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Archives of Environmental Contamination and Toxicology (v.58, #1)

Chemical and Biological Characterization of Estrogenicity in Effluents from WWTPs in Ria de Aveiro (NW Portugal) by Ana Sousa; René Schönenberger; Niels Jonkers; Marc J.-F. Suter; Shinsuke Tanabe; Carlos M. Barroso (pp. 1-8).

Effluents from wastewater treatment plants (WWTPs) are responsible for the input of estrogenic contaminants into aquatic ecosystems, leading to widespread effects in wildlife. In the present work, levels of estrone (E1), 17α- and 17β-estradiol (E2), 17α-ethinylestradiol (EE2), bisphenol A (BPA), and nonylphenol (NP) were quantified in effluents from WWTPs located in Ria de Aveiro (NW Portugal), as well as in the final effluent discharged into the Atlantic Ocean through the S. Jacinto submarine outfall. Reference sites, located at the entrance of the estuarine system and at the seaside, were also included. Samples were collected under summer (June 2005) and winter (February 2006) conditions. For the summer survey samples, estrogenicity and androgenicity were evaluated using the yeast estrogen screen (YES) and the yeast androgen screen (YAS) assay. Estrone levels varied from 0.5 to 85 ng/L in the summer survey and between

A Survey of Metal and Pesticide Levels in Stormwater Retention Pond Sediments in Coastal South Carolina by Kevin D. Crawford; John E. Weinstein; Ronald E. Hemingway; Thomas R. Garner; Gavin Globensky (pp. 9-23).

During the summer of 2007, sediment samples were collected from 16 stormwater detention ponds and 2 reference ponds located in coastal South Carolina. The sediments were analyzed for more than 30 pesticides with current and historical uses, six polybrominated diphenyl ethers (PBDEs), and seven metals. The results are compared with established screening assessment parameters, with copper found to be the contaminant of highest concern. Lead levels were found to correlate well with pond drainage area, while copper and zinc levels correlated with both pond drainage area and pond surface area. Chlorpyrifos levels were found to correlate with pond surface area. Our results also show that ponds draining commercial areas were likely to have higher levels of zinc and lead in the sediments compared to other pond classes.

Comparison of Pb(II) Immobilized by Bone Char Meal and Phosphate Rock: Characterization and Kinetic Study by Shibao Chen; Yibing Ma; Li Chen; Liqun Wang; Haitao Guo (pp. 24-32).

Adsorption isotherms and kinetics of aqueous Pb(II) by bone char meal (BM) and phosphate rock (PR) were investigated using batch experiments. Pb removal was characterized and effects of pH and reaction time on Pb removal efficiency by BM/PR were also investigated. Results indicated that Pb removal efficiency by BM and PR is mainly controlled by dissolution of phosphatic components associated with apatite, followed by subsequent precipitation of geochemically stable pyromorphite [Pb₁₀ (PO₄)₆(OH,Cl)₂]. Sorption kinetics results indicated that Pb sorption onto BM or PR was generally biphasic, with initially fast reactions followed by slow and continuous reactions. Pb removal efficiency by BM and PR increased with increasing pH except at a pH higher than 6.15. Sorption isotherms showed that BM has a much higher Pb removal rate than PR in solution; the greater capability of BM than of PR to remove aqueous Pb indicates its potential as another promising way to remediate Pb-contaminated media.

Interactive Effects of 1,4-Dichlorobenzene and Heavy Metals on Their Sorption Behaviors in Two Chinese Soils by Fuhong Sun; Qixing Zhou (pp. 33-41).

Organic contaminants and toxic heavy metals often coexist in contaminated soils. However, little information concerning the interactive effects of organic contaminants and heavy metals on their sorption behaviors on soils is available. In this study, the competitive sorption effects of 1,4-dichlorobenzene (1,4-DCB) and typical heavy metals, cadmium (Cd) and copper (Cu), on their sorption characteristics on two Chinese soils were examined using a batch equilibration method. The results indicated that the coexistence of Cd and Cu inhibited the sorption of 1,4-DCB on soils and the inhibitive effects increased with increasing concentrations of Cd and Cu. This can be attributed to the inner-sphere complexation of Cd and Cu to form hydration shells of dense water that directly compete for surface sorption sites with 1,4-DCB. Additionally, pore blockage is presumed to be an alternative mechanism for the inhibitive effect. The extent of inhibitive effects depends on not only the relatively chemical characteristics of metals but also on the types of soils. However, the concurrence of 1,4-DCB (50 mg/L) in soils has little effects on the sorption capacity of Cd and Cu due to their higher sorption affinity for the soils than 1,4-DCB. These findings indicated that the coexistence of metals can greatly affect sorption behaviors of hydrophobic organic compounds on soils and thus their fate, transportation, and bioavailability.

Responses of Antioxidant Systems After Exposition to Rare Earths and Their Role in Chilling Stress in Common Duckweed (Lemna minor L.): A Defensive Weapon or a Boomerang? by M. P. Ippolito; C. Fasciano; L. d’Aquino; M. Morgana; F. Tommasi (pp. 42-52).

