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1.
Estrogens appear to have a modulating effect on the expression of cytochrome P4501A (CYP1A) in fish. A number of in vivo studies have demonstrated that hepatic CYP1A expression in females decrease during sexual maturation when plasma levels of 17 beta-estradiol (E2) increase, or in cases when the fish in injected with E2. Since a number of environmental contaminants have weak estrogen-like activities, the question arises if these compounds are able to modulate CYP1A expression as well. In the present study, we used in vitro monolayer cultures of rainbow trout, Oncorhynchus mykiss, liver cells to compare concentration-dependent (10(-9) to 10(-5) M) effects of the natural steroid E2 and the non-steroidal xenoestrogen 4-tert-octylphenol (OP) on CYP1A-catalyzed 7-ethoxyresorufin-O-deethylase (EROD) activity. The concentration dependency of the estrogenic activity of the two test compounds was assessed by determination of hepatocellular vitellogenin (Vg) release into the culture medium. Exposure of hepatocytes to E2 concentrations of 10(-8) M and higher led to a significant inhibition of basal cellular EROD activity. On the contrary, exposure to OP did not result in an inhibition of EROD activity, even at OP concentrations (10(-6) M, 10(-5) M) which were associated with a significant induction of Vg synthesis.  相似文献   

2.
We exposed sexually maturing male rainbow trout (Oncorhynchus mykiss) to BDE-47 (a polybrominated diphenyl ether) and female rainbow trout to trenbolone (an anabolic steroid). Male trout were orally exposed for 17 days to 55 microg/kg/day BDE-47 and female trout continuously exposed for 60-77 days to a measured trenbolone water concentration of 35 ng/L. After the exposure, eggs and semen were collected and in vitro fertilization trials performed using a sperm:egg ratio of 300,000:1. In the BDE-47 study, eggs from control females were fertilized with semen from exposed males, while in the trenbolone study, eggs from exposed females were fertilized with semen from control males. All treatments were evaluated at two-three early developmental time-points representing first cleavage (0.5 day), embryonic keel (9 days), and eyed stages (19 days), respectively. The results indicated that BDE-47 exposure did not alter fertility as embryonic survival was similar between control and exposed groups. Trenbolone exposure also did not alter embryo survival. However, in the embryos fertilized with eggs from trenbolone exposed females, a noticeable delay in developmental progress was observed. On day 19 when eye development is normally complete, the majority of the embryos either lacked eyes or displayed under-developed eyes, in contrast to control embryos. This finding suggests steroidal androgen exposure in sexually maturing female rainbow trout can impact developmental timing of F1 offspring.  相似文献   

3.
Effects of cadmium exposure on plasma levels of calcitonin and free and protein-bound calcium were studied in vitellogenic female rainbow trout kept in brackish water (7%(.)). Fish were exposed to 100 μg cadmium litre−1 for four weeks. Exposure of female rainbow trout in the stage of vitellogenesis, with increased total plasma levels of calcium, resulted in a complex hypocalcemic response. Thus, hypocalcemia was found to be due to three different processes: (1) a decrease in the free plasma calcium, and a reduction in protein-bound calcium; due both to (2) decreased plasma levels of vitellogenin; and (3) a reduced binding of calcium to vitellogenin. These findings support the concept of an interference of cadmium with ionregulating tissues as a mechanism for hypocalcemia in rainbow trout. A direct effect on the vitellogenin-binding of calcium was also observed and reproductive function in the females was affected by decreased plasma levels of vitellogenin. In spite of the marked changes of plasma calcium in exposed fish, no significant effects on plasma calcitonin were observed, indicating a lack of a direct relationship between plasma calcium and calcitonin levels in rainbow trout.  相似文献   

