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Archives of Environmental Contamination and Toxicology (v.58, #3)

A Preliminary Assessment of Polycyclic Aromatic Hydrocarbon Distribution in the Kenting Coral Reef Waters of Southern Taiwan by Jing-O. Cheng; Ying-Ming Cheng; Te-Hao Chen; Ping-Chieh Hsieh; Meng-Der Fang; Chon-Lin Lee; Fung-Chi Ko (pp. 489-498).

This distribution and variation of 45 polycyclic aromatic hydrocarbons (PAHs) in the waters of Kenting coral reefs were investigated by using sensitive research-grade sampling and analytical methods. The concentration levels of total PAHs (including particulate and dissolved PAHs) in the Kenting waters were relatively low, ranging from 2.2 to 34.4 ng/L. There was no significant spatial difference in PAH concentrations among the sampling stations, even between the surface and bottom waters. The fraction of dissolved PAHs on average was greater than that of particulate PAHs due to the small amount of total suspended particles. The partition coefficients between the particulate and dissolved phase (log K _oc) were positively correlated with their log K _ow values (r ² = 0.67, n = 80), except for the more hydrophobic PAHs with log K _ow values greater than 5.91. Results of principal component analysis indicate that the patterns of particulate PAH compositions were similar through the Kenting waters in the fall and winter, whereas there was a significant spatial difference between the spring and summer samples, which might be affected by a combination of land runoff and coastal currents. PAHs along the Kenting coasts were dominated by low-molecular PAHs, which have higher solubility than high-molecular PAHs. Based on isomer ratios, analysis of the source of PAHs in the Kenting coastal waters demonstrated that the PAHs come mostly from petroleum and, to a lesser degree, from combustion.

Effect of Alteration Zones on Water Quality: A Case Study from Biga Peninsula, Turkey by Alper Baba; Orhan Gunduz (pp. 499-513).

Widespread and intense zones of silicified, propylitic, and argillic alteration can be found in the Çan volcanics of Biga Peninsula, northwest Turkey. Most of the springs in the study area surface out from the boundary between fractured aquifer (silicified zone) and impervious boundary (argillic zone). This study focuses on two such springs in Kirazlı area (Kirazlı and Balaban springs) with a distinct quality pattern. Accordingly, field parameters (temperature, pH, and electrical conductivity), major anion and cation (sodium, potassium, calcium, magnesium, chloride, bicarbonate, and sulfate), heavy metals (aluminum, arsenic, barium, chromium, cobalt, cupper, iron, lithium, manganese, nickel, lead, and zinc), and isotopes (oxygen-18, deuterium, and tritium) were determined in water samples taken from these springs during 2005 through 2007. The chemical analyses showed that aluminum concentrations were found to be two orders of magnitude greater in Kirazlı waters (mean value 13813.25 μg/L). The levels of this element exceeded the maximum allowable limits given in national and international standards for drinking-water quality. In addition, Balaban and Kirazlı springs are >55 years old according to their tritium levels; Kirazlı spring is older than Balaban spring. Kirazlı spring is also more enriched than Balaban spring based in oxygen-18 and deuterium values. Furthermore, Kirazlı spring water has been in contact with altered rocks longer than Balaban spring water, according to its relatively high chloride and electrical conductivity values.

Distribution of Organochlorine Pesticides in Intertidal and Subtidal Sediments in Coastal Wetland with High Tidal Ranges by Seung-Kyu Kim; Dong-Jin Kang; Kyung-Ryul Kim; Dong Soo Lee (pp. 514-522).

The present study aimed to understand the distribution characteristics of organochlorine pesticides (OCPs) in wetlands experiencing high tides and the manner in which these characteristics are affected by various factors, in particular, two distinct physical and topographical features (i.e., sub- and intertidal zones). For all OCPs except HCHs, the distribution levels were higher in the intertidal zone than in the subtidal zone. The spatial heterogeneity in the isomer compositional pattern, distribution levels, and correlation among individual OCPs were pronounced in the intertidal zone. Spatial homogeneity was observed within the subtidal zone, indicating that the effect of flushing and mixing was strong enough to diminish the potential local concentration peaks and unique composition pattern. It was evident that input paths and their strength impact the horizontal and transversal distribution of OCPs. The OCP group-specific discrepancy in spatial distribution suggested that (1) chlordane and chlorobenzenes were from a single dominant innermost terrestrial input path, (2) DDTs were from multiple terrestrial input paths, and (3) HCH was likely to be from the outer sea. The observations in this study imply that (1) benthic organisms could experience greater exposure in the intertidal basin than in the subtidal zone and (2) management measures of OCPs should be set after considering the tidal effect and the OCP-specific input paths.

Comparison of Polycyclic Aromatic Hydrocarbons Levels in Sludges from Municipal and Industrial Wastewater Treatment Plants by N. Kamil Salihoglu; Güray Salihoglu; Yücel Tasdemir; S. Sıddık Cindoruk; Didem Yolsal; Ruken Ogulmus; Gizem Karaca (pp. 523-534).

This study was carried out to investigate the concentrations and potential sources of the polycyclic aromatic hydrocarbons (PAHs) in sludge of 14 wastewater treatment plants (WWTPs). Sludge samples were collected from 2 municipal WWTPs, 11 industrial WWTPs, and 1 sanitary landfill leachate treatment plant within the city of Bursa, Turkey during the summer of 2008. Ultrasonication was applied for extraction and gas chromatography–mass spectrometry was used to analyze the PAH contents of the samples. Twelve of the 16 EPA-listed PAH compounds were determined. Total PAH concentrations (∑₁₂ PAHs) determined in all of the sludge samples ranged from 1,781 to 19,866 μg/kg dry matter (dm). The sum of 8 of the 11 EU PAHs varied between 1,481 and 17,314 μg/kg dm, and 3 of the samples exceeded the proposed EU limit for land application. One of the automotive industry sludges contained the highest level of PAHs, followed by one of the municipal sludges. The average sum of 5- and 6-ring PAH compounds in all of the sludge samples amounted to almost 65% of the total PAHs. The diagnostic ratios of specific PAHs were calculated to determine the dominant sources for the PAHs in the sludge samples.

In Vitro Cyclooxygenase Inhibition Assay for Evaluating Ecotoxicity of the Surface Water and Domestic Wastewater in the Tone Canal, Japan by Iwaki Nishi; Takaaki Komuro; Tsuyoshi Kawakami; Sukeo Onodera (pp. 535-542).

Cyclooxygenase (COX) plays an important role in eicosanoid metabolism. Nonsteroidal anti-inflammatory drugs (NSAIDs) function as COX inhibitors and are frequently detected in the aquatic environment. Here, we measured the in vitro COX-inhibiting activity of the surface water and domestic wastewater in the Tone Canal, Japan. The concentrations of several NSAIDs in the some samples were also determined using gas chromatography–tandem mass spectrometry for confirming the validity of the assay. The target compounds were extracted from the samples using a solid-phase extraction cartridge. A dose–response relationship between the inhibiting activity and sample volume were observed in the wastewater sample. The higher COX-inhibiting activities were observed in the wastewater sample, as compared with the samples of the surface water in the canal. These inhibiting activities reflected the trends of NSAIDs distribution in the canal. However, the inhibiting activities of the water samples could not be entirely explained by the NSAIDs that were selected for instrumental analysis in this study. Other compounds that were not measured by instrumental analysis in this study might contribute to the inhibiting activities. Therefore, the COX-inhibiting assay would be effective for evaluating inclusive ecotoxicity in the aquatic environment.

Perchlorate in Tap Water, Groundwater, Surface Waters, and Bottled Water From China and its Association with Other Inorganic Anions and with Disinfection Byproducts by Qian Wu; Tao Zhang; Hongwen Sun; Kurunthachalam Kannan (pp. 543-550).

Perchlorate is a potent thyroid hormone–disrupting compound. Drinking water is one of the major sources of human exposure to perchlorate. Little is known about the occurrence of perchlorate in waters from China. In this study, water samples (n = 300) collected from 15 locations in 13 provinces and municipalities were analyzed for the presence of perchlorate. In addition, other inorganic anions that commonly occur in water—iodide, bromide, and nitrate—and the disinfection byproducts, bromate, chlorate, and chlorite were determined by high-performance liquid chromatography interfaced with tandem mass spectrometry. Perchlorate was detected in 86% of the samples analyzed, at concentrations ranging from <0.02 to 54.4 μg l⁻¹ (mean ± SD 2.20 ± 6.39 μg l⁻¹; median 0.62 μg l⁻¹). Mean concentrations of perchlorate in tap water, groundwater, surface waters, and bottled water were 2.46, 3.04, 2.82, and 0.22 μg l⁻¹, respectively. Significant positive correlations were found between the concentrations of perchlorate and nitrate, perchlorate and chlorate, bromide and iodide, and nitrate and iodide.

