[Show abstract][Hide abstract] ABSTRACT: A multi-biomarker approach was developed to evaluate responses of European flounder (Platichthys flesus) in three contrasted estuaries over the English Channel: the Canche (pristine site), Tamar (heavy metals and PAHs contamination) and Seine (heavily pollution with a complex cocktail of contaminants). The condition factor and several biomarkers of the immune system, antioxidant enzymes, energetic metabolism and detoxification processes were investigated in young-of-the-year (0+) and one-year-old (1+) flounder. Results underlined the difference between the pristine site and the Seine estuary which showed a lower condition factor, a modulation of the immune system, a higher Cytochrome C oxidase activity, and an up-regulation of BHMT expression. The moderate biomarker responses in the Tamar fish could be linked to the specific contamination context of this estuary. Flounder life history traits were analyzed by otolith microchemistry, in order to depict how the fish use their habitat and thus respond to chemical stress in estuaries.
[Show abstract][Hide abstract] ABSTRACT: A multibiomarker approach was developed to evaluate the juvenile European flounder responses to a complex mixture of 9 polycyclic aromatic hydrocarbons (PAHs) and 12 polychlorinated biphenyls (PCBs). Exposure was performed through contaminated food pellets displaying: (1) PAH and PCB levels similar to those detected in the heavily polluted Seine estuary, respectively in sediments and in flatfish and (2) ten times these concentrations. Several biomarkers of the immune system (e.g., lysozyme concentration and gene expression of complement component C3 and TNF-receptor), DNA damage (e.g., Comet assay), energetic metabolism (e.g., activity of cytochrome C oxidase), detoxification process (e.g., cytochrome P450 1A1 expression level: CYP1A1; betaine homocysteine methyl transferase expression level: BHMT) were investigated after 14 and 29 days of contamination, followed by a 14-days recovery period. After 29 days of contamination, the detoxification activity (CYP1A1 expression level) was positively correlated with DNA damages; the increase of the BHMT expression level could also be related to the detoxification process. Furthermore, after the recovery period, some biomarkers were still upregulated (i.e., CYP1A1 and BHMT expression levels). The immune system was significantly modulated by the chemical stress at the two concentration levels, and the lysozyme appeared to be the most sensitive marker of the mixture impact.
Environmental Science and Pollution Research 12/2014; 21(24):13789-13803. DOI:10.1007/s11356-014-2563-y · 2.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The St. Lawrence River (SLR) is the second largest waterway in North America. The discharge of the City of Montreal wastewater treatment plant (WWTP) represents the largest volume of treated wastewaters being released into the river. It also ranks as the largest sewage treatment plant of its kind in North America. Over the last decade, intensive multidisciplinary research has focused on assessing the impacts of Montreal wastewater effluents on the SLR. We describe the major findings of these investigations, including the determination of the fate of contaminants, bioaccumulation in fish and invertebrates, ecotoxicological measurements of aquatic animal health, evaluation of endocrine disruption, parasitism in fish, and combined effects of multiple stressors on the SLR. Impacts of the effluents from the WWTP on aquatic organisms from the SLR are both toxicological and ecological, demonstrating the need for an integrated view of the impacts of municipal effluents on aquatic ecosystems.