4.
Methoxychlor (MXC) has been shown to possess estrogenic activity in mammals and fish. Although MXC does not appear to appreciably bind the mammalian estrogen receptor, its demethylated metabolites have been shown to be significantly more potent agonists and are believed responsible for estrogenic effects in mammals following exposure to this pesticide. To determine whether catfish were capable of MXC demethylotion, and, hence, activation to a more estrogenic compound, in vitro biotransformation studies were carried out using hepatic microsomes from mature male channel catfish. Hepatic microsomes catalyzed the NADPH-dependent formation of monodemethylated (mono-MXC) and bisdemethylated (bis-MXC) metabolites of MXC. Treatment with mono-MXC at 40% of the MXC dose in catfish significantly induced serum vitellogenin (Vg) levels compared to MXC. Estrogen receptor binding studies in catfish liver cytosol showed that a racemic mixture of the mono-MXC had approximately 43 times the affinity for the receptor than MXC, but was still over 1000-fold less potent that 17β-estradiol. These results demonstrate that catfish are capable of biochemically activating MXC to a more potent hepatic estrogen receptor agonist.  相似文献   

5.
The ability of 17α-ethinylestradiol (EE2) to elevate vitellogenin levels were investigated in male flounder Platichthys flesus and vitellogenin concentrations in flounders from the Danish coastal environment were determined. Male flounders were exposed to 17α-ethinylestradiol (EE2) via food or water. Average vitellogenin concentrations in the control fish ranged between 25 and 100 ng mL−1. Exposure to 5.1, 8.1 and 16.8 ng EE2 L−1 in water and 500 and 5000 ng EE2 kg−1 body weight (bw) every second day in the food increased the plasma vitellogenin concentration in a concentration and time dependent manner, whereas exposure to 2.7 ng EE2 L−1 in water for 21 d and 5 and 50 ng EE2 kg−1 bw for 12 days in the food did not. EE2 could be detected in liver and testes (but not in muscle) after exposure to 8.1 and 16.8 ng EE2 L−1 in the water and 5000 ng EE2 kg−1 bw in the food; the highest concentration was 6 ng g−1 wet weight in liver. The majority of the male flounders collected from nine coastal Danish sites from 1999 to 2004 had vitellogenin concentrations below 100 ng mL−1, and only at two sites moderate estrogenic inputs were indicated.  相似文献   

6.
The effects of isosafrol (ISF) or β-naphthoflavone (βNF) treatments on cytochrome P450 (P450) levels in rainbow trout liver were investigated using immunochemical and catalytic techniques. The discrepancies in catalytic activities and ELISA quantification of rainbow trout P4501A1 protein levels between ISF- and βNF-treated fish indicate that important differences exist between the responses induced by βNF and ISF treatments in the rainbow trout liver.  相似文献   

7.
Expression of flavin-containing monooxygenase(s) (FMO) correlates with salinity exposure in certain species of euryhaline fish, such as the rainbow trout, Oncorhynchus mykiss. The mechanism(s) by which salinity regulates FMO is unclear. Adult rainbow trout were infused through the dorsal aorta with either cortisol or urea. At 500 ng/ml, cortisol caused a significant increase in FMO-catalyzed thiourea oxidase activity in gill and liver microsomes. FMOI expression, however, was significantly increased by the high cortisol dose only in gill microsomes. The levels of TMAO and urea were not altered by cortisol. In the liver, urea infusion caused an increase in hepatic FMO activity. FMO expression and activity correlated with elevated tissue urea levels, but TMAO concentrations were not related. These results indicate that FMO expression and activity may be partially controlled by the osmoregulatory/stress hormone. cortisol, and concentrations of the organic osmolyte, urea, in the rainbow trout.  相似文献   

8.
Several environmentally relevant contaminants are characterized as xenoestrogens by virtue of their ability to induce responses similar to 17β-estradiol (E2). There is concern that exposure to these xenoestrogens may result in endocrine and, thus, reproductive disruption. The objective of the present research was to determine whether xenobiotics known to elicit estrogenic responses in mammals were estrogenic in channel catfish, and if these compounds were capable of altering normal physiological response to E2. Sexually immature catfish were exposed by intraperitoneal injection to E2, suspected xenoestrogens or combination doses of E2 and xenoestrogen. Appearance of vitellogenin (Vg) in serum was used as the bioindicator of estrogenicity; Vg was measured by ELISA 7 days after injection. The ED50 of E2 for the appearance of Vg in blood, 0.6 mg/kg, was used as the positive control. Synthetic estrogens, ethinylestradiol, mestranol and DES were effective in inducing Vg; the antiestrogen tamoxifen inhibited response to E2 when given in a combination dose. Treatment by both methoxychlor and p-nonylphenol resulted in Vg appearance in serum; the doses required were 300 times that of E2, and the vitellogenic response was less when compared to the E2-induced response. In addition, a combination dose of methoxychlor with E2 decreased the magnitude of the response to E2. Other mammalian xenoestrogens, o,p′-DDT, chlordecone, lindane and β-HCH, were not able to produce a vitellogenic response in this study.  相似文献   

