Plasma steroid levels in female flounder (Platichthys flesus) after chronic dietary exposure to single polycyclic aromatic hydrocarbons

The chronic effects of polycyclic aromatic hydrocarbons (PAHs) on ovary development, total hepatic lipids and plasma sex- and corticosteroid levels in female flounder (Platichthys flesus) were examined. Sexually mature feral female flounder were exposed via the diet to phenanthrene (0.5, 2.5 or 12.5 nmol/g food) or chrysene (0.4 nmol/g food) for 12 weeks, during the previtellogenic phase of the annual reproductive cycle. PAH exposure did not directly affect germ cell development since no structural and/or developmental differences were observed between control and exposed fish. On the contrary, all treatments resulted in altered plasma steroid levels. The most pronounced effect was the significant decrease in plasma 17β-estradiol to 19±11%, 27±7%, 63±20% and 61±12% in relation to control fish, respectively, in flounders exposed to 12.5, 2.5 or 0.5 nmol phenanthrene/g food and 0.4 nmol chrysene/g food. Impaired ovarian growth was not observed, most likely because experiments were ended before the period of vitellogenesis, even though a non-significant general decline in total hepatic lipids could be observed. Moreover, all exposed flounders, except fish fed with the highest amount of phenanthrene, showed a negative correlation between plasma 17β-estradiol and 17α-hydroxyprogesterone levels (r=−0.46). One possible explanation is that PAH action may be mediated by a specific inhibition of steroidogenic enzymes. These findings provide evidence that selected PAHs are antiestrogenic xenobiotics with the capability to impair female teleost reproductive function.     

外源激素对养殖牙鲆血浆睾酮和雌二醇含量的影响研究

雄性牙鲆单独注射促黄体激素释放激素类似物(LHRH-A)(每千克质量的注射量,下同)25μg/kg或人绒毛膜促性腺激素(hCG)1 000 IU/kg,血浆睾酮(T)含量在注射后6 h大幅度降低,12h时达到最低值,以后逐渐回升;LHRH-A与hCG联合注射[LHRH-A(12.5μg/kg)+hCG(500 IU/kg)],注射后6 h血浆T含量剧烈下降,24 h时达到最低点,以后缓慢回升;对照组雄鱼血浆T含量随着采样进行而呈现降低趋势.雌鱼单独LHRH-A后,血浆雌二醇(E₂)水平逐渐下降,24 h时达到最低值,30 h后略有回升;单独注射hCG后,血浆E₂含量缓慢下降,30 h达到最低值;LHRH-A与hCG联合注射,注射后6 h血浆E2含量逐渐下降,24 h时达到最低点,30 h时缓慢回升.对照组雌鱼血浆E₂水平也随着采样的进行而逐渐降低.结果表明:单独或联合注射LHRH-A和hCG降低了雌、雄牙鲆血浆E₂和T含量,与对照组结果相同.     

夏季珠江口水体中多环芳烃的分布、组成及来源

利用1999年7月对珠江口海域的调查资料,对该区表层海水中优控多环芳烃的分布、组成及来源进行了分析和讨论,结果表明:(1)夏季珠江口海域表层海水中14种溶解态多环芳烃[苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为63.8~171.7 ng/L,且沿着冲淡水流向呈降低趋势;(2)颗粒态中15种多环芳烃[萘、苊、芴、菲、蒽、荧蒽、芘、苯并(a)蒽、艹屈、苯并(b)荧蒽、苯并(k)荧蒽、苯并(a)芘、二苯并(a,h)蒽、苯并(g,h,i)艹北、茚并(1,2,3-cd)艹比]的质量浓度为60.7~186.7 ng/L,其分布与水体载沙量及悬浮颗粒物的性质、粒径有关,具有从河口内向外海降低的分布特征;(3)多环芳烃组成和特征参数比值的分析表明,珠江口海域高温裂解来源的多环芳烃在伶仃洋海区输入最多,且主要为人类活动中煤燃烧排放的,而在香港岛周围海区的输入则相对较少,且主要为油燃烧排放的;(4)与法国塞纳河及长江口等河口相比,珠江三角洲海域水体中存在高菲含量排放源。     