Extensive agriculture application of rare earth elements (REEs) in Far East countries might cause spreading of these metals in aquatic and terrestrial ecosystems, thus inducing a growing concern about their environmental impact. In this work the effects of a mix of different REE nitrate (RE) and of lanthanum nitrate (LA) on catalase and antioxidant systems involved in the ascorbate–glutathione cycle were investigated in common duckweed Lemna minor L. The results indicated that L. minor shows an overall good tolerance to the presence of REEs in the media. Treatments at concentrations up to 5 mM RE and 5 mM LA did not cause either visible symptoms on plants or significant effects on reactive oxygen species (ROS) production, chlorophyll content, and lipid peroxidation. Toxic effects were observed after 5 days of exposition to 10 mM RE and 10 mM LA. A remarkable increase in glutathione content as well as in enzymatic antioxidants was observed before the appearance of the stress symptoms in treated plants. Duckweed plants pretreated with RE and LA were also exposed to chilling stress to verify whether antioxidants variations induced by RE and LA improve plant resistance to the chilling stress. In pretreated plants, a decrease in ascorbate and glutathione redox state and in chlorophyll content and an increase in lipid peroxidation and ROS production levels were observed. The use of antioxidant levels as a stress marker for monitoring REE toxicity in aquatic ecosystems by means of common duckweed is discussed.

Response of Pistia stratiotes to Heavy Metals (Cr, Ni, and Zn) and Phosphorous by M. M. Mufarrege; H. R. Hadad; M. A. Maine (pp. 53-61).

The effects of Cr, Ni, Zn, and P exposure on the root anatomic structure, growth, and chlorophyll a concentration of Pistia stratiotes L. were studied. Plastic aquaria containing 50 g of wet plants and 5 L of pond water added with the contaminant(s) were disposed. The treatments were: (1) Cr, (2) Ni, (3) Zn, (4) P, (5) Cr + Ni + Zn, (6) Cr + Ni + Zn + P, and (7) control. Contaminant additions were done seven times. In each addition, concentrations of 1 mg of metals or 5 mg of P per liter of water were added. Chlorophyll a was an indicator more sensitive to Zn and Cr toxicity than the relative growth rate. Ni and Cr + Ni + Zn treatments were the most toxic ones, in which biomass and the root anatomical parameters (root length, cross-sectional areas [CSAs] of root, stele, and metaxylem vessels) decreased significantly. The addition of P to the treatment with combined metals attenuated the decrease in plant growth and root length, and caused a significant increase in CSAs of total metaxylem vessels, suggesting that P increased the tolerance of P. stratiotes to metals. This fact has important implications for the use of this macrophyte in constructed wetlands for industrial wastewater treatment.

Using Matrix Solid-Phase Microextraction (Matrix-SPME) to Estimate Bioavailability of DDTs in Soil to Both Earthworm and Vegetables by Hua Fang; Xiaoqiang Chu; Xiuguo Wang; Guohui Pang; Yunlong Yu (pp. 62-70).

This study was conducted to find an appropriate approach for the assessment of bioavailability of DDTs in soil to both earthworm and vegetables. Four chemical approaches—Soxhlet extraction with n-hexane, n-butanol agitation extraction, water agitation extraction, and matrix solid-phase microextraction (matrix-SPME)—were used to assess the relationships between the extractability of 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p′-DDE), 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethane (o,p′-DDT), 1,1-dichloro-2,2-bis(p-chlorophenyl) ethane (p,p′-DDD), and 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (p,p′-DDT) in soil and their amounts uptaken by the earthworm (Eisenia foetida), Chinese cabbage (Brassica campestris L. spp.), and cole (Brassica napus L.). These results indicated that the extractability and bioavailability of DDTs in soil decreased with time of aging. Correlation analysis showed that n-butanol extraction or 12-h matrix-SPME could be used to assess the bioavailability of DDTs to the earthworm, and Soxhlet extraction, n-butanol extraction, or 12-h matrix-SPME could be used to predict the bioavailability of DDTs to both Chinese cabbage and cole. As a solventless, time-efficient, and negligible-depletion technique, it could be concluded that matrix-SPME is a better approach to predict the bioavailability of DDTs to both the earthworm and vegetables, compared with Soxhlet extraction, n-butanol extraction, and water extraction.

Establishment and Succession of an Epibiotic Community on Chromated Copper Arsenate-Treated Wood in Mediterranean Waters by Hera Karayanni; Konstantinos Ar. Kormas; Simon Cragg; Artemis Nicolaidou (pp. 71-78).

Colonization and succession of an epibiotic animal community on chromated copper arsenate (CCA)-treated wood were studied for 18 months in the eastern Mediterranean (Saronikos Gulf, Aegean Sea). Pine wood panels, 200 × 100 × 25 mm, impregnated with CCA at retentions of 0, 12, 24, and 48 kg m⁻³ were used. The abundance or surface coverage of the most characteristic taxa (polychaetes, mollusca, crustacea bryozoa, sponges, ascidians) was measured in situ, while 12 months after submersion two panels of each retention were removed and examined in the laboratory. A total of 26 taxa were identified, among which polychaetes of the family Serpulidae dominated. The controls carried the largest number of species (17) but the lowest number of individuals. On panels with CCA retentions of 12 and 24 kg m⁻³, 14 and 16 species were observed, respectively, while at 48 kg m⁻³, only 9 species were found. Only the controls were affected by boring bivalves of the family Teredinidae and started to break up within 3 months of submersion. Statistically significant differences in barnacle and polychaete abundance were found between treated and untreated panels. There were no significant differences among panels treated at the three CCA loadings. Ordination by nonmetric multidimensional scaling showed a seasonal effect on the colonization of the treated panels, with the highest recruitment during the warmer months of the study.