Surface Water Concentrations and Loading Budgets of Pharmaceuticals and Other Domestic-Use Chemicals in an Urban Watershed (Washington, DC, USA) by Lirije Shala; Gregory D. Foster (pp. 551-561).

Pharmaceuticals and domestic-use chemicals (PDCs) are classes of emerging chemical contaminants thought to enter the aquatic environment primarily through wastewater treatment plant (WTP) discharges. The intent of this study was to quantify loadings of PDCs in an urban watershed. The watershed has two major branches but with wastewater discharge occurring in only one of the two major branches. Surface water from the Anacostia River (Washington, DC) was collected in base-flow and storm-flow regimes. Surface water was filtered to separate water and particles, and the PDCs were extracted from water with Oasis HLB solid-phase extraction cartridges and extracted from sediments using microwave-assisted extraction. The PDCs in the extracts were quantified using gas chromatography–mass spectrometry in the form of the trimethylsilyl ether derivatives. The most frequently detected PDC with the highest concentration was bisphenol-A in both branches of the Anacostia River watershed and the least frequently detected PDC was diclofenac. The overall median concentrations for all measured PDCs in surface water ranged from nondetectable to 54.9 ng/l. Alternatively, in the collected WTPs samples, naproxen was observed, with the highest concentration and the median concentrations in WTP effluent ranging from nondetectable to 276 ng/l. Estimates of PDC loadings for February 2006 from WTP effluent showed that <2% of the downstream load in the NE Branch was derived from WTP discharge. PDC sources other than WTP effluent appear to influence surface water concentrations in the urban Anacostia River watershed.

Impact of Urban Wastewater Discharges on the Sediments of a Small Mediterranean River and Associated Coastal Environment: Assessment of Estrogenic and Dioxin-like Activities by A. David; E. Gomez; S. Aït-Aïssa; D. Rosain; C. Casellas; H. Fenet (pp. 562-575).

The Mediterranean region includes many small coastal rivers about which little is known concerning organic contaminant loads in their sediment. This study was designed to assess organic contamination in one of these small coastal rivers (Lez River) and associated coastal sediments. Levels of alkylphenols (APs), polycylic aromatic hydrocarbons (PAHs), and polychlorinated biphenyls (PCBs) were measured in sediments of the Lez River and two coastal lagoons impacted by wastewater discharges. In parallel, sediment surrounding a recently built submarine outfall that discharges treated wastewater, from an area encompassing some 450,000 inhabitants, into the sea was monitored a year after the beginning of emission via the outfall. Finally, these sediments were characterized by screening estrogenic, PAH-like and dioxin-like activities using in vitro bioassays. Both chemical analyses and bioassays revealed that wastewater inputs were a source of organic contamination of sediments from the Lez and lagoons, which still persisted 2 years after the discharges were stopped. APs could explain a small proportion of the overall estrogenic activities (up to 31%), suggesting that other estrogenic compounds were also present in the sediments. PAHs explained a great share (83% on average) of the EROD induction potency of the extracts. This survey should be the first step in the long-term monitoring of these sites.

In Vitro and Immunological Assessment of the Estrogenic Activity and Concentrations of 17β-Estradiol, Estrone, and Ethinyl Estradiol in Treated Effluent from 45 Wastewater Treatment Plants in Victoria, Australia by M. Allinson; F. Shiraishi; S. A. Salzman; G. Allinson (pp. 576-586).

The project was conducted between May 2006 and September 2007, and involved the collection of effluent samples from 45 wastewater treatment plants (WWTPs). The 45 WWTPs included 16 lagoon-based plants and 29 with activated sludge-based processes. Permission was obtained from all the relevant water authorities to collect samples of final effluent at point of discharge to the environment, whether that was to a creek, a river, the ocean, or the land. Samples were collected on two occasions, namely, in August 2006 (winter) and late February–early March 2007 (summer), and subjected to a number of biological and chemical analyses, including toxicity tests, measurement of hormonal (estrogenic) activity using yeast-based bioassays, and measurement of specific hormonal concentrations using enzyme-linked immunosorbent assays (ELISAs). Almost all of the effluents examined showed estrogenic activity: in winter, no activity to 73 ng/l 17β-estradiol equivalents (EEQ); and in summer, no activity to 20 ng/l EEQ. On the whole, the levels of estrogenic activity observed were comparable with the range recently reported in Australia and New Zealand using human estrogen receptor-based assays (“not detected” to ~10 ng/l EEQ). The low/no bioassay response was confirmed by the chemical assessment of estradiol, estrone, and ethinyl estradiol concentrations by ELISA, which returned concentrations of these compounds for the most part below 10 ng/l.

Investigation of the Acute Toxic Effect of Chlorpyrifos on Pseudomonas putida in a Sterilized Soil Environment Monitored by Microcalorimetry by Huilun Chen; Jun Yao; Fei Wang; Yong Zhou; Ke Chen; Rensheng Zhuang; Gyula Zaray (pp. 587-593).

Chlorpyrifos (CPF) is moderately persistent in soils. In our study, microcalorimetry was introduced for the first time to explore the acute toxic effect of CPF on a Pseudomonas strain in sterilized soil. Firstly, it was determined by microcalorimetry that P. putida failed to degrade CPF. Then the acute toxicity of increasing concentrations of CPF to P. putida was determined by its temporal effects on metabolism and counts of colony forming units. Results revealed that the increase of CPF concentration could induce a decrease of the growth rate constant (k) and the total thermal effect (Q _T), representing an inhibiting action on P. putida. In addition, the colony forming units (CFU) for P. putida were counted. Results showed that the number of P. putida decreased with increasing CPF dose after 18 h of incubation in sterilized soil. Interestingly, the trend of the number of CFU was similar to the growth rate constant k, whereas the trend became irregular after 36 h of incubation. This indicated that P. putida resisted and also expresses high metabolic activity during the exponential growth phase of 18 h; thereafter the microorganisms showed a certain adaptation, even declining in number and activity.

Changes in Growth, Biochemical Components, and Antioxidant Activity in Aquatic Plant Wolffia arrhiza (Lemnaceae) Exposed to Cadmium and Lead by Alicja Piotrowska; Andrzej Bajguz; Beata Godlewska-Żyłkiewicz; Elżbieta Zambrzycka (pp. 594-604).

The present study investigated the biochemical response of aquatic plant Wolffia arrhiza (Lemnaceae) treated with lead (Pb) and cadmium (Cd) at a range of concentrations from 1 to 1000 μM. W. arrhiza has been identified as good scavenger of heavy metals from aqueous solution. Pb and Cd accumulation was found to be increased in a concentration- and duration-dependent manner. However, the highest biosorption of heavy metals was found in plants exposed to low levels (10 μM) of Cd and Pb in the nutrient medium. In observing the response to heavy-metal stress, we noted inhibited plant growth and decreased photosynthetic pigments, monosaccharides, and proteins. In addition, Cd was found to be more toxic to plants than Pb. Heavy metals also induced oxidative damage as evidenced by increased lipid peroxidation and hydrogen peroxide levels. In contrast, the deleterious effects resulting from the cellular oxidative state can be alleviated by enzymatic (catalase, ascorbate peroxidase, nicotinamide dinucleotide [NADH] peroxidase) and nonenzymatic (ascorbate, glutathione) antioxidant mechanisms activated in W. arrhiza plants exposed to Cd and Pb, especially at 10 μM. These results suggest that W. arrhiza is a promising bioindicator of heavy-metal toxicity.

Effects of Herbicides on Lemna gibba and Recovery from Damage After Prolonged Exposure by M. Mohammad; K. Itoh; K. Suyama (pp. 605-612).

To determine the potential impact of contaminants on the aquatic vascular plants Lemna sp., toxicity tests are usually conducted for a 4- to 14-day exposure, and the toxicity is usually expressed as EC50. However, the effects of longer exposure and the recovery potential after exposure to chemicals are other important factors which should be considered. We present the relative risks of a variety of exposure scenarios and recovery potentials from damage, using herbicides with different modes of action. Toxicity was assessed on the basis of both EC50 and relative growth rate (RGR) compared with untreated controls in exposure and recovery. The EC50 of atrazine was found to be 89 ppb, and its phytostatic concentrations were 1600 and 800 ppb for xposure periods of 14 and 28 days, respectively, and no phytocidal effects were observed up to 3200 ppb for a 28-day exposure. The RGR in recovery was not affected by the RGR in exposure, and regrowth was possible even after complete inhibition of growth for 28 days at the highest concentration tested. Alachlor, with an EC50 of 31 ppb, was phytostatic at 400 ppb for a 14-day exposure and phytocidal at 200 ppb for 21- and 28-day exposures. Paraquat, with an EC50 of 31 ppb, showed phytocidal rather than phytostatic effects. All phytostatic fronds could not grow in the recovery period, and the phytocidal concentration decreased with exposure period, from 80 ppb for a 7-day exposure to 20 ppb for 21- and 28-day exposures. The RGR of alachlor and paraquat in recovery was dependent on the RGR in exposure. In the case of cyclosulfamuron, phytostatic concentrations were 100 and 50 ppb for 7- and 14-day exposures, respectively. In the case of exposures longer than 21 days, however, it exhibited phytocidal activity at 10 ppb. The results of this study suggest that it is important to examine the effects of chemicals over a longer exposure period as well as the recovery potential from damage for reliable ecological risk assessment.