Relative potency of PAHs and heterocycles as aryl hydrocarbon receptor agonists in fish

The relative potency of polycyclic aromatic compounds as aryl hydrocarbon receptor (AhR) agonists in fish was determined using data on CYP1A induction or AhR binding for 74 polycyclic aromatic hydrocarbons (PAHs) and heterocycles in teleost, avian, or mammalian systems from 18 published papers. Each PAH was assigned a fish potency factor relative to the potency of 2,3,7,8-tetrachlorodibenzo-p-dioxin as an AhR agonist. Two and three ring unsubstituted PAHs were generally inactive in fish, avian, and mammalian systems. Benzo[k]fluoranthene and indeno[1,2,3-cd]pyrene were consistently the most potent PAHs, with fish potency factors of 0.001-0.002. Common structural features associated with higher potency PAHs included 4-6 rings containing fluoranthene or phenanthrene structures with an exposed bay region. These results show that PAHs can have similar potency as many dioxin-like PCBs, and AhR mediated toxicity should be considered in assessing the risks of PAHs in fish.     

Size at first maturity of two species of flounders in the Hauraki gulf,New Zealand

The length at first maturity of the sand flounder, Rhombosolea plebeia (Richardson), and the yellow‐belly flounder, Rhombosolea leporina Gunther, (Teleostei: Heterosomata) in the Hauraki Gulf was investigated by the incidence of sexually tnature and immature fish grouped at 1‐cm length intervals. All male sand flounders and yellow‐bellies were sexually mature at sizes considerably less than the legal minima of 22.9 cm (9 in.) and 25.4 cm (10 in.) respectively. About 95% of female sand flounders and about 15% of female yellow‐bellies matured before reaching the respective minimum legal sizes. Most (i.e., over 50%) female sand flounders and yellow‐bellies were mature when they reached lengths of 18 cm and 26 cm respectively.     

PAH-biodegradation potential of indigenous microorganisms: evidence from the respiratory activity of surface sediments in the Quanzhou Bay in China

Seven stations were established in the Quanzhou Bay (24.73°-24.96°N, 118.50°-118.70°E) in China on three cruises to determine the concentrations of polycyclic aromatic hydrocarbons (PAHs) and the numbers of PAH-degrading bacteria in surface sediments. Assessing the biodegradation poten- tial of indigenous microorganisms by measuring the respiratory intensity with the addition of PAHs in sediment samples was also one of the aims of this study. The results show that the total PAH concentrations of the sedimen...     

Habitat selection of stone and starry flounders in an estuary in relation to feeding and survival

Juveniles of both stone flounder Platichthys bicoloratus and starry flounder Platichthys stellatus utilize estuaries as nursery grounds. To understand their habitat selection and the functions of habitats such as food supply, we defined the seasonal distribution of recently settled fish of these species in shallow nursery areas and investigated their feeding habits in the Natori River estuary, Japan. Distribution of stone flounder was limited to the lower estuary (<3 km upriver from the mouth) and stone flounder were most abundant near the mouth. Recently settled starry flounder were first detected further upstream in areas characterized by low salinity <10 and by the absence of the predatory sand shrimp Crangon uritai. Early juvenile stone and starry flounders consumed mainly siphons of the bivalve Nuttallia olivacea and the mysid Neomysis awatschensis, respectively; however, 1- and 2-yr-old fish of both stone and starry flounders fed mainly on the bivalve siphons. These results indicate that habitat selections of juvenile stone and starry flounders enable utilization of preferred prey and predator avoidance, respectively, and that non-overlap of these species' habitats results in avoidance of inter-specific competition for food between these two species.     