Baseline Trace Metals in Seagrass, Algae, and Mollusks in a Southern Tyrrhenian Ecosystem (Linosa Island, Sicily) by Marcelo Enrique Conti; Beatrice Bocca; Marta Iacobucci; Maria Grazia Finoia; Mauro Mecozzi; Anna Pino; Alessandro Alimonti (pp. 79-95).

Trace elements were analyzed in organisms collected at five sampling stations along coastal areas of Linosa Island, Sicily (southern Tyrrhenian Sea, Italy). Concentrations of Cd, Cr, Cu, Pb, and Zn were measured in Posidonia oceanica L. Delile tissues, the two brown algae Padina pavonica (L.) Thivy and Cystoseira sp., and the two gastropod mollusks Monodonta turbinata Born and Patella caerulea L. Seawater samples were also collected at each site to assess soluble metal concentrations and to gain relevant information on their bioaccumulation ability. Data were processed by multivariate techniques, such as principal component analysis (PCA) and linear discriminant analysis on PCA factors. The scoreplots obtained showed that the pollutant distribution is more significantly correlated with species than with sites. For seaweeds, P. oceanica was associated with higher Cd, Cu, and Zn levels; Padina species had higher Cr concentrations, and Cystoseira had higher Pb levels. For mollusks, Monodonta species had high concentrations of Cu and Cr and Patella species were associated with Cd. Some general metal bioaccumulation patterns are described but no one sampling site was more contaminated than the others. The hypothesis of Linosa island serving as a reference ecosystem for baseline trace metal levels in southern Tyrrhenian areas is indeed supported by the statistical comparison among other southern Tyrrhenian ecosystems performed with Kruskall Wallis and Mann-Whitney tests. For P. oceanica leaves, P. pavonica, M. turbinata, and P. caerulea, this study confirms their usefulness as possible cosmopolitan biomonitors of trace metals in marine Mediterranean areas.

Acrosome Reaction of Sperm in the Mud Crab Scylla serrata as a Sensitive Toxicity Test for Metal Exposures by Ziping Zhang; Hua Cheng; Yilei Wang; Shuhong Wang; Fangjing Xie; Shaojin Li (pp. 96-104).

In order to test the sensitivity of the sperm cell of the mud crab Scylla serrata to heavy metals, the toxic effects of Ag⁺, Cd²⁺, Cu²⁺, and Zn²⁺ on the acrosome reaction (AR) were studied by artificially inducing the AR of sperm exposed to heavy metals, counting the AR rates by light microscopy, and observing structural changes in sperm by transmission electron microscopy. The AR in S. serrata occurs at two stages. The first stage (ARI) is the eversion of the subacrosomal material. The second stage (ARII) is the ejection of the acrosomal filament. The results showed the EC₅₀ values of the AR based on (ARI + ARII)% for Ag⁺, Cd²⁺, Cu²⁺, and Zn²⁺ were 10.02, 2.14, 13.69, and 2.21 μg/L, and the EC₅₀ values based on ARII % of Ag⁺, Cd²⁺, Cu²⁺, and Zn²⁺ were 1.96, 0.20, 1.46, and 0.34 μg/L. The order of toxicity is Cd²⁺ > Zn²⁺ > Cu²⁺ > Ag⁺ based on the percentage of reacted sperm at the second stage. Sperm cells exposed to heavy metals showed an increased rate of swelling, shape irregularities, and the acrosomal filament of some sperm cells was, crooked, ruptured, and even dissolved. The AR of the sperm cell from S. serrata is more sensitive to the tested heavy metals compared to sea urchin sperm cell toxicity tests.

Deposition and Air Concentrations of Permethrin and Naled Used for Adult Mosquito Management by Jerome J. Schleier III; Robert K. D. Peterson (pp. 105-111).

One of the most effective ways of managing adult mosquitoes that vector human and animal pathogens is the use of ultra-low-volume (ULV) insecticides. Because of the lack of environmental fate studies and concerns about the safety of the insecticides used for the management of adult mosquitoes, we conducted an environmental fate study after truck-mounted applications of permethrin and naled. One hour after application, concentrations of permethrin on cotton dosimeters placed at ground level 25, 50, and 75 m from the spray source were 2, 4, and 1 ng/cm² in 2007 and 5, 2, and 0.9 ng/cm² in 2008, respectively. One hour after application, concentrations of naled 25, 50, and 75 m were 47, 66, and 67 ng/cm² in 2007 and 15, 6.1, and 0 (nondetectable) ng/cm² in 2008, respectively. Deposition concentrations 12 h after application were not significantly different than 1 h after application for permethrin and naled either year. During 2007 and 2008 permethrin applications, two quantifiable air concentrations of 375 and 397 ng/m³ were observed 1 h after application. In 2007 and 2008, naled air concentrations ranged from 2300 to 4000 ng/m³ 1 h after application. There were no quantifiable air concentrations between 1 and 12 h after application in either 2007 or 2008 for both naled and permethrin. Environmental concentrations observed in this study demonstrate that models used in previous risk assessments were sufficiently conservative (i.e., the models overestimated environmental concentrations). However, we also demonstrate inadequacies of models such as AgDrift^® and AGDISP, which currently are used by the US Environmental Protection Agency to estimate environmental concentrations of ULV insecticides.