Phytotoxicity and Accumulation of Lead in Australian Native Vegetation by Dane T. Lamb; Hui Ming; Mallavarapu Megharaj; Ravi Naidu (pp. 613-621).

Lead (Pb) is a wide spread contaminant in the terrestrial landscape. It is highly detrimental to plant and animal life and possesses no known biologic function. Yet there is a paucity of reliable information available on the response of Australian and other plant species to Pb exposure at phytotoxic doses. In this study, the response of three Australian native grass species and two tree species to Pb in nutrient solution culture was investigated. Plants were exposed to average Pb concentrations ranging from 0.020 to 15.2 μM. The plant species included Acacia decurrens, Austrodanthonia richardsonii, Bothriochloa macra, Eucalyptus camaldulensis, and Dichanthium sericeum. Few foliar symptoms were evident in any plant species, although some discolouration in young leaves of E. camaldulensis was evident from 1 μM, and B. macra showed pronounced reddening at the highest treatments. The most tolerant plant species studied based on solution EC_{50, roots} (μM) results was B. macra (7.0 ± 0.2), followed by A. decurrens (3.9 ± 0.2), D. sericeum (2.9 ± 0.3), E. camaldulensis (1.1 ± 0.3), and A. richardsonii (0.4). A hazardous concentration value (HC₅) (n = 9) for soil solution was estimated to be 0.16 μM. A. richardsonii was highly sensitive to Pb and possessed little ability to restrict Pb translocation to its shoots. B. macra was able to tolerate high root (3924 mg kg⁻¹) and shoot (743.0 mg kg⁻¹) Pb concentrations. A. decurrens excluded Pb from it shoots. The high tolerance of A. decurrens to Pb and limited translocation to shoots indicates it may be useful in revegetation of Pb-contaminated soils.

Effects of Nutrient Pre-Exposure on Atrazine Toxicity to Vallisneria americana Michx. (Wild Celery) by Darrin D. Dantin; Ronald G. Boustany; Michael A. Lewis; Stephen J. Jordan; Rebecca F. Moss; Thomas C. Michot (pp. 622-630).

Accelerated eutrophication is common to many freshwater and marine environments and often co-occurs with the presence of anthropogenic chemicals. However, the toxic effects of common chemical stressors such as herbicides in the presence of elevated nutrients are not well understood for most aquatic flora, particularly vascular species. To provide insight, field-collected Vallisneria americana Michx. (wild celery) were sequentially exposed to three nutrient concentrations for 3 months and then to nominal 11 and 110 μg L⁻¹ atrazine for 96 h. Nutrient concentrations (combined NH₄ ⁺, NO₂ ⁻, NO₃ ⁻, PO₄ ⁻) were based on ambient concentrations in the St. Johns River (FL) and ranged from 0.013 to 0.668 mg L⁻¹. Nutrient pretreatment potentiated the toxicity of atrazine as determined by chlorophyll fluorescence activity. Electron transport rates (ETR) were significantly less (48–59%) for plants pretreated with low and ambient nutrient levels in the presence of an average of 107.5–128.1 μg L⁻¹ atrazine. Significant ETR reductions were also observed for plants exposed to an average of 11.4 μg L⁻¹ atrazine after exposure to nutrients three times the ambient concentration in the St. Johns River. The results indicate the importance of considering the presence of nutrients in chemical hazard assessments, particularly for phytotoxicants and nontarget vascular plants.

Levels and Sources of PAHs in Selected Sites from Portugal: Biomonitoring with Pinus pinea and Pinus pinaster Needles by Nuno Ratola; José Manuel Amigo; Arminda Alves (pp. 631-647).

Pine needle samples from two pine species (Pinus pinaster Ait. and Pinus pinea L.) were collected at 29 sites scattered throughout Portugal, in order to biomonitor the levels and trends of 16 polycyclic aromatic hydrocarbons (PAHs). The values obtained for the sum of all PAHs ranged from 76 to 1944 ng/g [dry weight (dw)]. Despite the apparent matrix similarities between both pine species, P. pinaster needles revealed higher mean entrapment levels than P. pinea (748 and 399 ng/g (dw) per site, respectively). The urban and industrial sites have the highest average of PAH incidence [for P. pinea, 465 and 433 ng/g (dw) per site, respectively, and for P. pinaster, 1147 and 915 ng/g (dw)], followed by the rural sites [233 ng/g and 711 ng/g (dw) per site, for P. pinea and P. pinaster, respectively]. The remote sites, both from P. pinaster needles, show the least contamination, with 77 ng/g (dw) per site. A predominance of 3-ring and 4-ring PAHs was observed in most samples, with phenanthrene having 30.1% of the total. Naphthalene prevailed in remote sites. Rainfall had no influence on the PAHs levels, but there was a relationship between higher wind speeds and lower concentrations. PAH molecular ratios revealed the influence of both petrogenic and pyrogenic sources.

Assessment of the Effects of the Pesticide Imidacloprid on the Behaviour of the Aquatic Oligochaete Lumbriculus variegatus by A. M. Sardo; A. M. V. M. Soares (pp. 648-656).

Contaminants, such as pesticides, can cause direct toxic effects when released into aquatic environments. Suitably sensitive species can help us understand and predict the impacts of such pollutants. Automated sediment toxicity testing and biomonitoring has grown rapidly, and biomonitoring instruments have proven appropriate for studying the effects of pollutants. A new approach in online biomonitoring, using the multispecies freshwater biomonitor was developed in the present study, using whole-sediment toxicity tests and behavioural responses of the freshwater oligochaete Lumbriculus variegatus. Endpoints, such as mortality and growth, were used to study the effects of the pesticide imidacloprid and to achieve a gradient of responses; exposures to contaminated sediments were performed over 10 days’ duration (short-term tests). High mortality was observed in the three highest concentrations of imidacloprid, and inhibition of behaviour was monitored along a gradient of pesticide concentration. Exposure to imidacloprid-contaminated sediments affected growth, behaviour, and avoidance in L. variegatus.

Toxicity of Metals to the Bivalve Tellina deltoidalis and Relationships Between Metal Bioaccumulation and Metal Partitioning Between Seawater and Marine Sediments by C. K. King; M. C. Dowse; S. L. Simpson (pp. 657-665).

The Australian benthic bivalve Tellina deltoidalis tolerates a wide range of sediment and water conditions, is easy to handle in the laboratory, and is a useful species for undertaking whole-sediment toxicity tests. The sensitivity of T. deltoidalis to metals was investigated in 10-day metal-spiked sediment exposures for Cd, Cu, Ni, Pb, and Zn and in water-only exposures for Cu and Zn. The survival of T. deltoidalis in 10-day exposures to metal-spiked sediments was 88–100% for Cd, Ni, Pb, and Zn concentrations of 75, 420, 1,000, and 4,000 mg/kg, respectively. The 4-day LC₅₀s for dissolved Cu and Zn were 0.18 and 13 mg/L, respectively. The 8-day LC₅₀ for Cu was 31 (24–34) μg/L. Cu and Zn concentrations in the tissues of T. deltoidalis increase linearly with increasing dissolved exposure concentration. In the Cu-spiked sediment and water exposures, the survival was negatively correlated with the Cu concentration in both the overlying water and in the tissues of T. deltoidalis. In contrast, particulate Cu concentrations were found to be a poor predictor of Cu bioaccumulation and toxicity for Cu-spiked sediments.

Morphological Response of Typha domingensis to an Industrial Effluent Containing Heavy Metals in a Constructed Wetland by H. R. Hadad; M. M. Mufarrege; M. Pinciroli; G. A. Di Luca; M. A. Maine (pp. 666-675).