4-tert-octylphenol and 17beta-estradiol applied by feeding to flounder Platichthys flesus: induction of vitellogenin and accumulation in tissues

In order to simulate a possible natural administration route of xenoestrogens male flounder Platichthys flesus were exposed via the diet to the alkylphenol 4-tert-octylphenol. Treatment with 4-tert-octylphenol (10, 50 and 100 mg OP kg(-1) BW) or 17beta-estradiol (0.05 mg kg(-1) BW) every second day during a period of 11 days resulted in a significant increase in plasma vitellogenin (Vtg) concentrations. The induction of Vtg was greatest in the fish receiving 50 mg OP kg-' BW. A significant accumulation of 4-tert-octylphenol was found in liver and muscle tissue of the OP treated groups. The tissue concentrations of 4-tert-octylphenol and the plasma vitellogenin concentration were positively correlated. The results show that 4-tert-octylphenol accumulates in liver and muscles of flounder P. flesus, and exerts estrogenic effects such as vitellogenin induction.     

Growth of two species of flounders in the hauraki gulf,New Zealand

Growth of the sand flounder, Rhombosolea plebeia (Richardson), and the yellow‐belly flounder, Rhombosolea leporina Günther, in the Hauraki Gulf is described. In both species females grow more quickly than males. At 2 years of age male and female sand flounders reach mean total lengths of 17 cm and 23 cm respectively, and male and female yellow‐belly flounders reach mean total lengths of 24 cm and 29 cm respectively.     

Spawning and fecundity of two flounder species in the Hauraki gulf,New Zealand

Spawning grounds and spawning times of the sand flounder, Rhombosolea plebeia (Richardson), and the yellow‐belly flounder, Rhombosolea leporina Gunther, in the Hauraki Gulf are described. The occurrence of female fish at different stages of ovarian development at different stations during the year, the distribution of eggs in the plankton, and changes in ovary weight during the year were used as indicators of spawning grounds and spawning times.

The sand flounder was found to spawn in the waters to the east of Waiheke and Ponui Islands, at the northern end of the Firth of Thames, from June to November.

Yellow‐belly flounders spawned during September, October, and November, slightly to the south of the sand flounder spawning grounds in a belt extending from Tapu, on the eastern side of the Firth of Thames, north‐westwards towards Ponui Island.

Fecundity of both species was approximately proportional to the weight of the fish, or to the length cubed, and less than proportional to the ovary weight. The mean fecundity of sand flounders in the Hauraki Gulf varied from approximately 100,000 eggs in a fish of 18 cm to 500,000 in a 30 cm fish. That of yellow‐belly flounders varied from approximately 250,000 eggs in a fish of 30 cm to 1.25 million in a 45 cm fish.     

菲、芘、苯并(a)芘单一暴露及分别与α-萘黄酮(ANF)联合暴露对海水青鳉(Oryzias melastigma)胚胎发育毒性效应的比较研究

为了解和探讨3~5环PAHs对海水鱼类胚胎发育的毒性及作用方式,比较研究了菲(phenanthrene,Phe)、芘(pyrene,Py)、苯并(a)芘(benzo(a)pyrene,BaP)单一暴露和三者各自与α-萘黄酮(α-naphthoflavone,ANF)联合暴露对海水青鳉(marine medaka, Oryzias melastigma)胚胎发育的毒性效应。胚胎体内EROD活性、发育畸形、孵化率和心律等毒性指标被测定,结果显示:Phe,Py和BaP对海水青鳉胚胎体内EROD活性的诱导能力大小为BaP>Py>Phe,各化合物对EROD诱导与发育畸形之间的关系较为复杂,除Phe所引起的EROD诱导与畸形指数之间呈显著相关(r=0.95,p=0.015)外,Py和BaP均无相关性;在100 μg/dm3 ANF影响下,CYP1A活性诱导被抑制,但胚胎发育的畸形指数被显著提高,ANF分别与Phe,Py和BaP的联合暴露对胚胎发育呈潜在的协同作用。本文研究初步表明,3~5环PAHs化合物对海水青鳉胚胎发育的毒性作用方式可能不同;CYP1A活性抑制在PAHs混合物对海水青鳉胚胎发育的毒性作用过程中未起到缓解毒性的作用,CYP1A抑制剂与PAH型CYP1A诱导剂的混合物对鱼类胚胎发育具有潜在的协同毒性作用,现有的PAHs混合物毒性风险评价方法可能低估了实际环境中PAHs的风险;海水青鳉早期生活阶段的心脏发育对PAHs混合物暴露较为敏感,可推荐其作为生物标志物指示PAHs或溢油污染。     