Evaluation of DNA Damage Induced by Environmental Exposure to Mercury in Liza aurata Using the Comet Assay by Carla Sofia Alves Pereira; Sofia Isabel Antunes Gomes Guilherme; Carlos Miguel Miguez Barroso; Luc Verschaeve; Mário Guilherme Garcês Pacheco; Sónia Alexandra Leite Velho Mendo (pp. 112-122).

Mercury (Hg) is one of the major aquatic contaminants even though emissions have been reduced over the years. Despite the relative abundance of investigations carried out on Hg toxicity, there is a scarcity of studies on its DNA damaging effects in fish under realistic exposure conditions. This study assessed the Hg genotoxicity in Golden grey mullets (Liza aurata) at Laranjo basin, a particularly contaminated area of Ria de Aveiro (Portugal) well known for its Hg contamination gradient. (1) Fish were seasonally caught at Laranjo basin and at a reference site (S. Jacinto), and (2) animals from the reference site were transplanted and caged (at bottom and surface), for 3 days, in two different locations within Laranjo basin. Using the comet assay, blood was analyzed for genetic damage and apoptotic cell frequency. The seasonal survey showed greater DNA damage in the Hg-contaminated area for all sampling seasons excluding winter. The temporal variation pattern of DNA lesions was: summer ≈ autumn > winter > spring. Fish caged at Laranjo also exhibited greater DNA damage than those caged at the reference site, highlighting the importance of gill uptake on the toxicity of this metal. No increased susceptibility to apoptosis was detected in either wild or caged fish, indicating that mercury damages DNA of blood cells by a nonapoptotic mechanism. Both L. aurata and the comet assay proved to be sensitive and suitable for genotoxicity biomonitoring in mercury-contaminated coastal systems.

Investigation of the Genotoxicity of Malathion to Freshwater Teleost Fish Channa punctatus (Bloch) Using the Micronucleus Test and Comet Assay by Ravindra Kumar; N. S. Nagpure; Basdeo Kushwaha; Satish K. Srivastava; W. S. Lakra (pp. 123-130).

Malathion [S-(1,2-dicarboethoxyethyl) O, O-dimethyl phosphorodithioate] is a widely used organophosphorus insecticide throughout the world. However, limited efforts have made to study its genotoxic effect in different fish tissues. The present investigation was aimed to assess the genotoxic potential of the pesticide to the freshwater teleost fish Channa punctatus at sublethal concentrations using the micronucleus test and comet assay. Initially, the 96-h LC₅₀ value of commercial-grade malathion (50% EC) was determined as 5.93 ppm in a semistatic system. Based on LC₅₀, three test concentrations (viz. sublethal I, sublethal II, and sublethal III) were determined to be 1.48, 0.74, and 0.59 ppm, respectively, and the fish specimens were exposed to these concentrations. Tissue samplings were done on days 0, 1, 3, 7, 15, 22 and 29 of malathion exposure for assessment of the induction of micronuclei (MN) frequency and DNA damage. The MN formation in the peripheral blood cells was found to be significantly higher (p < 0.05) in the treated specimens at all sampling intervals compared to the control. The MN frequency reached maximum on days 3 and 7 at sublethal I and II concentrations, respectively, followed by a nonlinear decline with the progression of the experiment. Similarly, significant effects (p < 0.05) of both concentration and time of exposure were observed on DNA damage in the gill, kidney, and lymphocytes. All of the tissues exhibited a concentration-dependent increase in DNA damage up to day 3, followed by a nonlinear decrease with the duration of exposure. A comparison of the extent of DNA damage among the tissues showed the sensitivity of gill tissue to malathion.

A Reproductive Toxicology Study of Phenanthrene in Medaka (Oryzias latipes) by Ching-Yi Horng; Hui-Chen Lin; Wenjau Lee (pp. 131-139).

Environmental contaminants, such as polycyclic aromatic hydrocarbons (PAHs), can disrupt the endocrine system and affect reproductive function of humans and wildlife. In this study, we exposed Japanese medaka (Oryzias latipes) to water-borne or food-spiked phenanthrene, an ubiquitous PAH, and investigated the chronic effects of the chemical on medaka reproduction. The results showed that phenanthrene significantly prolonged the time to hatch for embryos. Other parameters such as fecundity and fertility, organ-somatic ratios, hepatic vitellogenin production, and the histology of testes and ovaries were not different between the control and phenanthrene-treated groups. This suggests that the time to hatch in embryos might be a more sensitive biomarker for environmental contaminants.

Toxic Effects of Zinc on the Development, Growth, and Survival of Red Sea Bream Pagrus major Embryos and Larvae by Wei Huang; Liang Cao; Xiujuan Shan; Zhizhong Xiao; Qiyao Wang; Shuozeng Dou (pp. 140-150).

