Endocrine response of the freshwater teleost, Sarotherodon mossambicus (Peters) to dimecron exposure

The endocrine response in a freshwater teleost, Sarotherodon mossambicus (Peters) under dimecron (an organophosphate pesticide) toxicity was investigated by estimating the serum levels of T3 (triiodothyronine), T4 (thyroxine), cortisol, prolactin and insulin in control and sub-lethal (0.001 ml l(-1)) dimecron-exposed fish for 1, 6, 12, 24h and 5 days. In control S. mossambicus, the serum levels of T3 ranged from 0.80+/-0.01 to 0.82+/-0.01 ng ml(-1); T4 from 2.20+/-0.01 to 2.25+/-0.01 microg dl(-1); cortisol from 8.30+/-0.03 to 8.34+/-0.01 microg dl(-1); prolactin from 1.50+/-0.01 to 1.54+/-0.01 microg ml(-1); insulin from 9.70+/-0.01 to 9.76+/-0.01 microU ml(-1) up to a maximum period of 5 days maintained in pollutant-free tap water. Exposure of fish to sub-lethal concentration of dimecron caused varying changes in the levels of serum hormones studied. Based on the results obtained, it was concluded that (i) the fish adaptively maintains a probable low metabolic rate, as indicated by the reduced levels of thyroid hormone (T3) as well as the glucocorticoid hormone (cortisol), which could be considered advantageous for the fish to indirectly reduce the toxic impact of the pesticide, (ii) the elevated levels of prolactin in the fish under pesticide stress is indicative of a possible hydromineral regulatory effect of the hormone (probably by influencing specific organs such as gills and kidney) under pesticide toxicity, (iii) the increased insulin level in the fish under pesticide stress is indicative of its role in favouring an adaptive tissue glycogenesis besides a possible increased lipogenesis to sequester the pesticide residue thereby reducing the toxic effect of the pesticide and (iv) the prolonged exposure of the fish (for 5 days) to sub-lethal dimecron appeared to exhibit a uniform recovery response in the different hormonal levels of the fish.     

In situ studies on the distribution patterns and dynamics of microcystins in a biomanipulation fish--bighead carp (Aristichthys nobilis)

The distribution and dynamics of microcystins in various organs of the phytoplanktivorous bighead carp were studied monthly in Lake Taihu, which is dominated by toxic cyanobacteria. There was a good agreement between LC-MS and HPLC-UV determinations. Average recoveries of spiked fish samples were 63% for MC-RR and 71% for MC-LR. The highest MC contents in intestine, liver, kidney and spleen were 85.67, 2.83, 1.70 and 1.57 microg g-1 DW, respectively. MCs were much higher in mid-gut walls (1.22 microg g-1 DW) than in hind- and fore-gut walls (0.31 and 0.18 microg g-1 DW, respectively), suggesting the importance of mid-gut wall as major site for MC absorption. A cysteine conjugate of MC-LR was detected frequently in kidney. Among the muscle samples analyzed, 25% were above the provisional tolerable daily intake level by WHO. Bighead is strongly resistant to microcystins and can be used as biomanipulation fish to counteract cyanotoxin contamination in eutrophic waters.     

Biological response of high-back crucian carp (Carassius auratus) during different life stages to wastewater treatment plant effluent

This study presents the adverse effects of endocrine disrupting chemicals (EDCs) in effluent of wastewater treatment plants (WwTPs) on fish health. A study of chronic exposure to WwTPs effluent for 10 months was undertaken in high-back crucian carp (Carassius auratus) during different life stages, covering early-life-stage (ELS), prespawning period, and postspawning period. Condition factor (CF), gonadosomatic index (GSI), hepatosomatic index (HSI), and plasma vitellogenin (VTG) levels were employed as indicators to assess biological effects of effluent on this gynogenesis species. Meanwhile, some high-back crucian carp were caged in Demonstration Base of Biological Purification for Filter-feeding Fish (hereinafter, Demonstration Base), as WwTPs effluent exposure controls. In the meantime, a depuration study was carried out to determine whether or not the estrogenic effects caused by effluent exposure could be reduced after moving fish into EDCs-free water. CF, HSI, GSI, and plasma Vtg levels of high-back crucian carp caged in Demonstration Base were generally in accordance with seasonal change. Effluent exposure inhibited gonadal growth, reducing GSI in ELS while increasing it around spawning, sharpened liver burdens, increasing HSI, and induced abnormal Vtg expression in juvenile high-back crucian carp, augmenting Vtg concentrations in plasma. Around spawning period, Vtg in high-back crucian carp were mainly induced by endogenous estrogens, and EDCs in effluent had less influence on them. Staying in EDCs-free water for 30 days made high-back crucian carp recover from effects of previous effluent exposure, relieving inhibition of gonadal development and hypertrophy of liver as well as reducing Vtg induced by EDCs in effluent. The results revealed that high-back crucian carp in ELS are more sensitive to WwTPs effluent exposure. Additionally, the depuration study showed a clearance of the estrogenic effects caused by effluent.     

