were the major OC contaminants detected in 44 belugas biopsied in 1994-1998. SigmaPCB (the sum of 104 congeners) ranged from 2080 to 128,000 ng/g lipid in males (n=34; minimum estimated ages 8-22 years), and from 148 to 44,100 ng/g lipid in females (n=10; minimum estimated ages 7-22 years). The concentrations of PCBs and OC pesticides in the blubber of these whales overlapped those observed in stranded belugas from an earlier study, and demonstrated comparable age and sex-related trends. However, lower proportions of mirex, HCB, DDTs, and many of the highly chlorinated PCBs occurred in the biopsy samples compared to results for blubber from stranded carcasses. Most major OC compounds were present at lower concentrations in the biopsies, but this does not appear to be solely related to age differences between the two groups, or to emaciation in the stranded whales. Nor does it appear to be associated with the use of superficial biopsies, and the possible stratification of lipids and OCs in the blubber layer. Nevertheless, given these possible confounding factors, and the uncertainty in age estimates for the biopsied whales, the results point to the need for careful interpretation of biopsy results when comparing with data taken from the full depth of the blubber mantle in stranded whales. Taken together, results from both biopsied whales and previously studied stranded belugas indicate that PCB and OC pesticide contamination of St. Lawrence beluga whales may occur across a broader range of levels than previously thought, at least for males which formed the largest group in this study, possibly due to different degrees of dietary exposure. It also appears that measuring contaminant concentrations only in stranded whales, may overestimate OC levels in the population as a whole, especially for highly chlorinated OCs.     

Occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata)

A study was conducted in 1999 to determine the occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata). Concentrations of selected contaminants were measured in surface water, municipal effluent, sediments and mussels. Analyses were performed on 4-tert-octylphenol (4-t-OP), 4-n-nonylphenol (4-n-NP), nonylphenol polyethoxylates (NP(1-16)EO), nonylphenol-mono and di-ethoxycarboxylic acids (NP(1)EC and NP(2)EC), and octylphenol-mono and di-ethoxycarboxylic acids (OP(1)EC and OP(2)EC). Mussels (Elliptio complanata) taken from a reference lake were placed in cages and submerged for 62 days at two sites in the St. Lawrence River, 1.5 km upstream and 5 km downstream of the outfall of a municipal wastewater treatment plant. The results showed that many of the target chemicals were present in all matrices studied: in water, at ppt and ppb levels, and reaching ppm levels in sediments and mussels. Concentrations of these contaminants were higher in matrices sampled at the downstream site than in those drawn at the site upstream of the Montreal effluent outfall, especially in sediments. Likewise, the slight, but not significant, bioconcentration of certain alkylphenol polyethoxylates (AP(n)EO) in the mussels was more noticeable at the downstream site than at the upstream site.     

Organochlorine and heavy metal contamination in the eggs of the Spanish Imperial Eagle (Aquila (heliaca) adalberti) and accompanying changes in eggshell morphology and chemistry

Eggs of the Spanish Imperial Eagle (Aquila (heliaca) adalberti) contained small amounts of heavy metals and organochlorine contaminants, including DDE, aldrin, dieldrin and HCHs, PCBs. Levels of organochlorine, except DDE, have decreased from 1972 to 1984. This paper reports how the structural and chemical characteristics of modern shells differ from those of older shells, unaffected by pesticides. Also, fertile and infertile recent eggs are compared. A 12.6% decrease in eggshell thickness has occurred in conjunction with a slight decrease in degree of crystallisation, but no significant changes have been found in other structural variables. The difference in chemical composition of the shell between modern and old eggs is both qualitative (presence of chlorine, potassium, magnesium and copper in recent shells) and quantitative (increase in the percentage of phosphorus and sulphur). The only clear differences between fertile and infertile eggs were a greater increase of phosphorus content and altered crystallisation in the latter. No clear relation between the amount of the mineral constituents of the shell and morphological alterations was established. The levels of contaminants in the contents seemed to be independent of the chemical alterations of the eggshell. The breeding parameters of the overall population showed no significant changes after the introduction of pesticides. However, eggshell changes have occurred in this species, which is one of the rarest birds of prey of the world, and some breeding individuals have sufficiently thin shells to suspect reduced reproductive success.     