This study investigated the zinc toxicity to red sea bream Pagrus major embryos and larvae at 18 ± 1°C (33 ± 1‰ in salinity) under laboratory conditions. The acute toxicity tests indicated that zinc 48-h LC₅₀ to embryos and 96-h LC₅₀ to larvae were 4.3 (3.3–6.3; 95% confidence limits) and 10.1 (9.0–11.4) mg l⁻¹, respectively, suggesting that embryos were more sensitive than larvae to zinc exposure. The subchronic toxicity test, in which embryos and larvae were continuously exposed to 0, 0.1, 0.3, 0.5, 0.7, 1.0, 1.5, 2.0, and 2.5 mg Zn²⁺ l⁻¹ solutions for 10 days, demonstrated that waterborne zinc had distinctly toxic effects on the development, growth, and survival of red sea bream embryos and larvae. Zinc exposure at concentrations ≥ 0.5 mg l⁻¹ would lead to a low hatching rate (19–78%, vs. 98% in controls), high mortality (29–91%, vs. 10% in controls), and morphological abnormality (12–77%, vs. 0.3% in controls) in embryos and larvae, while it caused delay in time-to-hatch in embryos at concentrations ≥ 1.0 mg l⁻¹. These four biological parameters were zinc concentration dependent and could be effective bioindicators for evaluating the toxicity of zinc to the early life stage of this fish. Heartbeats of embryos (9–13 beats 10 s⁻¹) were relatively low and were not significantly influenced by zinc concentration, although they rose remarkably with elevated zinc concentration in larvae at the end of the test, particularly when it was ≥ 1.0 mg l⁻¹ (36–38, vs. 31 beats 10 s⁻¹ in controls). The total length (L_T) of the larvae at the end of the test was reduced by 12.2% and 15.6% in the 1.0 and 2.0 mg l⁻¹ solutions but did not vary significantly in other solutions in comparison with the controls. Heartbeat and L_T were less sensitive to zinc exposure and might not be good biological parameters for determining the toxicity of zinc to the early life stage of red sea bream.

Individual and Combined Effects of Heavy Metals on Serum Biochemistry of Nile Tilapia Oreochromis niloticus by Özgür Fırat; Ferit Kargın (pp. 151-157).

Changes in serum biochemistry in response to single- and combined-metal exposure were studied in a freshwater fish Oreochromis niloticus. Fish were exposed to 5.0 mg/L Zn, 1.0 mg/L Cd, and 5.0 mg/L Zn+1.0 mg/L Cd mixtures for 7 and 14 days to determine levels of biochemical parameters and metals in blood serum. The individual and combined effects of metals caused an increase in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and in levels of albumin, transferrin, ceruloplasmin, cortisol, glucose, and total protein, whereas they caused a decrease in cholesterol levels. At both exposure periods, increased ALT activity of fish exposed to Cd was higher compared with the Zn and Zn+Cd groups, respectively. The decreased cholesterol level was higher in the Cd alone, and for Cd in combination with Zn, than in Zn alone at 14 days. Zn or Cd levels increased in the blood serum of fish exposed to metals individually or in combination. When fish were exposed to the mixtures of Zn+Cd, concentrations of these metals in their serum were lower than in fish exposed to individual metals. One metal blocks or even antagonizes the gill epithelium absorption of the other and thereby limits the distribution of the metal in blood. The results indicate that biochemical parameters in fish blood can be used as an indicator of heavy-metal toxicity.

Influence of Sediment Contaminated with Untreated Pulp and Paper Mill Effluent on Winter Flounder, Pleuronectes americanus by R. A. Khan (pp. 158-164).

This study was conducted to ascertain the influence of sediment contaminated with pulp and paper mill effluent in a fjord on winter flounder, Pleuronectes americanus, based on a laboratory study. Flounder, captured from a pristine site, were exposed in a flow-through system for 16 weeks to sediment collected at 2, 5, 7, and 10 km from the outfall. A group of controls was placed in uncontaminated sediment. Mortality occurred almost exclusively in fish exposed to sediment taken from 2 km than from more distant sites. Additionally, the condition factor was lower, the liver was enlarged, and toxicopathic lesions in the liver and spleen were significantly greater in fish submerged in the sediment than in fish from the more distant locations or the controls. Two ectoparasites including a ciliate, Trichodina jadranica, and a monogenean, Gyrodactylus pleuronecti, were observed only in the control group, while a digenean in the digestive tract, Steringophorus furciger, was more abundant in fish exposed to sediment from sites more distant from the outfall and the controls than at 2 km. Comparison of these results with data from a previous gradient field study on biological variables in winter flounder, captured at 2, 5, 7, and 10 km down-current from the outfall, revealed an enlarged liver that was associated with elevated levels of detoxification of hepatic enzymes and prevalence of toxicopathic lesions in both the liver and the spleen; these were significantly greater in samples taken nearest to the outfall from the mill than at more distant sites. Moreover, two metazoan parasites, S. furciger (Digenea) and Echinorhynchus gadi (Acanthocephala), in the digestive were more abundant in samples taken at farther locations and also from the reference sites. These results, based on a laboratory study, are in agreement with previous observations that winter flounder exposed to sediment at the site nearest to the outfall, where high concentrations of toxic contaminants persisted, was greater than in the fish from the other locations.

Xenoestrogen Exposure and Effects in Bluegill from the Reedy River, South Carolina, USA by Pamela S. Truman; Peter van den Hurk (pp. 165-175).