Importance of natural sedimentation in the fate of microcystins

Sedimentation processes of microcystins (MCs), cyanobacterial toxins, were studied in three reservoirs located in Central Spain in which the cyanobacterial community was dominated by the genus Microcystis. MCs were detected in the sediment traps deployed in all reservoirs. In Santillana reservoir, MCs were identified in sediment traps even though they could not be found in the pelagial samples. In the other reservoirs studied, sedimentation rates for MC-containing particles during the bloom period ranged from 0.43 to 2.53 mg m⁻² d⁻¹. Interestingly, this very high sedimentation of toxic biomass is not exclusively related to decaying blooms or autumnal sedimentation due to a drop in water temperature. Instead, it seems that MC-containing colonies may be settling constantly during the bloom period and we were able to estimate that during such a Microcystis dominated bloom, around 4.5% of pelagial MCs may be involved in sedimentation. Further, these settling colonies seem to maintain good cell integrity and MCs seem not to be excreted massively. A certain loss of toxin content along the vertical settling may be attributed to minor losses due to cell lysis or to variations in MC cell quota explained by reduced production or internal consumption. Our results for the first time establish specific settling rates for MC-containing particles in freshwaters and definitely identify sedimentation as a major destination for these toxins. These data may contribute to improve managing strategies concerning risks associated with MCs.     

Endocrine and metabolic alterations in the mink (Mustela vison) due to chronic phytoestrogen exposure

Phytoestrogens are natural components of plant-based food items with beneficial health effects. The aim of the present study was to investigate the chronic effects of dietary phytoestrogens, genistein (8 mg kg(-1) day(-1)) and beta-sitosterol (50 mg kg(-1) day(-1)), on the weight regulation of the mink (Mustela vison). The parental generation was exposed from August 2002 to May-June 2003 to either beta-sitosterol or genistein, while the kits were exposed through gestation and lactation. Food consumption and body masses were monitored monthly. Plasma lipid, glucose, total protein and hormone (ghrelin, leptin, triiodothyronine and thyroxine) concentrations were measured from the parents in August 2002, January 2003 and at the end of the experiment in May-June 2003 when the kits were 21 days of age. Relative food intake was higher in the beta-sitosterol-exposed minks than in the control or genistein minks in September 2002. Plasma leptin and total protein concentrations were lower in the beta-sitosterol kits compared to the control kits. Furthermore, plasma ghrelin levels and liver phosphorylase activities of the mink kits were higher due to genistein exposure. In mink kits, exposure to both phytoestrogens reduced the plasma thyroxine concentrations. The kidney glycogen concentrations and the muscle phosphorylase activities of phytoestrogen-treated adult minks were elevated. The results of this study suggest that minks are sensitive to perinatal phytoestrogen exposure.     

Possible effects of persistent organochlorinated pollutants cocktail on thyroid hormone levels and pituitary-thyroid interrelations