AMBIO A Journal of the Human Environment 11/2014; DOI:10.1007/s13280-014-0577-9 · 2.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The discharge of organic waste from the petrochemical industry into the Mercier lagoons caused major groundwater contamination. The objective of this study was to determine the immunotoxic potential of three groundwater wells at increasing distance from the incinerator dumping site (1.17, 2.74 and 5.40 km). Rainbow Trout were exposed to increasing concentrations of water from three groundwater wells for 14 days. Immunocompetence was characterized by phagocytosis, mitogen-stimulated proliferation of lymphocytes, cell cycle analysis and apoptosis. A significant increase in innate (phagocytosis) and specific immune response (B lymphocyte proliferation) was observed in trout exposed to water collected from the well at 2.74 km. However, phagocytosis activity was suppressed in groups at 1.17 and 5.40 km. The proportion of lymphocytes in S phase was significantly increased in groups at 2.74 and 5.40 km, while lymphocytes in G0/G1 phase were decreased in all three exposure groups. Additionally, dexamethasone (DEX)-induced apoptosis of lymphocytes was significantly reduced in the group at 2.74 km, which suggests decreased lymphocyte turnover. Furthermore, the ratio of DEX-induced apoptosis/apoptosis was lower in the groups at 2.74 and 5.40 km. In summary, our experiments have shown that exposure to the mixture of organic compounds present in Mercier groundwater modulates phagocytosis and cell proliferation, disrupts the cell cycle and reduces the ratio of DEX-induced apoptosis/apoptosis. It is concluded that groundwater collected in the vicinity of an incinerator containment field could impact immunocompetence in fish.
[Show abstract][Hide abstract] ABSTRACT: Lyngbya wollei is a benthic filamentous cyanobacterium that produces a toxin analogous to the neurotoxic saxitoxin known as lyngbyatoxin (LYNGTX). Microcystis aeruginosa form blooms in the pelagic area of eutrophic lakes and produce a series of potent hepatotoxins-microcystins (MCYST). The aim of this study in vitro study was to examine the difference between the crude extracts of either M. aeruginosa or L. wollei toward the immune system of Elliptio complanata mussels. Freshly isolated hemolymph was plated and exposed to the crude extract of each species at LYNGTX or MCYST equivalent concentrations of 5, 10 and 25 μg/L for 18 h. Immunocompetence was characterized by following changes in hemocyte numbers, metabolic activity (viability), and phagocytosis. Hemocyte counts were not affected, indicating no turnover of hemocytes. Hemocyte metabolic activity was higher in cells exposed to crude extracts of L. wollei. Exposure to L. wollei extracts led to decreased pro-inflammatory precursors such as reactive oxygen species (ROS) and cyclooxygenase (COX) activities. Phagocytosis increased at 25 μg/L for both types of crude extracts. However, hemocytes exposed to crude extracts of M. aeruginosa produced more ROS and COX compared to hemocytes exposed to crude extracts of L. wollei. In conclusion, the data suggest that the crude extract of M. aeruginosa was more toxic than crude extract of L. wollei to mussel hemocytes.
[Show abstract][Hide abstract] ABSTRACT: Municipal effluents are known to impede the immune system of aquatic organisms. The purpose of this study was to examine the immunotoxicity of urban wastewaters before and after 6 treatment processes from 12 cities toward trout leucocytes. Freshly prepared trout leucocytes were exposed to increasing concentrations of solid phase (C18) extracts of wastewaters for 24 hr at 150C. Immunocompetence was determined by following changes in leucocyte viability and the proportion of cells able to ingest at least one (immunoactivity) and at least three (immunoefficiency) fluorescent beads. The influents were treated by six different treatment strategies consisting of facultative aerated lagoons, activated sludge, biological aerated filter, biological nutrient removal, chemically-assisted physical treatment and trickling filter/solid contact. Water quality parameters of the wastewaters revealed that the plants effectively removed total suspended solids and reduced the chemical oxygen demand. The results revealed that the effluents' immunotoxic properties were generally more influenced by the properties of the untreated wastewaters than by the treatment processes. About half of the incoming influents decreased leucocyte viability while 4 treatment plants were able to reduce toxicity. The influents readily increased phagocytosis activity for 8/12 influents while it was decreased in 4/12 influents. This increase was abolished for 4/12 of the effluents using treatments involving biological and oxidative processes. In conclusion, municipal effluents have the potential to alter the immune system in fish and more research will be needed to improve the treatments of wastewaters to better protect the quality of the aquatic environment.