Water quality in the Reedy River basin of Greenville, South Carolina, has been impacted by diverse and highly urbanized land uses. It has been demonstrated that urban runoff and point sources, such as effluent from wastewater treatment facilities, introduce organic pollutants and potentially endocrine disrupting compounds (EDCs) into the watershed. The objective of this study was to investigate the potential toxicological effects of EDCs that may be present in the Reedy River watershed using a set of biomarkers measured in indigenous fish to characterize the exposure and biological effects of these contaminants. Bluegill (Lepomis macrochirus) were collected during three different sampling seasons (spring, summer, and fall) from several sites along the length of the Reedy River and from an unimpacted site at Lake Robinson. Fish were analyzed for xenoestrogenic exposure (estrogenic effect of bile extracts) and effects (vitellogenin production in juvenile fish), which were compared to the hepatosomatic index as a general health parameter. Samples downstream of Greenville, especially downstream of the wastewater treatment facilities, were found to have significantly higher levels of estrogenic activity in bile extracts, which correlated well with elevated plasma vitellogenin concentrations relative to the specimens collected in reference sites. The results provide evidence that bluegill in the Reedy River were exposed to elevated concentrations of xenoestrogenic compounds and that these xenoestrogens were bioavailable, resulting in biological effects.

Trace Element Concentrations in Raillietina micracantha in Comparison to Its Definitive Host, the Feral Pigeon Columba livia in Santa Cruz de Tenerife (Canary Archipelago, Spain) by Jordi Torres; Pilar Foronda; Catarina Eira; Jordi Miquel; Carles Feliu (pp. 176-182).

The use of systems involving bird parasites as bioindicators of environmental pollution has been scarcely studied in comparison to other models involving fish and rodent parasites, which have been demonstrated as particularly adequate due to their bioaccumulation capacities. The present study evaluated the accumulation of nine trace elements in the cestode Raillietina micracantha and in its host Columba livia collected from the densely populated city of Santa Cruz de Tenerife (Canary Islands, Spain). Samples (kidney, liver, pectoral muscle, feathers, and R. micracantha) of 27 infected C. livia were selected for trace element analysis by inductively coupled plasma–mass spectrometry. Element levels in pigeon tissues revealed some degree of pollution in Santa Cruz de Tenerife, particularly by Pb and Zn. Pb and Mn mean concentrations were higher in R. micracantha than in the pigeon’s soft tissues, with subsequent high bioaccumulation factors for Pb (kidney = 15.38, liver = 10.38, muscle = 79.83) and Mn (kidney = 6.81, liver = 7.52, muscle = 19.89, feathers = 6.11), among others. The negative relations detected for As concentrations between liver and R. micracantha emphasize a possible role of the cestode in As detoxification in host tissues. Considering the obtained bioaccumulation factors, the system R. micracantha/C. livia is proposed as another promising bioindicator system to evaluate environmental toxic element exposure, particularly Pb and Mn, in areas where pollution levels are still relatively low and where both common species are present.

Polychlorinated Biphenyls and Biotransformation Enzymes in Three Species of Sea Turtles from the Baja California Peninsula of Mexico by K. L. Richardson; M. Lopez Castro; S. C. Gardner; D. Schlenk (pp. 183-193).

Concentrations of polychlorinated biphenyls (PCBs) as well as the expression patterns of cytochrome P450 (CYP) enzymes and glutathione-S-transferase (GST) activities were measured in livers of loggerhead (Caretta caretta), green (Chelonia mydas), and olive ridley (Lepidocheyls olivacea) sea turtles from the Baja California peninsula of Mexico. The mean concentrations of total PCBs were 18.1, 10.5, and 15.2 ng/g wet weight (ww) respectively for the three species and PCB 153 was the dominant congener in all samples. Total PCB concentrations were dominated by penta- and hexa-chlorinated biphenyls. The mean estimated TEQs were 42.8, 22.9, and 10.4 pg/g (ww) for loggerhead, green, and olive ridley, respectively, and more than 70% was accounted for by non-ortho PCBs. Western blots revealed the presence of hepatic microsomal proteins that cross-reacted with anti-CYP2K1 and anti-CYP3A27 antibodies but not with anti-CYP1A antibody. There were no significant differences in GST activities between species. Grouping congeners based on structure–activity relationships for CYP isoenzymes suggested limited activity of CYP1A contribution to PCB biotransformation in sea turtles. These results suggest potential accumulation of PCBs that are CYP1A substrates and provide evidence for biotransformation capacity, which differs from known animal models, highlighting the need for further studies in reptiles, particularly those threatened with extinction.

Effects of Exposure to Heavy Metals on Viability, Maturation, Fertilization, and Embryonic Development of Buffalo (Bubalus bubalis) Oocytes In Vitro by S. Nandi; P. S. P. Gupta; S. Selvaraju; S. C. Roy; J. P. Ravindra (pp. 194-204).

