Plasma biochemical responses of the omnivorous crucian carp (Carassius auratus) to crude cyanobacterial extracts

Healthy crucian carp (Carassius auratus) were treated by intraperitoneal (i.p.) injection of crude cyanobacterial extracts at two doses, 50 and 200 mu g MC-LR equiv kg(-1) BW. High mortality (100%) was observed within 60 h post injection in the high-dose group. In the treated fish, activities of four plasma enzymes, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), all showed substantial increases, with both dose and time-dependent effects. These increases of enzyme activity indicate severe impairment occurred in the liver of crucian carp over time. Plasma concentrations of energy-related biomolecules including glucose (GLU), cholesterol (CHO), triglyceride (TG), and total protein (TP) showed marked changes in the high-dose group, possibly a nutritional imbalance correlated with the liver injury caused by intraperitoneal exposure to crude cyanobacterial extracts.

Relationships among the contents of total phenolics, soluble carbohydrate, and free amino acids of 15 aquatic macrophytes

In order to examine how carbon and nitrogen status of a macrophyte may affect its total phenolics (TP) production, the contents of free amino acids (FAA), soluble carbohydrate (SC) and TP were examined in leaves of seven submersed, four floating-leaved, and four emergent macrophytes. The floating-leaved and emergent macrophytes had much higher contents of SC and TP than the submersed macrophytes. The contents of FAA were not significantly different among the submersed, floating-leaved, and emergent macrophytes. Correlations among the contents of FAA, SC, and TP indicated that the production of TP was more dependent on the SC content than on the FAA content.

The role of glutathione metabolism in tolerance of tobacco BY-2 suspension cells to microcystin-RR

This study was undertaken to investigate the role of the glutathione-involved detoxifying mechanism in defending the tobacco BY-2 suspension cells against microcystin-RR (MC-RR). Analysis showed that exposure of the cells to different concentrations of MC-RR (0.1, 1 and 10 mu g/mL) for 0-6 days resulted in a time and concentration-dependent decrease in cell viability and increase in reactive oxygen species (ROS) content. Reduced glutathione (GSH) and total glutathione (tGSH) content as well as glutathione reductase (GR), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) activities significantly increased after 3-4 days exposure in the highest two concentration treated groups, while decreased until reaching the control values except for GPX at day 6. Oxidized glutathione (GSSG) content markedly increased compared with control in high concentration MC-RR treated group after 6 days exposure. The GSH/GSSG ratio was much higher than control in 10 mu g/mL MC-RR treated group at day 4, but after 6 days exposure, the ratios in all treated groups were lower than that of the control group.

PCBs and OCPs in human milk and selected foods from Luqiao and Pingqiao in Zhejiang, China

This study was conducted to measure the levels of 23 PCB congeners and 6 organochlorine pesticides (OCPs) in human milk and three food types collected from Luqiao and Pingqiao in Zhejiang Province, China. An effort was also made to explore the potential health risk for the mothers and breast-fed infants living in these two localities. Luqiao was selected as the sampling site because it is the largest place for the disassembly of obsolete transformers and electrical waste in China. Pingqiao, located 100 kin NW of Luqiao, is not known to be a place for any electronic or electrical waste and hence was chosen as the control site. Both localities are important agricultural places in the province. The organochlorines were measured in the samples using the GC-PECD technique. Micro-EROD bioassay method was also used as a complement of the chemical analysis to estimate the TEQ levels of dioxin-like PCBs in human milk. The data showed that the human milk, rice, hen egg, and fish samples from Luqiao were more heavily contaminated with PCBs than those from Pingqiao, suggesting that the mothers and their breast-fed infants in Luqiao tended to receive greater exposure to PCBs than those living in Pingqiao. The OCP levels in the two localities were found comparable, suggesting that the major source of contamination with these pesticides was from their agricultural uses. Significant correlation (R-2 = 0.87, P < 0.001) of PCB TEQs was found between the bioassay and chemical analysis method, suggesting that micro-EROD is an effective method for comprehensive determination of TEQ levels in human milk. Comparison with literature data showed that the PCB levels in milk samples from Luqiao were significantly higher than those from localities in other Chinese provinces and comparable to those in developed or industrialized countries. (c) 2007 Published by Elsevier B.V.

