Mechanisms of imposex induction in the mud snail,Ilyanassa obsoleta: TBT as a neurotoxin and aromatase inhibitor

The occurrence of imposex, imposition of male sex characteristics on female snails, has been extensively documented throughout the world. Tributyltin (TBT) and other organotins have been causally linked to imposex induction at levels as low as 2 ng/l. There are several proposed mechanisms of action. First, TBT has been shown to be neurotoxic and to accumulate in snail ganglia. Peptide hormones control sexual differentiation in gastropods, and one hypothesis is that TBT acts as a neurotoxin to abnormally release the peptide hormone Penis Morphogenic Factor (PMF). However, PMF has not been characterized to date. The neuropeptide APGWamide significantly induces imposex in the mud snail, Ilyanassa obsoleta, at 10(-16) moles sub-cutaneous (SQ) injection over 2 weeks, and could be the PMF in this species. A second hypothesis is that TBT inhibits aromatase activity leading to increased testosterone levels and decreased estradiol. In vitro studies with snail digestive gland microsomes showed that TBT-dosed snails not exhibiting imposex had a 52% reduction in aromatase activity. Although the role of vertebrate sex steroids is not known in gastropods, it is possible that the combination of changes in peptide and steroid hormones may lead to imposex induction at extremely low doses of TBT.     

Increasing imposex in populations of Bolinus brandaris (Gastropoda: Muricidae) in the north-western Mediterranean

The muricid gastropod Bolinus brandaris exhibited imposex, a phenomenon linked to the use of organotin biocides. Several stages of imposex development (1–5) could be distinguished and were documented with SEM-photographs for the first time in this species. Additional alterations of the genital tract in females (blocked vulva preventing copulation and deposition of egg capsules) and males (excrecences on the penis and vas deferens) were shown. Imposex intensity on the Catalan coast was very high. The frequency of imposex off Barcelona and Vilanova i la Geltrú was 100%, with all females showing stage 4. Imposex frequency off Sant Carles de la Ràpita was 99.7% and the VDS index scored 3.9. Despite regulations of TBT-containing antifouling paints in Spain since 1990, the present study revealed that frequency and intensity of imposex have increased on the Catalan coast, at least since 1997. However, at present, imposex does not apparently affect the population dynamics of B. brandaris in the studied area, since no serious consequences on female reproduction (only one case of female sterilization) were detected. Gonadal activity and oogenesis were not suppressed or supplanted by spermatogenesis. Sex ratios were not male-biased. Possible sources of organotin biocides are examined.     

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.     

Community diversity of bacteria in digestive tract of mud snail (Bullacta exarata Philippi) and its rearing shoal

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The effect of tributyltin upon steroid titres in the female dogwhelk, Nucella lapillus, and the development of imposex

Exposure of female Nucella lapillus to tributyltin (TBT) in seawater at a concentration of 40 ng Sn/litre led to accumulation of TBT in the tissues, and an increase in penis length compared to control animals. Investigation of the steroid concentrations by radioimmunoassay revealed a significant increase in testosterone after 28 days exposure to TBT. No significant effect was noted upon the amounts of progesterone or estradiol-17β. When testosterone was injected into female N. lapillus in the absence of TBT, penis length was found to increase significantly. These observation suggest that there may be an association between the change in testosterone titre in response to exposure to TBT and the development of imposex in the dogwhelk.     

锯缘青蟹大颚器的显微和超微结构

采用组织切片和电镜技术,对锯缘青蟹大颚器的形态结构进行了观察.大颚器位于大颚肌几丁质腱的后侧基部.腺细胞胞质丰盈,质膜大量内陷和作不同程度的卷绕,内含大量具管状脊的线粒体和滑面内质网.卵巢成熟时大颚器腺细胞内线粒体增多,少数线粒体形态异样,高尔基体易见,散布的小管状滑面内质网(tSER)少,高度汇合的tSER增多,大小形状不同的空泡数量明显增加,且近质膜分布,质膜内陷和卷绕程度加剧.表明此时锯缘青蟹大颚器的合成与分泌能力增强,其生理功能与卵巢发育密切相关.     