[Show abstract][Hide abstract] ABSTRACT: Il est aujourd’hui prouvé que les organismes sauvages subissent des perturbations du système immunitaire. Une réduction de certaines activités fonctionnelles du système immunitaire peut altérer la résistance de l'hôte aux pathogènes dans l'environnement. Plusieurs perturbations sont causées par des produits pharmaceutiques présents dans les effluents municipaux rejetés dans l'environnement aquatique. Bien qu'étant moins persistant que les PCBs, la consommation accrue de ces composés et leur décharge constante dans le milieu naturel exigent une évaluation des risques sur les espèces sauvages, soumises à ces rejets. Le but de cette étude est de déterminer l'effet immunotoxique et génotoxique des produits pharmaceutiques sur les cellules immunitaires de phoques communs (Phoca vitulina) in vitro. Une lignée cellulaire de lymphomes B (11B7501) et des cellules mononucléées de sang périphérique (PBMC) de phoques en captivité ont été exposés aux produits pharmaceutiques. Les composés étudiés sont des analgésiques (ibuprofène, naproxène), des substances psychoactives (carbamazépine, paroxétine), des antibiotiques (érythromycine, sulfaméthoxazole, triméthoprime), le 17α-éthynylestradiol, le gemfibrozil et de la caféine. Nos résultats ont montré qu'aux concentrations trouvées dans les eaux de surface, les composés n'entrainaient pas d'effets significatifs sur la phagocytose ni sur la transformation lymphoblastique avec la ConA. A des concentrations plus élevées, les expériences avec la lignée cellulaire ont révélé des effets à la fois sur la prolifération lymphoblastique, sur le cycle cellulaire et sur l'apoptose, et ce pour plusieurs composés. Aucun changement significatif n’a été observé pour la phagocytose, dans la gamme de concentrations testée. En conclusion, la prolifération lymphoblastique semble être la méthode la plus sensible pour détecter un effet immunotoxique des produits pharmaceutiques chez le phoque.
[Show abstract][Hide abstract] ABSTRACT: Cyanobacteria have often been described as nutritionally poor for herbivorous organisms. To gain additional information on the potential impacts of invertebrates feeding on cyanobacteria, we fed Elliptio complanata mussels with two types of algae: Anabaena flos-aquae (cyanobacteria) and Pseudokirchneriella subcapitata (green algae). Physiological parameters were examined at the energy status, immune system and oxidative stress levels. Energy status was examined by following the rate of electron transport activity in mitochondria (a measure of cellular energy expense) and lipid/sugar stores in the visceral mass. The cyanobacteria were not actively producing toxins. Based on the digestive gland index, the mussels fed equally on either regime. However, the energy status in mussels fed A. flos-aquae revealed that the total sugar was lower in the digestive gland, whereas mitochondrial electron transport activity (MET), once corrected against the digestive gland somatic index, showed increased energy expenses. Acetylcholinesterase activity and lipid peroxidation (LPO) were also higher in mussels fed with A. flos-aquae compared with mussels fed with P. subcapitata. LPO was correlated by mitochondrial activity in both the digestive gland and gills, suggesting that oxidative stress resulted from metabolic respiration. Immunocompetence (phagocytic activity, natural killer cell-like activity, haemocyte count and viability) and humoral level of lysozyme were not affected in mussels by the algae or cyanobacteria regime. Moreover, the xenobiotic conjugating enzyme, glutathione S-transferase, hemoprotein oxidase and vitellogenin-like proteins were not affected in mussel organs via ingestion of A. flos-aquae. Our study suggests that ingestion of cyanobacteria leads to increased energy expenses, oxidative stress and increased acetylcholine turnover in mussels.
[Show abstract][Hide abstract] ABSTRACT: In summer 2001, the St. Lawrence River (Québec, Canada) experienced perhaps the largest massive fish kill of its history, with more than 25 000 carp (Cyprinus carpio) found dead. This experiment therefore investigated the effects of heat stress on the gene transcription level of heat shock protein 70, cytochrome c oxidase and mitochondrial superoxide dismutase expression as well as on phagocytosis of kidney cell suspensions and animal behaviour. Our study suggests that in summer 2001, elevated water temperatures could have contributed to the fish kill through immunosuppression during an already stressful spawning period.