The aim of the present study was to examine the effect of heavy metals, cadmium and lead, on buffalo oocyte viability and in vitro development. Oocytes were aspirated from ovaries of slaughtered buffaloes. Only viable and metabolically active oocytes with more than three layers of cumulus cell layers and homogeneous ooplasm were selected. Effects of nine concentrations (0, 0.005, 0.05, 0.5, 1.0, 1.5, 2.5, 5, and 10 μg/mL) of cadmium or lead on buffalo oocyte viability, morphological abnormities, maturation, and embryonic development in vitro were studied. Oocytes were cultured for 24 h and then checked for viability (0.05% trypan blue staining for 2 min), morphological abnormalities, and reduction assay by MTT test in experiment 1. The doses of cadmium and lead causing 100% oocyte death (1-day culture) were determined (experiment 2). In experiment 3, viable oocytes were matured in vitro in media containing different levels of cadmium or lead and then inseminated in vitro with frozen-thawed spermatozoa, and the resultant cleaved embryos were cultured in a control embryo culture medium for 8 days. In experiment 4, oocytes were cultured in control oocyte maturation medium, then fertilized, and the resultant embryos were cultured in media containing different levels of cadmium or lead for 8 days. The number of cells in the trophectoderm and inner cell mass (ICM) and the total cell counts (TCN) of blastocysts derived by in vitro culture of two- to four-cell-stage embryos (produced in control medium) in media containing 0, 0.005, 0.05, 0.5, and 1.0 μg/mL of cadmium or lead were analyzed by differential staining technique (experiment 5). Cadmium and lead were found to have a dose-dependent effect on viability, morphological abnormities, maturation, cleavage and morula/blastocyst yield, and blastocyst hatching. A significant decline in viability of oocytes was observed at 1.0 mg/mL cadmium or lead compared to the control group. The doses of cadmium and lead causing 100% oocyte death (1-day culture) were 18 and 32 μg/mL, respectively. Cadmium and lead at 1.0 and 2.5 μg/mL, respectively, caused a significant reduction of maturation of oocytes compared to the lower concentrations. No cleavage or morulae/blastocysts were produced when the oocytes/embryos were cultured in media containing 2.5 and 5.0 mg/mL of either cadmium or lead, respectively. Similarly, no morulae/blastocysts were produced from cleaved embryos cultured in media containing 2.5 and 5.0 μg/mL cadmium and lead, respectively. The developmental block, degeneration, and asynchronous divisions were higher in embryos exposed to cadmium than in those exposed to lead. TCN and number of cells in ICM were significantly lower in blastocysts derived from two- to four-cell-stage embryos cultured in media containing heavy metals. In conclusion, cadmium and lead lowered the viability and development of buffalo oocytes but at a concentration higher than that estimated in the body fluids and environment. Cadmium was found to be more ovotoxic than lead.

Excretion of PFOA and PFOS in Male Rats During a Subchronic Exposure by Lin Cui; Chun-yang Liao; Qun-fang Zhou; Tong-mei Xia; Zhao-jun Yun; Gui-bin Jiang (pp. 205-213).

Perfluorinated compounds (PFCs), a class of synthetic surfactants that are widely used, have become global environmental contaminants because of their high persistence and bioaccumulation. An increasing number of studies have described the pharmacokinetics of PFCs following in vivo exposure, however, few papers have focused on the excretion of these compounds during a period of consecutive exposure. In this study, the excretions of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) in male Sprague–Dawley rats gavaged consecutively for 28 days were investigated and compared. The faster elimination rate in urine compared to feces indicated that urinary excretion is the primary clearance route in rats for either PFOA or PFOS. During the first 24 h after administration of PFOA (5 and 20 mg/kg body weight/day), about 24.7–29.6% of the oral dose was excreted through urine and feces, while for PFOS, the excretion amounts were only 2.6–2.8% of the total gavaged doses (5 and 20 mg/kg body weight/day). The excretion rates of both PFCs increased with increasing exposure doses. The higher elimination rate of PFOA through excretion indicated its lower accumulation in rats, thus inducing possible lower toxicities compared to PFOS.

Intake and Potential Health Risk of Polycyclic Aromatic Hydrocarbons Associated with Seafood Consumption in Korea from 2005 to 2007 by Hyo-Bang Moon; Hye-Seon Kim; Minkyu Choi; Hee-Gu Choi (pp. 214-221).

Concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) were measured in 26 types of seafood commonly consumed by the general Korean population. The concentrations of total and carcinogenic PAHs ranged from 12.3 to 243 ng/g dry weight and from 0.21 to 18.4 ng/g dry weight, respectively, similar to those reported in other countries. The lower-molecular–weight aromatics, such as naphthalene, phenanthrene, and fluorine, predominated in the seafood samples. Estimated intakes of PAHs by seafood consumption for the general population, for men, and for women were 15.3, 16.7, and 13.8 ng/kg body weight/d, respectively. Squid, mackerel, and yellow croaker contributed to the highest intakes of PAHs. Among the 8 age groups investigated, children age <2 years and 3 to 6 years had high intakes of PAHs compared with adult groups. Dioxin toxic equivalents (TEQs) of PAHs were calculated using a TEQ proposed in the literature, and the intake was estimated to be 0.91 pg TEQ/kg body weight/d, which indicates that PAHs contributed the most (43%) to total TEQ intake. The estimated excess cancer risk values from seafood consumption for the general population (2.85 × 10⁻⁶), for men (2.93 × 10⁻⁶), and for women (2.68 × 10⁻⁶) slightly exceeded the guideline value (1.0 × 10⁻⁶) for potential cancer risk.

Effects of Repeated Seafood Consumption on Urinary Excretion of Arsenic Species by Volunteers by Byung-Sun Choi; Seong-Jin Choi; Dong-Won Kim; Mingai Huang; Na-Young Kim; Kyung-Su Park; Choong-Yong Kim; Hyo-Min Lee; Young-Na Yum; Eui-Sik Han; Tae-Seok Kang; Il-Je Yu; Jung-Duck Park (pp. 222-229).