黄河三角洲关键种泥螺Bullacta exarata对沉积物-水体界面溶解氧和营养盐通量的影响

泥螺自2001因养殖目的引入黄河三角洲以来,随着分布范围的扩大和种群数量的增长,已成为当地潮间带的关键种。泥螺给当地民众带来较高收入,但其对底栖生态系统的潜在影响还不清楚。为了明确该种群的生物扰动对底栖微藻和微生物的初级生产,以及对水体-沉积物界面营养盐交换的影响,开展了该项实验。结果表明,在光照条件下,泥螺能显著影响沉积物-水体界面的溶解氧DO通量;在黑暗及无生物处理组的恢复期,泥螺能显著增加两界面间氨氮的通量。底栖微藻和小型生物在黑暗条件下也能增加氨氮的通量;但在有光照条件下,上述微藻和小型底栖动物对沉积物-水体界面间溶解氧和氨氮通量的影响不明显。实验结果为更深入理解泥螺对生态系统的影响提供重要理论支持。     

黄河三角洲潮间带泥螺重金属含量与生物质量评价

泥螺(Bullacta exarata)作为黄河三角洲区域成功引入的滩涂养殖种类, 为当地获得了巨大的经济效益, 但对其生态影响还了解不多。为了解该区域泥螺体内重金属含量并评价其生物质量, 于2012年8月对该区域进行泥螺和沉积物样品采集调查和分析。采用富集系数K、残留量指数I以及金属污染指数I_MP进行分析。结果表明, 黄河三角洲沉积物及泥螺体内6种重金属含量均相对较低, 多数符合国家海洋沉积物质量标准第一类。泥螺对不同重金属的富集系数K有明显差别, 其中对Cu 和Cd 的富集能力最强。残留量指数I富集系数K及金属污染指数I_MP均表明, 黄河三角洲潮间带泥螺对重金属的富集程度均较低, 说明该海域泥螺未受到重金属离子的污染。根据国家生物质量标准分析, 黄河三角洲潮间带泥螺的生物质量总体较好, 生物体内重金属含量总体上与海水中重金属含量无显著的相关性。     

生物胺对锯缘青蟹精巢发育的调控作用

采用活体注射和体外培养方法,以精巢指数、精巢生精小管的成熟节段比例和促雄腺B型细胞比例为量化指标,观察3种生物胺对雄性锯缘青蟹生殖神经内分泌的调控作用.活体注射实验表明:5-HT组锯缘青蟹精巢发育和促雄腺分泌活动具有显著的促进作用;DA组精巢发育受到抑制,但促雄腺细胞的分泌活动不受影响;OA组与同步对照组差异不显著.离体实验表明:5-HT刺激了脑和胸神经团的分泌活动,从而促进促雄腺分泌活动;OA和DA对脑和胸神经团分泌活动没有明显影响.视神经节的生殖内分泌活动几乎不受3种生物胺的影响.本实验首次报道了体外条件下生物胺对雄性甲壳动物生殖神经内分泌的调控作用,支持了5-HT首先刺激脑和胸神经团分泌GSH,再通过促雄腺活动从而促进精巢发育的观点.     

牙鲆芳香化酶Cyp19a多克隆抗体制备及其应用

芳香化酶Cyp19a在鱼类性别决定和性别分化中起关键作用,通过调控体内雌、雄激素的转化来影响鱼类性别表型形成。为深入研究Cyp19a在牙鲆(Paralichthys olivaceus)性腺分化和发育过程中的表达规律及作用机制,本文从牙鲆cDNA中克隆获得1557bp的cyp19a基因编码序列,并成功构建了原核重组表达质粒。经体外重组与纯化获得较高纯度的重组Cyp19a蛋白;以此作为抗原免疫兔子,制备多克隆抗体。用间接酶联免疫吸附剂测定(ELISA)技术检测抗血清效价,评估其免疫原性。结果表明免疫结束后得到的抗体血清效价超过了1∶50000,且纯化后抗体具有较好活性。Western Blot(WB)检测结果表明Cyp19a兔多抗可以特异性识别牙鲆重组Cyp19a蛋白和内源性Cyp19a蛋白。蛋白水平的雌雄性腺差异表达分析显示,牙鲆Cyp19a蛋白在卵巢中高表达,在精巢中微量表达。利用Foxl2和Dmrt1重组蛋白处理牙鲆性腺分化期幼鱼可分别显著上调和下调Cyp19a的表达(P<0.05)。综上所述,本研究成功制备了牙鲆Cyp19a多克隆抗体并进行了应用,为深入研究牙鲆等鱼类性别分化机制提供有力工具。     

Seasonal abundance, distribution and recruitment of mud crabs (Scylla spp.) in replanted mangroves