Typha domingensis had become the dominant species after 2 years of operation of a wetland constructed for metallurgical effluent treatment. Therefore, the main purpose of this study was to investigate its ability to tolerate the effluent and to maintain the contaminant removal efficiency of the constructed wetland. Plant, sediment, and water at the inlet and outlet of the constructed wetland and in two natural wetlands were sampled. Metal concentration (Cr, Ni, and Zn) and total phosphorus were significantly higher in tissues of plants growing at the inlet in comparison with those from the outlet and natural wetlands. Even though the chlorophyll concentration was sensitive to effluent toxicity, biomass and plant height at the inlet and outlet were significantly higher than those in the natural wetlands. The highest root and stele cross-sectional areas, number of vessels, and biomass registered in inlet plants promoted the uptake, transport, and accumulation of contaminants in tissues. The modifications recorded accounted for the adaptability of T. domingensis to the conditions prevailing in the constructed wetland, which allowed this plant to become the dominant species and enabled the wetland to maintain a high contaminant retention capacity.

Comparison of Bulk Sediment and Sediment Elutriate Toxicity Testing Methods by Herman J. Haring; Mark E. Smith; James M. Lazorchak; Philip A. Crocker; Abel Euresti; Melissa C. Wratschko; Michael C. Schaub (pp. 676-683).

Numerous methods exist for assessing the potential toxicity of sediments in aquatic systems. In this study, the results from 10-day bulk sediment toxicity test methods using Hyalella azteca and Chironomus tentans were compared to results from 96-h Pimephales promelas and Ceriodaphnia dubia renewed acute toxicity tests conducted using elutriate samples prepared from the same sediments. The goal of the study was to determine if the results from the elutriate tests were comparable to those obtained from the bulk sediment tests. Of the 25 samples analyzed, 16 were found to be toxic to at least one of the species tested, in either elutriate or bulk sediment tests. The C. tentans 10-day bulk sediment test was the most sensitive, with 12 sediment samples exhibiting toxicity to this species, whereas the H. azteca bulk sediment test and C. dubia 96-h elutriate test were the least sensitive, exhibiting toxicity in only 7 of the 25 sediments tested. The P. promelas elutriate test found 8 of the 25 sediments to be toxic. Based on the total number of sites found to show toxicity, results from testing indicate 96-h elutriate tests show a level of sensitivity comparable to 10-day bulk sediment tests in assessing toxicity quantitatively. However, the methods did not always find toxicity at the same sites, suggesting that the ability of elutriate tests to predict toxicity (quantitatively) is not statistically correlated with bulk sediment methods. This would indicate that a suite of toxicity test methods would provide the most complete measure of site condition; however, in circumstances where bulk sediment testing is not feasible, elutriate tests can provide a practical and credible alternative for toxicity assessment.

Assessment of Antifouling Biocides Contaminations in Maizuru Bay, Japan by Sayaka Eguchi; Hiroya Harino; Yoshikazu Yamamoto (pp. 684-693).

The concentrations of organotin compounds in the aquatic environment of Maizuru Bay and their spatial distribution are discussed. The concentrations of tributyltin (TBT) compounds in water samples ranged from 0.001 to 0.002 μg l⁻¹, and monobutyltin compounds were the dominant species among the butyltin compounds. TBT concentrations in Maizuru Bay are low compared with other coastal waters of Japan. Drastic differences in TBT concentrations were not observed among the Maizuru Bay sites. Phenyltin compounds were not detected in the water samples. Concentrations of TBT and triphenyltin (TPT) in sediment from Maizuru Bay ranged, respectively, from 0.9 to 11 μg kg⁻¹, from 0.2 to 17 μg kg⁻¹ dry weight (dw). TBT concentrations in Maizuru Bay were lower than those in other coastal areas of Japan. TPT concentrations were greater than TBT concentrations in the fishing port. The concentrations of TBT and TPT in blue mussels (M. galloprovincialis) from Maizuru Bay were in the range of 2.4 to 9.3 μg kg⁻¹ and 0.2–13 μg kg⁻¹ wet weight (ww), respectively. A tolerable average residue level (TARL) was estimated at 74.8 μg kg⁻¹ from a tolerable daily intake (TDI) of 0.25 bis(tributyltin)oxide μg kg⁻¹ body weight day⁻¹. TBT concentrations detected in blue mussel samples were lower than the TARL values. The acceptable concentration of TPT, which were calculated using acceptable daily intake instead of TDI, was 127 μg kg⁻¹. Concentrations of TPT in blue mussel samples were also lower than the TARL. TBT compounds in blue mussel samples were at similar levels among the various sampling sites, indicating that TBT is not currently being used in ship hull paints; the ratios of degradation products of TBT and TPT were greater than those of the parent compounds. Concentrations of alternative biocides in water samples were also investigated in the bay. Although Sea-Nine 211, M1, and Pyrithiones were not detected, Diuron and Irgarol 1051 were detected at 0.010–0.257 and at 0.002–0.018 μg l⁻¹, respectively. Concentrations of Diuron were great in the shipping route and near the shipyard, whereas the concentration of Irgarol 1051 was great at the fishing port. Concentrations of Diuron and Irgarol 1051 in sediment from Maizuru Bay ranged, respectively, from <0.08 to 12, from <0.08 to 9.8 μg kg⁻¹ dw, respectively. Despite being a semi-enclosed bay, we found that sediment in Maizuru Bay is not contaminated by alternative biocides to the degree found in other coastal areas. Copper concentrations of sediment were at ordinary levels, and those of blue mussels were slightly lower than those reported previously in other coastal areas of Japan. In both the sediment and blue mussels, there was no correlation between the presence of copper and antifouling biocides.

Effects of Elevated Nitrate Concentration on Mortality, Growth, and Egestion Rates of Gammarus pseudolimnaeus Amphipods by Robert S. Stelzer; Brandon L. Joachim (pp. 694-699).

There is relatively little known about nitrate toxicity in aquatic invertebrates, especially sublethal effects of elevated nitrate. We exposed the amphipod Gammarus pseudolimnaeus to elevated nitrate concentrations (up to 128 mg NO₃-N/L) in a laboratory experiment and measured mortality, growth rates, egestion rates, molting, and C:N ratio. Nitrate concentration did not affect mortality, egestion rate, molting, and C:N ratio. Amphipod growth decreased slightly with increasing nitrate concentration based on the results of a linear regression, but a one-way ANOVA suggested that mean growth rates were not different among nitrate treatments. We suggest that additional research is needed on the sublethal effects of elevated nitrate concentration on aquatic invertebrates, particularly for those taxa that have demonstrated sensitivity to other chemical stressors.

Concentrations and Accumulation Features of Organochlorine Pesticides in the Baiyangdian Lake Freshwater Food Web of North China by Guocheng Hu; Jiayin Dai; Bixian Mai; Xiaojun Luo; Hong Cao; Jianshe Wang; Fengchao Li; Muqi Xu (pp. 700-710).

Organochlorine pesticides (OCPs), such as hexachlorocyclohexane (HCH) and dichlorodiphenyltrichloroethane (DDT), are ubiquitous anthropogenic environmental contaminants. They are persistent, broad-spectrum toxicants that accumulate in the food web with potential risks to the ecosystem and human health. HCHs were the predominant contaminants in Baiyangdian Lake, North China. Concentrations of HCHs and DDTs ranged from 58 to 563 ng/g lipid weight (lw) and 21 to 401 ng/g lw, respectively, for aquatic biota samples. The highest levels of HCHs and DDTs were observed in muscles of yellow catfish. The mean concentrations of OCPs were 4.6 ng/L for water, 95 ng/g dry weight (dw) for aquatic plants, and 14 ng/g dw for sediments. Among the isomers and metabolites, α-HCH and p,p′-1,1-di(p-chlorophenyl)-2,2-dichloroethylene (p,p′-DDE) were the predominant congeners in biota samples. Correlations between log lipid-normalized concentrations of HCHs and DDTs and trophic levels (TLs) based on analysis of stable isotopes of nitrogen confirmed that persistent organic pollutants were magnified in the Baiyangdian Lake food web. Significant positive relationships were found for α-HCH and p,p′-DDT and their trophic magnification factors, which were 1.6 and 1.7, respectively. These results provide evidence of biomagnification of persistent organic pollutants (POPs) in freshwater food webs.

Polycyclic Aromatic Hydrocarbon Contamination of Recent Sediments and Marine Organisms from Xiamen Bay, China by Qingzhao Li; Xian Zhang; Changzhou Yan (pp. 711-721).