In polluted district of Michalovce in East Slovakia (POLL) and two districts with background pollution (BCGR) 2046 adults (834 males and 1212 females aged 20-75 years) were examined. Serum levels of thyrotropin (TSH), free thyroxine (FT4), total triiodothyronine (TT3) and antithyroperoxidase antibodies (TPOab) were estimated by electrochemiluminiscent assay and also these of 15 polychlorinated biphenyl congeners (PCBs), p,p'-DDE, p,p'-DDT, hexachlorobenzene (HCB) and hexachlorocyclohexane were measured by high resolution gas chromatography/mass spectrometry. In addition, also dioxins, furans, coplanar- and mono-ortho-PCBs as well as selected hydroxylated and methylsulphonated PCBs and DDE metabolites were measured by appropriate methods based on gas chromatography/mass spectrometry principle. In POLL significantly higher levels of all organochlorines were found than these in BCGR. When pooled values from both areas were stratified in terms of PCBs level and treated as continuous variables, positive association of PCBs with FT4 and TT3 was found, the latter two being also mutually associated. However, within the category of PCBs level <530 ng/glipid (n=232) the association between PCBs and both the FT4 (p<0.09) and TT3 (p<0.03) was negative and any association of these was not found within the category of PCBs level of 531-1000 ng/g (n=691). In contrast, in the category of 531-2000 ng/g (n=1307) positive association appeared between PCBs and FT4 (p<0.001) as well as TT3 (p<0.05). Highly significant association of PCBs with FT4 (p<0.001) was further found in the categories with PCBs level of 1001-101414 ng/g (n=1307) and 2001-101414 (n=1123), while significant association with TT3 was observed only in the category of 531-2000 ng/g. Such findings suggest possible threshold level in positive effect of PCBs on FT4 and TT3 level which seems to be individual and located somewhere around the PCBs level of 1000 ng/g. However, highly significant negative association of both FT4 and TT3 with TSH was found in each of above indicated PCBs categories. Considerable difference in FT4 and TT3 level between large groups of subjects with the same range of PCBs level was also found suggesting different individual susceptibility to the effects of organochlorines. Among a total of 26 cases from POLL with very low TSH level (<0.5 mU l(-1)) 13 cases showed very high level of PCBs, FT4 and TT3, thus supporting a hypothesis on a novel sporadic form of high PCBs related peripheral subclinical hyperthyroidism possibly resulting from the long-term disruption of equilibrium between bound and free thyroxine in plasma by high PCBs level followed by a striking inhibition of TSH release from the pituitary.     

Disruption of thyroid hormone binding to sea bream recombinant transthyretin by ioxinyl and polybrominated diphenyl ethers

A number of chemicals released into the environment share structural similarity to the thyroid hormones (THs), thyroxine (T(4)) and triiodothyronine (T(3)) and it is thought that they may interfere with the thyroid axis and behave as endocrine disruptors (EDs). One of the ways by which such environmental contaminants may disrupt the TH axis is by binding to TH transporter proteins. Transthyretin (TTR) is one of the thyroid hormone binding proteins responsible for TH transport in the blood. TTR forms a stable tetramer that binds both T(4) and T(3) and in fish it is principally synthesized in the liver but is also produced by the brain and intestine. In the present study, we investigate the ability of some chemicals arising from pharmaceutical, industrial or agricultural production and classified as EDs, to compete with [I(125)]-T(3) for sea bream recombinant TTR (sbrTTR). Ioxinyl, a common herbicide and several polybrominated diphenyl ethers were strong inhibitors of [I(125)]-T(3) binding to TTR and some showed even greater affinity than the natural ligand T(3). The TTR competitive binding assay developed offers a quick and effective tool for preliminary risk assessment of chemicals which may disrupt the thyroid axis in teleost fish inhabiting vulnerable aquatic environments.     

Bioaccumulative characteristics of hexabromocyclododecanes in freshwater species from an electronic waste recycling area in China

Hexabromocyclododecanes (HBCDs) are now emerging ubiquitous contaminants due to their wide usage, persistence and toxicities. To investigate the bioaccumulative characteristics of HBCDs, sediments, Winkle (Littorina littorea), crucian carp (Carassius carassius) and loach (Misgurnus anguillicaudatus) were collected from two streams near an E-waste dismantling site in China, and HBCD exposure test was then conducted on Chinese rare minnow. The concentration of HBCDs was 14 ng g⁻¹ dry weight in sediments, 186, 377 and 1791 ng g⁻¹ lipid weight in winkle, crucian carp and loach, respectively. γ-HBCD was found to be the dominant diastereoisomer in the sediments (63% of total HBCDs). However, α-HBCD was selectively accumulated in the biotic samples and contributed to 77%, 63% and 63% of total HBCDs in winkle, crucian carp and loach, respectively. Moreover, an enrichment of (−)-enantiomers of α- and γ-HBCD were found in the winkle. The reverse results were observed in the crucian carp and loach. Similar observations of diastereoisomeric and enantiomeric composition were obtained in Chinese rare minnow with those found in the crucian carp and loach. These results indicate that the freshwater species from the streams are contaminated by HBCDs. α-HBCD can be selectively accumulated in organisms and the accumulative characteristics are enantioselective among species.     

