High incidence of deformity in aquatic turtles in the John Heinz National Wildlife Refuge

The John Heinz National Wildlife Refuge is subject to pollution from multiple sources. We studied development of snapping turtle (Chelydra serpentina) and painted turtle (Chrysemys picta) embryos from the refuge from 2000 through 2003. Mean annual deformity rate of pooled painted turtle clutches over four years ranged from 45 to 71%, while that of snapping turtle clutches ranged from 13 to 19%. Lethal deformities were more common than minor or moderate deformities in embryos of both species. Adult painted turtles had a higher deformity rate than adult snapping turtles. Snapping turtles at JHNWR had high levels of PAH contamination in their fat. This suggests that PAHs are involved in the high level of deformities. Other contaminants may also play a role. Although the refuge offers many advantages to resident turtle populations, pollution appears to place a developmental burden on the life history of these turtles.     

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.     

Environmental contamination and developmental abnormalities in eggs and hatchlings of the common snapping turtle (Chelydra serpentina serpentina) from the Great Lakes-St Lawrence River basin (1989-1991)

During 1989-1991, we assessed developmental abnormalities in embryos and hatchlings from eggs of the common snapping turtle (Chelydra serpentina serpentina). Eggs were collected and artificially incubated from eight sites in Ontario, Canada and Akwesasne/New York, USA. In eggs from the same clutches we measured 20 organochlorine pesticides, 48 polychlorinated biphenyl (PCBs) congeners including 6 non-ortho PCBs, 8 polychlorinated dibenzodioxins (PCDDs), 14 polychlorinated dibenzofurans (PCDFs) and total mercury. We found a significant increase in abnormal development with increasing polychlorinated aromatic hydrocarbon exposure in eggs, particularly PCDD and PCDF concentrations. In contrast, the risk of abnormality was not significantly higher as toxic equivalent concentrations increased in eggs. We also found significant 7-ethoxyresorufin O-deethylase and Cytochrome P4501A responses in livers of hatchling turtles from Lake Ontario relative to hatchlings from a clean, inland site whereas we did not find any evidence of porphyria in the hatchlings from either site.     

Benzo[a]pyrene exposure influences the cardiac development and the expression of cardiovascular relative genes in zebrafish (Danio rerio) embryos

It is reported that the most abundant polycyclic aromatic hydrocarbons (PAHs) in weathered crude oils are cardiotoxic. However, the action mechanism of PAHs on vertebrate cardiovascular development and disease is unclear. In the present study, the cardiac morphology and functioning of zebrafish embryos exposed to benzo[a]pyrene [B(a)P], as a high-ring PAHs, for 72 h were observed and determined. The results showed that B(a)P exposure resulted in cardiac developmental defects in zebrafish embryos. Significant changes in expression level of multiple genes potentially critical for regulating the B(a)P-induced cardiovascular developmental defects were also found. A gene network regulating cardiac development perturbed by B(a)P exposure was identified and established by computational analysis and employment of some databases. The information from the network could provide a clue for further mechanistic studies explaining molecular events regulating B(a)P-mediated cardiovascular defects and consequences.     

Organochlorine, PCB, PAH, and metal concentrations in eggs of loggerhead sea turtles (Caretta caretta) from northwest Florida, USA

Composite samples of unhatched and physically unaltered loggerhead sea turtle, Caretta caretta, eggs collected from 20 nests along northwest Florida were analyzed for organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and metals. Chemical analyses revealed that turtle eggs contained detectable amounts of metals, PAHs, and PCBs. Only one OCP, p,p'-DDD, was detected, and its presence was restricted to eggs from two nesting sites. None of the PCB concentrations exceeded the Food and Drug Administration's (FDA) action limit. Concentrations of dioxin-like PCB congeners, 105, 118, and 126, and total PCBs were also detected and are contributors to the toxic burden of loggerhead sea turtle eggs. Concentrations of PAHs, 1,2,5,6-dibenzanthracene, 1-methyl naphthalene, C1-naphthalene and naphthalene were variable at nesting sites. Comparison of mean metal burdens in eggs from different beaches suggested that no uniform geographic gradients exist. Presence of OCPs, PCBs, PAHs and metals and their additive or synergistic toxicity is a concern to loggerhead sea turtle eggs; however, additive or synergistic impacts for loggerhead sea turtles are largely undocumented.     

