Influence of a Toxic <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> Strain on Glutathione Synthesis and Glutathione-S-Transferase Activity in Common Carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> (Teleostei: Cyprinidae)

We evaluated the effects of aqueous extracts of the cyanobacterium-producing microcystin (MC), Microcystis aeruginosa (strain RST9501), on detoxification capacity and glutathione (GSH) synthesis in liver, brain, gill, and muscle—as well as apoptotic protease (calpain) activity in liver and brain—in the common carp Cyprinus carpio (Teleostei: Cyprinidae). Experimental groups were defined as follows: (1) control (CTR); (2) carp treated with an aqueous extract from the toxic cyanobacteria M. aeruginosa in a final MC concentration of 25 μg/kg (MC 25); and (3) carp treated with an aqueous extract from the toxic cyanobacteria M. aeruginosa in a final MC concentration of 50 μg/kg (MC 50). Carp were gavaged with a cyanobacterial aqueous solution or MilliQ water (CTR group). The experiment was conducted for period of 48 h comprising two gavages with a 24-h interval between them. Some of the parameters analyzed in liver, brain, gill, and muscle included activity of the enzymes glutathione-S-transferase (GST), glutamate cysteine ligase (GCL), glutathione reductase (GR), and GSH concentration. We also evaluated GST pi concentration by Western blot as well as calpain activity in liver and brain samples. In carp liver from the MC 50 group, we observed a decrease in GST and GCL activity, which was accompanied by a decreased GSH concentration. In addition, liver calpain activity was highly induced in carp at both MC doses. Thus, MC ingestion affected the liver antioxidant status through decreasing the GSH concentration and the activity of the enzyme involved in its synthesis (GCL). It also decreased the MC detoxification capacity of the liver because total GST activity decreased, a result that cannot be ascribed to GST pi levels. Because GSH acts as an uncompetitive inhibitor of calpain, its decrease should explain the higher activity of this apoptotic enzyme. The main goal of this study was to show that a decrease in GSH concentration is related to decreased activity of GCL, the limiting enzyme involved in GSH synthesis. Because MCs are phosphatase inhibitors and GCL is allosterically inhibited by phosphorylation, the cellular hyperphosphorylation state induced by MC exposure could act as a modulator factor for antioxidant defenses.     

Total and Methyl Mercury Concentrations in Antarctic Toothfish (<Emphasis Type="Italic">Dissostichus mawsoni</Emphasis>): Health Risk Assessment

The concentrations of total mercury (THg) in different organs of the Antarctic toothfish (Dissostichus mawsoni) collected from CCAMLR research blocks in Subarea 88.3 and Division 58.4.1 off the coast of Antarctica were determined. The results revealed THg concentrations of 0.165?±?0.095 mg/kg (0.023–0.454 mg/kg, wet weight) in the Antarctic toothfish. In muscle, methyl mercury (MeHg) accounted for approximately 40% of the THg. In a comparison analysis, muscle and liver tended to bioaccumulate the highest levels of THg, and both THg and MeHg contents showed correlations with fish length and weight. Compared with international guidelines, fish contained 2.5–6.4% and 4.0–10.3% of the provisional tolerable weekly intake for THg recommended by the Joint FAO/WHO Expert Committee on Food Additives and the tolerable weekly intake for MeHg proposed by the European Food Safety Authority, respectively. These results suggest that consumption of the Antarctic toothfish presents no health risk to humans.     

Glyphosate,Alachor and Maleic Hydrazide have Genotoxic Effect on <Emphasis Type="Italic">Trigonella foenum</Emphasis>-<Emphasis Type="Italic">graecum</Emphasis> L.