The abundance and distribution of mud crabs were studied in a replanted mangrove forest in Buswang, Aklan, Philippines. Two fishing gears, lift nets and bamboo traps, were used to monitor relative abundance of Scylla spp. populations from March 2002 to December 2003 inside the mangrove forest. A third gear, a stakenet set across a creek, was used to monitor crabs migrating out of the mangroves during the ebb tide. Scylla olivacea formed 99.3% and 70.3% of the catch in the mangrove and the stakenet, respectively. The percentage of Scylla tranquebarica increased from <1% in the mangrove catches to 29% in the stakenet. Scylla serrata was present at very low levels in both catches. The lack of modal progression in the size–frequency plots and the year-round catch rate of gravid females suggested that recruitment was constant throughout the year. Even though relative abundance decreased over the study period indicating that the stock is being over-exploited, mud crab production is more than equivalent to that of most natural mangroves.     

Occurrence of gonadtropins like substance in the thoracic ganglion mass of the mud crab, Scylla paramamosain(Crustacea: Decapoda: Brachyura)

The identification and localization of vertebrate follicle stimulating hormone (FSH) and luteinizing hormone (LH) in crustaceans may further elucidate the regulation mechanisms in arthropod repro-duction.Using immunocytochemical staining techniques,this study has localized vertebrate FSH-like and LH-like substances in neurons in the subesophageal and thoracic ganglia from the thoracic ganglion mass (TGM) of Scylla paramamosain (Crustacea:Decapoda:Brachyura).Enzyme-linked immunosorbent assay (ELISA) has shown that the concentrations of both FSH-and LH-like sub-stances increased markedly in the TGM during the vitellogenic stage compared with that in the previtellogenic stage.These results indicate that substances resembling the vertebrate FSH and LH are present in S.paramamosain,and they may be involved in the development of the ovary as well as in ovulation.     

Risk assessment of TBT in the Japanese short-neck clam (Ruditapes philippinarum) of Tokyo Bay using a chemical fate model

A risk assessment of Tributyltin (TBT) in Tokyo Bay was conducted using the Margin of Exposure (MOE) method at the species level using the Japanese short-neck clam, Ruditapes philippinarum. The assessment endpoint was defined to protect R. philippinarum in Tokyo Bay from TBT (growth effects). A No Observed Effect Concentration (NOEC) for this species with respect to growth reduction induced by TBT was estimated from experimental results published in the scientific literature. Sources of TBT in this study were assumed to be commercial vessels in harbors and navigation routes. Concentrations of TBT in Tokyo Bay were estimated using a three-dimensional hydrodynamic model, an ecosystem model and a chemical fate model. MOEs for this species were estimated for the years 1990, 2000, and 2007. Estimated MOEs for R. philippinarum for 1990, 2000, and 2007 were approximately 1–3, 10, and 100, respectively, indicating a declining temporal trend in the probability of adverse growth effects.A simplified software package called RAMTB was developed by incorporating the chemical fate model and the databases of seasonal flow fields and distributions of organic substances (phytoplankton and detritus) in Tokyo Bay, simulated by the hydrodynamic and ecological model, respectively.     

中国海汇螺科Potamididae的分类学研究II．拟蟹守螺属、锥蟹守螺属、望远蟹守螺属及笋光螺属

继中国沿海汇螺科Potamididae分类学研究I后,又完成了汇螺科的分类学研究II。本文主要论述了汇螺科中拟蟹守螺属Cerithidea、锥蟹守螺属Cerithideopsis、望远蟹守螺属Telescopium及笋光螺属Terebralia共4属9种,其中1个中国新纪录种为邱氏拟蟹守螺Cerithideaquoyii(Hombron&Jacquinot,1848)。文中对其形态特征、生活习性和地理分布等进行了研究论述,并对以往鉴定有误和使用混乱的种名进行了修订。目前为止,共整理分类出中国沿海汇螺科5属18种。为了解中国汇螺资源情况、生物多样性和生态学研究提供基础资料。     