Organochlorine,PCB, PAH,and metal concentrations in eggs of loggerhead sea turtles (Caretta caretta) from northwest Florida,USA

Abstract

Composite samples of unhatched and physically unaltered loggerhead sea turtle, Caretta caretta, eggs collected from 20 nests along northwest Florida were analyzed for organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and metals. Chemical analyses revealed that turtle eggs contained detectable amounts of metals, PAHs, and PCBs. Only one OCP, p, p'‐DDD, was detected, and its presence was restricted to eggs from two nesting sites. None of the PCB concentrations exceeded the Food and Drug Administration's (FDA) action limit. Concentrations of dioxin‐like PCB congeners, 105, 118, and 126, and total PCBs were also detected and are contributors to the toxic burden of loggerhead sea turtle eggs. Concentrations of PAHs, 1,2,5,6‐dibenzanthracene, 1‐methyl naphthalene, Cl‐naphthalene and naphthalene were variable at nesting sites. Comparison of mean metal burdens in eggs from different beaches suggested that no uniform geographic gradients exist. Presence of OCPs, PCBs, PAHs and metals and their additive or synergistic toxicity is a concern to loggerhead sea turtle eggs; however, additive or synergistic impacts for loggerhead sea turtles are largely undocumented.     

Long-term disruption of growth,reproduction, and behavior after embryonic exposure of zebrafish to PAH-spiked sediment

A natural sediment spiked with three individual polycyclic aromatic hydrocarbons (PAHs; pyrene, phenanthrene and benzo[a]pyrene) was used to expose zebrafish embryos and larvae during 4 days. The total PAH concentration was 4.4 μg g^?1 which is in the range found in sediment from contaminated areas. Quantification of metabolites in the larvae after exposure confirmed the actual contamination of the larvae and indicated an active metabolism especially for pyrene and benzo[a]pyrene. After a transfer in a clean medium, the larvae were reared to adulthood and evaluated for survival, growth, reproduction, and behavior. Measured endpoints revealed a late disruption of growth (appearing at 5 months) and a trend toward a lower reproductive ability. Adults of embryos exposed to sediment spiked with PAHs displayed lethargic and/or anxiety-like behaviors. This latter behavior was also identified in offspring at larval stage. All together, these effects could have detrimental consequences on fish performances and contribution to recruitment.     

Polycyclic aromatic hydrocarbons in crude oil-contaminated soil: A two-step method for the isolation and characterization of PAHs

A two-step analytical method is developed for the isolation and characterization of polycyclic aromatic hydrocarbons (PAHs) in crude oil contaminated soil. In the first step, those crude oil components were isolated which are easily mobilized with water from the contaminated soil (determination of groundwater pollution potential). In the second step, the fraction containing the remaining crude oil compounds was extracted using toluene. After the cleanup of the fractions, both fractions were analyzed using high-performance liquid chromatography (HPLC). The HPLC of the toluene extracted fraction shows that along with the sixteen priority pollutants from the US-EPA list, many other polycyclic aromatic hydrocarbons (PAHs) are present as well. It is evident from the chromatograms that a significant amount of PAHs are present as is also the case in the fractions eluted by water. The described method allows the determination of total organic pollutants from crude oil, some of them being potential groundwater contaminants. The major part of the total pollutants could not be mobilized by water and therefore remains in the soil, which was extracted in the second step.     

Comparative study of different exposure routes on the biotransformation and genotoxicity of PAHs in the flatfish species, Scophthalmus maximus

In this study, laboratory experiments were carried out in order to come to a better understanding of the fate of polycyclic aromatic hydrocarbons (PAHs) in the marine environment and especially on their bioaccumulation, biotransformation and genotoxic effects in fish. Juveniles of turbot (Scophthalmus maximus) were exposed to PAHs through different routes via (1) a mixture of dissolved PAHs, (2) a PAH-polluted sediment and (3) an oil fuel elutriate. Fish were exposed 4 days followed by a 6-day depuration period. In each experiment, PAH concentrations in the seawater of the tanks were analysed regularly by gas chromatography coupled with mass spectrometry. Muscle and liver samples were also analysed for parent PAH levels and PAH bioconcentration factors were calculated. Biotransformation was evaluated by measuring the levels of PAH metabolites in fish bile. Genotoxicity was assessed by the alkaline comet assay. Regardless of exposure route, the parent PAH concentrations in the liver and muscle showed a peak level 1 day after the beginning of the exposure, followed by a decrease up to the background level towards the end of the experiment, except for the exposure to dissolved PAHs for which levels were relatively low throughout the study. As a consequence, no bioaccumulation was observed in fish tissues at the end of the experiment. In contrast, regardless of exposure routes, a rapid production of biliary metabolites was observed throughout the whole exposure experiment. This was especially true for 1-hydroxypyrene, the major metabolite of pyrene. After 6 days of recovery in clean water, a significant decrease in the total metabolite concentrations occurred in bile. Fish exposed through either route displayed a significant increase in DNA strand breaks after 4 days of exposure, and significant correlations were observed between the level of biliary PAH metabolites and the level of DNA lesions in fish erythrocytes. Overall results indicate that exposure to either a mixture of dissolved PAHs, a PAH-contaminated sediment or a dispersed oil fuel elutriate leads to biotransformation and increase in DNA damage in fish. The quantification of PAH metabolites in bile and DNA damage in erythrocytes appear to be suitable for environmental monitoring of marine pollution either in the case of accidental oil spills or sediment contamination.     