Prenatal exposures to persistent organic pollutants as measured in cord blood and meconium from three localities of Zhejiang, China

Prenatal exposures to persistent organic pollutants were assessed using the levels of PCBs and organochlorine pesticides (OCPs) measured in cord blood and meconium samples from Luqiao and two other localities of the Zhejiang province in China. Luqiao is a town with the largest site for disassembly of PCB-containing obsolete transformers and electrical waste in China. The other two localities Pingqiao (100 km NW of Luqiao) and Lin'an (500 km NW of Luqiao) are towns without known electronic or electrical waste sites. A total of 23 PCB congeners (including 12 dioxin-like) and 6 OCPs were measured using the traditional GC-mu ECD technique. Micro-EROD bioassay was additionally used to measure TCDD-based TEQ levels of the 12 dioxin-like PCBs. Significant correlations were found between the TEQs measured by the two methods, supporting the application of micro-EROD as a practical toot for complementing the chemical analysis. The data showed that beta-HCH, p,p'-DDE, and 6 PCB congeners (101, 138 153, 180, 183, and 187) were the predominant pollutants, with PCB 138 being the best indicator (predictor) for total PCB levels. Cord blood and meconium from Luqiao have higher levels of PCBs than those from the other two localities, suggesting that a disassembly site for electronic and electric waste would provide an environment for greater exposure to these chemicals. The cord blood or meconium levels of beta-HCH, though likewise considerably high, were comparable in the three localities. Similar findings were observed for p,p'-DDE. Pollution by these OCPs might have come from past use of agricultural pesticides in the three localities. (c) 2007 Published by Elsevier B.V.

Effects of solar ultraviolet radiation on photosynthesis of the marine red tide alga Heterosigma akashiwo (Raphidophyceae)

In order to assess the short- and long-term impacts of UV radiation (LTVR, 280-400 nm) on the red tide alga, Heterosigma akashiwo, we exposed the cells to three different solar radiation treatments (PAB: 280-700 rim, PA: 320-700 nm, R 400-700 nm) under both solar and artificial radiation. A significant decrease in the effective quantum yield () during high irradiance periods (i.e., local noon) was observed, but the cells partially recovered during the evening hours. Exposure to high irradiances for 15, 30, and 60 min under a solar simulator followed by the recovery (8 h) under dark, 9 and 100 mu mol photons m(-2) s(-1) of PAR, highlighted the importance of the irradiance level during the recovery period. Regardless the radiation treatments, the highest recovery (both in rate and total Y) was found at a PAR irradiance of 9 mu mol photons m(-2) s(-1), while the lowest was observed at 100 mu mol photons m(-2) s(-1). In all experiments, PAR was responsible for most of the observed inhibition; nevertheless, the cells exposed only to PAR had the highest recovery in any condition, as compared to the other radiation treatments. In long-term experiments (10 days) using semi-continuous cultures, there was a significant increase of UV-absorbing compounds (UVabc) per cell from 1.2 to > 4 x 10(-6) mu g UVabc cell(-1) during the first 3-5 days of exposure to solar radiation. The highest concentration of UVabc was found in samples exposed in the PAB as compared to PA and P treatments. Growth rates (mu) mimic the behavior of UV-absorbing compounds, and during the first 5 days mu increased from < 0.2 to ca. 0.8, and stayed relatively constant at this value during the rest of the experiment. The inhibition of the Y decreased with increasing acclimation of cells. All our data indicates that H. akashiwo is a sensitive species, but was able acclimate relatively fast (3-5 days) synthesizing UV-absorbing compounds and thus reducing any impact either on photosystem 11 or on growth. (c) 2006 Published by Elsevier B.V.

Methylmercury, total mercury and total selenium in four common freshwater fish species from Ya-Er Lake, China

Snakehead fish (Ophiocephalus argus cantor), silver carp (Hypophthalmichthys molitrtix), crucian carp (Carassius carassius), and common carp (Cyprinus carpio) are four common freshwater fish species in China. In this study, the level of methylmercury (MeHg), total mercury (T-Hg), and total selenium (T-Se) in muscle samples of these four fish species from Ya-Er Lake, China, were analyzed using atomic fluorescence spectrometry coupled with high-performance liquid chromatography, and inductively coupled plasma mass spectrometry. The concentrations of MeHg in all the fish species were significantly correlated with those of T-Hg. Higher T-Hg and MeHg concentrations had accumulated in the snakehead fish, which is a strongly predatory fish, than in the other three species. The concentration ratios of MeHg and T-Hg in the muscles of these four fish species were almost equal. Conversely, there was negative correlation between the concentrations of T-Hg and T-Se, which implies that there is a competition between these two elements with respect to bioaccumulation. It is noteworthy that of all the muscle samples tested, the level of T-Hg exceeded the maximum allowable limit in fish [0.4 mg kg(-1) (w/w) recommended by the World Health Organization] in 38.46% of those of the silver carp, 18.18% of those of the crucian carp, and 100% of those of snakehead fish. These results show that the consumption of contaminated fish is a potential threat to human health and that necessary preventive measures to safeguard public health should be emphasized.