Reduction of organochlorine residues in Goshawk eggs (Accipiter gentilis) from Northern Germany (1971-2002) and increasing eggshell index

We investigated unhatched eggs (n = 57) of Goshawks (Accipiter gentilis) from the period 1988--2002 and compared them with eggs (n = 134) from 1971--1978. Mean concentrations of PCBs, sigmaDDT and HCB in the earlier period were (mg kg(-1) lipid weight) 50.1, 126.6, and 117.7, while the values from the later period were statistically significantly lower: 27.7, 4.1, 0.1, respectively. Nevertheless, the concentration of PCB is relatively constant during the last 8-10 years. The variability of residues was lower within clutches than among clutches. No age-dependent accumulation in eggs could be demonstrated. A statistically significant lower shell index was measurable for the period 1971--1978, while this index during 2001--2002 was relatively high as it was during the late 18th century. We are suggesting a threshold value for effect concentrations on shell index. Residues of the DDT-group had an influence on the number of fledged young per nest site.     

Levels and temporal trends (1983-2003) of persistent organic pollutants (POPs) and mercury (Hg) in seabird eggs from Northern Norway

The main objective of this study was to investigate possible temporal trends of persistent organic pollutants (POPs) and mercury in eggs of herring gulls (Larus argentatus), black-legged kittiwakes (Rissa tridactyla), common guillemots (Uria aalge) and Atlantic puffins (Fratercula arctica) in Northern Norway. Eggs were collected in 1983, 1993 and 2003. Egg concentrations of POPs (PCB congeners IUPAC numbers: CB-28, 74, 66, 101, 99, 110, 149, 118, 153, 105, 141, 138, 187, 128, 156, 157, 180, 170, 194, 206, HCB, alpha-HCH, beta-HCH, gamma-HCH, oxychlordane, trans-chlordane, cis-chlordane, trans-nonachlor, cis-nonachlor, p,p'-DDE, o,p'-DDD, p,p'-DDD, o,p'-DDT and p,p'-DDT) and mercury were quantified. Generally, POP levels decreased between 1983 and 2003 in all species. No significant temporal trend in mercury levels was found between 1983 and 2003.     

Temporal trend (1988-2008) of hexabromocyclododecane enantiomers in herring gull eggs from the German coastal region

Levels of α-, β-, and γ-hexabromocyclododecane (HBCD) were determined in pooled eggs from herring gulls (Larus argentatus) sampled on three bird sanctuaries near the German North Sea coast between 1988 and 2008 (Mellum and Trischen) and the German Baltic Sea coast between 1998 and 2008 (Heuwiese) and archived by the German Environmental Specimen Bank. Pressurized fluid extraction, gel permeation chromatography, and LC-MS/MS using ¹³C₁₂-labelled isotope standards and a chiral column were applied. α-HBCD was the dominating diastereomer and ranged between 3.7 and 107 ng g⁻¹ lw while β- and γ-HBCD were throughout close to LOQ. The highest α-HBCD concentration was found in eggs from Mellum sampled in the year 2000. Interestingly, HBCD in eggs from the three islands displayed similar time courses with levels increasing to a peak contamination around 2000 and decreasing levels ever since. Chiral signatures of α-HBCD in eggs differed among the islands but indicated a preferential enrichment of the first eluting enantiomer (−)-α-HBCD.     

Sediments as monitors of heavy metal contamination in the Ave river basin (Portugal): multivariate analysis of data

The concentrations of heavy metals (Cd, Cr, Cu, Ni, Pb, Zn) were determined in river sediments collected at the Ave river basin (Portugal) to obtain a general classification scenery of the pollution in this highly polluted region. Multivariate data analysis techniques of clustering, principal components and eigenvector projections were used in this classification. Five general areas with different polluting characteristics were detected and several individual heavy metal concentration abnormalities were detected in restricted areas. A good correlation between the overall metal contamination determined by multivariate analysis and metal pollution indexes for all sampling stations was obtained. Some preliminary experiments showed that the metal concentrations normalised to the volatile matter content in the sediment fraction with grain size <63 microm seems to be an adequate method for assessing metal pollution.     

Blood biomarkers and contaminant levels in feathers and eggs to assess environmental hazards in heron nestlings from impacted sites in Ebro basin (NE Spain)

Blood biomarkers and levels of major pollutants in eggs and feathers were used to determine pollution effects in nestlings of the Purple Heron Ardea purpurea and the Little Egret Egretta garzetta, sampled on three Ebro River (NE Spain) areas: a reference site, a site affected by the effluents of a chlor-alkali industry and the river Delta. The two impacted heron populations showed mutually different pollutant and response patterns, suggesting different sources of contamination. In the population nesting near the chlor-alkali plant, elevated levels of hexachlorobenzene (HCB) and polychlorobiphenyls (PCBs) in eggs, and mercury in feathers in A. purpurea chicks were related with reduced blood antioxidant defenses and increased levels of micronuclei. In Ebro Delta, high levels of plasmatic lactate dehydrogenase in A. purpurea chicks and high frequency of micronuclei in blood of both species were tentatively associated with intensive agricultural activities taking place in the area. These results provide the first evidence of a biological response in heron chicks to the release of pollutants at a chlor-alkali plant.     