Effects of naphthalene and phenanthrene on visual and prey capture endpoints during early stages of the dourado Salminus brasiliensis

Naphthalene (NAP) and phenanthrene (PHE) effects on Salminus brasiliensis, a carnivorous freshwater fish, were investigated using behavioral tests. Larval stages of S. brasiliensis were exposed to water concentrations of 0, 0.04 mg/l, 0.20mg/l and 0.50mg/l for naphthalene and 0, 0.01 mg/l, 0.05 mg/l and 0.1mg/l for phenanthrene during two developmental phases. The prey fish Prochilodus lineatus were not exposed. Visual acuity was measured at the end of phase 2 in individual S. brasiliensis, which were also tested at the end of each phase for number of attacks on prey, number of prey captured, prey capture efficiency, and distance swam. Vision was impaired by PHE exposure, as acuity angles increased in exposed fish. At Stage I 2.3+/-1.2 prey were captured with 46% efficiency in controls compared to 0.4+/-0.3 prey captured with 13.4% efficiency in fish exposed to 0.05 mg PHEl(-1), the LOEL for both endpoints. At Stage II 4.0+/-1.1 preys were captured in controls compared to 2.5+/-0.9 preys captured in fish exposed to 0.01 mg PHEl(-1), the LOEL. Stage II control fish captured prey with 70% efficiency compared to 30% efficiency at Stage II fish exposed to 0.05 mg PHEl(-1), the LOEL. Distance swam was not affected by either NAP or PHE exposure. The exposure of larval stages of S. brasiliensis to realistic water concentrations of PHE impairs foraging skills and could affect recruitment of the species.     

Determining the sensitive developmental stages of intersex induction in medaka (Oryzias latipes) exposed to 17 beta-estradiol or testosterone.

Certain environmentally persistent compounds can adversely affect reproduction by acting as steroid hormone agonists or antagonists. The goal of the present study was to determine the developmental stage most susceptible to exogenous hormone (estradiol and testosterone) exposure using a small teleost model. In the first (pilot study) of two experiments, medaka (Oryzias latipes), at varying developmental stages, were bath-exposed to 5 micrograms/l 17 beta-estradiol for 24 h. At 5 months of age, fecundity, fertility and embryo and larval viability (reproductive success) were investigated in control and exposed groups. Fish at 1, 1.5, 2 and 5.5 months of age were also sampled, processed and examined histologically for gonadal alteration. No significant differences in mortality, gonadal morphology, body weight, sex-ratio or time to maturity were seen between control and exposed fish. At 5 months, however, when exposure groups were compared to controls, significant differences were seen in reproductive success and viability of offspring. A second experiment exposed embryo stage 10, and 1-, 7- and 21-day-old larvae for 6 days to 15 micrograms/l 17 beta-estradiol or 100 micrograms/l testosterone. No significant differences were seen at 5 months in mortality, body weight, or time to sexual maturity. However, sex-ratios were significantly biased toward female in the stage 10, 1- and 7-day post-hatch estradiol exposure groups. No significant changes in sex-ratio were associated with testosterone exposure at any developmental stage. Further, intersex gonads were observed in fish from all groups exposed to 15 micrograms/l estradiol. Only those fish exposed as newly hatched fry or at 1 week post-hatch displayed intersex gonads following 100 micrograms/l testosterone exposure. Data from these experiments show that newly hatched fry are that life stage most sensitive to hormone exposure and the most appropriate to use in determining effects of known endocrine-disrupting compounds.     