[Show abstract][Hide abstract] ABSTRACT: In this study, we assessed in vitro the effects of PCDD/Fs on the NK-like cell activity in Eisenia andrei earthworms using flow cytome-try for analysis. NK-like coelomocytes isolated from E. andrei and used as effectors were exposed to various concentrations of PCDDs/Fs mixture, C1 (6.25¥10 -3 ng 2378-TCDD/mL), C2 (12.5¥10 -3 ng 2378-TCDD/mL) and C3 (25¥10 -3 ng 2378-TCDD/mL), before adding them to human tumoral cells (K562) used as targets. We evaluated the percentage of targets lysed by Nk-like cells. The results showed a significant stimulation of the NK-like activity at C3 when PCDD/Fs were not removed from effectors before contact with tar-gets, while no effects were noted when the effectors were washed (PCDD/Fs removed) or fixed. Assessment of the viability of the targets (K562), exposed alone and separately from effectors, to the three concentrations of PCDD/Fs, C1, C2 and C3, showed that all these concentrations were cytotoxic for K562. Results suggest that PCDD/Fs concentrations tested in this assay may be considered too low to induce suppressive effects on the immune function such as the NK-like activity in E. andrei earthworms.
[Show abstract][Hide abstract] ABSTRACT: A multibiomarker approach was developed on juvenile Atlantic tomcod (Microgadus tomcod) to evaluate the pertinence of this approach for low-cost screening assessment of the environmental quality of various coastal sites within estuaries. Several biometric indices and biomarkers (ethoxyresorufin-O-deethylase (EROD) activity, metallothionein concentration, and immune responses) were investigated on immature and maturing tomcods (≤31 months) collected in four environmentally contrasted sites in the St. Lawrence Estuary (SLE). Simultaneous examination of various age classes provides the opportunity to detect short-term responses in sensitive young-of-the-year fish (e.g., EROD induction) and longer-time effects associated with chronic exposure and bioaccumulation (e.g., metallothionein induction). Principal component analysis was helpful to discriminate between responses possibly related to contaminant exposure (EROD, metallothionein) and responses that could be affected by upstream-downstream gradient (immune response, biometric indices). Measurement of a battery of biomarkers in young tomcods at several sites along the shore of the SLE is a low-cost screening investigation useful to identify hot spots requiring further investigation with chemical analysis and additional reference sites.
Environmental Science and Pollution Research 11/2012; 20(2). DOI:10.1007/s11356-012-1285-2 · 2.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Municipal wastewaters are major sources of pollution for the aquatic biota. The purpose of this study was to determine the levels of some pharmaceutical products and the immunotoxic potential of a municipal wastewater aeration lagoon for the treatment of the domestic wastewaters of a small town with wastewater inputs from a 400-bed hospital complex. Endemic mussels were collected, caged and placed in the final aeration lagoon and at sites 1 km upstream and 1 km downstream of the effluent outfall in the receiving river for a period of 14 days. The results showed that the final aeration lagoon contained high levels of total coliforms, conductivity and low dissolved oxygen (2.9 mg/L) as well as detectable amounts of trimethoprim, carbamazepine, gemfibrozil, and norfloxacin at concentrations exceeding 50 ng/L. The lagoon effluent was indeed toxic to the mussel specimens, as evidenced by the appearance of mortality after 14 days (10% mortality), decreased mussel weight-to-shell-length ratio and loss of hemocyte viability. The number of adhering hemocytes, phagocytic activity, total nitrite levels and arachidonic cyclooxygenase activity were significantly higher in mussels placed in the final aeration lagoon. A multivariate analysis also revealed that water pH, conductivity, total coliforms and dissolved oxygen were the endpoints most closely linked with phagocytic activity, the amount of adhering hemocytes and loss of hemocyte viability. In conclusion, exposure of mussels to treated aerated lagoon wastewater is deleterious to freshwater mussels where the immune system is compromised.