Arsenic (As) is a known human carcinogen and widely distributed in the environment. The main route of As exposure in the general population is through food and drinking water. Seafood harvested in Korea contains high-level organoarsenics such as arsenobetaine, arsenocholine, and arsenosugars, which are much less harmful than inorganic arsenics. However, for those who eat large amounts of seafood it is important to understand whether seafood consumption affects urinary levels of inorganic As metabolites such as arsenite, arsenate, monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA). In this study we investigated urinary As metabolites (inorganic As, MMA[V], DMA[V]) and some biological indexes such as AST, GSH, GPX, lipid peroxidation, and uric acid in volunteer study subjects (seven males and nine females). Total urinary As metabolites were analyzed by the hydride generation method, followed by arsenic speciation using HPLC with ICP-mass spectrometry. Study subjects refrained from eating seafood for 3 days prior to the first urine collection and then ingested seafood daily for 6 consecutive days. The first voided urine of the morning was collected from each subject the first day of the consecutive 6 days of seafood ingestion but prior to the first seafood meal. The first voided urine of the morning was also collected on days 1, 2, 3, 4, 5, 6, 7, 10, and 14 after seafood ingestion. The daily mean intake of total As was 6.98 mg, comprised of 4.71 mg of seaweed (67%), 1.74 mg of flat fish (25%), and 0.53 mg of conch (8%). We observed a substantial increase in total urinary As metabolites for subjects consuming seafood from day 1, which recovered to control level at day 10. The increase in total urinary As metabolites was attributed to the increase in DMA, which is a more harmful metabolite than organoarsenics. However, no significant changes in response biological indexes were observed. These results suggest that it is necessary to evaluate As metabolism when assessing the exposure to inorganic As and potential chronic health effects of seafood consumption in Korea.

Comparison of Organochlorine Pesticide Levels in Human Adipose Tissue of Inhabitants from Veracruz and Puebla, Mexico by Stefan M. Waliszewski; R. Valencia Quintana; C. A. Corona; M. Herrero; K. Sánchez; H. Aguirre; I. A. Aldave; S. Gomez Arroyo; R. Villalobos Pietrini (pp. 230-236).

Since the discovery of insecticide properties of DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) and HCH (hexachlorocyclohexane), they have provided great benefits to humans in sanitary actions to combat the spread of infection-borne disease vectors. Public Health Programs in Mexico used DDT and HCH until 1999 as the insecticides of choice to control disease-transmitting organisms. Because of their persistence and accumulative properties, organochlorine pesticides bioconcentrate in lipids of the human body, reflecting the rate of environmental exposure. Eighty human abdominal adipose tissue samples from Veracruz and 80 samples from Puebla were analyzed and the obtained results were compared among both populations. The results from Veracruz showed higher contamination levels (mg/kg on lipid base) compared to Puebla: β-HCH, 0.072 vs. 0.029; pp′DDE (Dichlorodiphenyldichloroethylene), 2.364 vs. 0.726; op’DDT, 0.022 vs. 0.025; pp′DDT, 0.192 vs. 0.061; and Σ-DDT, 2.589 vs. 0.806. The population from Veracruz and from Puebla divided by sex, origin, and cause of death presented no statistical differences. The comparison between sexes (women and men groups) at Veracruz and Puebla indicated significantly higher levels in Veracruz and statistical significant differences. Calculating possible risks (odds ratios, OR), pp′DDE (OR = 5.04) and op′DDT (OR = 2.93) revealed significantly higher risk for the Veracruz population. The study indicated prolonged DDT exposure of Mexicans caused by the past sanitary use and persistence of its residues in soils and air.

Effects of Dietary Selenomethionine on Cutthroat Trout (Oncorhynchus clarki bouvieri) Growth and Reproductive Performance Over a Life Cycle by Ronald W. Hardy; Libbie L. Oram; Gregory Möller (pp. 237-245).

A 2.5-year feeding trial was conducted in which cutthroat trout (Oncorhynchus clarki bouvieri) were fed either a basal diet (1.2 μg Se/g diet) or the basal diet supplemented with 2, 4, 6, 8, or 10 μg selenomethionine/g diet from 1 g weight to maturation. After 44 weeks of feeding, a subsample of fish was removed from dietary treatment groups and fed the basal diet for an additional 32 weeks. Concentrations of Se in whole fish and eggs increased in proportion to dietary Se intake, but no differences in growth, feed intake, survival, or egg hatchability were observed among dietary groups. Cranial–facial deformities in second-generation offspring were less than 6% in all treatment groups except for fish fed the diet supplemented with 4 μg selenomethionine/kg, for which a 9.2% incidence was observed. Fish switched from selenomethionine-supplemented diets to the basal diet lost Se, calculated as μg Se lost/g weight gain, at 1.01, 2.84, 4.42, and 4.42 for dietary treatment groups 3, 4, 5, and 6, respectively. Results suggest no toxicity of dietary selenomethionine up to 10 μg/g supplemented diet and that with total life-cycle exposure, cutthroat trout increase Se excretion to maintain whole-body concentrations below toxic levels.

Erratum to: Effects of Dietary Selenomethionine on Cutthroat Trout (Oncorhynchus clarki bouvieri) Growth and Reproductive Performance Over a Life Cycle by Ronald W. Hardy; Libbie L. Oram; Gregory Möller (pp. 246-246).

Erratum to: Biochemical Response of the Copepod Tigriopus japonicus Mori Experimentally Exposed to Cadmium by Ming-Hua Wang; Gui-Zhong Wang (pp. 247-247).

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Archives of Environmental Contamination and Toxicology (v.58, #1)