Food and feeding habits of juvenile flounder Platichthys flesus (L.), abd turbot Scophthalmus maximus L. in the åland archipelago, northern Baltic Sea

The food choice of juvenile flounder (Platichthys flesus) and turbot (Scophthalmus maximus) was studied in the northern Baltic Sea during the years 1988, 1989, 1994 and 1995. The diet included organisms from 30 species/taxa in flounder (n = 306) and 10 species/taxa in turbot (n = 41). Flounder 45 mm mainly consumed meiofauna (dominating taxon: Harpacticoida, Copepoda) and larger fish (46–101 mm) consumed macrofauna (dominating taxa: Oligochaeta, Amphipoda and Chironomidae). In terms of biomass, macrofauna dominated for all sizes of flounders, and meiofauna was important only for the smallest fish. A strong seasonal variation could be detected in the diet. In spring, macrofauna dominated for all size classes of fish (only fish > 30 mm were caught in spring), while in summer and autumn meiofauna dominated the diets for fish 45 mm in size. Juvenile turbot (22–88 mm) consumed macrofauna and small fish. Turbot 30 mm consumed mainly amphipods, while > 30 mm turbot consumed mysid shrimps, amphipods and fish.The ontogenetic shift from meio- to macrofauna-sized prey in flounders occurs at a larger fish size in the northern Baltic Sea than reported in other areas, possibly depending on the increased relative importance of meiofauna in the northern Baltic. The seasonal variation in the diet could be due to seasonally changing abundances in the zoobenthos, or for the small fish (1-group, spring), to switching from meio- to macrofauna in order to optimize their energy gain. The 0-group flounders consumed meiofauna for a long period, possibly due to a learning-process or simply due to easy availability of meiofauna. Turbot has a much larger mouth gap than flounders, thus allowing them to consume macrofauna from the beginning of their benthic life.     

Reproductive life history stages sensitive to oil and naphthalene in Atlantic croaker

The reproductive effects of chronic exposure to water-soluble fractions (WSF) of diesel fuel oil and naphthalene in female Atlantic croaker during critical periods of their reproductive life history were investigated. Fish were repeatedly exposed to WSF of oil (2.5 or 5%) or naphthalene (0.5 or 1.0 ppm) for 5 or 8 weeks. Both pollutants blocked sexual maturation in some fish and impaired ovarian recrudescence in others. The majority of the oocytes in exposed fish were undeveloped and widespread oocyte atresia was evident at higher concentrations of these polycyclic aromatic hydrocarbons (PAH). The disruption of ovarian growth and oocyte development in croaker after PAH exposure was associated with alterations in reproductive endocrine function at multiple sites on the hypothalamus-pituitarygonadal axis. A third period of increased sensitivity to xenobiotics was observed during final oocyte maturation.     

Inhibition by polycyclic aromatic hydrocarbons of ATPase activities in Sebastiscus marmoratus larvae: relationship with the development of early life stages

Sebastiscus marmoratus larvae were exposed to waterborne polycyclic aromatic hydrocarbons (PAHs) containing 3-5 rings, benzo[a]pyrene (BaP), pyrene (Py) or phenanthrene (Phe), respectively at 0.01, 0.1 and 1 μg L?1. Cumulative mortality, frequency of dorsal curvature and rate of pericardial and yolk sac edema in larvae treated for 8 days were significantly increased in a dose-dependent manner. All three PAHs resulted in reduction of the lower jaw, and the extent of reduction increased with increasing concentrations of the PAHs. Na+/K+-ATPase and Ca2+-ATPase activity in larvae exposed to the three PAHs were all significantly inhibited in a dose-dependent manner. Analysis using the Pearson correlation coefficient indicated a significant correlation between the rate of the dorsal curvature and edema and the inhibition of Na+/K+-ATPase and Ca2+-ATPase activity, suggesting that the developmental defects caused by PAHs were related to their inhibition of Na+/K+-ATPase and Ca2+-ATPase activity.     