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Exposure of spermatozoa to duroquinone may impair reproduction of the common carp (Cyprinus carpio) through oxidative stress

Toxicity of many waterborne organic contaminants to aquatic organisms is mediated through oxidative damages resulting from the production of reactive oxygen species (ROS). Using duroquinone as a model ROS inducer, we carried out in vitro and in vivo experiments to test the hypothesis that reproduction in common carp (Cyprinus carpio) can be impaired through oxidative damage of their spermatozoa. In vitro exposure of fish spermatozoa to 0, 12.5, 25, 50, 100 and 200 mu M duroquinone for 2 h showed a significant increase in the level of ROS in a dose-dependant manner. Sperm motility was significantly reduced in all exposure groups, but lipid peroxidation (LPO) and DNA strand break (measured by comet assay) were only enhanced at 50 mu M and above. A significant decrease in subsequent hatching rate was recorded in all the exposure groups, despite fertilization rate was not affected. In the in vivo experiment, spermatozoa were collected 24 and 72 h after fish received intra-peritoneal injections of 1.0 and 10 mg kg(-1) body weight duroquinone. DNA damage was clearly evident in spermatozoa of all treatment groups after 72 h exposure, and ROS was significantly enhanced in the high concentration group. LPO however, remained unchanged in both treatment groups. The overall results of both our in vitro and in vivo experiments demonstrated that duroquinone can induce ROS production in spermatozoa, which may impair sperm quality and subsequently reproductive success through oxidative stress. (c) 2006 Elsevier B.V. All rights reserved.

Reproductive effects of prenatal exposure to nonylphenol on zebrafish (Danio rerio)

Mature female and male zebrafish were separated and exposed to nonylphenol (NP) at 0.1, 1, 10, 50, 100 and 500 mu g/L, respectively, for 3 weeks. Gonadosomatic index (GSI) in both sexes and vitellogenin (VTG) induction in males was measured as the bioindicators for the impairment to the parents. The results indicated that 50 mu g/L of NP was the non-observed effect concentration (NOEC) for GSI and VTG induction. Afterwards, the 50 mu g/L NP exposed females and males, and the control females and males were cross-wise pair-bred in the control water for one week to examine the reproductive effects. The embryonic cathepsin D (CAT D) activity, eggshell thickness, fecundity, hatching rate and malformation (vertebral column flexure) rate of offspring were determined in the four pair-bred groups. While endpoints remained unchanged in the groups with exposed males, prenatal exposure of females to 50 mu g/L of NP resulted in the impairment of reproduction in groups with exposed females including inhibition of CAT D activity (P < 0.05), decrease of eggshell thickness (by 23.6%) and elevation of malformation rate (P < 0.001). These results suggested NP could induce reproductive damage to zebrafish at NOEC for parents. The results also imply that alterations of CAT D activity and eggshell thickness may be more sensitive biomarkers to indicate the reproductive effects caused by endocrine disrupting chemicals. (c) 2005 Elsevier Inc. All rights is reserved.

Partial and total replacement of fishmeal with poultry by-product meal in diets for gibel carp, Carassius auratus gibelio Bloch

Triplicate groups of gibel carp Carassius auratus gibelio Bloch (initial body weight: 4.89 g) were fed for 8 weeks at 24.8-30.8 degrees C with nine isonitrogenous and isoenergetic diets. The control diet (F1) used white fishmeal (FM) as the sole protein source. In the other eight diets (F2-F9), 40.5-100% of FM protein was substituted by poultry by-product meal (PBM) at 8.5% increments. The specific growth rate (SGR), feed efficiency ratio, protein efficiency ratio, protein retention efficiency and energy retention rate for fish fed PBM diets (F2-F9) were all higher, but not always significantly, than those for fish fed F1. All apparent digestibility coefficients for fish fed PBM diets were lower than those for fish fed F1. Fish fed F1 had a significantly higher hepatosomatic index value than fish fed PBM diets (P < 0.05). No significant (P > 0.05) effect of diet was found in whole-body moisture and fat content. Whole-body protein and energy content for fish fed PBM diets were slightly higher than that for fish fed F1. The optimal replacement level of FM by PBM was estimated by second-order polynomial regression to be 66.5% in protein.

Total and subcellular distribution of trace elements in the liver of a mother-fetus pair of Dall's porpoises (Phocoenoides dalli)

Total and subcellular hepatic Zn, Cu, Se, Mn, V, Hg, Cd, and Ag were determined in a mother-fetus pair of Dall's porpoises (Phocoenoides dalli). Except for higher fetal Cu concentration, all maternal elements were higher. Elements existed mostly in the cytosol of both animals except in the case of maternal Ag in the microsome and fetal Cu and Ag in the nuclei and mitochondria. In the maternal cytosol, Zn, Mn, Hg, and Ag were present in the high-molecular-weight substances (HMW); Se and V were present in the low-molecular-weight substances (LMW); Cu and Cd were mostly sequestered by metallothionein (MT). In the fetal cytosol, Zn, Se, Mn, Hg, Cd, and Ag were present in the HMW and V in the LMW, while Cu and Ag were mostly associated with MT. MT isoforms were characterized using the HPLC/ICP-MS. Two and four obvious peaks appeared in the maternal and fetal MT fractions, respectively. The highest elemental ion intensities were at a retention time of 7.8 min for the mother, and for the fetus the peak elemental ion intensities occurred at a retention time of 4.3 min, suggesting that different MT isoforms may be involved in elemental accumulation in maternal and fetal hepatocytosols. (C) 2003 Elsevier Ltd. All rights reserved.

