Residues of 14C-4n-nonylphenol in mosquitofish (Gambusia holbrooki) oocytes and embryos during dietary exposure of mature females to this xenohormone

In order to assess in fish the maternal transfer of alkylphenolic compounds to the progeny, the identification and quantification of the labelled compounds present in oocytes and embryos was conducted after dietary exposure of mature female mosquitofish to 14C-4n-nonylphenol during vitellogenesis and embryogenesis respectively. Radioactivity found in bile and liver extracts accounted for 0.9-0.6 and 0.2-0.1% of ingested radioactivity for females exposed during vitellogenesis and embryogenesis respectively. The amount of extractable radioactivity present in oocytes and embryos was 0.19 and 0.07% of the ingested dose respectively. The radio-HPLC profiles obtained from bile, liver, oocyte and embryo extracts were similar. They showed the presence of 4n-NP-glucuronide as the major metabolite and traces of unchanged 4n-NP. The other metabolites corresponded to 8-hydroxynonylphenol, 9-(4-hydroxyphenyl)-nonanoic acid and para-hydroxybenzoic acid which is the final product of the alkyl chain oxidation. Our results indicate that exposure of ovoviviparous female fish to 4-NP during vitellogenesis and embryogenesis leads to the contamination of the progeny by 4-NP and its metabolites.     

o,p′-DDT induction of vitellogenesis and its inhibition by tamoxifen in Nile tilapia (Oreochromis niloticus)

In order to investigate the mechanism by which o,p′-DDT disrupts endocrine functioning of Nile tilapia in vivo, the estrogenicity of o,p′-DDT was investigated in conjunction with 17β-estradiol (E₂) and tamoxifen. Mature, male tilapia were treated intraperitoneally with o,p′-DDT (60 mg/kg, one dose) or E₂ (5 mg/kg, four doses) in the presence or absence of tamoxifen (5 mg/kg, six doses) for 12 days and then plasma vitellogenin (Vtg) (measured as alkaline-labile phosphorous), E₂, and testosterone (T) were measured. Vtg levels were increased dramatically by E₂ (1744±171 μg/ml) and moderately by o,p′-DDT (82±15 μg/ml) compared with controls (23±3.5 μg/ml). Tamoxifen alone had no effect on Vtg production, but inhibited both E₂ and o,p′-DDT stimulated vitellogenesis. T levels were reduced with E₂ administration (1688±383 pg/ml) and declined further with the combined treatment of E₂ and tamoxifen (281±70 pg/ml), compared with controls (6558±1438 pg/ml). Tamoxifen or o,p′-DDT alone did not affect T levels, but their combined treatment did (2069±647 pg/ml). The results of this study suggest that o,p′-DDT is weakly estrogenic in male tilapia, and that this activity may be mediated through the estrogen receptor.     

An aromatase inhibitor and tamoxifen decrease plasma levels of o,p-DDT and its metabolites in Nile tilapia (Oreochromis niloticus)

The objective of this study was to determine if tamoxifen or an aromatase inhibitor (4-hydroxyandrostenedione; 4-OHA) affected plasma concentrations of o,p^′-DDT and its metabolites, o,p^′-DDD and o,p^′-DDE, in mature tilapia. Male and female tilapia were injected 6 times intraperitoneally with o,p^′-DDT (40 mg/kg) alone or in combination with 4-OHA (2 mg/kg) over a 12 day period. An additional group of male fish was injected with tamoxifen (5 mg/kg) plus o,p^′-DDT. At the end of the treatment period, plasma samples were extracted and analyzed by GC/ECD. Females injected only with o,p^′-DDT had significantly higher levels of o,p^′-DDT compared with males. Interestingly, females and males treated concomitantly with o,p^′-DDT and 4-OHA or tamoxifen had significantly lower concentrations of plasma o,p^′-DDT (about 50%) compared with fish treated with only o,p^′-DDT. These initial results suggest that an interaction between endocrine-active compounds occurs in vivo in tilapia and may involve alterations in metabolism of o,p^′-DDT.     