Surface marine sediments from 12 sites within Xiamen Bay (XMB) and marine organisms (clam, crab, and fish) from a heavily polycyclic aromatic hydrocarbon (PAH)-contaminated site at Yuandang Lagoon were sampled and analyzed for 16 priority PAH content by gas chromatography/mass spectrometry. The total PAH concentrations in sediments ranged from 203.7 to 1590.5 ng/g, with an average value of 670.0 ng/g. Overall, the total PAH concentrations were relatively lower in the east coastal zone and significantly higher in Yuandang Lagoon. These concentrations were intermediate in comparison with those observed in other estuaries in China. The sources of PAH inputs to sediments in XMB were quantitatively determined by principal components analysis with multiple linear regression. The results showed that, on average, vehicle emissions, petroleum spills, and coal combustion contributed to 41, 36, and 23% of the total PAHs, respectively. Analysis of the PAH composition pattern in marine organisms showed that the total PAHs levels of fishes were lower than those of the clam and crab. The PAH burden of the crab and calm was characterized by the non-negligible occurrence of high-molecular-weight compounds, whereas the fish PAH pattern was dominated by the lower-molecular-weight compounds. In addition, the carcinogenic PAH benzo[a]pyrene was detected in all samples. Molecular indices based on isomeric PAH ratios differentiated the uptake pathway among organisms. The water soluble matter and the petroleum seemed to be the most favorable uptake pathway in fish from Yuandang Lagoon, whereas the crab and clam shared a mix uptake by the water-soluble and the particulate matter.

Toxic Effect and Biochemical Study of Chlorfluazuron, Oxymatrine, and Spinosad on Honey Bees (Apis mellifera) by Entsar I. Rabea; Hoda M. Nasr; Mohamed E. I. Badawy (pp. 722-732).

Under laboratory conditions, the comparative effects of two insect growth regulators, chlorfluazuron and oxymatrine, and spinosad as a biopesticide were examined on honey bee workers (Apis mellifera L.). Separate groups of bees were left for 24 h to feed on 50% sucrose solution containing different concentrations of the tested insecticides, and the lethal concentration that caused 50% mortality (LC₅₀) was estimated. The inhibitory effects on acetylcholinesterase (AChE) and adenosine triphosphatase (ATPase) activities as biochemical indicators were determined in vivo after 24 h in head, thorax, and abdomen of surviving bees obtained after treatments with a view to explore the possible mode of action of these compounds. Results indicated that exposure to spinosad showed toxicity to honey bees with LC₅₀ value of 7.34 mg L⁻¹, followed by oxymatrine (LC₅₀ = 10.68 mg L⁻¹), while chlorfluazuron was the least acutely toxic of the tested compounds (LC₅₀ = 2,526 mg L⁻¹). Oxymatrine and spinosad at the same tested concentrations (2.5, 5, 10, and 20 mg L⁻¹) significantly inhibited AChE activity in different organs of honey bee workers, and high inhibition percentage was obtained with the enzyme isolated from the thorax. However, chlorfluazuron at 400, 1,000, 2,000, and 4,000 mg L⁻¹ caused high inhibition of AChE activity isolated from the head (39.65% and 44.22% at 2,000 and 4,000 mg L⁻¹, respectively). In addition, the toxic effects of the tested compounds on activity of ATPase indicated that spinosad caused the highest inhibitory effect in different organs compared with oxymatrine at the same concentrations, and high inhibition was found with ATPase isolated from the head. The results also indicated that oxymatrine was the least active compound for inhibition of AChE and ATPase.

Impact of the Use of Fluvalinate on Different Types of Beeswax from Spanish Hives by Sabine Adamczyk; Regina Lázaro; Consuelo Pérez-Arquillué; Susana Bayarri; Antonio Herrera (pp. 733-739).

Acaricides are applied in agriculture as phytosanitary products against pests and in apiculture to control the bee parasite Varroa destructor. Poor apicultural practices could result in an accumulation of residues in honeybees, in the environment, and in beeswax and other bee products by migration from the wax comb into stored honey through a process of diffusion and consequently constitute a potential risk for humans. In this study, six different types of beeswax samples were analysed for the determination of residues of fluvalinate, coumaphos, and bromopropylate and its metabolite 4,4′-dibromobenzophenone, all of which are the most commonly acaricides used by Spanish beekeepers against V. destructor. The analytic method consists of solid-phase extraction on a SPE Florisil cartridge and high-performance liquid chromatography separation using a photo diode array detector. The results show that fluvalinate residues were detected in 36.3% of samples, ranging from 1.2 to 6.6 μg/g wax. Residues of coumaphos, bromopropylate, and 4,4′-dibromobenzophenone were not found to be greater than their detection limits. This study indicates that the analysis of these compounds in beeswax samples could be used as bioindicators of fluvalinate sanitary treatment and handling practices applied by beekeepers.

Herbicide Formulation with Glyphosate Affects Growth, Acetylcholinesterase Activity, and Metabolic and Hematological Parameters in Piava (Leporinus obtusidens) by Joseânia Salbego; Alexandra Pretto; Carolina Rosa Gioda; Charlene Cavalheiro de Menezes; Rafael Lazzari; João Radünz Neto; Bernardo Baldisserotto; Vania Lucia Loro (pp. 740-745).

The teleost fish Leporinus obtusidens (piava) was exposed to different concentrations of Roundup, a commercial herbicide formulation containing glyphosate (0, 1, or 5 mg L⁻¹), for 90 days. Acetylcholinesterase (AChE) activity was verified in brain and muscle. Hepatic and muscular metabolic parameters as well as some hematological parameters were determined. The results showed that brain AChE activity was significantly decreased in fish exposed to 5 mg L⁻¹ Roundup, whereas muscular AChE activity was not altered. Both Roundup concentrations significantly decreased liver glycogen without altering the muscle glycogen content. Hepatic glucose levels were reduced only in fish exposed to 5 mg L⁻¹ Roundup. Lactate levels in the liver and muscle significantly increased in fish exposed to both Roundup concentrations. Hepatic protein content remained constant at 1 mg L⁻¹ but increased at 5 mg L⁻¹ Roundup. In the muscle however, protein content decreased with increasing exposure concentration. The herbicide exposure produced a decrease in hematological parameters at both concentrations tested. The majority of observed effects occur at environmental relevant concentrations, and in summary, the results show that Roundup affects brain AChE activity as well as metabolic and hematologic parameters of piavas. Thus, we can suggest that long-term exposure to Roundup causes metabolic disruption in Leporinus obtusidens.

Influences of Sex, Habitat, and Seasonality on Heavy-Metal Concentrations in the Burrowing Crab (Neohelice Granulata) From a Coastal Lagoon in Argentina by María Ornela Beltrame; Silvia G. De Marco; Jorge E. Marcovecchio (pp. 746-756).

Cadmium, chromium, copper, and manganese concentrations were determined in hepatopancreas of both sexes as well as in eggs at different embryonic development stages of the burrowing crab Neohelice granulata (Brachyura, Varunidae) from Mar Chiquita Coastal Lagoon, a biosphere reserve from Argentina, during a period of 1 year, to assess the bioaccumulation of metals associated with sex and seasonality. Furthermore, metal levels in associated superficial sediment samples were also determined. Two different “cangrejales,” one in a mudflat and another one in a salt marsh, were studied. The results showed high concentrations of copper within the hepatopancreas, which was considered a strong reflection of high exposure of N. granulata to this metal. Metal accumulation in hepatopancreas from both study sites and sexes did not present significant differences, as did seasonality. In this sense, both spring and summer metal levels in hepatopancreas were the greatest. Eggs did not present differences in metal accumulation, with the exception of manganese, between sites or between crabs at different embryonic stages. For this metal, eggs from female crabs inhabiting mudflats showed higher levels than those from inhabiting salt marshes. Moreover, eggs in the late embryonic stage also showed the highest manganese concentrations. Metal levels in sediments, however, were similar in both sites. These are the first results of metal level in biota and sediments in this particular environment. Such results could be used as a baseline for the monitoring of metal levels in future studies in Mar Chiquita Coastal Lagoon.

Modulatory Effects of Deltamethrin on Antioxidant Defense Mechanisms and Lipid Peroxidation in Carassius auratus gibelio Liver and Intestine by Diana Dinu; Diana Marinescu; Maria Cristina Munteanu; Andreea Cristina Staicu; Marieta Costache; Anca Dinischiotu (pp. 757-764).

Pyrethroids, such as deltamethrin, are toxic substances that lead to generation of reactive oxygen species, which harm living organisms. We assessed the level and patterns of imbalance evolved by a single dose of 2 μg/L deltamethrin on the lipid peroxidation (LPO) and the antioxidant defense system of Carassius auratus gibelio liver and intestine, and monitored the recovery dynamics of these parameters during a 14-day post-exposure period. LPO and antioxidative defense mechanisms displayed different responses in the investigated tissues. Sudden increase of LPO in the liver, persisting at this elevated level throughout the test period, was observed on the third day post-exposure, while in the intestine significant enhancement of this parameter was recorded from the seventh day. Reduced glutathione (GSH) showed a transient increase in the liver, and was depleted in the intestine by the second day of exposure, with signs of recovery by the end of the experimental tenure. In the liver of fish a temporary inhibition of superoxide dismutase (SOD) and catalase (CAT) activity, and activation of glutathione peroxidase (GPX), glutathione transferase (GST), and glutathione reductase (GR) enzymes was observed, with maximum thresholds recorded on the third and second days, respectively. In the intestine a relevant increase in CAT and GST activity up to the second day and almost complete recovery by the end of the experiment was recorded, while for GR a continuous enhancement was apparent.

