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Imposex in Rock Whelks Thais and Ocenebra Species (Mollusca, Neogastropoda, Muricidae) from Gujarat Coast

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Imposex in Rock Whelks Thais and Ocenebra Species (Mollusca, Neogastropoda, Muricidae) from Gujarat Coast Indian Journal of Marine Sciences Vol. 31(4), December 2002, pp. 321-328 Imposex in rock whelks Thais and Ocenebra species (Mollusca, Neogastropoda, Muricidae) from Gujarat coast A. Tewari, C. Raghunathan, H. V. Joshi & Yasmin Khambhaty Central Salt and Marine Chemicals Research Institute, Bhavnagar-364 002, Gujarat, India Revised 3 October 2001, revised 19 July 2002 Imposex in Thais bufo, T. rudolphi, T. tissoti and Ocenebra bombayana was observed from Saurashtra coast of Gujarat. The maximum percentage of imposex out of total sample size (44.80 and 46.55%) were observed at Porbandar in T. bufo and T. rudolphi respectively while T. tissoti and O. bombayana at Mahuva showed 10.29 and 9.72% of imposex respectively. The percentage conversion of potential females to imposex was minimum at Diu and maximum at Porbandar. The number of ships operated from Porbandar and Diu directly correlated with severity of CPFII and percentage of imposex at these places. The results indicate that severity of imposex in T. rudolphi could be utilized as a bioindicator of TBT contamination in the marine environment. The Relative Penis Size Index(RPSI) ranged from 3.24 at Veraval to 16.50 at Porbandar in T. bufo while in T. rudolphi it ranged from 44.26 at Okha and 55.96 at Veraval. It was 21.19 and 14.26 for T. tissoti and O. bombayana respectively. The results indicate that RPSI may not be a foolproof indicator of severity of imposex in these organisms. Therefore a new index has been developed which is termed as Conversion of Potential Females to Imposex Index (CPFII) to calculate the potential females of a given area of a population. It seems the causative agents in seawater is neither toxic nor growth inhibitory at all the places of study but it interferes with the reproductive mechanisms and morphogenesis in these four species. The imposex in these organisms and its relation to TBT concentration in the marine environment is discussed. [ Key words: Imposex, neogastropods, Thais, tributyltin ] Imposex is the superimposition of male sex organs, prohibited from the year 2003 onwards10. Vessels penis and vas deferens on the female own genital must be free of these coatings10 altogether by 2008. system in gonochoristic gastropods1. It is also called These proposals are part of a stepwise approach by pseudohermaphroditism. In advance cases the imposex IMO, whereby all antifoulants which exhibit may bring out reproductive failure and consequential potentially harmful effects in the marine environment population decline1,2. The reproductive failure leads to will eventually be banned10. sterility in females and in extreme cases, the whole Imposex has been described in 118 species of neo- population vanishes from that area. The imposex is gastropods, belonging to 63 genera11. But, informa- reported to be caused by organotin compounds like tion on imposex in gastropods from Indian coastal tributyltin (TBT), triphenyltin3 etc. which are used as waters is not available. However, the effect of TBT on anti-fouling paints in ships, fishing nets and the toxicity of some bivalves under laboratory condi- mariculture cages4,5. The imposex occurs at levels of tions has been reported12. TBT contamination as low as parts per trillion, Muricid gastropods especially Chicoreus, Thais, however, it can be caused by agents other than TBT Murex etc. are commercially important as regular diet including parasitic trematode larvae6 and other and other products. Since it has got widespread eco- important pollutants such as copper compounds7 and nomic implications, the present study was taken to nonylphenol8. However the effect of copper on examine the severity of imposex on Gujarat coast, imposex has not been proved9. All ships including which is the second largest industrialized state of Indian ships use the antifouling paint containing TBT India. as there is no regulation as on date in India and several other countries to ban the use of TBT in Materials and Methods antifouling paints. International Maritime Organiza- The specimens of rock whelk Thais bufo (Lamarck) tion (IMO) is expected to endorse a recommendation and Thais rudolphi (Lamarck) were collected from from its Maritime Environmental Protection Committee Gujarat coastline--Okha (lat.22° 30′N, long.69° 03′E), that the application of TBT based coatings should be Dwarka (lat.21° 15′N, long. 68°41'E), Porbandar 322 Indian J. Mar. Sci., Vol. 31, No. 4, December 2002 (lat.21° 39′N, long. 69° 38′E), Mocha (lat.21° 20′N, long. 69° 53′E), Veraval (lat.20° 54′N, long. 69° 53′E) and Diu (lat.20° 43′N, long. 70° 47′E). Thais tissoti (Petit) and Ocenebra bombayana (Melvill) were collected from Mahuva (lat.21° 05′N, long. 71° 47′E) (Fig. 1). They were collected from the intertidal region of the rocky coasts during October and November 2000. Specimens of each species were randomly collected during low tide in considerable numbers for imposex studies. Soon after collection, the epiphytic flora and fauna on the surface of the shell were thoroughly removed and washed with clean seawater. The animals were preserved in ice cold Fig. 1 ⎯ Location of sampling sites condition and transported to laboratory where the morphometric measurements such as shell length and of Potential Female to Imposex Index (CPFII). The weight were measured by vernier caliper and analyti- formula for the index is : cal balance respectively. The morphometric meas- Number of imposex animals CPFII = ×100 urements were made with a view to identify the Number of females + Number of growth stage at which imposex occurs. The shell of imposex animals the rock whelks were cracked carefully and the soft The density of four species at all sampling stations tissues were gently removed for the determination of were also estimated from their natural habitat along sex and imposex. Only reproductively matured indi- 2 viduals were studied during the investigation. Sex was the intertidal belt by 0.5 m quadrate having 10 × 10 cm determined primarily by the presence of bright yellow sub quadrates. Data were collected at every 5 m inter- ovaries in the case of females and orange testes for val starting from 0.32 m at Okha and Dwarka, 0.21 m males and both of these are located ventrally in the at Porbandar and Mocha, 0.45 m at Veraval, 0.52 m at coiled section adjacent to the digestive gland on the Diu and Mahuva (taken as baseline) above the zero of posterior of the animal. In addition to that sex was the chart datum extending towards highest high tide also confirmed by the presence of capsule gland, mark. The density in terms of numerical abundance sperm ingesting gland and albumen gland on the ante- and biomass per unit area was determined. The stan- rior to the ovary for females and prostate gland for dard deviation and correlation coefficient were deter- males in the same position. Imposex was identified in mined by Microsoft Excel software. females by the formation of vas deferens and devel- The turbidity was determined by turbidity meter opment of penis behind the right cephalic tentacle. (Model HANNA HI 93703). The number of ships The length and width of the penis of males and im- operating from Okha (near Dwarka), Porbandar (near posex specimens were measured from its tip to junc- Mocha), Mul Dwarka (near Veraval and Diu) and Pi- tion with the wall to 0.1 mm using an eye piece pavav (near Mahuva) during different years were col- graticule mounted on a binocular stereozoom micro- lected from the concerned port authorities. scope. The penises were then excised at their bases and dried to constant weight at 60ºC and the dried Results tissue was weighed to the nearest 10 µg using an ana- The length, weight, percentage occurrence of lytical balance. The intensity of imposex development males, females and imposex and CPFII of T. bufo and in animals were calculated by Relative Penis Size In- T. rudolphi in different stations is presented in Table 1. dex(RPSI) using the following formula4; The maximum percentage of imposex out of total sample size(44.80 and 46.55 %), were observed at Mean length of imposex penis 3 Porbandar in T. bufo and T. rudolphi respectively RPSI = ×100 Mean length of male penis 3 (P < 0.01, r = 1 for both) (Fig. 2). However the mini- mum percentage in these organisms (11.90 and A new index has been developed to calculate the po- 13.76 % respectively) were observed at Diu (P < 0.01, tential females of a given area of population con- r = 1). The CPFII was minimum at Diu and maximum verted to imposex. This index is termed as Conversion at Porbandar. The least CPFII (22.22 and 25.86) in Tewari et al.: Imposex in rock whelks 323 T. bufo (P < 0.05, r = 0.62) and T. rudolphi (P < 0.01, and CPFII in O. bombayana were 9.72 % and 16.07 r = 1) respectively were observed at Diu and highest respectively. The length and weight of O. bombayana (71.80 and 71.05 respectively) were recorded at were least in imposex animals and highest in males Porbandar. The number of ships operated from (P < 0.05, r = 0.62). Porbandar and Diu directly correlated with severity of The mean length, width and dry weight of penis in CPFII and percentage of imposex at these places normal and imposex animal as well as Relative Penis (Tables 1, 2). The weight and length of males, Size Index (RPSI) for T. bufo and T. rudolphi are pre- females and imposex of T. rudolphi were always sented in Table 4 and for T. tissoti and O. bombayana higher than T. bufo at all the stations studied (P < 0.01, in Table 5.
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