Bioaccumulation, depuration and biotransformation of 4,4'-dibromodiphenyl ether in crucian carp (Carassius auratus)

Polybrominated diphenyl ethers (PBDEs) are extensively used as a class of flame retardants and have become ubiquitous environmental pollutants. Significant biotransformation of some PBDEs via reductive debromination has been observed. However, little is known about the fate of lower brominated BDEs in fish. In this study, the tissue distribution, excretion, depuration and biotransformation of 4,4′-dibromodiphenyl ether (BDE 15) were investigated in crucian carp (Carassius auratus) which were exposed to spiked water solution at different concentrations for 50 d, followed by a 14-d depuration period. Bioaccumulation parameters were calculated and the results showed that BDE 15 was mainly concentrated in the gill and liver. In particular, five biotransformation products of BDE 15 in carp were identified using GC-MS/MS. Besides two debrominated metabolites, three of the metabolites were mono-OH-BDE 15, diOH-BDE 15 and bromophenol. Our results unequivocally suggested that BDE 15 oxidation did occur via the formation of hydroxylated (OH-) metabolites in crucian carp exposed in vivo. These findings will be useful for determination of the metabolic pathways of PBDEs in freshwater fish, especially about their oxidation metabolism.     

Bioavailability of 2,3,7,8-tetrachlorodibenzo-p-dioxin from municipal incinerator fly ash to freshwater fish

The bioavailability of 2,3,7,8-TCDD from municipal incinerator fly ash to freshwater fish was determined. It was observed that carp exposed to fly ash containing all 22 TCDD isomers, or the solvent extract of the fly ash, retain only 2,3,7,8-TCDD. Exposures with fly ash appears to follow a dose response relationship for bioconcentration, however, the bioavailability of 2,3,7,8-TCDD was not directly related to the level of 2,3,7,8-TCDD in fly ash for two fly ash samples studied.     

Fish from industrially polluted freshwater as the main source of organochlorinated pollutants and increased frequency of thyroid disorders and dysglycemia

In a certain area of Michalovce district in East Slovakia, heavy industrial pollution by polychlorinated biphenyls (PCBs) developed in 1955-1984 and very high PCB levels in environmental and human samples are still persisting. Recently, a total of 2045 adults from this and the surrounding background pollution area have been examined using questionnaire data, thyroid volume by ultrasound (ThV), urinary iodine and serum levels of 15 PCB congeners, hexachlorobenzene (HCB), 2,2'-2-bis(4-chlorobiphenyl)-1,1-dichloroethylene (DDE), 2,2'-bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT), alpha-, beta- and gamma-hexachlorocyclohexane (HCH), thyrotropin (TSH), free thyroxine (FT4), anti-thyroperoxidase antibodies (TPOab) and fasting glucose. As based on our previous findings of strikingly high level of PCBs in fish from high pollution area (e.g. mean level of 375430 ng/g lipid) and considerably lower, but still relatively high level in background pollution area (e.g. mean PCB level of 5150 ng/g), the information on the frequency of fish meals and approximate annual consumption of fish from local waters was obtained by questionnaires. The association of contaminated fish consumption with very high blood levels of PCBs, DDE and HCB and increased ThV as well as with increased frequency of positive TPOab, high values of FT4 and impaired fasting glucose (IFG) was found. These associations were also confirmed in 16 marital pairs from high pollution area with very high PCB level in both members associated with high fish consumption. It was concluded that, due to persistent heavy pollution of waters, soil and food chain namely by PCBs, but also by pesticides (e.g. DDE and HCB) resulting from their previous extensive use in agriculture, the fish from local waters still remains the most important source of these toxic pollutants which results in considerable adverse health effects.     