Distribution and biological impact of dioxin-like compounds in risk zones along the Ebro River basin (Spain)

The aim of this investigation was to evaluate the environmental impact associated to PCDDs/Fs and dioxin-like PCBs in the Ebro River basin. Sediments and fish from several species were sampled at three sites with different historical pollution records, including the Barbastro area with different industrial activities, and the Flix and Monzón sites, associated to heavy organochlorine compound pollution. Seventeen toxic PCDDs/Fs and 12 dioxin-like PCBs were analyzed by GC-MS. The results obtained indicated significant accumulation of dioxin-like PCBs, but not PCDDs/Fs, in sediments and fish at the Flix site compared to the other sites. Concomitantly, cytochrome p450 1A (CYP1A) expression, a known indicator for pollution by dioxins and dioxin-like PCBs, was significantly elevated in barbel (Barbus graellsii) from the Flix site, compared to the population from the Barbastro site. CYP1A expression correlated with the concentration of dioxin-like PCBs in the fish fat, whereas no significant correlation was found with PCDDs/Fs concentrations. Our data suggest a significant biological impact at the Flix site, closely related to the presence of dioxin-like PCBs, whereas the PCDDs/Fs contribution to this impact appears to be non-significant, at least in the studied sites.     

Decrease of PCDD/F levels in human blood from Germany over the past ten years (1989-1998)

744 whole blood samples of normal subjects from Germany collected in 1989-1998 have been analyzed for polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDF) by capillary gas chromatography/high resolution mass spectrometry. Over the examined time period a continuous decrease of the PCDD/F concentrations in human blood was observed. The mean levels found were 43.7 pg I-TEq/g (lipid basis) in 1989 and 20.7 pg I-TEq/g (lipid basis) in 1996/98 [median: 42.2 and 19.4]. The reduction to about the half was found for most congeners. Each one-year subset of the collective and the entire collective shows a positive correlation of the PCDD/F blood levels with age for most of the congeners, the sum values and the calculated toxicity equivalents. For statistical evaluation a multiplicative model was used: Concentration = A x Age(B). The correlation is mostly pronounced for lower chlorinated PCDD and for 2,3,4,7,8-PentaCDF. The PCDD/F concentrations in human blood in relation to the year of examination and the age of the subjects can be described by a linear model: I-TEq [pg/g (lipid basis)] = 6176 - 3.097 x Year + 0.6482 x Age or by a multiplicative model: I-TEq [pg/g(lipid basis)] = 10(89.08-0.04415 x Year + 0.008468 x Age).     

Mercury (Hg) in fish consumed by the local population of the Jaguaribe River lower basin,Northeast Brazil

The knowledge of Hg concentrations in fish is of considerable interest since these organisms are a major source of protein to coastal human populations and fishing communities. The main source of human exposure to Hg contamination occurs through the consumption of fish. In this paper, we compare Hg concentration in 13 fish species from Jaguaribe River lower basin and an adjacent coastal region in the northeastern coast of Brazil. We sampled fish from three stretches of the river: fluvial, estuarine, and marine regions. We tested the hypothesis that Hg concentration in muscle tissue vary according to species, location, and trophic level. Significant differences were observed among species and trophic level, but these could not be observed among the regions studied. As expected, the highest concentrations were observed in carnivorous fish (5.6–107.5; 26.9?±?18.8 ng g^?1). Hg concentrations observed in this study are similar to those observed in regions of low environmental contamination. We estimated Hg intake to vary between 0.02 and 0.22 ng Hg kg body weight^?1 week^?1, for the average body weight of 56.7 kg, which was considered as low exposure and therefore, a low risk to consumers of fish from the regions studied.     

Persistent organochlorines in beluga whales (Delphinapterus leucas) from the St Lawrence River estuary--I. Concentrations and patterns of specific PCBs, chlorinated pesticides and polychlorinated dibenzo-p-dioxins and dibenzofurans

