疣荔枝螺(Thais clavigera)性畸变现象的解剖学和组织学研究

以解剖学和组织学的方法研究了有机锡污染生物指示种——疣荔枝螺的正常雄、雌和性畸变个体的生殖系统。结果表明，性畸变个体除具有正常的雌性器官外，还有输精管或阴茎。当性畸变程度严重时，性畸变不育个体的卵囊腺组织出现破损，并具有不规则块状的不育卵囊和未排出的正常卵囊。虽然性畸变个体的雄性器官具有表现出雄性功能的潜能，但由于缺少精巢或完整的前列腺，不可能发展成为具有雄性功能的个体，这与性逆转有着本质上的区别。有机锡污染引起的性畸变已对疣荔枝螺种群生存构成了潜在的威胁，深化此类负面生态效应的研究是人类面临的重要课题。     

方斑东风螺性畸变现象的解剖学和组织学研究

以解剖学和组织学方法研究了方斑东风螺未成熟和成熟的正常雌、雄个体以及性畸变个体的生殖系统结构.性畸变个体除具有正常的雌性器官外,还有输精管或阴茎.在正常雄性输精管的个体发育过程中再现了系统发育的过程,而且在性畸变个体输精管的形成过程和发育过程中组织学结构特征的变化与正常雄性输精管的个体发育过程的相似.性畸变雌体的性腺发育受阻.目前,性畸变现象仅存在于成熟个体,但这并不表明未成熟个体中不会发生性畸变现象.具有较高食用价值的方斑东风螺因有机锡污染可能会对人类健康不利,应引起人们的关注.     

The Mollusc Thais haemastoma -An Exhibitor of 'Imposex' and Potential Biological Indicator of Tributyltin Pollution

Abstract. In the Azores the gastropod Thais haemastoma shows varying degrees of imposex, the induction of male sex characters in the female. It is inferred that imposex in T. haemastoma is caused by tributyltin (TBT) associated with boats using organotin-based antifouling paints. The two indices - relative penis size and vas deferens sequence - employed previously for Nucella lapillus , were used without alteration. T. haemastoma is only the third species reported as exhibiting 'effective sterilisation' of the female. Unlike Nucella lapillus, T. haemastoma has a planktonic larva, allowing populations to be sustained even when local TBT pollution is high. This makes T. haemastoma potentially a better indicator species, as monitored populations can be maintained by the influx of larvae. T. haemastoma may thus provide a cheap method of assessing the extent of TBT contamination in warm waters, particularly in the Mediterranean and Atlantic Islands. Preliminary examination of material from a marina in Spain showed a high incidence of 'sterile' animals.     

An easy, non-destructive, probabilistic method to evaluate the imposex response of gastropod populations

Endocrine disruption in marine populations is a growing concern. One of the best documented examples is imposex, the development of male characteristics in female gastropods, including the development of a penis and/or vas deferens, by exposure to organotins, mainly tributyltin. This syndrome has been an effective and widely used biomarker of organic tin contamination for almost 20 years. However, established imposex evaluation protocols sacrifice all the animals and repeated assessments can result in depletion of populations. To preserve animal lives as much as possible and to have a fast screening of areas subjected to organotin impacts, a non-destructive approach was tested. This approach uses reversible narcotization and a statistical criterion to evaluate the extent of imposex in the populations, instead of imposex indexes, which are based on evaluation of sacrificed, fixed animals. The data consist of two categories: males plus masculinized females in one category and normal females in another. The probability is then calculated that this sample could have been drawn from a normal population. This new method was tested in two contrasting areas of Brazil and with two different species, and the results compared to the conventional evaluation. Narcotization was completely reversed with a survival rate of 98%. Impacted sites could be identified with this technique in the same way as with the conventional one. The only limitation was that female sterility could not be verified. This disadvantage could well be compensated by the minimal mortality in the studied populations.     

Imposex and surface sediment speciation: a combined approach to evaluate organotin contamination in Guanabara Bay, Rio de Janeiro, Brazil

An integrated biological-chemical survey of organotin compounds was carried out in Guanabara Bay, the second most important Brazilian harbor complex. The biological survey revealed high levels of imposex in Stramonita haemastoma populations. Inside the bay values of relative penis length index from 42.7 to 107.6 and vas deferens sequence index from III to VI were found, while organisms collected outside the bay had values ranging from <0.1 to 35.2 and from 0 to II. None of the females sampled inside the bay were normal and imposex was found in all stations. Surface sediments in the bay are contaminated by tributyltin (10-522 ng/g d.w.) and triphenyltin (<3.9-39.4 ng/g d.w.) with greater concentrations close to shipyards and marinas. The observed predominance of parent compounds (TBT and TPT) is commonly found when recent inputs occur, but may also indicate slow degradation processes in the anoxic conditions of these sediments. To our knowledge, this is the first report of a combined imposex-sediment approach to evaluate organotin contamination in marine environments of South America, and also the first report of TPT detection in environmental matrices in this region.     