9.
Recent studies demonstrating feminization of effluent-exposed wild-caught male fish in the UK have prompted much research regarding the estrogenic activity of effluent from municipal sewage treatment plants (MSTPs). To investigate the estrogenicity and cytochrome P450 1A (CYP1A) induction potency of MSTP effluent, two species of fish, adult male mummichogs, Fundulus heteroclitus, and juvenile sunshine bass, Morone saxatilis x Morone chrysops, were exposed to un-chlorinated effluent (75% effluent, 25% seawater) from a large MSTP in Yonkers, NY, USA. After a 21-day static-daily (75%) renewal exposure, significant elevations over controls were observed in levels of vitellogenin (VtG) in plasma (1730%) and liver (131%) in effluent-exposed sunshine bass. In contrast, hepatic VtG was not elevated in mummichogs; plasma VtG was not measured in this species. Effluent exposure elevated hepatic CYP1A protein (140-145%) and ethoxyresorufin-O-deethylase (EROD) activity (408-598%) in both species. These findings suggest ontogenetic and/or species differences in response to estrogenic compounds in MSTP effluent. Furthermore, the elevation of CYP1A in response to sewage effluent exposure indicates the presence of additional compounds that may alter xenobiotic and/or steroid biotransformation in fish.  相似文献   

10.
The estrogenicity of several xenobiotics was evaluated using in vivo vitellogenin (Vtg) synthesis in immature rainbow trout as a biomarker. 17β-estradiol, DES and ethinyl estradiol were tested as positive controls. The xenobiotic compounds tested were technical nonylphenol, bisphenol A, butylbenzylphthalate (BBP) and dibutylphthalate (DBF). Measurements of the Vtg concentration was performed with a direct sandwich ELISA. 17β-estradiol, DES and ethinyl estradiol caused up to 100 000-fold increases in the Vtg-levels. Nonylphenol and bisphenol A had the highest estrogenic potency of the xenobiotics increasing the vitellogenin level approximately 300-fold while BBP was only weakly estrogenic, increasing the concentration about 3 times. DPB did not raise the vitellogenin contration above the detection limit.  相似文献   

11.
The aim of this study was to examine the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, given intraperitoneally) alone and together with bleached kraft pulpmill effluents (BKME) on fish biotransformation reactions and testosterone levels in mature male rainbow trout (Salmo gairdneri Richardson). The results show extremely high induction in liver and kidney monooxygenase (PSMO) activities in 3 weeks caused by TCDD alone or TCDD together with BKME. BKME alone did not cause significant effects. 7-Pentoxyresorufin O-dealkylase activity, previously shown to be an indicator of phenobarbital-type induction, was shown to be highly inducible by TCDD in fish. Plasma testosterone levels, although decreased in another subacute exposure to BKME, showed only slight changes due to TCDD and TCDD + BKME.  相似文献   

12.
In this project we investigated the ecotoxicological effects of endocrine disrupters in a four-year survey of the Mediterranean population of swordfish (Xiphias gladius). In the Mediterranean environment, top predators, such as swordfish, accumulate high concentrations of polyhalogenated aromatic hydrocarbons (PHAHs) and toxic metals, potentially incurring high toxicological risk. The effects of organochlorines and trace elements (Hg, Cd and Pb) in 192 swordfish specimens, caught in the Strait of Messina, Sicily, Italy, were investigated using vitellogenin (Vtg), zona radiata proteins (Zrp) and CYP1A (BPMO, EROD) activities. Vtg and Zrp were found to be dramatically induced in some adult male specimens, suggesting that this species is highly exposed to estrogens in the Mediterranean Sea. A role of organochlorines in this induction phenomenon is suggested by the statistically significant correlations between Zrp in plasma and PCB concentrations in muscle (p<0.032) and Vtg in plasma and PCB concentrations in liver (p<0.034) of male specimens. Levels of trace elements in liver were in the following ranges: Hg 1–22, Cd 1–28 and Pb 0–1.6 ppm d.w. These data indicate potential reproductive alterations in large pelagic fish and suggest the need for continuous monitoring to avoid reductions in the population of this fish species of high commercial and ecological interest.  相似文献   