Dynamics of microcystins-LR and -RR in the phytoplanktivorous silver carp in a sub-chronic toxicity experiment

A sub-chronic toxicity experiment was conducted to examine tissue distribution and depuration of two microcystins (microcystin-LR and microcystin -RR) in the phytoplanktivorous filter-feeding silver carp during a course of 80 days. Two large tanks (A, B) were used, and in Tank A, the fish were fed naturally with fresh Microcystis viridis cells (collected from a eutrophic pond) throughout the experiment, while in Tank B, the food of the fish were M. viridis cells for the first 40 days and then changed to artificial carp feed. High Performance Liquid Chromatography (HPLC) was used to measure MC-LR and MC-RR in the M. viridis cells, the seston, and the intestine, blood, liver and muscle tissue of silver carp at an interval of 20 days. MC-RR and MC-LR in the collected Microcystis cells varied between 268-580 and 110-292 microg g(-1) DW, respectively. In Tank A, MC-RR and MC-LR varied between 41.5-99.5 and 6.9-15.8 microg g(-1) DW in the seston, respectively. The maximum MC-RR in the blood, liver and muscle of the fish was 49.7, 17.8 and 1.77 microg g(-1) DW, respectively. No MC-LR was detectable in the muscle and blood samples of the silver carp in spite of the abundant presence of this toxin in the intestines (for the liver, there was only one case when a relatively minor quantity was detected). These findings contrast with previous experimental results on rainbow trout. Perhaps silver carp has a mechanism to degrade MC-LR actively and to inhibit MC-LR transportation across the intestines. The depuration of MC-RR concentrations occurred slowly than uptakes in blood, liver and muscle, and the depuration rate was in the order of blood>liver>muscle. The grazing ability of silver carp on toxic cyanobacteria suggests an applicability of using phytoplanktivorous fish to counteract cyanotoxin contamination in eutrophic waters.     

Effects of cyanobacteria producing microcystins on seed germination and seedling growth of several agricultural plants

The effects of cyanobacteria aqueous extracts containing Microcystin-LR (MC-LR) on the seed germination and growth of Pisum sativum, Lens esculenta, Zea mays and Triticum durum were investigated. Experiments were carried out on a range of doses of the extract (equivalent to 0, 1.6, 2.9, 5.8, 8.7 and 11.6 mu g MC-LR/mL). The results confirm that these plants were sensitive to cell-free extracts of a toxic Microcystis and that germination inhibition was dose dependent. One-way analysis of variance (ANOVA) showed that P. sativum is the most sensitive tested species with a 97% germination rate reduction and L. esculenta was the most resistant. At the 8th day, the exposure to the microcystins (MC) resulted in a significant decrease of plant epicotyls length, roots length and a net inhibition of lateral root formation. It is concluded that MC could affect also terrestrial plants seedling germination and growth. Therefore, the use of water for irrigation contaminated by MC could exert negative biochemical effects on seed and plant metabolism which might influence the agricultural crops.     

Reprint of: Concentrations, distribution, and bioaccumulation of synthetic musks in the Haihe River of China

Seven typical synthetic musks (SMs) in the samples from the surface water, sediment and fish of the Haihe River were measured. The SM concentrations in the sediment and surface water of the Haihe River were significantly lower than those in the Dagu Drainage River and Chentaizi Drainage River (p < 0.05). Along the flow direction, the SM concentrations in surface water and sediment tended to increase from the upstream to the downstream of Dagu Drainage River. The Bioaccumulation factors (BAFs) of galaxolide (HHCB) and tonalide (AHTN) were calculated at high levels in the muscles of crucian carp, common carp, and silver carp. Most of the biota-sediment accumulation factors (BSAFs) for HHCB and AHTN were higher than 1.7, suggesting magnification possibly exist in the musk bioaccumulations of the three fishes in the Haihe River. No significant differences in HHCB/AHTN ratios were observed among the water, fish, and sediment samples (p > 0.05). However, the HHCB/AHTN values in the Haihe River were much lower than those in the Dagu Drainage River and Chentaizi Drainage River (p < 0.05). Compared with several typical persistent organic pollutants (POPs), the musk concentrations were higher or comparable in the Haihe River.     