Increasing imposex in populations of Bolinus brandaris (Gastropoda: Muricidae) in the northwestern 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.     

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.     

Contamination of organotin compounds and imposex in molluscs from Vancouver,Canada

Gastropods and bivalves were collected at 15 sites at Vancouver and Victoria, Canada between 24 May and 7 June, 1999, to establish tissue concentrations of butyltin and phenyltin compounds, to record imposex symptoms in gastropods, and to assess the present status of organotin contamination around Vancouver. No neogastropods (such as Nucella lima) were found around Vancouver. Neogastropod populations could have been extirpated by severe TBT contamination in Vancouver, as relatively high concentrations of TBT were detected in tissues of Mytilus trossulus from Vancouver, and the neogastropods distributed in Vancouver might be sensitive to TBT. Recovery from imposex, however, was observed in neogastropod populations from three sites at Victoria and Mission Point. TBT contamination has continued around Vancouver, arising from continuous use of TBT in antifouling paints for vessels larger than 25 m in length; however, TBT has decreased around Victoria and Mission Point. Different patterns of TBT accumulation in tissue were observed among the bivalve species from Vancouver. The highest TBT concentration detected in Tresus capax suggested some possible adverse effects. TBT was the most predominant butyltin component in almost all bivalve specimens surveyed, suggesting a low rate of TBT metabolism. Phenyltin compounds were not detected in any molluscan specimens in this study.     

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.     

Tributyltin and triphenyltin induce spermatogenesis in ovary of female abalone,Haliotis gigantea

Two-month flow-through exposure experiments of tributyltin (TBT) and triphenyltin (TPhT) were conducted with abalone, Haliotis gigantea. Nominal concentrations of 100 ng TBT/l and 100 ng TPhT/l caused significant spermatogenesis in ovaries of exposed females. There were also significantly more contracted primary oocytes observed in females exposed to either TBT or TPhT than controls. The incidence of two types of unknown cells was also significant in females exposed to TPhT. No significant histological changes were observed in testis of exposed males. This ovarian spermatogenesis caused by TBT and/or TPhT resembles gastropod imposex. Remarkably high concentrations of TBT and TPhT were observed in the head (including central nervous system ganglia), compared to muscles concentrations. Accumulation of TBT and TPhT in the head may disturb reproductive hormonal regulators through neuropeptides released from ganglia. This, as well as possible aromatase inhibition, may be one of the inducers for spermatogenesis in the abalone ovaries.     

性畸变在三丁基锡污染的生物监测中的应用

该文概述三丁基锡在海洋防附着涂料中的应用及其环境效应，综述因TBT污染而导致某些螺类雌性个体中发育出雄性性征的性畸变现象发生情况，探讨性畸变在TBT污染生物监测中的应用。并以香港地区性畸变的发生为例，论证性畸变在生物监测中的应用，同时比较香港及世界其他地区性畸变的程度。     

Imposex in the rock shell, Thais clavigera, as evidence of organotin contamination in the marine environment of Korea

Imposex was measured in the rock shell, Thais clavigera, from the coast of the Korean peninsula. Frequency of imposex was 0% at two reference sites, but at 47 out of 61 sites, frequency of imposex was recorded as 100%. The degree of imposex was relatively high at the sites near a harbor and a shipyard. Tributyltin (TBT) and triphenyltin (TPT) concentrations in T. clavigera ranged from 5 to 508 ng/g and from 3 to 2460 ng/g, respectively. A significant positive relationship was found between degree of imposex and organotin concentration, whereas a significant negative relationship was obtained between female-to-male sex ratio and the degree of imposex. During a field transplantation of T. clavigera from a pristine area to a port, TBT and TPT were accumulated in T. clavigera, and imposex was induced. T. clavigera shows considerable potential as a bioindicator species of the adverse effects of TBT and TPT contamination.     

海产腹足类性畸变现象的形态特征

本文选取我国东南沿海的重要港口海口、北海、湛江、汕头和厦门港等作为海产腹足类性畸变现象的现场调查区，结果发现有11种海产腹足类存在性畸变。文中对分布广泛、易采集、性畸变现象普遍的疣荔枝螺的性畸变特征的解剖观察结果进行了详细的描述，同时对其他几种海产腹足类性畸变特性作了对比。根据研究结果将疣荔枝螺性畸变雌体的阴茎和输精管的畸变程度划分为5个阶段。     

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.     

