Imposex in Three Marine Gastropod Species in Chile and Potential Impact on Muriciculture

A survey of imposex was conducted between the Bay of San Vicente, Concepción, Chile (36° 50′) and the Bay of Renocavi, Puerto Montt, Chile (41° 35′), to determine if imposex is occurring in an area of collection and cultivation of the gastropod, Chorus giganteus. Imposex was recorded in three chilean muricids C. giganteus, Xanthochorus cassidiformis and Nucella crassilabrum complex. Percentage of imposex in females sampled ranged from 0% to 100%. The degree of expression of imposex was not so severe as to elicit sterility in the individuals surveyed. Incidence of imposex was as high as 47% in areas of harvest of C. giganteus for reproductive and cultivation studies for commercial production of this species. A six percent incidence of imposex was observed in Nucella proximal to the facility for cultivation of C. giganteus. Gastropods of the genus Nucella are used in other countries to monitor incidence of imposex and have demonstrated sensitivity toward tributyltin (TBT), the associated causal agent. Since sex of individuals in muriciculture is determined by the presence or absence of a penis, imposex may lead to erroneous classification of males and females. The incidence and severity of imposex in C. giganteus and related species may have a negative impact on sustainable cultivation and harvest of gastropods in Chile.     

Imposex in Morula Granulata as Bioindicator of Tributyltin (TBT) Contamination in the Dampier Archipelago, Western Australia

This study was undertaken to examine the use of imposex in the tropical neogastropod whelk Morula granulata as bioindicator of tributyltin (TBT) contamination. Samples were collected from sites throughout the Dampier Archipelago; both impact sites close to intensive shipping activity and remote control sites. TBT in this area originates mainly from commercial vessels using the Port of Dampier that are >25 m in length and exempt from legislation controlling the use of TBT in antifouling paints. A field survey was undertaken in July 1997; 100 M. granulata were taken at each of 18 sites with varying vessel activity, and examined for imposex. The level of TBT contamination in the Archipelago was low; ranging from <0.3 to 25 ng Sn l⁻¹ in water,<0.3 to 80 ng Sn g−¹ in oyster tissue (Saccostrea cuccullata – prey of M. granulata), and <0.3 to 33 ng Sn g⁻¹ in whelks. Percentage of imposex in M. granulata ranged from 0% to 57%, and was correlated with distance to vessel activity (r=0.646, p<0.05). The only significant relationship of imposex with butyltin concentrations was with TBT in oysters (r=0.826, p<0.05), and dibutyltin (DBT) in whelks (r=0.783, p<0.05). It is postulated that this is because of interspecies variation in ability to metabolize TBT to DBT and MBT (monobutyltin), as well as induction of imposex in earlier life stages, so that imposex in adults could be a reflection of prior contamination. M. granulata is a suitable bioindicator of TBT contamination in the Dampier Archipelago, which could be used throughout the Indo-Pacific given its abundance in the region.     

Effects of Animal Size on Concentrations of Metallothionein and Metals in Periwinkles Littorina littorea Collected from the Firth of Clyde, Scotland

Different sizes of Littorina littorea were collected from four areas in the Firth of Clyde, Scotland. Their metallothionein (MT) and heavy metal concentrations were analysed using the Silver Saturation Method and Atomic Absorption Spectrophotometry, respectively. Concentrations of MT, Cd and Zn (as μg g⁻¹ dry soft-body weight) generally decreased with an increase in size of L. littorea. MT concentrations were better correlated with Cd than with Zn or Cu concentrations. Nevertheless, MT and the metals in periwinkles (as μg individual⁻¹) increased significantly with increasing size. Concentrations of MT and the metals among the sampling areas were compared at a standardised soft-body weight (10 mg). The results from Discriminant Analysis based on all metal concentrations indicate that Largs is different from the other areas and characterised by high Fe concentrations in L. littorea. The problems and differences in using either soft-body weight or shell length as independent variable for size-standardisation are discussed.     