In the present study effects of herbicides glyphosate (GP), alachlor (AL) and maleic hydrazide (MH) is studied on mitotic cells of Trigonella foenum-graecum L. Seeds of T. foenum-graecum L. treated with a series of concentrations ranging from 0.1%, 0.2%, 0.3%, 0.4% and 0.5% for 1, 2 and 6 h and their effect on mitotic index and chromosomal aberrations was studied. The results indicate that these herbicides reduced mitotic index in dose-dependent manner. In addition, increase in the percentage of abnormal mitotic plates was observed in herbicide treated groups which was both concentration and time dependent. Commonly observed abnormalities were c-mitosis, laggards, bridges, stickiness, c-anaphase, precocious separation, un-equal distribution and fragments. The result of the present investigation indicates that commonly used herbicides GP, AL and MH have significant genotoxic effect on T. foenum-graecum plant.     

Mercury and Methylmercury Accumulation and Excretion in Prairie Voles (<Emphasis Type="Italic">Microtus ochrogaster</Emphasis>) Receiving Chronic Doses of Methylmercury

Methylmercury cation (MeHg) and divalent mercury (Hg⁺⁺) accumulation in liver, kidney, and brain were quantified in prairie voles (Microtus ochrogaster) at 0, 3, 6, and 12 weeks during chronic exposure to aqueous MeHg. Dose groups received deionized water or aqueous solutions containing 9, 103, or 920 ng MeHg/ml. Our study presents temporal patterns of Hg⁺⁺ and MeHg concentrations in organ tissues and makes inter-tissue comparisons at each time point to illustrate the accumulation and distribution of Hg species during the study. MeHg was accumulated in tissues for 3 weeks and then concentrations plateaued. Mercury accumulated in brain, liver, and kidney to average concentrations of 510 ng/g, 180 ng/g, and 3400 ng/g, respectively. MeHg and Hg⁺⁺ concentrations were roughly equivalent in liver, kidney, and urine. MeHg concentrations in brain tissue were 2 to 20 times the concentrations of Hg⁺⁺. Regression analysis was also used to demonstrate the utility of urinalysis as an indicator of Hg⁺⁺ and MeHg concentrations in organ tissue (p < 0.001).     

Biochemical Response of the Copepod <Emphasis Type="Italic">Tigriopus japonicus</Emphasis> Mori Experimentally Exposed to Cadmium

The response of the copepod (Tigriopus japonicus Mori) to cadmium (Cd) additions was investigated under laboratory-controlled conditions in a 12-day exposure. Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione-S-transferase (GST), acetylcholinesterase (AchE), reduced glutathione (GSH), the ratio of reduced to oxidized glutathione (GSH/GSSG), and metallothionein (MT) were analyzed for Cd treatments (0, 10, 20, 40, and 100 μg/L) after exposure for 1, 4, 7, and 12 days. Additionally, thiobarbituric reactive species assay was used to evaluate lipid peroxidation (LPO) of the copepod after the 12-day exposure. The results indicated that Cd treatments significantly influenced the biochemical indexes (SOD, GPx, GST, AchE, GSH, and GSH/GSSG) after certain exposure times. Exposure to Cd induced LPO in the treated copepods, hinting that the copepods had suffered from oxidative damage. During exposure, the Cd initiated an induced MT synthesis in the copepods by day 7, which peaked at day 12 and which was probably responsible for Cd detoxification. Thus, Cd exposure significantly affected the detoxification process and antioxidant system of this copepod, and T. japonicus could be used as a suitable bioindicator of exposure to Cd using SOD, GPx, GST, LPO, and GSH/GSSG as biomarkers.     

<Emphasis Type="Italic">Chlamydia</Emphasis><Emphasis Type="Italic">trachomatis</Emphasis> and placental inflammation in early preterm delivery

Chlamydia trachomatis may infect the placenta and subsequently lead to preterm delivery. Our aim was to evaluate the relationship between the presence of Chlamydia trachomatis and signs of placental inflammation in women who delivered at 32 weeks gestation or less. Setting: placental histology and clinical data were prospectively obtained from 304 women and newborns at the Erasmus MC-Sophia, Rotterdam, the Netherlands. C. trachomatis testing of placentas was done retrospectively using PCR. C. trachomatis was detected in 76 (25%) placentas. Histological evidence of placental inflammation was present in 123 (40%) placentas: in 41/76 (54%) placentas with C. trachomatis versus 82/228 (36%) placentas without C. trachomatis infection (OR 2.1, 95% CI 1.2–3.5). C. trachomatis infection correlated with the progression (P = 0.009) and intensity (P = 0.007) of materno-fetal placental inflammation. C. trachomatis DNA was frequently detected in the placenta of women with early preterm delivery, and was associated with histopathological signs of placental inflammation.     