Effect of Parathion-Methyl on Amazonian Fish and Freshwater Invertebrates: A Comparison of Sensitivity with Temperate Data by Andreu Rico; Rachel Geber-Corrêa; Paola S. Campos; Marcos V. B. Garcia; Andrea V. Waichman; Paul J. van den Brink (pp. 765-771).

Parathion-methyl is an organophosphorous insecticide that is widely used in agricultural production sites in the Amazon. The use of this pesticide might pose a potential risk for the biodiversity and abundance of fish and invertebrate species inhabiting aquatic ecosystems adjacent to the agricultural fields. Due to a lack of toxicity data for Amazonian species, safe environmental concentrations used to predict the ecological risks of parathion-methyl in the Amazon are based on tests performed with temperate species, although it is unknown whether the sensitivity of temperate species is representative for those of Amazonian endemic species. To address this issue, the acute toxic effect (LC₅₀–96 h) of parathion-methyl was assessed on seven fish and five freshwater invertebrate species endemic to the Amazon. These data were used to compare their pesticide sensitivity with toxicity data for temperate species collected from the literature. The interspecies sensitivity was compared using the Species Sensitivity Distribution (SSD) concept. The results of this study suggest that Amazonian species are no more, or less, sensitive to parathion-methyl than their temperate counterparts, with LC₅₀ values ranging from 2900 to 7270 μg/L for fish and from 0.3 to 319 μg/L for freshwater arthropods. Consequently, this evaluation supports the initial use of toxicity data of temperate fish and freshwater invertebrate species for assessing the effects of parathion-methyl on Amazonian freshwater ecosystems.

CYP1A Expression in Caged Rainbow Trout Discriminates Among Sites with Various Degrees of Polychlorinated Biphenyl Contamination by Ben F. Brammell; J. Scott McClain; James T. Oris; David J. Price; Wesley J. Birge; Adria A. Elskus (pp. 772-782).

It has become increasingly apparent that resident fish can develop resistance to chemicals in their environment, thus compromising their usefulness as sentinels of site-specific pollution. By using a stream system whose resident fish appear to have developed pollutant resistance (Brammell et al., Mar Environ Res 58:251–255, 2005), we tested the hypothesis that the pollutant-inducible biomarker, cytochrome P4501A (CYP1A), as measured in field-caged juvenile rainbow trout (Oncorhynchus mykiss), would reflect relative pollution differences between reference and polychlorinated biphenyl (PCB)-contaminated sites. Trout were caged in the Town Branch/Mud River system (Logan County, KY), a stream system undergoing remediation for PCBs. Fish were held in remediated (Town Branch), unremeditated (Mud River), and reference sites for 2 weeks during spring 2002. At the end of this period, gill and hepatic CYP1A expression were measured. To evaluate the relative PCB exposure of caged trout and provide a reference point against which to calibrate CYP1A response, PCB levels were quantified in sediments from each site. Hepatic CYP1A expression in caged trout clearly detected the presence of PCBs in the Town Branch/Mud River stream system. Sediment PCB levels and hepatic CYP1A expression in caged trout produced identical pollution rankings for the study sites. Gill CYP1A expression, although suggestive of site differences, was not statistically different among sites. Unlike resident fish, which failed to show site differences in hepatic CYP1A expression in this waterway (Brammell et al. 2005), caged fish proved to be a sensitive discriminator of relative PCB contamination in this system. In summary, we determined that CYP1A expression in caged fish reflected relative in situ pollutant exposure. The exposure paradigm confirmed that 2 weeks was a sufficient caging period for evaluating CYP1A response in this species at these temperatures (13–19°C). In addition, these studies demonstrate that tissue-specific CYP1A expression can provide insights into likely routes of exposure. We conclude that CYP1A expression in caged trout is a reliable and inexpensive first-pass determination of relative environmental pollutant exposure and bioavailability in aqueous systems.

Effects of a Chemically Polluted Discharge on the Relationship Between Fecundity and Parasitic Infections in the Chub (Leuciscus cephalus) from a River in Southern England by N. J. Morley; H. H. Costa; J. W. Lewis (pp. 783-792).

In fish there is a close association between endocrine and immune function that during seasonal changes in reproductive status can affect the fishes susceptibility to parasitic infection. The effects of endocrine-disrupting pollution on this association are not known. The present study investigates the effects of a chemically polluted (ammoniacal nitrogen, zinc, bromide) discharge in Cranleigh Waters, a river in southern England, on the fecundity and parasitism of chub, Leuciscus cephalus, over an 18-month period. In the polluted site, the gonado-somatic index (GSI) of fish was lower in females and higher in males compared to an unpolluted upstream site, indicating an androgenic effect on reproduction. Parasite species richness, especially the intestinal helminths Proteocephalus torulosus and Pomphorynchus laevis, was increased in fish in the polluted site. However, the occurrence of the two most common parasite species in the river—Myxobulus sp. 1, a gill myxozoan, and Diplostomum sp., a trematode found in the eye—was unchanged between polluted and unpolluted river sites. Nevertheless, there was a significant relationship between GSI and the prevalence of these two parasite species in the unpolluted site but not the polluted site suggesting that endocrine disruption of reproduction did not lead to an increase in host susceptibility. The reasons for this are discussed.

Variable Vitellogenin Response of Japanese Medaka (Oryzias latipes) to Weekly Estrogen Exposure by K. Erica Marsh; Gordon Paterson; Christy M. Foran; Erin R. Bennett (pp. 793-799).

Japanese medaka (Oryzias latipes) is a valuable model organism in reproductive and developmental toxicity testing. The purpose of this experiment is to assess the response of medaka to aquatic estrogen exposure over the course of 1 year. Each week, three pairs of adult male medaka were exposed separately for 4 days (100% static renewal daily) to 17β-estradiol at a nominal level of 25 μg/l, with a fourth pair of fish exposed separately to an ethanol control. Vitellogenin (VTG) induction was observed each week, with hepatic and plasma VTG levels significantly higher (P < 0.001) than reported for ethanol control specimens. A significant (P < 0.001) increasing trend was observed for plasma VTG results over the duration of the study, whereas a decreasing trend (P = 0.030) of hepatic VTG was evident. A Durbin–Watson test, however, did not demonstrate any serial autocorrelation of hepatic (d = 1.180) or plasma (d = 1.311) VTG levels over the duration of the study. Time-series transformations of the hepatic and plasma VTG data did not reveal any significant seasonal or behavioral patterns. However, significant intermittent peaks in VTG production were observed in both tissue types during the study. These data indicate that some consideration must be taken to time long-term medaka exposures (>20 weeks) in order to eliminate any influence of cyclic changes on plasma VTG response. Alternatively, hepatic cytosolic measurement of VTG appears to show a more sensitive response to aquatic estrogen exposure.

Bioaccumulation and Biomagnification of Organochlorines in a Marine Food Web at a Pristine Site in Iceland by Halldora Skarphedinsdottir; Karl Gunnarsson; Gudmundur A. Gudmundsson; Erick Nfon (pp. 800-809).

Organochlorine (OC) bioaccumulation and biomagnification were studied in a marine food web at a pristine site in Iceland. The species studied were the gastropod and grazer chink shell (Lacuna vincta), the filter feeding bivalve blue mussel (Mytilus edulis), the predators butterfish (Pholis gunnellus), and the seabird black guillemot (Cepphus grylle), all sampled and analysed in 1996–1997. Individual OC levels were generally low in chink shell and blue mussels, somewhat elevated in the fish, and an order of a magnitude higher in the top predator black guillemot, except for ΣHCH (hexachlorocyclohexane isomers) and Σchlordane levels, which were similar in all organisms, ranging from 10 to 36 ng/g lipid weight (lw). In the molluscs and fish, mean concentrations of ΣPCB (polychlorinated biphenyl) ranged from 111 to 377 ng/g lw, ΣDDT (dichlorodiphenyltrichloroethane) ranged from 19 to 65 ng/g lw, and HCB (hexachlorobenzene) ranged from 21 to 30 ng/g lw. The levels of same OCs in the black guillemot were on average 2352, 361, and 283 ng/g lw, respectively. The OC tissue concentrations in blue mussel and black guillemot are comparable to levels in Arctic and sub-Arctic regions, but OC levels in blue mussel tissue were an order of magnitude lower than found in the North Sea and the Baltic Sea. The relative composition of OCs were generally similar among species with the PCB congeners emerging as the most abundant compounds with levels an order of magnitude higher than the other compounds in all species. Food web magnification factors (FWMFs) were determined for the OCs by using trophic levels determined from δ¹⁵N. FWMFs >1, indicating biomagnification, were found for ΣPCB, penta- or higher chlorinated PCBs (e.g., PCB 101, 118, 138, 153, 180), β-HCH, HCB, ΣDDT, p,p-DDE, and transnonachlor. The highest FWMF was observed for PCB 180 at FWMF = 5.8.