Largemouth bass (Micropterus salmoides) and striped mullet (Mugil cephalus) as vectors of contaminants to human consumers in northwest Florida

The health benefits of regular consumption of fish and seafood have been espoused for many years. However, fish are also a potential source of environmental contaminants that have well known adverse effects on human health. We investigated the consumption risks for largemouth bass (Micropterus salmoides; n = 104) and striped mullet (Mugil cephalus; n = 170), two commonly harvested and consumed fish species inhabiting fresh and estuarine waters in northwest Florida. Skinless fillets were analyzed for total mercury, inorganic arsenic, polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/F), polychlorinated biphenyls (PCBs), and organochlorine pesticides. Contaminant levels were compared to screening values (SV) calculated using U.S. Environmental Protection Agency (EPA) recommendations for establishing consumption advisories. Largemouth bass were found to contain high levels of total mercury at all sampling locations (0.37–0.89 ug/g) and one location exhibited elevated total PCBs (39.4 ng/g). All of the samples exceeded Florida fish consumption advisory trigger levels for total mercury and one location exceeded the U.S. EPA SV for total PCBs. As a result of the high mercury levels, the non-cancer health risks (hazard index–HI) for bass were above 1 for all locations. Striped mullet from several locations with known point sources contained elevated levels of PCBs (overall range 3.4–59.3 ng/g). However, total mercury levels in mullet were low. Eight of the 16 mullet sampling locations exceeded the U.S. EPA SV for total PCBs and two locations exceeded an HI of 1 due to elevated PCBs. Despite the elevated levels of total PCBs in some samples, only two locations exceeded the acceptable cancer risk range and therefore cancer health risks from consumption of bass and mullet were determined to be low at most sampling locations.     

Fine-scale distribution of sardine (Sardina pilchardus) eggs and adults during a spawning event

High-resolution acoustic and ichtyoplankton sampling with a ‘continuous under-way fish egg sampler (CUFES)’ was performed in two regions of approximately 100 square nautical miles off southern Iberian Peninsula, with the aim of studying the small scale distribution of sardine (Sardina pilchardus) adults and eggs during a spawning event. Very dense patches (246 eggs m^− 3) of recently spawned eggs with dimensions (up to 3 nautical miles wide) significantly larger than daytime sardine schools were present in both regions. Egg staging and ageing showed very little intra-sample variation, indicating a synchronous spawning period at dusk. The internal structure of the patches evaluated by variography showed very low internal variability, as if they consisted of a single unit. This hypothesis is confirmed by the acoustic finding of large sardine shoals with similar dimensions to those of the patches after sunset and throughout the night. During that period, adults were found near or in contact with the bottom, suggesting that spawning occurred at depth. A distinct patch of older eggs was found in both areas, but with a few nautical miles of horizontal separation. Their characteristics (a larger area, lower egg densities and a more irregular shape) indicate that these patches were exposed to dispersion and ‘stirring’ by physical forces, reshaping their initial appearance, while mesoscale water circulation could have displaced the core of the patches away by several kilometres within a day.     

Metallothionein and heavy metal levels in rainbow trout (Salmo gairdneri) during exposure to cadmium in the water

Rainbow trout were exposed to 200 ppb cadmium in the water during four months at 5°C. The liver, kidney and gills were analyzed for cadmium, copper, zinc and metallothionein. Cadmium accumulated in all three organs and reached the highest concentration in the kidney. The zinc and copper concentrations were not altered during the experiment. The metallothionein concentration was significantly higher in liver of exposed fish than in control fish after three months. The kidneys reached significantly elevated levels of metallothionein in the exposed group after four months. These results demonstrate that metallothionein is induced by cadmium after exposure to the metal via water.     

Influence of diet on growth, condition and reproductive capacity in Newfoundland and Labrador cod (Gadus morhua): Insights from stable carbon isotopes (δC)

Cod populations in Newfoundland and Labrador waters have shown differing growth, condition and recruitment since near-universal declines in these properties during the cold period of the late 1980s and early 1990s. To assess the influence of variable prey communities on these parameters, we compared cod energetics and diet in populations off Labrador and the northeast and south coasts of Newfoundland. Many properties were highest in the southern group(s) and lowest in the northern group(s), including growth, somatic condition, liver index and age-at-maturity. Most differences could be explained by variations in diet, as measured by stomach contents and stable carbon isotopes (δ¹³C). The diet of Labrador cod consisted almost entirely of northern shrimp (Pandalus borealis), and these cod displayed the most benthic δ¹³C signatures. Northeast cod had a more varied diet that included capelin and other fish, but still had mostly benthic δ¹³C signatures, suggesting the importance of benthic prey like shrimp in this population. South coast cod exhibited the most varied diet, including capelin (Mallotus villosus), zooplankton, crabs and other fish, and had the most pelagic δ¹³C signatures. Among and within populations, the benefits of a more pelagic diet in medium-sized (30–69 cm) cod included higher somatic condition, higher liver index (lipid stores) and greater spawning potential (decreased incidence of atresia). It is hypothesized that major rebuilding of Newfoundland and Labrador cod stocks will require a return to a system that supports mostly pelagic feeding (i.e. capelin) in cod.     