Imposex levels in the dogwhelk Nucella lapillus (L.)--continuing improvement at high latitudes

In 1990, restrictions on the use of tributyltin (TBT)-based antifouling paints were implemented in Iceland. A previous study showed that the level of imposex in the dogwhelk, Nucella lapillus, in Icelandic waters had decreased significantly between 1992 and 1998. In this study, we repeated the survey on imposex in N. lapillus at 33 locations from the Icelandic coast in 2003. The results indicated that both Vas Deferens Sequence Index (VDSI) and Relative Penis Size Index (RPSI) had further declined in 13 locations since 1998. Among these 13 sites, RPSI was reduced to zero in five cases. While improvements from 1992/1993 to 1998 were seen in reduced levels of imposex near both large and small harbours, the pattern from 1998 to 2003 was somewhat different, with improvement mainly observed near smaller harbours. No significant changes in imposex levels near larger harbours occurred over this period. Although the imposex levels still remain high near the large harbour complexes in Reykjavík and Hafnarfjörður, it is evident that regulations, including the use of less toxic antifouling paints and community action, have lead to substantial improvements in the marine environment of Iceland. International Maritime Organisation’s ban on the application of TBT after 2003 is apparently necessary to allow further improvements in larger harbours. The environmental effects of new antifoulants replacing TBT need to be further evaluated.     

Imposex in the indigenous Nassarius kraussianus (Mollusca: Neogastropoda) from South African harbours

Nassarius kraussianus populations in the Durban and Richards Bay harbours (along the eastern seaboard of southern Africa) and in the Knysna lagoon (southern Cape coast) showed imposex development, confirming bioavailability of tributyltin (TBT) in these waterways. Incidence of imposex (which ranged from 29% to 100%) and relative penis length of females (RPL; which ranged from 1.3% to 55.2%) were markedly variable among populations from different sites in each waterway, indicative of localised effects of TBT contamination. Shell length of the snails was apparently not correlated with contamination level, as assessed from imposex measurements. While population imposex features are easy to determine and often provide a sensitive biomarker of TBT exposure, the discussion outlines reservations when extrapolating these to general ecological situations. Additionally, this study represents the first published report on the topic for sub-Saharan, African harbours, some of which seem remarkably less affected than suggested for other world regions.     

Imposex levels in the dogwhelk Nucella lapillus (L.)—continuing improvement at high latitudes

In 1990, restrictions on the use of tributyltin (TBT)-based antifouling paints were implemented in Iceland. A previous study showed that the level of imposex in the dogwhelk, Nucella lapillus, in Icelandic waters had decreased significantly between 1992 and 1998. In this study, we repeated the survey on imposex in N. lapillus at 33 locations from the Icelandic coast in 2003. The results indicated that both Vas Deferens Sequence Index (VDSI) and Relative Penis Size Index (RPSI) had further declined in 13 locations since 1998. Among these 13 sites, RPSI was reduced to zero in five cases. While improvements from 1992/1993 to 1998 were seen in reduced levels of imposex near both large and small harbours, the pattern from 1998 to 2003 was somewhat different, with improvement mainly observed near smaller harbours. No significant changes in imposex levels near larger harbours occurred over this period. Although the imposex levels still remain high near the large harbour complexes in Reykjavík and Hafnarfjörður, it is evident that regulations, including the use of less toxic antifouling paints and community action, have lead to substantial improvements in the marine environment of Iceland. International Maritime Organisation’s ban on the application of TBT after 2003 is apparently necessary to allow further improvements in larger harbours. The environmental effects of new antifoulants replacing TBT need to be further evaluated.     

Metallothioneins and trace metals in the dogwhelk Nucella lapillus (L.) collected from Icelandic coasts

Different sizes of the dogwhelk Nucella lapillus were collected from eight locations along the southwest and north coasts of Iceland. Concentrations of total metallothioneins (MTs) and heavy metals (Cd, Cu, Cr, Mn, Ni, and Zn) were analysed using the silver saturation method and inductively coupled plasma-atomic emission spectrometry. The level of tributyltin (TBT) contamination was also assessed using imposex indices, the vas deferens stage index (VDSI) and relative penis size index (RPSI). Gufunes N. lapillus presented the highest values of VDSI (4.0) and RPSI (11.1), followed by Grenivík individuals (VDSI = 3.0; RPSI = 0.9), while the Strandakirkja population showed the lowest VDSI (0.3) and zero RPSI. At a standardised size (0.25 g dry soft-body weight), Grenivík N. lapillus exhibited significantly higher concentrations of all metals whereas overall metal concentrations were significantly lower in individuals from Strandakirkja and Garðskagi compared to other study sites. Partial correlation analyses with size correction indicated that MT concentrations were better correlated with Cd and Cu concentrations than with other metals. At the standard size, the pattern of MT concentrations in N. lapillus from different sites was, however, very different from those of metal profiles. Such discrepancies between the patterns of MT and metals in N. lapillus might be explained by the fact that MT induction could be influenced by various factors such as temperature, dietary metal intake, growth rate and co-existence of other MT-inducing chemicals.     