Nonessential Metals in Striped Marlin and Indo-Pacific Sailfish in the Southeast Gulf of California, Mexico: Concentration and Assessment of Human Health Risk by M. F. Soto-Jiménez; F. Amezcua; R. González-Ledesma (pp. 810-818).

Seventeen sailfish (Istiophorus platypterus; 166–246 cm long) and 13 striped marlin (Tetrapturus audax; 159–254 cm long) specimens from the southern Gulf of California were analyzed for As, Cd, Hg, and Pb concentrations. More than 20% of the As and Cd levels exceeded the guideline levels of the World Health Organization (WHO), the U.S. Food and Drug Administration, and the European Union. About 65–90% of the specimens exceeded the limit value for Hg. Pb levels in billfish were significantly lower than guideline values. The intakes of As and Cd through billfish consumption by the population represented 20–40% of the provisional tolerable weekly intake (PTWI) values as recommended by the WHO. Levels of Hg exceeded the PTWI value. Pb intake represented <5% of the PTWI through these species in all age and gender groups. The target hazard quotients (THQs) for Cd and Pb were <1 in each group of interest. In contrast, THQ values based on As and Hg showed large variations, with the highest values for children, then pregnant women, and, finally, adults. Values of THQ >1 indicate risk for As and Hg; the risk from As was estimated assuming that 20% of the total As was inorganic. These metals can affect the nervous system and intellectual and physical development of unborn children and infants. A national moratorium on billfish consumption in México is recommended for children (0–6 years old), pregnant women, and women planning pregnancy.

Larval Responses of Three Midwestern Anurans to Chronic, Low-Dose Exposures of Four Herbicides by Bethany K. Williams; Raymond D. Semlitsch (pp. 819-827).

Low levels of agricultural herbicides often contaminate surface water and might persist throughout the growing season, potentially acting as stressors on aquatic organisms. Although low-dose, chronic exposures to agrochemicals are likely common for many nontarget organisms, studies addressing these effects using end-use herbicide formulations are rare. We exposed three common species of tadpoles to conservative levels of atrazine, S-metolachlor, and glyphosate end-use herbicide formulations throughout the larval period to test for survival differences or life-history trait alterations. Exposure to the glyphosate product Roundup WeatherMax^® at 572 ppb glyphosate acid equivalents (a.e.) resulted in 80% mortality of western chorus frog tadpoles, likely as a result of a unique surfactant formulation. Exposure to WeatherMax^® or Roundup Original Max^® at 572 ppb a.e. also lengthened the larval period for American toads. Chronic atrazine and S-metolachlor exposures induced no significant negative effects on survival, mass at metamorphosis, or larval period length at the levels tested. These results highlight the importance of explicitly tying chronic tests to the natural environment and considering contributions of surfactant/adjuvant components to end-use formulation toxicities, even between very similar products.

Oxidative Stress Biomarkers in the Digestive Gland of Theba pisana Exposed to Heavy Metals by M. A. Radwan; K. S. El-Gendy; A. F. Gad (pp. 828-835).

The in vivo toxic effects of sublethal treatment of 40 and 80% of 48-h LD₅₀ of topically applied trace metals [copper (Cu), lead (Pb), and zinc (Zn)] on oxidative stress biomarkers in the digestive gland of Theba pisana were examined. Oxidative individual perturbations were assessed by measuring nonenzymatic (glutathione; GSH) and enzymatic (catalase, CAT; glutathione peroxidase, GPx; and glutathione-S-transferase, GST) antioxidants in digestive gland of the snails. Lipid peroxidation (LPO) was also evaluated as a marker of cell damage. The results indicated that the copper ion was the most potent metal against this snail, followed by zinc and lead, for which the corresponding LD₅₀ values were 37.88, 261.72, and 652.55 μg/snail, respectively. The no-observed effect concentration (NOEC) values for Cu, Zn, and Pb were 10, 50, and 500 μg/snail, respectively, and the corresponding lowest-observed effect concentration (LOEC) values were 50, 100, and 1000 μg/snail. All trace metals resulted in a significant increase in the level of LPO, whereas a significant decline in the content of GSH was observed when compared with untreated controls. Treatment with both sublethal doses of the metals caused significant increase in CAT activity, except in the case of 40% LD₅₀ Zn and 80% LD₅₀ Cu, which exhibited no alteration in CAT when compared to control animals. GPx was significantly increased in snails exposed to 40% LD₅₀ Cu and Pb as well as 80% LD₅₀ Cu. However, an opposite effect was observed in snails exposed to 80% LD₅₀ Pb and in either 40 or 80% LD₅₀ of Zn-intoxicated animals. Treatment with Pb at two sublethal doses significantly increased GST activity, whereas treatment the animal with Cu caused significant inhibition in this enzyme. Snails exposed to 40% LD₅₀ Zn showed significant enhancement of GST, whereas snails exposed to 80% LD₅₀ showed ignificantly reduced GST activity. Biphasic responses were observed for CAT, GPx, and GST activities in snails exposed to Cu, Pb, and Zn, respectively. This study suggests that upregulation of the antioxidant enzyme activities, elevation of LPO, and the reduction in GSH content is related to oxidative stress in this species that could be useful as biomarkers for the evaluation of contaminated terrestrial ecosystems.

Toxicity of Octahydro-1,3,5,7-Tetranitro-1,3,5,7-Tetrazocine (HMX) in Three Vertebrate Species by Mark S. Johnson; Craig A. McFarland; Matthew A. Bazar; Michael J. Quinn Jr.; Emily May LaFiandra; Larry G. Talent (pp. 836-843).

The explosive, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine or high-melting explosive (HMX), has been found in soils in areas used for testing and training by the military. Many of these areas contain habitat for valued wildlife species. In an effort to better understand the environmental consequences from exposure, a reptilian (western fence lizard [Sceloporus occidentalis]), an amphibian (red-backed salamander [Plethodon cinereus]), and a mammalian species (rabbit [Oryctolagus cuniculus]) were exposed to HMX under controlled laboratory conditions. Lizards and rabbits were exposed to HMX by way of corn oil through gavage, and salamanders were exposed to HMX in soil. Two deaths occurred from acute oral exposures to lizards to 5000 mg HMX/kg BW. Histological and gross pathologic assessment suggested gut impaction as a possible cause of death. Salamanders exposed to concentrations of HMX in soil ≤1970 mg HMX/kg soil for 10 days did not show adverse effects. Rabbits, however, showed neurologic effects manifested as hyperkinetic events with convulsions at >24 h after oral exposures. An LD₅₀ for rabbits was calculated as 93 mg/kg (95% confidence interval 76–117). A subacute 14-day testing regime found a lowest observed effect level of 10 mg/kg-d and a no observed adverse effect level of 5 mg/kg-d based on hyperkinesia and seizure incidence, although changes suggesting functional hepatic alterations were also found. These data suggest that physiologic differences between species, particularly in gastrointestinal structure and function, can affect the absorption of HMX and hence lead to marked differences in toxicity from exposure to the same compound.

Trace Element Concentrations (Mercury, Cadmium, Copper, Zinc, Lead, Aluminium, Nickel, Arsenic, and Selenium) in Some Aquatic Birds of the Southwest Atlantic Coast of France by Magali Lucia; Jean-Marc André; Karine Gontier; Nicolas Diot; Jesus Veiga; Stéphane Davail (pp. 844-853).

Trace elements (mercury [Hg], cadmium [Cd], copper [Cu], zinc [Zn], lead [Pb], aluminium [Al], nickel [Ni], arsenic [As], and selenium [Se]) were investigated using inductively coupled plasma–mass spectrometry in liver, kidney, muscle, and feather of aquatic birds wintering or inhabiting the wetlands situated on the Southwest Atlantic coast of France. A majority of greylag geese, red knots, and grey plovers were collected from among hunter-shot animals. The relation between residue concentrations, age (juvenile vs. adult), and sex was investigated. Trace elements were lower than threshold levels of toxicity, except for Pb. Greylag geese sampled could be considered Pb-poisoned. These consequential levels of contamination could be the result of the ingestion of Pb-shot from ammunition used in hunting areas they crossed during migration. Cd accumulation increased with age, whereas Pb levels in feathers were lower in adult birds in connection with moulting. As was influenced by sex. Female birds displayed higher concentrations in liver and feathers than did male birds.