Biochemical and histological responses in mussel (Mytilus edulis) exposed to North Sea oil and to a mixture of North Sea oil and alkylphenols

Several environmental chemicals are suspected to be responsible for adverse health effects on the reproductive system in various organisms. During this work, environmentally relevant concentrations of North Sea oil were used alone or in combination with alkylphenols and additional PAH to study the effect on vitellogenin-like protein expression and gonadal development in mussels. North Sea oil (0.5 ppm) induced the expression of phospho-proteins in both sexes indicating that some compounds are oestrogen-mimics. This induction was not seen in samples dosed with the mixture but signs of toxic effects were observed in the gonads. Indeed, numerous degenerating ovarian follicles in females and foci, similar to vertebrate melanomacrophage centres, were observed in testes.     

Rates of oxygen consumption and tolerance of hypoxia and desiccation in Chinese black sleeper(Bostrichthys sinensis) and mudskipper(Boleophthalmus pectinirostris) embryos

1Introduction Afossorialmodeoflifehasbeenadoptedbynu merousfishes(Gibson,1982).Burrowingmodeof lifeclearlyoffersanumberofadvantages,butburro wingfishmustalsobeadaptedtocopewiththespe cialproblemsthatthislifestylepresents.Fishthat constructburrowsininterti…     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...     

Identification of transcriptional effects of ethynyl oestradiol in male plaice (Pleuronectes platessa) by suppression subtractive hybridisation and a nylon macroarray

Suppression subtractive hybridisation (SSH) was used to generate cDNA libraries representing genes differentially expressed in response to ethynyl oestradiol (EE2) exposure in liver from male plaice (Pleuronectes platessa) previously analysed for vitellogenin (VTG) induction. Characterisation of the cDNA clones identified many as VTG (2 genes) and zona radiata proteins (ZRP) (3 genes), but 40 encoded other proteins, with more than half cryptic. Further analysis identified 85 non-redundant clones suitable for array on nylon membrane. Radiolabelled cDNAs were prepared from hepatic mRNA from EE2 treated plaice (0 and 21 days) and hybridised with the arrayed clones. Analysis of the data showed that 11/17 novel, 21/22 VTG, 13/14 ZRP, 2/2 liver aspartic proteinase (LAP) and 8/10 other mRNAs were up-regulated by EE2 exposure.     

凡纳滨对虾水孔蛋白-4的cDNA克隆及盐度胁迫对其肝胰腺mRNA表达水平的影响

为了探索水孔蛋白(Aquaporin,AQP)在凡纳滨对虾(Litopenaeus vannamei)渗透压调节过程中作用,本研究通过RACE方法获得克隆的凡纳滨对虾AQP(命名为LvAQP4)的c DNA全长序列,并分析了盐度胁迫对其肝胰腺m RNA表达的影响。结果发现:LvAQP4 c DNA序列全长为1 048 bp,其中包括75 bp的5￠UTR,187 bp的3￠UTR和786 bp的ORF。根据ORF序列推导出LvAQP4编码261个氨基酸,预测其分子质量为27.85 k Da,等电点为8.11。推导的氨基酸序列与其他甲壳物种的AQP相似度为48.8%到97.3%。进化分析显示LvAQP4属于AQP1-like亚族、AQP4类。定量PCR(q PCR)检测到LvAQP4在不同组织中均有表达,其中鳃的表达量最高,肝胰腺、肌肉、脑和眼柄中也较高,而血淋巴、肠、胃和胸神经节中表达量则较低。在高盐(盐度40)刺激下,凡纳滨对虾的肝胰腺LvAQP4 m RNA表达量会随着时间推移而上升,到6 h达到最高,而后逐步下降。而在低盐(盐度4)刺激下,凡纳滨对虾的肝胰腺LvAQP4 m RNA表达量并无明显变化。在原代分离和培养的肝胰腺细胞中,培养液中加入额外的Na Cl会剂量依赖地提高LvAQP4 m RNA表达水平。同样,通过在培养液中额外加入蔗糖提高培养液渗透压,也会剂量依赖地提高LvAQP4 m RNA表达水平,但作用不如Na Cl明显。这些结果表明盐度与肝胰腺LvAQP4的表达量有关,LvAQP4对凡纳滨对虾的渗透压调节具有非常重要的作用。     