13.
Metallothionein has previously been shown to be regulated during the period of exogenous vitellogenesis (Olsson et al. (1987); Fish Physiol, Biochem., 3(1), 39–47). Following estradiol injection of rainbow trout it has been shown that hepatic metallothionein remains at basal levels until the vitellogenin mRNA levels begin to decline and Zn is released from high-molecular-weight proteins (Olsson et al. (1989); Biochem.J., 257, 555–559), In the present study the effects of estradiol treatment on the metal-inducibility of metallothionein have been investigated. Estradiol-treated fish that were injected with low doses of cadmium or zinc did not respond by induction of metallothionein.  相似文献   

14.
Rainbow trout hepatocytes were isolated by a two step perfusion technique and maintained either in monolayer culture for 5 days or in aggregate culture for 30 days. Cytochrome P450 content decreased from day 0 to day 5 in both culture systems, and then was preserved at the same level after one month in aggregated cells. 7-Ethoxyresorufin O-deethylase (EROD) and UDP-glucuronosyl transferase were not significantly different in freshly isolated cells and in 30-day aggregated hepatocytes, whereas a substantial increase in glutathione S-transferase was observed. Two-day exposure of cells to β-naphthoflavone led to a significant increase in EROD activity in both culture systems, especially after one month of aggregation (10-fold increase). According to these results, aggregate culture of rainbow trout hepatocytes seems to be a promising in vitro model to investigate the biotransformation pathways in fish and their regulation by endogenous and exogenous compounds.  相似文献   

15.
Nitroaromatic compounds frequently contaminate aquatic systems and may, therefore, impact fish. However, a known pathway of nitroaromatic toxicity in mammals, that of nitroaromatic stimulated superoxide (O2−) production, has yet to be addressed in fish. In this study we investigated this pathway in three species of freshwater fish-channel catfish (Ictalurus punctatus), largemouth bass (Micropterus salmoides) and rainbow trout (Salmo gairdneri)-exposed to nitrofurantoin (NF), p-nitrobenzoic acid (PNBA) and m-dinitrobenzene (MDNB). Our results indicate that these nitroaromatics elicit a stimulation of O2− production by fish liver fractions. Additionally, results suggests a similarity between, fish and mammals in the nitroreductases which mediate nitroaromatic activation and subsequent O2− production. These findings indicate a potential toxic consequence of fish exposure to nitroaromatic contaminants in aquatic systems.  相似文献   

16.
Temporary removal of aquatic plants in Lake Parkinson, a small, eutrophic dune lake, resulted in a number of changes to the population of stocked rainbow trout (Salmo gairdnerii). During each summer the lake stratified and low oxygen levels limited the distribution of trout to shallow (0–4 m) surface waters. In the first summer following weed removal the numbers of black shags (Phallacrocorax carbo) counted at the lake increased, and their predation resulted in a decline in trout density. However, the growth rate and condition of the trout population then exceeded that of trout present before weed removal. During the second summer after weed removal a cladoceran bloom was followed by low phytoplankton levels and high ammonia concentrations. A prolonged calm compounded this situation with the result that oxygen levels in bottom and surface waters dropped below 2 ppm. These low oxygen levels eliminated the trout population, but other fish species present survived. Elimination of aquatic plants affected the population dynamics of other fish species in the lake with potential implications for the trout. The experiment demonstrated the importance of weed beds in maintaining a stable fish community in lakes such as Lake Parkinson.  相似文献   

17.
Five lakes, varying from 10 to 100 hectares in area and each containing an existing population of Salmo gairdneri, were gill‐netted and stocked with tagged fish each year from 1960 to 1965 to determine the effect of regular stocking on the size and numbers of fish present. There was little natural spawning of trout in most of the lakes. Growth in length and weight of tagged fish was computed and an annual estimate of population calculated from the netting returns. Appendices give details of the statistical analyses of the data.