[Show abstract][Hide abstract] ABSTRACT: The earthworms Eisenia andrei were used to study the toxicity of PCDD/Fs mixtures to earthworms during 28 day of exposure. The experiments were performed on artificial soils contaminated with dioxins at levels of C1 (0.1 ng 2, 3, 7, 8-TCDD/g soil), C2 (1 ng 2, 3, 7, 8-TCDD/g soil) and C3 (1.5 ng 2, 3, 7, 8-TCDD/g soil). Effects of PCDD/Fs on survival, growth rate and immune responses; phagocytosis and NK-like cell activity, were determined. No mortality was observed at the lowest concentration (C1), while mortalities of 10 and 100% were noted at the highest concentrations tested C2 and C3, respectively. A significant reduction in growth rate was obtained at C2 and no effects at C1. Additionally, an inhibition of phagocytic activity and efficiency was observed at higher concentrations. In contrast, an enhancement of NK-like cell activity was shown at lower concentrations. Based on our results, we hypothesize that the PCDD/Fs mixtures tested at levels equal or higher to C2 (1 ng 2378-TCDD/g soil), lead to adverse effects on biotic potential and immune functions in E. andrei earthworms.
[Show abstract][Hide abstract] ABSTRACT: Seven mother-pup pairs of harp seal (Phoca groenlandica) were sampled during the lactation season near Magdalen Islands, QC, Canada. The blubber and serum of pups and mothers as well as the milk of mothers were analyzed for polybrominated diphenyl ethers and several chlorinated persistent organic pollutants (POPs) to evaluate their transfer prior to weaning. Principal component analysis demonstrated significant variations in contaminant distribution patterns between seal tissues of mothers and pups. The selectivity in the transfer of POPs between mothers and pups appeared mainly driven by their log K(ow) (octanol/water partition coefficient). The most selective transfer step for the POPs examined occurred between the blubber and the serum of the mother. A novel approach to examine temporal changes of POP concentrations in tissues of seals which take into account the contamination of mothers was developed. A general decrease of POP concentrations in pup tissues throughout lactation may suggest a growth dilution. A concomitant rise of POP concentrations was observed in maternal blubber and serum and in milk as lactation progresses. The intensive offloading of contaminants via lactation constitutes a major but selective excretory route for reproductive female seals and also a significant route of exposure for suckling pup seals.
Science of The Total Environment 02/2012; 417-418:98-107. DOI:10.1016/j.scitotenv.2011.11.084 · 4.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 2 Département de santé environ-nementale et santé au travail, Université de Montréal, Quebec; 3 Département des sciences de l'environnement de l'univer-sité du Québec à Montréal Abstract The immunotoxicological effects of poly-chlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/Fs) mixtures on Eisenia andrei earth-worms have never been studied. In this work we investigated these effects both for in vitro and in vivo exposure, using the viability and the phagocytic activity of coelomocytes as immunological biomarkers and the flow cytom-etry was used for analysis. The in vitro expo-sure revealed a cytotoxic effect of PCDD/Fs mixture (C2) containing 50¥10 -3 ng/mL of 2, 3, 7, 8-TCDD and an induction of the phagocytic capacity at the mixture (C1) containing 25¥10 -3 ng/mL of 2, 3, 7, 8-TCDD. In the in vivo filter paper exposure, the immunocompetence of earthworms was assessed after 3 h-exposure to mixtures of PCDD/Fs at the levels of C1, C2, C3 and C4 containing about; 0.05, 0.3, 0.5 and 0.83 ng of 2, 3, 7, 8-TCDD/cm², respectively. Morphological observations showed an exces-sive secretion of mucus and body surface lesions in worms exposed to higher concentra-tions (C3 and C4), which revealed that these organisms were affected by PCDD/Fs either through skin and/or by feeding. The levels of the extruded cell yield decreased significantly at all the concentrations tested. However, the cell viability was shown to be unaffected by PCDD/Fs concentrations. It was also shown, that exposure to the highest PCDD/Fs concen-trations; C2, C3 and C4 inhibited both phago-cytic activity and efficiency.