条纹锯■对高温胁迫及胁迫后恢复方式适应的初步研究

研究了质量为148.88 g±17.29 g和16.32 g±2.23g两种规格的条纹锯(Centropristis striata)鱼种对高温的耐受性及高温胁迫后恢复方式的适应.结果表明:大规格鱼种的高温半致死温度为26.80℃,最适温度上限为20.38℃.小规格鱼种的高温半致死温度为29.26℃,最适温度上限为25.11℃.依据实验鱼种耗氧率的恢复情况可以断定:在暴露于高温半致死温度48 h或72 h条件下,直接恢复方式略优于梯度恢复方式;而暴露24 h 条件下,两种恢复方式的作用无明显差异.高温半致死温度暴露下,要避免条纹锯达到半数死亡则积温不能超过(1064.67±72.01)℃?h.     

Environmental estrogens interact with and modulate the properties of plasma sex steroid-binding proteins in juvenile Atlantic salmon (Salmo salar)

Environmental chemicals may modulate the endocrine system through interaction with plasma sex steroid-binding proteins (SBP) and SBP-regulated processes. Some of these chemicals, which are known to interact with the estrogen receptor (ER), were found to bind competitively to the Atlantic salmon (Salmo salar) SBP and potentially disrupt the endocrine function of these proteins. Furthermore, both weakly acting (di-n-butyl phthalate) and potent estrogen mimics (ethynylestradiol), were able to induce a substantial up-regulation of circulating levels of SBP in vivo. Interestingly, modulation of SBP-levels was found to be a more sensitive endpoint than chemically induced interference with classical ER-mediated mechanisms for weakly acting estrogen mimics like di-(n-butyl) phthalate. Interference with the endocrine function of SBPs may thus introduce a novel mechanism for endocrine disruption, and give additional answers to the question why some weakly acting xenoestrogens are causing “estrogen-like” reproductive disturbances in developing males.     

三种典型POPs对紫贻贝不同组织DNA损伤的比较研究

采用彗星电泳技术,比较研究了Aroclor 1254、BaP、DDT三种典型POPs对紫贻贝(Mytilus edulis)鳃、性腺、消化腺细胞的DNA损伤效应的差异,以期为进一步评价这三种典型POPs遗传毒理机制及其预警监测提供科学依据.结果表明,Tail DNA%是评价DNA损伤的理想指标,另外,在急性毒性试验中,...     

Changes in the tissue concentrations and contents of calcium, copper and zinc in the shore crab Carcinus maenas (L.) (Crustacea: Decapoda) during the moult cycle and following copper exposure during ecdysis

The tissue concentrations and contents of calcium, copper and zinc were determined at different stages in the moult cycle of the shore crab Carcinus maenas. The crabs were dissected into the following tissues: the midgut gland, the gill, the ‘rest tissue’ (the latter defined as the left over tissue, i.e. muscles and exoskeleton), and the haemolymph, and analysed as such. One group of crabs was maintained in clean seawater. In these crabs, calcium was lost from haemolymph during premoult and apparently stored in the midgut gland. At ecdysis approximately 90% of the whole body calcium content was lost with the exuviae while the haemolymph calcium content doubled. During early postmoult calcium stored in the midgut gland was used in combination with calcium absorbed from the surrounding seawater for a rapid calcification of the new exoskeleton. Both the copper and zinc concentrations declined in the ‘rest tissue’ compartment during premoult. At ecdysis the haemolymph copper and zinc contents increased 35 and 86%, respectively, from premoult to the newly moult stage. At the same time, however, the copper and zinc haemolymph concentrations decreased. During postmoult, copper (25%) and zinc (45%) were lost from haemolymph and mainly excreted but a minor fraction (5–10%) was stored in the midgut gland.A second group of crabs was maintained in copper-contaminated seawater (0.1 mgl⁻¹) during ecdysis and postmoult. Following exposure of the animals to raised ambient levels of copper, the calcium and zinc contents of the midgut gland were reduced while the copper contents of the midgut gland and most other tissues were increased in comparison to crabs held in clean seawater.     