Blubber samples from beluga whales (Delphinapterus leucas) in the St Lawrence River estuary were analysed for PCB congeners (ortho- and non-ortho-substituted) and other persistent organochlorines as well as chlorinated dibenzo-dioxins/furans (PCDD/Fs). Major individual components (mean concentrations > 1 microg g(-1)) were 4,4'-DDE, -DDD and -DDT, T12 (a toxaphene-related compound), trans-nonachlor, oxychlordane, mirex, HCB, tris(p-chlorophenyl) methane and dieldrin. Concentrations of SigmaPCBs (8.3-412 microg g(-1)), SigmaDDT (3.36-389 microg g(-1)) and mirex (0.18-6.8 microg g(-1)) were particularly elevated relative to other odontocetes in Canadian waters. SigmaDDT, PCBs (as Aroclor), mirex and T12 concentrations were positively correlated with age of adult females (> 10 years) but only weakly, or not significantly, correlated with age of adult males. PCDD/Fs were present at low ng kg(-1) levels and consisted mainly of penta- and hexachlorofurans, and hepta- and octachlorodioxin. CB126 (3,3',4,4',5-PCB) was the most prominent non-ortho-substituted PCB congener in beluga blubber. Total TCDD toxic equivalents averaged 330 ng kg(-1) in females and 1400 ng kg(-1) in males and were dominated by CB126, and the mono-ortho-substituted congeners CB105 and CB118. Biomagnification factors (BMFs) for mirex and SigmaPCB from fish to beluga ranged from 11 to 16, and were similar to BMFs in Arctic animals, indicating that elevated levels in St Lawrence animals are a consequence of relatively high levels of recalcitrant organochlorines in prey of the beluga in the St Lawrence river system.     

PCBs in wild mussels (Mytilus galloprovincialis) from the N-NW Spanish coast: current levels and long-term trends during the period 1991-2009

Concentrations of polychlorinated biphenyls (PCBs) were determined in wild mussels (Mytilus galloprovincialis) from seven areas of the N-NW Spanish coast, during the period 1991-2009. The studied area is comprised of highly productive ecosystems which support important commercial fishing and shellfishing activities. The seven PCBs indicator congeners recommended by ICES (IUPAC nos. 28, 52, 101, 118, 138, 153 and 180), were analysed by gas chromatography with an electron capture detector (GC-ECD). Lowest average concentrations of PCBs were recorded in Arousa (1.04 μg kg(-1) wet weight), a low populated area with no industry; while the highest levels were found in areas near cities and/or showing a high industrial activity such as: Bilbao Ziérbena (139 μg kg(-1) ww) or A Coru?a (55 μg kg(-1) ww). The hexachlorinated congeners CB153 and CB138 dominated the profiles in all wild mussel populations, although a geographical pattern was found in which the relative contributions of the lower chlorinated congeners were higher in the west coast (Rías of Vigo, Pontevedra and Arousa), which may reflect the occurrence of recent inputs of those compounds in the area. Finally, the application of the Mann-Kendall's test demonstrate the existence of a significant decrease in PCBs levels since 1991 up to 1996 in the studied area, whereas either a slow decrease or stable concentrations were registered over the past decade.     

Tris (4-chlorophenyl) methane and tris (4-chlorophenyl) methanol in marine mammals from the Estuary and Gulf of St. Lawrence

Levels of tris (4-chlorophenyl) methanol (TCPM) and its presumed precursor tris (4-chlorophenyl) methane (TCPMe) are reported in marine mammals from the Estuary and Gulf of St. Lawrence, Canada. These compounds were measured in blubber samples of seals and whales using ion trap mass spectrometry (MS/MS) detection. Detectable concentrations of both TCPM and TCPMe were observed in all of the samples analysed. Concentrations of these compounds varied with species ranging from 1.7 to 153 and from 1.3 to 50.6 ng/g lipid wt. for TCPM and TCPMe, respectively. TCPM was from 1.3 to 10 times more concentrated than TCPMe. The highest levels of both TCPM and TCPMe were observed in adult male beluga whales (Delphinapterus leucas) from the St. Lawrence Estuary, while adult female beluga whales from the same area showed levels similar to those in the seals examined. Among the four seal species investigated, TCPM and TCPMe levels were the highest in grey (Halichoerus grypus) and hooded (Cystophora cristata) seals, and lowest in harp seals (Phoca groenlandica). Intermediate levels were found in harbour seals (Phoca vitulina); however, their concentrations might be underestimated considering the younger mean age of these animals. Ratios of both 4,4'-DDE/sigma DDT and TCPM/sigma TCP were very similar between animals from the same species. Strong correlations between sigma TCP and sigma DDT were also observed for each species of mammals, most likely indicating that both sigma TCP and sigma DDT are bioaccumulated in marine mammals. The relationships between sigma DDT and sigma TCP also demonstrate that sigma TCP are less bioaccumulated than sigma DDT by the marine mammal species examined.     

Integrated spatial health assessment of yellow perch (Perca flavescens) populations from the St. Lawrence River (QC,Canada), part B: cellular and transcriptomic effects

Environmental Science and Pollution Research - Multi-biological level assessments have become great tools to evaluate the health of aquatic ecosystems. Using this approach, a complementary study...