Imposex levels and concentrations of organotin compounds (TBT and its metabolites) in Nassarius nitidus from the Lagoon of Venice

Specimens of Nassarius nitidus were collected in seven stations of the Venice Lagoon to assess the levels of tributyltin (TBT) and its metabolites monobutyltin and dibutyltin in the tissues and monitor their effect on organisms, in particular the phenomenon of imposex (superimposition of male sexual characteristics on females). The following values of population indices were found: vas deferens sequence: 1.2+/-0.7-4.0+/-0.5; relative penis length: 6-47%. The least impacted station was situated in the northern part of the Lagoon, where females without imposex were found and Butyltin (BuTs) concentrations in the organisms (average sum of BuTs=43+/-14 ngSng(-1)w.) were significantly lower than in the other stations (range of average sum of BuTs: 101+/-22-217+/-27 ngSng(-1)d.w.). Population indices were found to be related to the TBT content in the tissues. In particular VDSI had a significant logarithmic correlation: r=0.95, n=8, p<0.05.     

A Comparison of Growth Biomarkers for Assessing Sublethal Effects of Cadmium on a Marine Gastropod, Nassarius festivus

The effect of cadmium on the growth of an intertidal marine gastropod, Nassarius festivus, was investigated using a number of biomarkers namely conventional measurements in terms of increase in shell length or body weight, scope for growth (SfG) and RNA/DNA ratio. After eight days of cadmium exposure, the no observable effect concentration (NOEC) and lowest observable effect concentration (LOEC) for the rate of increase in wet weight were 1.00 and 1.38 mg/l, respectively, while those for shell length did not show any significant difference from the control until day 16. After 16 days of cadmium exposure, the LOEC for both shell length and wet weight measurements was 0.22 mg/l. The 8-day SfG test was sensitive to cadmium stress, showing a significant decrease at 0.16 mg/l cadmium when compared to the control. The NOEC and LOEC of RNA/DNA ratio determination were 0.52 and 0.93 mg/l, respectively. An assessment based on sensitivity and technical complexity suggests that Scope for Growth is the most sensitive growth biomarker, followed by the RNA/DNA ratio, and then the conventional growth measurement based on shell size and body weight.     

Imposex levels and concentrations of organotin compounds (TBT and its metabolites) in Nassarius nitidus from the Lagoon of Venice

Specimens of Nassarius nitidus were collected in seven stations of the Venice Lagoon to assess the levels of tributyltin (TBT) and its metabolites monobutyltin and dibutyltin in the tissues and monitor their effect on organisms, in particular the phenomenon of imposex (superimposition of male sexual characteristics on females). The following values of population indices were found: vas deferens sequence: 1.2 ± 0.7–4.0 ± 0.5; relative penis length: 6–47%. The least impacted station was situated in the northern part of the Lagoon, where females without imposex were found and Butyltin (BuTs) concentrations in the organisms (average sum of BuTs = 43 ± 14 ng Sn g⁻¹ w.) were significantly lower than in the other stations (range of average sum of BuTs: 101 ± 22–217 ± 27 ng Sn g⁻¹ d.w.). Population indices were found to be related to the TBT content in the tissues. In particular VDSI had a significant logarithmic correlation: r = 0.95, n = 8, p < 0.05.     

Butyltin compounds,sterility and imposex assessment in Nassarius reticulatus (Linnaeus, 1758), prior to the 2008 European ban on TBT antifouling paints,within Basque ports and along coastal areas

Levels of imposex (superimposition of male characters, upon females) and the presence of sterile females are assessed in the gastropod Nassarius reticulatus (L.), at 35 locations in the Basque Country. At 18 of these localities, organotin bioaccumulation (tributyltin (TBT); dibutyltin; monobutyltin) was evaluated, using isotope dilution GC–ICP–MS. Higher imposex levels and TBT body burden were found in confined harbours, with large vessel traffic or the presence of an offshore fleet. The highest values were found for the port of Pasaia, near to a shipyard (a relative penis size index of up to 92%, and TBT body burden up to 405 ng g^?1 – given on a dry weight basis as Sn). Imposex effects were evaluated also in parasitized females, at some of the sampling sites. The percentage of presence of c-type Vas Deferens sequence stages (i.e., with vas deferens beginning at the oviduct opening) was higher in parasitized females. In turn, there was an absence of highly convoluted oviduct in parasitized females.     