基于鳃的microRNA转录组研究中国蛤蜊对重金属镉的响应

中国蛤蜊（Mactra chinensis）是一种重要的经济贝类,分布于我国的辽宁和山东。近年来,生境恶化和过度捕捞导致我国蛤蜊资源量减少。我们利用Illumina Hiseq-2500平台对中国蛤蜊鳃的microRNA转录组进行测序,运用差异表达寻找对镉刺激有响应的功能microRNA。结果显示对照组获得了14 415 256条clean reads,实验组获得了15 570 111条clean reads。在这些reads中一共存在14 584 077小RNA,包括1 898 035条unique小RNA,其中对照组和实验组共有187 859条unique小RNA。两个组的小RNA文库的片段长度分布是相似的,大部分小RNA长度分布在26~27 nt。在对照组文库中最丰富的片段长度是27 nt,其次是28 nt、26 nt和23 nt。在实验组文库中,数量最丰富的片段长度是26 nt,其次是27 nt、28 nt和23 nt。经过对序列前体以及结构特征的分析,发现这两个组中一共有50个microRNA,其中已知的是38个,新发现的是12个。对照组和实验组microRNA含量最丰富的片段长度是23 nt。通过差异表达分析,5个microRNA基因具有显著性差异且从对照组到实验组的表达量是上调的,其余45个没有显著性的差异。把这50个序列与中国蛤蜊转录组进行比对,一共找到了542个靶基因。5个具有表达差异的基因一共比对到11个靶基因。把这11个靶基因与NCBI数据库比对,4个靶基因在COG中得到注释,1个靶基因在GO中得到注释,6个靶基因在KEGG中得到注释,11个靶基因在nr数据库中得到注释,注释到的基因有泛素蛋白连接酶E3,Wnt信号通路,G蛋白信号调控等。     

The respiratory response of juvenile mud crabs, Rhithropanopeus harrisii to variations in salinity and following short-term exposure to halowax 1099®, a polychlorinated naphthalene (PCN)

Juvenile mud crabs Rhithropanopeus harrisii were exposed for five days to Halowax 1099, a polychlorinated naphthalene (approximately 1:1 ratio of tri- and tetrachlorinated isomers). Exposure concentrations were 0, 20, 50 or 100 μg/litre. The exposure salinity was 15%. At the end of the exposure period, respiratory rates of the crabs were determined at 15% S. The respiratory rates were also determined for control and exposed animals which were osmotically shocked by a move from 15% S to either 5 or 25% S immediately before respirometry. Exposure to PCNs always caused the mean respiratory rates of juvenile crabs to increase over control values. The respiratory response of crabs to PCN exposure was similar at the steady-state salinity (15% S) and after hyperosmotic shock (25% S). However, the mean rate for animals exposed to PCNs and given a hyposmotic shock was significantly higher than that for controls and for PCN-exposed animals at the other two salinities. Since hyperosmotic regulation of body fluids is most active at low salinities in this species, we suggest that the PCNs interfere with hyperosmotic regulation and reduce the efficiency of metabolic compensation for hyposmotic shock. The higher energetic cost of acclimation to transient reductions in salinity would be potentially detrimental to wild populations in impacted areas.     

Influences of diel and tidal cycles on fish assemblage in eelgrass (Zostera marina) bed of southern Korea during autumn

Short-term tidal and diel variations of autumn fish assemblage in a Zostera marina bed were investigated using 3 h interval samplings for 24 h in both spring and neap tide using a small beam trawl. A total of 1 346 fishes belonging to 19 species were collected at spring tide, whereas 1 115 fishes belonging to 17 species were at neap tide. The common fish species were Nuchequula nuchalis, Acanthogobius flavimanus, Takifugu niphobles, Acentrogobius pflaumii, and Pholis nebulosa with the former three species dominating at spring tide, while the latter two species being abundant at neap tide. Diel variation in abundance was significant with higher abundance at night than day, but there were no significant differences between spring and neap tides, and between ebb and flood tides (three-way ANOVAs). Diel variation in the abundance of fishes may be influenced by tidal range and cycle, and day-night differences of food availability and behaviors of fishes indirectly. Non-metric multidimensional scaling (nMDS) ordination and analysis of similarity (ANOSIM) results revealed significant differences in species compositions both between day and night, and between spring and neap tide. Eelgrass beds are highly productive marine ecosystem, and thus, our results will contribute to conservation of seagrass ecosystem in the study area.