Chronic Effects of <Emphasis Type="Italic">Pinus radiata</Emphasis> and <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Kraft Mill Effluents and Phytosterols on <Emphasis Type="Italic">Daphnia magna</Emphasis>

Two kraft pulp mill effluents were compared in terms of their chronic toxicity to Daphnia magna. One resulted from pulping Pinus radiata and the other came from a parallel processing of Pinus radiata and Eucalyptus globulus (mixed kraft pulp mill effluent). The concentration of phytosterols found in the mixed kraft pulp mill effluent was higher than in the effluent from Pinus radiata, with values of 0.1082 and 0.02 μg/L, respectively. The phytosterols per se are responsible for 12.9% and 8.1% of the deviation from the natural shape, while the kraft pulp mill effluents account for 25.6%–27.8% of shape deviation. The role of β-sitosterol and stigmasterol is discussed in relation to endocrine disruption.     

Effects of <Emphasis Type="Italic">trans</Emphasis>-10,<Emphasis Type="Italic">cis</Emphasis>-12 conjugated linoleic acid on cholesterol metabolism in hypercholesterolaemic hamsters

Background Conjugated linoleic acid (CLA) has received great attention in recent years because of its pleiotropic biological activities, but considerably fewer studies have been published addressing its role in serum lipids and atherosclerosis compared to other topics covered. Aims of the study The aim of the present study was to assess the effects of the trans-10,cis-12 isomer of CLA on cholesterolaemia and on several metabolic pathways involved in cholesterol metabolism in hamsters. Methods Animals were fed atherogenic diets supplemented with 0.5% linoleic acid, 0.5% trans-10,cis-12 CLA or 1.0% trans-10,cis-12 CLA, for 6 weeks. Serum lipoproteins were separated by FPLC. Cholesterol in serum and liver, as well as triacylglycerols and phospholipids in liver were assessed by spectrophotometry. 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGCoAR), acyl-coenzyme A:cholesterol acyltransferase (ACAT) and cholesteryl ester hydrolase (CEH) activities were measured by radiometry, and LDL receptors were determined by Western blot. Results trans-10,cis-12 CLA feeding did not modify food intake nor final body weight. Although serum total cholesterol remained unchanged, when cholesterol fractions were analyzed a significant decrease in VLDL-cholesterol was observed in CLA-fed animals, without changes in HDL-cholesterol or LDL-cholesterol. trans-10,cis-12 CLA decreased cholesterol ester content and increased free cholesterol in liver. The activity of HMGCoAR was not modified. In contrast, ACAT activity was reduced by both CLA doses and CEH was increased by the high CLA dose. LDL receptors were significantly reduced by trans-10,cis-12 feeding when expressed as arbitrary units per mg of protein, however, the total receptor mass remained unchanged. Conclusions These results suggest that, under the present experimental conditions, trans-10,cis-12 CLA feeding reduces cholesterol esterification in liver and decreases the minority serum VLDL-cholesterol fraction, but it does not show a hypocholesterolaemic effect. A dose–response effect was not observed.     

Impact of <Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis>-Dimethylformamide from Domestic Effluents on River Waters

The impacts of N,N-dimethylformamide (DMF) from domestic effluents on rivers were evaluated. The hourly DMF concentrations in the inflows (0.21–0.52 μg/L in mean) and the outflows (0.18–0.62 μg/L in mean) of two sewage treatment plants did not show any characteristic variations, and DMF was not removed at both plants. The monthly variations of DMF in the outflows of four sewage treatment plants (0.16–0.62 μg/L in mean) and three rivers (0.06–0.11 μg/L in mean) into which the outflows of the plants were discharged did not show any significant variations. The domestic DMF load per person was calculated to be 0.25 mg/d/person. The percent DMF loads of domestic effluents in the rivers were estimated to be below (8%–32%).     