盐度对拟穴青蟹幼蟹存活、生长和Na+/K+-ATP 酶活性的影响

研究了盐度(6、13、20、27和33)对拟穴青蟹(Scylla paramamosain)幼蟹生长发育和Na+/K+-ATP酶活性的影响。结果表明,盐度对拟穴青蟹幼蟹甲宽和体质量特定生长率、C3蜕壳间期、甲宽增量(C1至C2和C2至C3)及Na+/K+-ATP酶活性有显著影响(P<0.05),但对幼蟹存活率、C2蜕壳间期及C3至C4甲宽增量无显著影响(P>0.05)。盐度13处理组特定生长率最高、蜕壳间期最短,盐度6处理组特定生长率最低,但与盐度20、27和33处理组无显著差异(P>0.05),而盐度33处理组蜕壳间期最长;第一次蜕壳后盐度20处理组甲宽增量最大,后两次蜕壳盐度13处理组均是最大;盐度处理0.5、4、7和11 d时,Na+/K+-ATP酶活性分别表现为盐度6>13>20>27>33、6>13>27>20>33、6>13>20>27>33和6>13>33>20>27的趋势,低盐环境下Na+/K+-ATP酶活性的升高有助于拟穴青蟹幼蟹对低渗环境的适应。     

The blooms of a cyanobacterium, Microcystis cf. aeruginosa in a severely polluted estuary, the Golden Horn, Turkey

The distribution of toxic cyanobacterium Microcystis cf. aeruginosa in the severely polluted Golden Horn Estuary was studied from 1998 to 2000. Microcystis persisted at the upper estuary where the water circulation was poor and values ranged between 2.9 × 10⁴ and 2.7 × 10⁶ cells ml⁻¹ throughout the study. Simultaneously measured physical (salinity, temperature, rainfall and secchi disc) and chemical parameters (nutrients and dissolved oxygen) were evaluated together with Microcystis data. Although the Microcystis blooms generally occur in summer due to the increase in temperature, the blooms were recorded in winter in the present study. The abundance of Microcystis depended on the variations in salinity and both blooms were recorded below S = 2. A moderate partial correlation between Microcystis abundance and salinity was detected in the presence of temperature, dissolved oxygen and precipitation data (r = −0.561, p = 0.002). The M. cf. aeruginosa abundance was low in the summer when the salinity was higher than winter. A remarkable increase in the eukaryotic phytoplankton abundance following the improvements in the water quality of the estuary occurred, whilst the Microcystis abundance remained below bloom level.     

Spatial and temporal variation of Palaemon adspersus, Palaemon elegans, and Crangon crangon (Decapoda: Caridea) in the southern Black Sea

In order to characterize the seasonal, bathymetric, and spatial distribution of the species, Palaemon adspersus, Palaemon elegans, and Crangon crangon, shrimps were sampled with a beam trawl in four stations at depths between 1 and 30 m from February 2002 to January 2004 on the Sinop Peninsula coasts of the southern Black Sea. One-way analysis of similarity (ANOSIM) results demonstrated that the caridean composition was significantly different (p < 0.001) between seasons and between sampling areas. No significant relationship of the caridean composition was evident between depth zones ranging from 1 to 30 m. These three carideans occurred together in 28.5% of the sampling occasions. Different seasonal migration pattern was evident for all the three species. Palaemon adspersus migrated inshore during relatively higher water temperatures, whereas C. crangon density decreased in the shallow waters during the same period, and P. elegans population was mostly observed at depth zones of 5–10 m and was only observed in the 30 m depth zone in winter. The abiotic factors that characterize the coexistence of these three carideans were primarily determined by the habitat types and bottom structures.     

Trophic relations of capelin Mallotus villosus and polar cod Boreogadus saida in the Barents Sea as a factor of impact on the ecosystem

The purpose of the study is to assess the role of trophic relations of the dominant pelagic fishes capelin and polar cod in the Barents Sea with regard to distribution and accessibility as prey for the Atlantic cod in warm years (2004–2005). Unlike in the previous period, during these warm years a dramatic increase of the polar cod population resulted in a northwards expansion of the feeding grounds where overlapping of polar cod and capelin concentrations was observed. This caused an increased competition for copepods, which are the main food item for young fish. In the areas dominated by polar cod the shortage of copepods forced immature capelin to switch to the chaetognath Sagitta, which affected their fatness negatively.During the warm years the feeding grounds of Atlantic cod also expanded, to a large degree caused by the shortage of their main food, the capelin. In 2004–2005 the cod formed feeding concentrations in the north and northeast Barents Sea where they fed on the capelin. In this area the consumption of polar cod by cod increased, and in some local areas the polar cod practically replaced the capelin in the diet of cod. In general polar cod in the diet of Atlantic cod were more important in the northern than in the southern part of the Barents Sea. The fatness of cod was extremely low during the whole spring–summer period (until August), and after the feeding period the fatness index of the Atlantic cod became lower than the average long-term autumn value.