Fish production has been correlated with the trophic status of the lakes as indicated by their temperature and oxygen content. There was no clear relationship between lake size and trout production but trout production decreased as the environment became more eutrophic. Results also showed a close inverse relationship between numbers supported and average weight achieved by the trout. It is concluded that the angling potential of local lakes can be maintained despite the eutrophication resulting from intensive agricultural development of the catchments. These lakes will produce large‐sized fish if the trout population is kept relatively small.

Loss in weight in brown and rainbow trout as a result of spawning activities in local lakes and streams was found to be similar. Both male and female fish lost between 22 and 40 percent of their weight over a spawning period. This probably accounted for the observed low‐condition factor among older fish in the lake populations.  相似文献   

18.
Vitellogenin (Vtg) is a yolk protein precursor that has been identified as a sensitive biomarker for exposure to estrogenic compounds. We evaluated specific monoclonal and polyclonal antibodies for reactivity with plasma Vtg from two Australian Perciformes, the tropical barramundi (Lates calcarifer) and the temperate black bream (Acanthopagrus butcheri). Blood plasma from 17beta-estradiol exposed (E2) male barramundi (20 mg kg(-1)) and male black bream (2.5-5.0 mg kg(-1)) were sent to Biosense Laboratories (Norway) for cross-reactivity testing using their extensive anti-Vtg antibody selection. Indirect ELISA results determined barramundi plasma displayed the highest binding affinities to ND-3G2 (monoclonal-Mab) and PO-1 (polyclonal-Pab). Black bream was most cross-reactive with ND-1C8 (Mab) and PO-2 (Pab). Next, plasma was assessed for Vtg induction in E2-dosed (5 mg kg(-1)), hatchery-reared barramundi and black bream versus a non-injected control group. Vtg production was assessed by Western blot and indirect ELISA using ND-3G2 and ND-1C8 Mabs, respectively. A prominent band was identified in the range of 100-200 kDa for all female black bream and for all E2-treated barramundi and black bream males, which was confirmed as Vtg by Western blot. Indirect ELISA results for barramundi demonstrated highly significant differences in E2-dosed fish as compared to control fish (Student t, P<0.001). E2 male black bream were significantly different than control males (Student t, P<0.001) and control and E2 females displayed highly significant differences (Student t, P<0.001). These results indicate that exposure to 17beta-estradiol induces significant Vtg production in males of the two Australian Perciformes, with potential use as a biomarker for exposure to estrogenic compounds.  相似文献   

19.
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20.
This study was conducted as an initial investigation of 'differential response' in one of the main sentinel organisms used for monitoring programmes in United Kingdom estuaries, the flounder Platichthys flesus. It has been hypothesised that monitoring using species with a wide geographical spread and limited migration, such as flounder, might result in the comparison of different genetic stocks and certainly of populations with differing early life stage contaminant exposure histories. Furthermore, it is probable that these pre-exposure and genetic differences could manifest themselves in an ability to respond differently to contaminant exposure, so-called 'differential response'. It is important that the extent and nature of this response is understood, if we want to be able to fully interpret the monitoring data from such programmes. During this study, flounder were collected from four separate sources; wild caught fish from the estuaries of the Rivers Alde, Mersey and Tyne, and farmed flounder from Port Erin Farm, Isle of Man. Under controlled laboratory conditions, groups of fish from each source were exposed to water-borne concentrations of the synthetic oestrogen ethynylestradiol (EE2) at a nominal concentration of 50 ng/l. Plasma was taken from each male fish after 6 and 10 days exposure and analysed for the presence of vitellogenin (VTG) using an ELISA technique. Significant levels of VTG induction were evident in fish from all sources after both 6 and 10 days exposure. Flounder from the Mersey were the only fish with significantly elevated initial background levels of VTG (day 0) and this appeared to be reflected in that these specimens showed the highest induction response after day 6. However, after day 10, fish from all other sites had a slightly higher mean VTG than those from the Mersey which showed significantly (p < 0.05) lower mean plasma VTG. It is suggested that other differential responses may have been masked by the use of a high dose of EE2 which produced maximum induction in nearly all fish. The findings of the study are discussed in terms of implications for further research into the differential response issue and how the initial plasma VTG figures contribute to a time-series from the Mersey, Tyne and Alde estuaries.  相似文献   

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