Total Mercury and Methylmercury Residues in River Otters (<Emphasis Type="Italic">Lutra canadensis</Emphasis>) from Wisconsin

The Wisconsin Department of Natural Resources (WDNR) collected trapper-caught river otter (Lutra canadensis) from 3 distinct areas of Wisconsin (north, central, and south). Otter carcasses were collected from a total of 12 counties during the trapping seasons of 2003 and 2004. Liver, kidney, muscle, brain, and fur tissue was collected for mercury (Hg) analysis. Analysis of variance identified collection zone as the significant factor for differences in tissue Hg levels, with a pattern of decreasing Hg concentrations from north to south (p < 0.0001). This trend was apparent in all tissue types analyzed. Strong relationships were observed between Hg concentrations in all tissues. Likewise, highly significant (p < 0.0001) relationships were found to exist between Hg concentrations in otter fur and Hg concentrations of internal organs (brain, muscle, kidney, and liver). Although these data suggest that Hg concentrations are related among tissues, they do not suggest uniform distribution of Hg throughout the tissues. The results suggest that Hg accumulates at higher concentrations in fur followed by liver, kidney, muscle, and brain. Analysis of a subset of samples for methylmercury (MeHg) revealed that MeHg made up a greater percentage of total Hg in brain and muscle compared to liver and kidney tissue. Although a gradient in tissue concentrations was observed from north to south, none of the tissue concentrations reached levels known to cause toxicity in either otter or mink.     

Toxicity of Chiral Pesticide <Emphasis Type="Italic">Rac-</Emphasis>Metalaxyl and <Emphasis Type="Italic">R-</Emphasis>Metalaxyl to <Emphasis Type="Italic">Daphnia magna</Emphasis>

Chirality in pesticides has become a challenge because of enantiomers’ different toxicities to non-target organisms. Acute and chronic toxicities of Rac-metalaxyl and R-metalaxyl to Daphnia magna were determined and compared. The 48-h LC50 for Rac- and R-metalaxyl to Daphnia magna were 51.5 and 41.9 mg/L. In a 14-day chronic test, the lowest-observed-effective concentration (LOEC) and no-observed-effective concentration (NOEC) of Rac-metalaxyl were 2 and 1 mg/L, respectively, whereas those of R-metalaxyl were 1 and 0.1 mg/L. Body length, days-to-first-brood and number of broods per female were significantly (p < 0.05) affected by R-metalaxyl at >1.0 mg/L, but affected by Rac-metalaxyl at ≥2.0 mg/L.     

Cadmium Accumulation and Antioxidant Responses in <Emphasis Type="Italic">Sparus aurata</Emphasis> Exposed to Waterborne Cadmium

Cadmium (Cd), a nonessential trace element, is rapidly accumulated by most living organisms and subsequently exerts its toxicity at different molecular levels. This study exposed gilthead sea bream (Sparus aurata) to waterborne 0.1 mg/l Cd for 11 days and investigated the Cd accumulation pattern, lipid oxidation, and response of antioxidant defences. At the end of the experiment, mean Cd concentrations in gills and liver, the organs most prone to metal accumulation, were 209.4 and 371.7 ng/g ww, respectively. Muscle did not show any Cd retention during the 11 days of exposure. In liver, the cytosolic fraction of the metal was chelated into the nontoxic form by metallothionein (MT), a specific Cd-inducible protein. Zn and Cu concentrations were not influenced by Cd exposure. Glutathione (GSH) concentrations and the antioxidant enzyme activities of GSH reductase and GSH peroxidase showed an overall decreasing trend. In addition, lipid and aqueous hydroperoxide levels did not show any significant variation. Oxidative stress indirectly generated by Cd seems to be compensated for by the different biochemical systems tailored to decrease cellular damage. In particular, the negative effects of Cd accumulation in tissues were probably counteracted by the induction of MT.     