Vitellogenin as biomarker for estrogenicity in flounder Platichthys flesus in the field and exposed to 17α-ethinylestradiol via food and water in the laboratory

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.     

Accumulation of polycyclic aromatic hydrocarbons from contaminated soil by Kandelia candel

The purpose of this study was to investigate the accumulation behavior of two typical PAHs,phenanthrene(PHE) and fluoranthene(FLA),uptaken by the mangrove Kandelia candel in various treated sandy soils.The experiment lasted for three months.Soil and plant samples were collected after 3,6,9 and 13 weeks of the incubation and the levels of PAHs were analyzed using a GC/MS(HP6890-5975B).The results showed that the root accumulations of PHE and FLA were elevated with the increase of the PAH concentrations in the soil.With prolonged exposure time,PHE and FLA concentrations of the roots increased significantly in comparison with the control.PHE and FLA concentrations of root after 13 weeks of treatment ranged from 0.28 to 4.36 μg·g^-1 and 0.79 to 35.76 μg·g^-1,respectively.The calculated mean values of the root concentration factors were 0.58 for PHE and 0.71 for FLA.This indicated that root accumulation of FLA was greater than PHE,which would result from the higher octanol-water partition coefficient(K_ow) of FLA.These results underline the ability of mangroves to remove PAHs from contaminated marine substrates and hence their potential use in bioremediation and the removal of PAHs from the marine food chain.     

Sex-specific biochemical and histological differences in gonads of sea urchins (Psammechinus miliaris) and their response to phenanthrene exposure

Female and male individuals of the same species often differ with respect to their susceptibility to toxicant stress. In the present study, sea urchins (Psammechinus miliaris) of both sexes were exposed to high (150 μg L?1) and environmentally relevant (5 μg L?1) concentrations of phenanthrene over 10 days. While food intake was significantly decreased following exposure to 150 μg L?1 phenanthrene, histological indices (lipofuscin accumulation, fibrosis, oocyte atresia), energetic status (energy charge, sum adenylates, AMP/ATP ratio) as well as ascorbate levels in the gonads showed either little or no effect upon phenanthrene exposure. However, most parameters (vitamin C, energy charge, sum adenylates, AMP/ATP ratio, ATP and ADP concentrations, lipofuscin content, fibrosis) significantly differed between male and female animals. This study illustrates the difficulties to identify toxic injury in reproductive tissue as it may be superimposed by gametogenesis and spawning of gametes.     

Evaluation of non-specific immune functions in dab,Limanda limanda L., following short-term exposure to sediments contaminated with polyaromatic hydrocarbons and/or polychlorinated biphenyls

Experimental exposure tanks were set up containing 'clean' marine sediment spiked with set doses of polyaromatic hydrocarbons (PAH), or polychlorinated biphenyls (PCB), or both. Reference tanks contained only 'clean' sediment. Female dab Limanda limanda L., exposed to these sediments for 7 days showed no differences in serum total protein concentration, serum lysozyme activity or body growth. Their kidney leucocytes were assayed for extracellular production of reactive oxygen species of the phagocytic respiratory burst. No significant differences in superoxide anion (O2-) production were found between fish exposed to the 'spiked' sediments and those on the 'clean' reference sediment. The production of hydrogen peroxide (H2O2) was, however, reduced in the fish held on the PAH and PAH/PCB spiked sediments. H2O2 levels for fish exposed to PCBs alone, did not differ significantly from those of control fish. Our results suggest that the decrease in H2O2 production was due to exposure to PAHs, rather than to PCBs.