Selenium Accumulation in Sea Ducks Wintering at Lake Ontario by Michael L. Schummer; Shannon S. Badzinski; Scott A. Petrie; Yu-Wei Chen; Nelson Belzile (pp. 854-862).

Numbers of wintering sea ducks, including buffleheads (Bucephala albeola; BUFF), common goldeneyes (Bucephala clangula; COGO), and long-tailed ducks (Clangula hyemalis; LTDU), increased substantially at Lake Ontario after Dreissenid mussels (Dreissena bugensis and D. polymorpha) colonized the Great Lakes. Invertebrates, including Dreissenid mussels, are major diving duck prey items that can transfer some trace elements, such as selenium (Se) to higher trophic levels. Se can be problematic for waterfowl and it often has been detected at elevated levels in organisms using the Great Lakes. There are, however, few data on hepatic Se concentrations in sea ducks, particularly during the winter at Lake Ontario. In this study, we evaluated interspecific differences and temporal trends in hepatic Se concentrations among BUFF (n = 77), COGO (n = 77), and LTDU (n = 79) wintering at Lake Ontario. All three species accumulated Se throughout winter, but COGO did so at a higher rate than did BUFF and LTDU. Overall, Se concentrations were higher in LTDU [ $$ {ar{ ext{x}}} = 22.7$$ ; 95% CI = 20.8–24.8 μg/g dry weight (dw)] than in BUFF ( $$ {ar{ ext{x}}} = 1 2. 3 $$ ; 95% CI = 11.6–13.1 μg/g dw) and COGO ( $$ {ar{ ext{x}}} = 1 2.0 $$ ; 95% CI = 10.7–3.5 μg/g dw) throughout the winter. Se concentrations were deemed elevated (>33 μg/g dw) in 0%, 5%, and 19% of BUFF, COGO, and LTDU, respectively. Presently there are no data on Se toxicity end points for these species, so it is unclear how acquiring concentrations of these magnitudes affect their short- and long-term health or reproduction.

Organochlorine and PBDE Concentrations in Relation to Cytochrome P450 Activity in Livers of Forster’s Terns (Sterna forsteri) and Caspian Terns (Hydroprogne caspia), in San Francisco Bay, California by Garth Herring; Joshua T. Ackerman; Collin A. Eagles-Smith; Terrence L. Adelsbach; Mark J. Melancon; Katie R. Stebbins; David J. Hoffman (pp. 863-873).

We measured halogenated organic contaminants (HOCs) [polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), and dichloro-diphenyl-trichloroethane (DDT)] and P450 [e.g., ethoxyresorufin-O-dealkylase (EROD)] stress in livers from Caspian tern (Hydroprogne caspia) adults and Forster’s tern (Sterna forsteri) adults and chicks in San Francisco Bay (SFB). Penta BDEs and tetra PBDEs composed 46–66% of ∑PBDE in terns. PCB homologues di, tri, penta, hexa, and hepta composed 93–95% of ∑PCBs and p′p-DDE composed 82–98% of all ∑DDTs. We found similar concentrations of ∑PBDEs [mean micrograms per gram wet weight (ww) ± standard error = 0.4 ± 0.1], ∑PCBs (5.9 ± 1.6), and ∑DDTs (0.6 ± 0.1) among species, sexes, and regions. However, concentrations were higher in Forster’s tern adults than chicks (∑PBDEs = 0.4 ± 0.1 and 0.1 ± 0.1; ∑PCBs = 7.08 ± 2.4 and 2.4 ± 1.4; ∑DDTs = 0.5 ± 0.1 and 0.1 ± 0.1; respectively), and there was a nonsignificant trend of elevated ∑PBDEs and ∑PCBs for adult Forster’s terns in the Central South Bay and Lower South Bay portions of SFB. Combined Forster’s tern and Caspian tern ∑DDTs bioaccumulated similarly to selenium, but not mercury, and there was a nonsignificant but positive trend for ∑PBDEs and ∑PCBs bioaccumulation with mercury. P450 protein activity was higher in adult Forster’s terns than Caspian terns, higher in Central South Bay than in Lower South Bay, and higher in adult Forster’s terns than in chicks.

Changes in Sperm Characteristics and Induction of Oxidative Stress in the Testis and Epididymis of Experimental Rats by a Herbicide, Atrazine by Sunny O. Abarikwu; Adebukola C. Adesiyan; Titilola O. Oyeloja; Matthew O. Oyeyemi; Ebenezer O. Farombi (pp. 874-882).

To study the effects of atrazine on reproductive functions and testicular and epididymal antioxidant defense, rats were exposed to 0, 120, or 200 mg/kg body weight atrazine orally for 7 and 16 days. Animals exposed to the high-dose atrazine had their body weights, feed intake, and reproductive organs weights significantly reduced, whereas testicular weights remain unaffected independent of the dose used. In comparison to control, glutathione (GSH) and glutathione-S-transferase (GST) activities were elevated in the high-dose group, whereas the activity of superoxide dismutase (SOD), catalase (CAT); ascorbate (AA), and malondialdehyde (MDA) levels and hydrogen peroxide production were unchanged in the testis during the 7-day-exposure protocol. When atrazine treatment was increased to 16 days, GSH levels remained unchanged, but lipid peroxidation levels were significantly increased in both the testes and epididymides. This corresponded to the significant diminution in the activities of GST and SOD. CAT activities were unaffected in the testes and then dropped in the epididymides. γ-Glutamyl transferase (γ-GT) activities increased during both studies, whereas AA levels remained unaffected (p < 0.05). Atrazine exposure has a dose-dependent adverse effect on the testicular and epididymal sperm numbers, motility, viability, morphology, and daily sperm production. Although the testes of the atrazine-treated animals appear normal, few tubules had mild degeneration with the presence of defoliated cells. Likewise, no perceptible morphological changes were observed in the epididymis. The results suggest that atrazine impairs reproductive function and elicits a depletion of the antioxidant defense system in the testis and epididymis, indicating the induction of oxidative stress.

Validation and Application of a Method for the Determination of Total Chromium in Rat Tissues by Inductively Coupled Plasma Mass Spectrometry by Keith E. Levine; Matthew D. Stout; Glenn T. Ross; Amal S. Essader; Frank X. Weber; Peter M. Grohse; Reshan A. Fernando; Lisa S. Milstein; Michelle J. Hooth; Bradley J. Collins (pp. 883-891).

The validation of a method for the determination of chromium (Cr) in F-344/N rat tissues by inductively coupled plasma-mass spectrometry is described. Samples were analyzed after a rapid, open-vessel microwave digestion procedure. Performance of the method was evaluated using kidney tissue across a concentration range of 0.50–5.00 μg Cr/g tissue. Data for method linearity, accuracy, precision, digest stability, and storage stability are presented along with limits of detection and quantitation data. Data from a method cross-validation for B6C3F1 mouse kidney tissue are also presented. After validation, the method was applied to analyze samples collected in support of two chronic toxicity and carcinogenesis studies conducted by the National Toxicology Program.

A Longitudinal Study of Trends in Blood Dioxins and Dioxin-Like Compounds Levels in Residents from Two Locations in Japan During 2002–2006 by Hiroyuki Furuya; Fujio Kayama; Manabu Hasegawa; Masaki Nagai; Takaichiro Suzuki (pp. 892-900).

To investigate the recent changes in background exposure to dioxin-related compounds, the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in the blood of individual residents in Japan were repeatedly examined. Twenty-six subjects (mean age, 52.7 ± 1.8) from District A and 23 subjects (mean age, 56.1 ± 1.2) from District B were longitudinally analyzed from 2002 to 2006. District A is a rural area in Osaka prefecture, and District B is an urban area in Saitama prefecture. For the longitudinal analysis of these concentrations, we used the generalized estimating equation (GEE). The PCDD + PCDF concentrations decreased significantly between 2002 and 2006 (p < 0.0001), but the concentrations of dl-PCBs declined to a smaller degree over the 5-year period. A time × body mass index (BMI) interaction suggests that the reduction rate of blood PCDDs + PCDFs and dl-PCB concentrations in subjects with a BMI greater than the 75th percentile is lower than that of subjects with a BMI less than the 75th percentile from 2002 to 2004 over the 5-year period. From other reports on the dietary intake of PCDDs, PCDFs, and dl-PCBs in Japan, the total amount of daily dioxin intake decreased, but the percentage of dl-PCBs increased yearly in the first half of the 2000s. The reason for the difference in the 5-year change in blood concentrations between PCDDs +PCDFs and dl-PCBs may be explained by the yearly change in the dietary dioxin intake.

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Archives of Environmental Contamination and Toxicology (v.58, #3)