DNA Damage Induced by Copper on Erythrocytes of Gilthead Sea Bream <Emphasis Type="Italic">Sparus aurata</Emphasis> and Mollusk <Emphasis Type="Italic">Scapharca inaequivalvis</Emphasis>

Heavy metal pollution in rivers and its impact on aquatic ecosystems is a dynamic process. Fish are ideal indicators of heavy metal contamination in aquatic systems because they occupy different trophic levels and are of different sizes and ages. In particular, copper is an essential trace metal for living organisms and it is present in all natural waters and sediments. In this paper, we report data on the effect of copper on DNA erythrocytes from the teleost gilthead sea bream Sparus aurata and the bivalve mollusk Scapharca inaequivalvis. In particular, the effect of 0.1 ppm of Cu²⁺ on the nucleated erythrocytes was analyzed using the comet assay. This test is a promising tool for estimation of DNA damage at the single cell level. The data obtained show that the in vivo treatment with 0.1 ppm of copper increased the susceptibility of DNA to be damaged. Exposure to Cu²⁺ produces a more evident effect on Sparus aurata, as all three comet parameters significantly increased (tail length, tail intensity, and tail moment). The higher comet parameters measured in Scapharca inaequivalvis compared to Sparus aurata could be due to the difference in stability of the respective hemoglobins. The comet assay could represent a useful test to evaluate the biological consequences of environmental contamination by metals on marine organisms.     

Kinetics of Cadmium Uptake and Subcellular Partitioning in the Earthworm <Emphasis Type="Italic">Eisenia fetida</Emphasis> Exposed to Cadmium-Contaminated Soil

To understand the bioaccumulation kinetics of cadmium (Cd) at the subcellular level, toxicokinetics and subcellular fractionation of Cd were determined for the terrestrial earthworm Eisenia fetida exposed to Cd-contaminated red soil. The bioaccumulation factor was 1.74 after exposure to soil containing 1 mg Cd/kg dry weight for 21 days. Four distinct compartments with different Cd-binding affinities were obtained by sequential centrifugations, including the cytosolic fraction (G), the organelles fraction (F), the granular fraction (D), and the tissue, cell membrane, and intact cell fraction (E). Most of the accumulated Cd in the organisms existed as fraction G. Cd bound to the tissue and cell membrane fraction (E) was <8% of total Cd. Only approximately 3% of Cd existed in fraction F, and <1% was found in fraction D. Accumulation and elimination kinetics of Cd in different fractions were accurately described by one-compartment models, and kinetic parameters (uptake rate constant k ₁ and elimination rate constant k ₂ ) were derived. Fractions G and F were responsible for the linear accumulation pattern during 3 weeks of exposure, whereas fractions D and E showed a nonlinear uptake curve, and steady state was achieved after 7 and 14 days of exposure, respectively.     

Fertilizer Industry Effluent Induced Biochemical Changes in Fresh water Teleost, <Emphasis Type="Italic">Channa striatus</Emphasis> (<Emphasis Type="Italic">Bloch</Emphasis>)