Endocrine effects of sublethal exposure to persistent organic pollutants (POPS) on silver carp (Hypophthalmichthys molitrix)

Silver carp (Hypophthalmichthys molitrix) samples were collected from five selected sites that represent diverse levels of downgraded persistent organic pollutants (POPs) contamination in Ya-Er Lake in October 1999. Hepatic ethoxyresorufin-O-deethylase (EROD) and UDP glucuronosyltransferase (UDPGT) activities, hepatosomatic index (HSI), hepatic retinoids, serum thyroid hormones were measured. It was found that hepatic retinol and serum free 3,5,3'- tetraiodothyronine (FT3) significantly increased (P < 0.01) when both hepatic EROD and UDPGT activities significantly declined (P < 0.01) from pond 1 to 5 with decrease in the degree of pollution. This significant negative correlation (P < 0.01) suggests that the persistent organochlorinated contaminants could induce hepatic EROD and UDPGT activities, alter retinoid and thyroid hormone homeostasis, and finally lead to the reduction of retinol and FT3, the two biologically active forms of retinoids and thyroid hormone in silver carp of Ya-Er Lake. (C) 2002 Elsevier Science Ltd. All rights reserved.

Contamination and distribution of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) in agriculture fields in Ya-Er Lake area, China

The contamination and distribution of polychlorinated dibeinizo-p-dioxins and dibenzofurans (PCDD/Fs) from two agricultural fields of a heavily polluted lake area in China (Ya-Er Lake) are presented. The vertical distribution pattern of total PCDD/Fs in soil cores reveals that the maximum concentration was in the layer of 20-30 cm. The concentrations in the top layer of soil at the two sites were similar (17.48 ng/kg at Site 1 and 18.10 ng/kg at Site 2), but the maximum concentration of Site 1 (120.8 ng/kg) was two times higher than that of Site 2 (64.39 ng/kg). The maximum concentration of PCDD/Fs in mud cores in rice fields (0-50 cm) at Sites 1 and 2 was in the layer of 0-10 cm. The maximum PCDD/F concentration in the top layer in mud at Site 1 (203.1 ng/kg) was higher than that at Site 2: (143.3 ng/kg). Significant correlations were found between the mind PCDD/Fs and the organic carbon content (R = 0.9743, P< 0,05 at Site 1; R = 0.9821, P< 0.05 at Site 2), the two variables being highly correlated (R = 0.9049, P< 0.05, at Site 1; R = 0.9916, P< 0.05 at Site 2). All correlation coefficients were significant at the 95% level. Concentrations were highly correlated with organic carbon, indicating that sorption to organic carbon was the dominant mechanism. Using principal component analysis, the homologue profiles of soil, mud, and plants (rice and radish) were compared. The PCDD/F patterns in plants were found not to be correlated to those in soil and mud. This suggests that atmospheric deposition may be the main source of PCDD/Fs in rice grain. However, mixed exposure involving uptake mechanisms and atmospheric deposition is considered main the source of PCDD/F pollution in radishes. (C) 2002 Elsevier Science (USA).

Cytological and biochemical alterations in Carassius auratus hepatocytes from exposure to sediment containing dioxins and related compounds

Cytological and biochemical alterations of crucial carp (Carassius auratus) hepatocytes were characterized after exposure to sediments from a lake contaminated with dioxins and other industrial chemicals. Carp were exposed in 20 L water containing 25, 50, or 100 g of contaminated sediment for 2 and 4 weeks. Ultrastructural changes in the liver were characterized by severe enlargement of hepatocytes. Alterations in the cell. included formation of condensed and irregular cell nucleus, polynuclei, dispersed heterochromatin, enlargement of the nucleolus, and degeneration of the nucleus. Mitochondrial numbers were reduced and cristae were deformed. Myelin figures and lysosomes were increased, and sometimes cell organelles and cell matrix were totally lost after 4 weeks of exposure. The ultrastructural alterations were correlated with exposure time and sediment concentrations. Hepatosometic index was significantly increased in experimental groups at 2 and 4 weeks as compared with the control group. EROD enzyme activities were strongly induced in liver. A trend from rough endoplasmic reticulum (RER) to SER was observed. Our results suggest that the dioxin-like compounds bound by sediment were bioavailable to C. auratus and cause sublethal effects.