Concentrations, distribution, and bioaccumulation of synthetic musks in the Haihe River of China

Seven typical synthetic musks (SMs) in the samples from the surface water, sediment and fish of the Haihe River were measured. The SM concentrations in the sediment and surface water of the Haihe River were significantly lower than those in the Dagu Drainage River and Chentaizi Drainage River (p < 0.05). Along the flow direction, the SM concentrations in surface water and sediment tended to increase from the upstream to the downstream of Dagu Drainage River. The Bioaccumulation factors (BAFs) of galaxolide (HHCB) and tonalide (AHTN) were calculated at high levels in the muscles of crucian carp, common carp, and silver carp. Most of the biota-sediment accumulation factors (BSAFs) for HHCB and AHTN were higher than 1.7, suggesting magnification possibly exist in the musk bioaccumulations of the three fishes in the Haihe River. No significant differences in HHCB/AHTN ratios were observed among the water, fish, and sediment samples (p > 0.05). However, the HHCB/AHTN values in the Haihe River were much lower than those in the Dagu Drainage River and Chentaizi Drainage River (p < 0.05). Compared with several typical persistent organic pollutants (POPs), the musk concentrations were higher or comparable in the Haihe River.     

Distribution characteristics of polychlorinated biphenyls in crucian carp (Carassius auratus) from major rivers in Korea

Polychlorinated biphenyls (PCBs) levels in crucian carp were determined at 20 locations along four major river systems, several small-scale rivers and a wetland in Korea. Twenty-eight congeners, ranging from tri- to hepta-CBs were detected. A gas chromatograph with a mass selective detector was used to quantify the individual PCB congeners. The objectives of this study were to investigate the levels of contamination of PCBs in freshwater fish and to observe the pattern of their distribution. The sampling locations were chosen among 31 sampling sites that are currently used as environmental residue checkpoints by the Korean Ministry of Environment. Concentrations of individual congeners ranged from not detectable (n.d.) to 0.75 ng g(-1) on a wet weight basis. The total concentrations of PCBs at each site ranged from n.d. to 5.41 ng g(-1) of wet weight. The most heavily contaminated site was the Nakdong estuary located near the Shinpyung-Janglim factory district. The PCB 153 and 138 were the principal congeners and penta- and hexa-chlorinated biphenyls comprised the main congener groups.     

Nationwide study of dioxins in the freshwater fish Carassius auratus (gibelio) langsdorfii (crucian carp) in Japan: concentrations and estimation of source contribution ratios

We investigated dioxin concentrations in freshwater fish in Japan by standardizing species to detect subtle decreasing trends of dioxin concentrations in the future with the reinforcement of regulations. The fish studied were crucian carp (Carassius auratus (gibelio) langsdorfii), an omnivorous species. Fish and sediments were collected from 14 rivers and lakes located in remote areas, agricultural areas, and small and large cities throughout Japan. The total toxic equivalent (TEQ) dioxin concentrations at the three remote sites were about 20% of the concentrations at the other 11 sites, which all had similar concentrations. The average concentrations in fish collected from these 11 sites were 0.69pgTEQg(-1) wet wt. (95% CI 0.52-0.85) or 57.3pgTEQg(-1) fat (95% CI 47.9-66.7). There were notable differences in congener profiles of polychlorinated p-dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) between sampling areas. A chemical mass balance method was used to estimate the proportions of dioxin contributions from different sources. The sampling areas were classified into four groups on the basis of the predominant source of dioxin contamination: a combustion group, a chlornitrofen herbicide group, a chlornitrofen and pentachlorphenol herbicide group, and a remote group. The congener profiles of dioxin like polychlorinated biphenyls (DL-PCBs) were almost the same in all areas and close to those in Kanechlor, which indicates that pollution from PCB products has spread throughout Japan. From samples collected during the spawning season, it was determined that about 20% of the dioxins in adult females were transferred to the eggs, resulting in higher dioxin concentrations in adult males than in females. Biota-sediment accumulation factors (BSAFs) of 2,3,7,8-substituted PCDDs/DFs for crucian carp were larger than those of non-2,3,7,8-substituted congeners, and BASFs decreased with increasing number of chlorines. The BSAFs of DL-PCBs were 10 times greater than those of PCDDs/DFs, and BSAFs of mono-ortho type DL-PCBs were higher than those of non-ortho types.     