The industrial activities pose threat to the life of aquatic organisms in many ways. This research communication presents an account of the impact of fertilizer industry effluent upon the levels of protein and the activity of lactate dehydrogenase (EC 1.1.1.28, LDH), a terminal key enzyme in glycolytic pathway, in different organs of a fresh water teleost fish, Channa striatus (Bloch). The fish exposed to different sublethal concentrations of fertilizer industry effluent (3.5, 4.7 and 7.0% v/v) equivalent to 1/20th, 1/15th and 1/10th of LC₅₀ value (70% v/v) for varying treatment periods (96 h and 15 days) exhibited decrease in the level of protein (8–76%) in different organs of the effluent treated fish. At highest effluent concentration (7% v/v) treatment for short (96 h) or long (15 days) duration, the liver of the fish registered significant (p < 0.001) decrease (62–76%) in protein content as compared to control, whereas other organs of the fish showed only 38–52% decrease in the level of protein. The industrial effluent also caused marked reduction in the activity of LDH in different fish tissues when compared to the control. The treatment of fish with 7% effluent concentration for 96 h caused 78% decrease (p < 0.001) in the LDH activity in fish muscle whereas after 15 days the effect was maximum in fish brain as it exhibited 86% decrease (p < 0.001) in LDH activity as compared to control. The effect of effluent on the activity of LDH and protein content in different body tissues of the fish was dependent on concentration and duration of exposure. The significant reductions in the activity of LDH and level of protein in fish tissues due to treatment with the fertilizer industry effluent indicated the possibility of impairments in energy metabolism and protein turnover, respectively, in C. striatus.     

Toxicity of Fuel Oil Water Accommodated Fractions on Two Marine Microalgae, <Emphasis Type="Italic">Skeletonema costatum</Emphasis> and <Emphasis Type="Italic">Chlorela spp</Emphasis>

In this paper, the acute toxicity of four fuel oils including F120, F180, F380 and No.-20 was evaluated by exposing the marine microalgae Chlorela spp. (Chlorophyta) and Skeletonema costatum (Bacillariophyta) in the fuel oil water accommodated fractions (WAF). The bioassay showed that F180 WAF was the most toxic to both microalgae. The 96 h EC₅₀ value of F180 WAF for Skeletonema costatum and Chlorela spp. was 9.41 and 13.63 mg/L expressed in concentration of total petroleum hydrocarbons, respectively. WAFs of F120, F180 and F380 were more toxic to Skeletonema costatum than to Chlorela spp. In contrast, No.-20 WAF did not show significant toxicity for both Skeletonema costatum and Chlorela spp.     

Impact of Distillery Soil Leachate on Haematology of Swiss Albino Mice (<Emphasis Type="Italic">Mus musculus</Emphasis>)

This study reports significant alterations in various haematological parameters such as red and white blood corpuscles counts, haemoglobin content and packed cell volume in adult Swiss albino mice orally administered with diluted distillery soil leachate (5%–20%) for 30 days. Soil leachate also affected red blood cell morphology (poikilocytosis). The haematology of exposed mice improved in the reversal groups. Present study infers contamination potential of distillery soil leachate in the groundwater.     

<Emphasis Type="Italic">Sasa veitchii</Emphasis> extract protects against carbon tetrachloride-induced hepatic fibrosis in mice

Background

The current study aimed to investigate the hepatoprotective effects of Sasa veitchii extract (SE) on carbon tetrachloride (CCl₄)-induced liver fibrosis in mice.

Methods

Male C57BL/6J mice were intraperitoneally injected with CCl₄ dissolved in olive oil (1 g/kg) twice per week for 8 weeks. SE (0.1 mL) was administered orally once per day throughout the study, and body weight was measured weekly. Seventy-two hours after the final CCl₄ injection, mice were euthanized and plasma samples were collected. The liver and kidneys were collected and weighed.

Results

CCl₄ administration increased liver weight, decreased body weight, elevated plasma alanine aminotransferase, and aspartate aminotransferase and increased liver oxidative stress (malondialdehyde and glutathione). These increases were attenuated by SE treatment. Overexpression of tumor necrosis factor-α was also reversed following SE treatment. Furthermore, CCl₄-induced increases in α-smooth muscle actin, a marker for hepatic fibrosis, were attenuated in mice treated with SE. Moreover, SE inhibited CCl₄-induced nuclear translocation of hepatic nuclear factor kappa B (NF-κB) p65 and phosphorylation of mitogen-activated protein kinase (MAPK).

Conclusion

These results suggested that SE prevented CCl₄-induced hepatic fibrosis by inhibiting the MAPK and NF-κB signaling pathways.