Stramonita haemastoma as a bioindicator for organotin contamination in coastal environments

Stramonita haemastoma was investigated as a suitable bioindicator of TBT and TPhT contamination in the tropical Atlantic Ocean by: 1. Imposex induction in healthy females after inoculation with TBT and TPhT in the laboratory; and 2. Determining incidence of imposex in S. haemastoma collected from areas with various levels of tributyltin (TBT) and triphenyltin (TPhT) and determining the concentrations of these chemical in its tissues and that of its prey, the mussell Perna pernas. Imposex intensities and organotin concentrations in tissues showed good correlation, indicating S. haemastoma as a reliable bioindicator of TBT and TPhT contamination in coastal waters. Body burden threshold of TBT and TPhT for imposex induction was estimated to be 10-20 ng g(-1).     

Exploring the use of caged Nucella lapillus to monitor changes to TBT hotspot areas: a trial in the River Tyne estuary (UK)

Caged dogwhelks (Nucella lapillus) and mussels (Mytilus edulis) were placed in situ at sites along the River Tyne (UK) in order to investigate the potential for monitoring any change in tributyltin (TBT) mediated effects. After 6 months the development of imposex was investigated in the dogwhelks and concentrations of TBT were determined in the whole tissues of both species. Sites within the estuary showed a high degree of imposex, whilst those at sites North and South of the estuary mouth exhibited significantly lower induction. Concentrations of TBT measured in the dogwhelk and mussel tissues showed the same pattern. The success of the caging technique, occurrence of imposex at other UK sites and the need for further monitoring at TBT impacted locations in the light of current international controls on the use of TBT-based antifouling paints on sea-going vessels are discussed.     

The impact of tributyltin from large vessels on dogwhelk (Nucella lapillus) populations around Scottish oil ports

Surveys of imposex in dogwhelk (Nucella lapillus) populations around oil terminals in Sullom Voe (Shetland) and Scapa Flow (Orkney) have confirmed that the effects of tributyltin (TBT) released from large vessels in coastal waters can be recognised. The impact of large vessels can be separated from the effects of other inputs of TBT in the areas studied. The intensity of the effects was greater in Shetland than in Orkney, reflecting the higher exposure to TBT resulting from the greater volume of traffic using the Shetland terminal, and the more restricted water exchange in that area. A large proportion of the female dogwhelks in Sullom Voe were unable to reproduce. Females in Scapa Flow were still able to reproduce, except for a small proportion close to two fish farm sites where TBT net-treatments were used prior to 1987.     

三种织纹螺性畸变的发展过程与雄性结构效应

从2000年3月到2002年10月对中国沿海海产腹足类性畸变的大范围调查过程中．在广东省汕头港、湛江港和厦门港分别采集到西格织纹螺(Nassarius siquijorensis)、胆形织纹螺(Nassarius thersites)和节织纹螺(Nassarius hepaticus)等3个种，解剖观察发现3种织纹螺的雌性个体都发生了性畸变现象。根据织纹螺性畸变的发展过程，对目前的性畸变发展划分体系作了新的修改和补充。通过对比性畸变个体与正常雄性个体的特征．探讨了雄性个体结构特征在性畸变过程中的效应。     

Specific accumulation of organotin compounds in tissues of the rock shell,Thais clavigera

Concentrations of organotin compounds (butyltins and phenyltins) were determined in gonad, accessory sex organs, penis, digestive gland, kidney, radula with sac, oesophagus with crop, stomach, hypobranchial gland, rectum, mantle, osphradium, ctenidium, heart, salivary gland, head ganglia and muscle of imposex-exhibiting female and male rock shells (Thais clavigera), by gas chromatography with flame photometric detection (GC-FPD). Different tissue distributions were observed between butyltin and phenyltin compounds. More than 1000 ng TBT/g wet wt. were observed in ovary, digestive gland, kidney, heart, ctenidium, osphradium, stomach, head ganglia and penis, of both imposex-exhibiting females and males. More than 1000 ng TPT/g wet wt. were found in almost all tissues of both sexes. Approximately one-third or more of total TBT was accumulated in the digestive glands of both females and males, respectively. Meanwhile, approximately 40–50% and one-half of total TPT accumulated in the digestive glands of females and males, respectively.     

Imposex still evident in eastern Australia 10 years after tributyltin restrictions

A survey of imposex in Thais orbita (Neogastropoda) along the New South Wales (NSW) coast, Australia, found imposex was still widespread 10 years after a partial ban on tributyltin (TBT) based antifouling paints. Transplant experiments also revealed that imposex was induced in T. orbita over a period of 9 weeks in Sydney Harbour, NSW. A comparison of the imposex data with that collected shortly after the introduction of restrictions on TBT revealed a general decline in imposex frequency occurring at open coast sites, an exception being the site at South West Rocks. Sites located within harbour/bay areas did not display a decline in imposex frequency but a decline in the severity of imposex was apparent. The observed decrease in imposex values in T. orbita may reflect a general decrease of butyltin contamination within the coastal environment. However, the persistence of contamination "hotspots" within harbours and bays remains a concern.