Effects of four chlorobenzenes on serum sex steroids and hepatic microsome enzyme activities in crucian carp, Carassius auratus

Four chlorobenzenes (chlorobenzene, 1,3-dichlorobenzene, 1,4-dichlorobenzene, p-chloro-methylbenzene) were administrated to the crucian carps (Carassius auratus) by peritoneal injections in the laboratory for 30 days. Serum testosterone and 17 beta-estradiol concentrations were detected using radioimmunology assay (RIA), and the activities of two hepatic microsome enzymes, glutathione s-transferase (GST) and UDP-glucuronosyltransferase (UDPGT), were measured using the modified methods as described by Habig and Owens. Results showed that the four chlorobenzenes caused significant increases in serum testosterone concentration in the crucian carps (P < 0.05) compared to the controls, but they caused no significant effect on 17 beta-estradiol level. All test chemicals caused a change in hepatic GST activity in crucian carps, with significant increases in enzyme activity (P < 0.05). Chlorobenzene, 1,3-dichlorobenzene and p-chloro-methylbenzene resulted in a marked inhibition to UDPGT activity in crucian carp (P < 0.05) except 1,4-dichlorobenzene. The changes in hepatic microsome enzyme activities may have resulted in the alterations of serum sex steroids levels in the crucian carps. The results indicated that these four chlorobenzenes may result in the changes of endocrine functions and may affect the reproductive success of this and other species.     

Effects of cyanobacteria producing microcystins on seed germination and seedling growth of several agricultural plants

The effects of cyanobacteria aqueous extracts containing Microcystin-LR (MC-LR) on the seed germination and growth of Pisum sativum, Lens esculenta, Zea mays and Triticum durum were investigated. Experiments were carried out on a range of doses of the extract (equivalent to 0, 1.6, 2.9, 5.8, 8.7 and 11.6 μ g MC-LR/mL). The results confirm that these plants were sensitive to cell-free extracts of a toxic Microcystis and that germination inhibition was dose dependent. One-way analysis of variance (ANOVA) showed that P. sativum is the most sensitive tested species with a 97% germination rate reduction and L. esculenta was the most resistant. At the 8th day, the exposure to the microcystins (MC) resulted in a significant decrease of plant epicotyls length, roots length and a net inhibition of lateral root formation. It is concluded that MC could affect also terrestrial plants seedling germination and growth. Therefore, the use of water for irrigation contaminated by MC could exert negative biochemical effects on seed and plant metabolism which might influence the agricultural crops.     

Arsenic contamination in the freshwater fish ponds of Pearl River Delta: bioaccumulation and health risk assessment

This study investigated the extent of arsenic (As) contamination in five common species of freshwater fish (northern snakehead [Channa argus], mandrarin fish [Siniperca chuatsi], largemouth bass [Lepomis macrochirous], bighead carp [Aristichthys nobilis] and grass carp [Ctenopharyngodon idellus]) and their associated fish pond sediments collected from 18 freshwater fish ponds around the Pearl River Delta (PRD). The total As concentrations detected in fish muscle and sediment in freshwater ponds around the PRD were 0.05–3.01 mg?kg^?1 wet weight (w. wt) and 8.41–22.76 mg?kg^?1 dry weight (d. wt), respectively. In addition, the As content was positively correlated (p?<?0.05) to total organic carbon (TOC) contents in sediments. Biota sediment accumulation factor (BSAF) showed that omnivorous fish and zooplankton accumulated higher concentrations of heavy metals from the sediment than carnivorous fish. In addition, feeding habits of fish also influence As accumulation in different fish species. In this study, two typical food chains of the aquaculture ponds were selected for investigation: (1) omnivorous food chain (zooplankton, grass carp and bighead carp) and (2) predatory food chain (zooplankton, mud carp and mandarin fish). Significant linear relationships were obtained between log As and δ ¹⁵N. The slope of the regression (?0.066 and ?0.078) of the log transformed As concentrations and δ ¹⁵N values, as biomagnifications power, indicated there was no magnification or diminution of As from lower trophic levels (zooplankton) to fish in the aquaculture ponds. Consumption of largemouth bass, northern snakehead and bighead carp might impose health risks of Hong Kong residents consuming these fish to the local population, due to the fact that its cancer risk (CR) value exceeded the upper limit of the acceptable risk levels (10^?4) stipulated by the USEPA.