Aspects of Assaying Sediment Toxicity in Irish Estuarine Ecosystems

The toxicity of surface sediments collected from eight estuarine sites around the Irish coast was assessed in March 1997 using a whole sediment bioassay. Two molluscan species, Tapes semidecussatus (1 + age class) and Scrobicularia plana (unknown age) were exposed to test sediments for 21 days. Survival, morphometric, behavioural and histochemical measurements were taken during the 21 day bioassay. 100% mortality occurred in both species in sediments from one site in Dublin Bay within 18 days of initial exposure. 100% survival of T. semidecussatus was observed in all but four of the test sediments to which it was exposed during this study. The animal condition and wet tissue weight of T. semidecussatus decreased for all sediments during the bioassay. 95% survival of S. plana was observed after 21 days exposure to Youghal (reference) sediment. Condition and wet tissue weight of S. plana increased in all test sediments during the bioassay, however, there was significant variation in wet tissue weight of this species on day 25 of the bioassay in all test sediments. There was significant variation in the burrowing behaviour of S. plana during the bioassay. Lipofuscin accumulation was observed in the digestive diverticula of T. semidecussatus exposed to all test sediments with the exception of Youghal (reference). The potential for both species as bioassay organisms is discussed.     

Seasonal changes in imposex and tissue burden of butyltin compounds in Thais clavigera populations along the coastal area of Mirs Bay, China

We assessed the current status of tributyltin (TBT) contamination of Thais clavigera (Gastropoda) along the coastal area of Mirs Bay, China for the first time. The snail samples were collected from 10 different sites in the summer (June) and winter (December) of 2006, respectively. They were analyzed for imposex status, i.e. relative penis size index (RPSI) and vas deferens sequence index (VDSI), followed by quantification of butyltins in their tissues. Most of the collected females suffered from imposex, but the level of butyltin contamination varied with the distance from Yantian Port, which is currently the fourth busiest container port in the world. The tissue concentration of TBT varied with season. For a particular site, the winter samples in general contained much higher concentration of TBT than the summer samples. RPSI, VDSI and organotin concentrations were higher in T. clavigera collected from sites closer to the Port, such as the Seafood Street and Kat O. Both RPSI and VDSI were positively correlated with the tissue burden of tributyltin. VDSI exhibited little seasonal variability, whereas RPSI showed marked seasonal variability, with lower values in the summer samples. The current results will serve as an important reference for long-term monitoring of butyltin contamination in this area.     

武汉后湖长角涵螺的周年生产量

采用两种方法对后湖长角涵螺(Alocinma longicornis)的周年生产量进行了测算,结果表明,两种方法得到的生产量较为吻合.生产量的带壳湿重和去壳干重分别是:体长频率法,2.22 g/(m2·a),0.111 g/(m2·a);瞬时生长率法,2.177 g/(m2·a),0.109 g/(m2·a).对应的P/B系数为4.0,5.4.     

The significance of imposex levels and TBT contamination in the red whelk, Neptunea antiqua (L.) from the offshore Irish Sea

The extent to which open seas and oceanic waters have been contaminated by TBT is unclear since most sampling programmes have concentrated on inshore areas, particularly near harbours. Accordingly, the red whelk, Neptunea antiqua L. (Mollusca: Prosobranchia), a sublittoral and dioecious prosobranch species, was selected to investigate the potential effects of TBT-induced imposex in the central western Irish Sea. The presence of imposex was noted in > 90% of all female whelks that were examined on a monthly basis over a 12-month period. However, the proliferation of the vas deferens was not considered severe enough to affect the reproductive capabilities of the population. Low concentrations of butyltins were detected in the soft tissues of all whelks examined, ranging from 0.009 to 0.112 microg g(-1) Sn dry wt There was no significant correlation between the documented imposex stages and total butyltin contamination, indicating that the species is not a reliable bioindicator species of the effects of this contaminant.     