Dissolution and removal of PAHs from a contaminated soil using sunflower oil

This study reports on the feasibility of remediation of polycyclic aromatic hydrocarbon (PAH) contaminated soils using sunflower oil, an environmentally-friendly solvent. Batch experiments were performed to test the influence of oil/soil ratio on the remediation of PAH contaminated soil, and to test the mass transfer behaviors of PAHs from soil to oil. An empirical model was employed to describe the kinetics of PAH dissolution and to predict equilibrium concentrations of PAHs in oil. PAH containing oil was regenerated using active carbon. Results show that dissolution of PAHs from a Manufactured Gas Plant (MGP) soil at oil/soil ratios of one or two were almost the same. Nearly all PAHs (81-100%) could be removed by sunflower oil dissolution. Mass transfer coefficients for low molecular PAHs namely fluoranthene, phenanthrene and anthracene were one or two orders of magnitude higher than those for high molecular PAHs with 4-6 rings. Ninety milliliters of PAH containing oil could be regenerated by 10 g active carbon in a batch reactor. Such a remediation procedure indicates that sunflower oil is a promising agent for the removal of PAHs from MGP soils. However, further research is required before the method can be used for in situ remediation of contaminated sites.     

Screening of hydrophobic DNA adducts in flounder (Platichthys flesus) from the Baltic Sea

Neoplasia and other histopathological lesions in flounder (Platichthys flesus) liver have been investigated in several European sea areas, including the Baltic Sea. Several studies have been able to link neoplasm epizootics in fish with the exposure to genotoxins such as polycyclic aromatic hydrocarbons (PAHs). The level of hydrophobic DNA adducts in tissue DNA reflects the exposure of the organism to PAHs.Using hydrophobic DNA adduct levels as biomarkers, possible PAH exposure was assessed in flounder from 10 different sites in the Baltic Sea, collected during the years 1995–1997. The results show that the overall levels of hepatic DNA adducts were low and, in general, the chromatograms appeared clean. The highest levels of DNA adducts were found at two sites in the southern Baltic Sea. There were no statistically significant differences in adduct levels between the sites. Our results indicate that flounder from studied off shore sites of the Baltic Sea had not been exposed to a greater extent to large polycyclic hydrophobic hydrocarbons in their environment.     

Monitoring contaminants from oil production at sea by measuring gill EROD activity in Atlantic cod (Gadus morhua)

An ex vivo gill EROD assay was applied in Atlantic cod (Gadus morhua) as a biomarker for waterborne CYP1A-inducing compounds derived from oil production at sea. Exposure to nominal concentrations of 1 ppm or 10 ppm North Sea crude oil in a static water system for 24 h caused a concentration-dependent gill EROD induction. Further, exposure of cod for 14 days to environmentally relevant concentrations of produced water (PW, diluted 1:200 or 1:1000) from a platform in the North Sea using a flow-through system resulted in a concentration-dependent induction of gill EROD. Crude oil (0.2 ppm) from the same oil field also proved to induce EROD. Finally, gill EROD activity in cod caged for 6 weeks at 500-10 000 m from two platforms outside Norway was measured. The activities in these fish were very low and did not differ from those in fish caged at reference sites.     

Possible chemical causes of skeletal deformities in natural populations of Aphanius fasciatus collected from the Tunisian coast

This study attempt to quantify and identify skeletal deformities in natural populations of Aphanius fasciatus collected from the Tunisian coast and tends to found a possible relationship between these anomalies and several types of pollutants presents in the environment. Fish were collected from one reference area (S1: coast of Luza) and three polluted areas (S2: industrialized coast of Sfax, S3: coast of Khniss and S4: Hamdoun’Oued). Various patterns of skeletal deformities were diagnosed using double staining technics, and the levels of heavy metals (Cd, Cu and Zn), various polycyclic aromatic hydrocarbons (PAHs) and estrogenic compounds were determined in water and sediment from different sites. Spatio-temporal variation of the spinal deformities frequencies in A. fasciatus show that high incidence of spinal anomalies has been recorded in population collected from S2 in comparison to three other ones. Morphological results indicated that skeletal deformities of adult A. fasciatus were grouped into 13 categories that described abnormalities on spines, vertebrae, arcs and mandibles. A total of 1025 abnormalities were quantified. The results of chemical analysis showed that the levels of heavy metals and PAHs were significantly higher in S2 than in S1, S3 and S4. High level of estrogenic activity was observed only in S4. A possible correlation between environmental exposures to a mixture of pollutants in coastal waters in S2 and spinal deformities in A. fasciatus was suggested.     

Heavy metals and PAHs in an open fishing area of the East China Sea: Multimedia distribution,source diagnosis,and dietary risk assessment

This study involved a comprehensive investigation of heavy metals and polycyclic aromatic hydrocarbons (PAHs) in seawater, sediments, and seafoods in an important open fishing area connecting China and Japan. Cu, Pb, Cd, Cr, As, and PAHs were detected in all the studied environmental media. Specifically, Pb and low molecular weight PAHs (LMW-PAHs) were observed at high levels, due to intensive ship activities and accidental crude oil spills. Additionally, source diagnosis of PAHs suggested a potential impact from combustion. Bioaccumulation factors (BAFs) and biota-sediment accumulation factors (BSAF) suggested moderate or low bioaccumulation potentials of the analytes from water and sediments, respectively. Noncarcinogenic risk assessment, based on target hazard quotients (THQs), indicated a potential low risk by consuming the studied seafoods, while the incremental lifetime carcinogenic risk assessment (ILCRs) suggested “acceptable risk” or “potential risk.” However, it should be noted that there have been possibilities that the carcinogenic risk could be elevated due to bioaccumulation of the PAHs in the seafoods after long-term exposure. In summary, it is necessary to evaluate the seafood security carefully in this open fishing area due to the potential health risks.

     

Identification of PAHs Sources in Bivalves and Sediments 5 Years After the Sea Prince Oil Spill in Korea

On 23 July 1995, the oil tanker Sea Prince ran aground near Sori Island, off the South Coast of Korea and spilled 5040 tons of crude and fuel oil into the marine environment. The effects of the Sea Prince oil spill on the marine environment have been investigated since 1996. The main objectives of this study were to find out the residual effects of beached oil and transport of dispersed oil into the subtidal area. Twenty-four PAHs were analyzed and principal component analysis was performed to elucidate weathering status, bioaccumulation pattern, and input sources. There were signs of bioconcentration of oil-derived PAHs in mussels of stranded oil remained sites. However, environmental factors overwhelmed these so that all the bivalves studied showed similar pattern in the last two sampling campaigns. There was no significant evidence of transport of oil-derived PAHs into the subtidal environment. However, one station showed an exceptionally high concentration (923 ng/g dry weight), which implies the limited input of particle-bound PAHs into this confined area.     

EROD activity and antioxidant defenses of sea bass (Dicentrarchus labrax) after an in vivo chronic hydrocarbon pollution followed by a post-exposure period

Chronic concentrations of polycyclic aromatic hydrocarbons (PAHs) have been commonly detected in international estuaries ecosystems. Reliable indicators still need to be found in order to properly assess the impact of PAHs in fish. After an in vivo chronic exposure to hydrocarbons, the enzymatic activity of 7-ethoxyresorufin O-deethylase (EROD) and the antioxidant defense system were assessed in sea bass, Dicentrarchus labrax. A total of 45 fish were exposed to the water-soluble fraction of Arabian crude oil, similar to a complex pollution by hydrocarbons chronically observed in situ, while 45 other control fish sustained the same experimental conditions in clean seawater. Fish samples were made after a 21-day exposure period and after a 15-day recovery period in clean fresh water. Throughout the experiment, liver EROD activity was significantly higher in contaminated fish than in control fish. In addition, nonenzymatic (total glutathione) and enzymatic (GPx, SOD, and CAT) antioxidant defense parameters measured in liver were not significantly different in fish. Furthermore, in gills, glutathione content had significantly increased while SOD activity had significantly decreased in contaminated fish compared to controls. On the other hand, CAT and GPx activities were not affected. Chronic exposure to PAHs disturbing the first step (SOD) and inhibiting the second step (GPx and CAT) could induce oxidative stress in tissues by the formation of oxygen radicals. After the postexposure period, there was no significant difference between control and contaminated fish in any of the antioxidant defense parameters measured in gills, attesting to the reversibility of the effects.     

Identification of PAHs Sources in Bivalves and Sediments 5 Years After the Sea Prince Oil Spill in Korea

On 23 July 1995, the oil tanker Sea Prince ran aground near Son Island, off the South Coast of Korea and spilled 5040 tons of crude and fuel oil into the marine environment. The effects of the Sea Prince oil spill on the marine environment have been investigated since 1996. The main objectives of this study were to find out the residual effects of beached oil and transport of dispersed oil into the subtidal area. Twenty-four PARs were analyzed and principal component analysis was performed to elucidate weathering status, bioaccumulation pattern, and input sources. There were signs of bioconcentration of oil-derived PAHs in mussels of stranded oil remained sites. However, environmental factors overwhelmed these so that all the bivalves studied showed similar pattern in the last two sampling campaigns. There was no significant evidence of transport of oil-derived PAHs into the subtidal environment. However, one station showed an exceptionally high concentration (923 ng/g dry weight), which implies the limited input of particle-bound PAHs into this confined area.     

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.