Responses of mud snails from low and high imposex sites to sex pheromones

Imposex, male secondary sexual characteristics in female snails, is a morphological indicator of sub-lethal exposure to organotin compounds. The relation between imposex and behavioral responses to sex pheromones was studied. Responses of snails to sex specific pheromones were determined in laboratory assays. Females and males from a low imposex site and females, imposex females and males from a high imposex site were tested. The snails from the low imposex site showed the expected strong behavioral dimorphism with females responding to male pheromone and males responding to female pheromone. In assays using snails from the high imposex site, female and imposex females showed no dimorphism in response to pheromones and males showed a relatively weak dimorphism. As there was no breeding or egg capsule deposition at the high imposex site, we conclude that even the morphologically normal snails were behaviorally and reproductively compromised.     

Spatial and temporal changes of TBT pollution along the Portuguese coast: inefficacy of the EEC directive 89/677

Nucella lapillus imposex (superimposition of male characters onto prosobranch females) and organotin body burden were surveyed in the Portuguese coast in 1997 and 2000. Time comparisons of the available data on Nucella lapillus imposex in the Portuguese coast indicate a global increase of TBT pollution in recent years, revealing the inefficacy of the 1993 legislation that bans the use of TBT paints on small boats (<25 m). The absence or scarcity of the species inside harbours, which are the most polluted sites in Portugal, is probably a consequence of extinction due to female sterilisation by TBT.     

Histopathological effects of petroleum hydrocarbons and heavy metals on the American oyster (Crassostrea virginica) from Tabasco, Mexico

Oyster tissues may be affected by the concentration of ions in the water (i.e. salinity) and by contaminants such as petroleum hydrocarbons and heavy metals. Oyster populations from three coastal lagoons (Mecoacan, Camen and Machona) in the Mexican state of Tabasco, in the southern Gulf of Mexico, were sampled for pollution studies during June, September and November 1992 and May 1993. No statistically significant relationships were found between the concentration of contaminants in the body tissues of organisms and their shell length, soft tissue weight and particulate matter; however, a significant correlation was found between condition index and salinity (r = −0.72). Generally, the lowest weights, shell lengths and indices of condition were found in Mecoacan. The concentrations of cadmium and zinc were inversely related to salinity (r = −0.52 and r = −0.32, respectively), so a riverine input is suspected. On average, 63% of the individual oysters showed histopathological lesions, which can be related to salinity and to the concentrations of cadmium and the unresolved fraction of hydrocarbons (UCM). The response of each of the tissues analysed was different. The percentage of individuals with damage in the digestive diverticulum increased linearly with UCM (r = 0.71), but in a saturation-response fashion (Y = B₀ − B₀/(1 + X^B1) r = 0.66) with cadmium. The percentage of individuals with damage in the gills increased linearly with weight (r = 0.68), cadmium (r = 0.60) and UCM (r = 0.60). The lesions in the connective tissue decreased linearly (r = −0.82) with salinity, but increased in a saturation-response way (r = 0.83) with cadmium. Finally, the percentage of individuals with lesions in the digestive tube decreased linearly with salinity (r = −0.59). Only the damage to the gills and digestive diverticula were dependent on gonadal maturity, while damage to the connective tissue was dependent on the sex of the individual.     

Seasonal variations of imposex indices and butyltin concentrations in the rock shell Thais clavigera collected from Hong Kong waters

This study investigated butyltin contamination in Hong Kong waters using the rock shell Thais clavigera as a biomonitor. We collected rock shells from nine coastal sites in both winter 2005 and summer 2006 to examine the spatial and seasonal patterns. The relative penis size index (RPSI) and vas deferens sequence index (VDSI) were determined following a standard protocol, while the tissue concentrations of butyltins (i.e., mono-, di- and tri-butyltin) were measured using gas chromatography. The results showed that imposex was particularly severe in T. clavigera collected from Victoria Harbour, Pak Sha Wan and Sai Kung Pier where shipping activities were frequent. Both imposex indices had a positive correlation with tissue concentration of tributyltin in T. clavigera. While VDSI exhibited little seasonal variation, RPSI of winter specimens was significantly higher than that of summer specimens. Thus, sampling season should be standardised when using RPSI as a biomarker.