Accumulation of Tributyltin (TBT) and Intersex Occurrence as TBT-contaminated indicator on the marine snail  Littoraria sc...
The role of molluscs in control & monitoring programme of marine environmental quality  Toxic Substances Discharge as resi...
INTERSEX: <ul><li>Pathological alteration in the oviduct of littorinids, and </li></ul><ul><li>Replacement of female by ma...
Littorina littorea Littorina scabra species complex <ul><li>Reid (1986) divided and reallocated this  </li></ul><ul><li>sp...
OBJECTIVES 1.  Accumulation of TBT to understand two different pathways of TBT accumulation (via TBT-contaminated water & ...
MATERIALS AND METHODS 1.  Accumulation of TBT <ul><li>Littoraria scabra </li></ul><ul><li>collected at intertidal area </l...
MATERIALS AND METHODS 2.  Intersex Occurrence <ul><li>Measurement on Intersex Stage has been done by calculating  </li></u...
MATERIALS AND METHODS <ul><li>Intersex Occurrence </li></ul>Littoraria scabra <ul><li>Shell length :  17.46 – 20.16 mm </l...
RESULTS Visual Observation on  L. scabra  contaminated to TBT at different pathways yes (before 9 days) yes, very rough pr...
RESULTS Qualitative measurement of TBT residue accumulated in the body (soft part) of  L. scabra (continued) no  :  undete...
Intersex Determination in  Littoraria scabra RESULTS (continued) ISI > 1.0 indicate that at least some of the females in t...
Locations ISI Values Intersex Intensity Comparison Qualitative residue of TBT in body of  L. scabra RESULTS (continued) no...
DISCUSSION Organotin (TBT) Availability <ul><li>Water </li></ul><ul><li>Sediment & food </li></ul>Body system of Littorari...
CONCLUSION <ul><li>TBT compound was accumulated in both pathways (via TBT- </li></ul><ul><li>contaminated water and TBT-co...
ACKNOWLEDGEMENT I am greatly indebted to the DANIDA sponsored Tropical Marine Mollusc Programme (TMMP) for the possibility...
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Accumualtion of TBT and Intersex Occurence on Marine Snails Littoraria Scabra

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Presented in the International Conference & Workshop XI of “Tropical Marine Mollusc Programme”, sponsored by DANIDA at Kodaikal, Rameswaran and Tuticorin-Tamilnadu, India. 28 September – 8 Oktober, 2000.

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Accumualtion of TBT and Intersex Occurence on Marine Snails Littoraria Scabra

  1. 1. Accumulation of Tributyltin (TBT) and Intersex Occurrence as TBT-contaminated indicator on the marine snail Littoraria scabra (Gastropoda: Mollusca) Laboratory of Toxicology and Marine Pharmaceutics Faculty of Fisheries & Marine Sciences Sam Ratulangi University, Manado, North Sulawesi, Indonesia By Markus T. Lasut TBT-contaminated indicator Presented in : International Conference & Workshop XI of “Tropical Marine Mollusc Programme”, sponsored by DANIDA at Kodaikal, Rameswaran and Tuticorin-Tamilnadu, India. 28 September – 8 Oktober, 2000
  2. 2. The role of molluscs in control & monitoring programme of marine environmental quality Toxic Substances Discharge as residues Introduced in purposes <ul><li>MARINE ENVIRONMENT </li></ul><ul><li>Effects: </li></ul><ul><li>Biological, </li></ul><ul><li>Instability, </li></ul><ul><li>Reduce species </li></ul><ul><li>BIVALVIA </li></ul><ul><li>Shell thickening </li></ul><ul><li>GASTROPODS </li></ul><ul><li>Imposex, Intersex </li></ul>CONTROL & MONITORING PROGRAMME Organotin Tributyltin (TBT) <ul><li>Measurement on </li></ul><ul><li>the concentration </li></ul><ul><li>Biomonitor using </li></ul><ul><li>MOLLUSCS </li></ul>Accumulation As antifouling paints INTRODUCTION INTERSEX MOLLUSCS
  3. 3. INTERSEX: <ul><li>Pathological alteration in the oviduct of littorinids, and </li></ul><ul><li>Replacement of female by male organs </li></ul><ul><li>Has been proven (together with imposex) as a highly </li></ul><ul><li>sensitive biomarker for the determination of the degree </li></ul><ul><li>of environmental organotin and especially tributyltin (TBT) </li></ul><ul><li>Atlantic, North Sea, Baltic Sea </li></ul><ul><li>Littorina littorea </li></ul>(continued) INTRODUCTION Littorina littorea
  4. 4. Littorina littorea Littorina scabra species complex <ul><li>Reid (1986) divided and reallocated this </li></ul><ul><li>species into 16 species of the genus Littoraria </li></ul><ul><li>1 species of the genus Littoraria is L. scabra </li></ul>(continued) INTRODUCTION Littoraria scabra
  5. 5. OBJECTIVES 1. Accumulation of TBT to understand two different pathways of TBT accumulation (via TBT-contaminated water & TBT-contaminated sediment and food) 2. Intersex occurrence to measure and compare intersex stage between two different locations Residues of TBT accumulated in the body of the animals were measured qualitatively using Tin-Layer Chromatography technique
  6. 6. MATERIALS AND METHODS 1. Accumulation of TBT <ul><li>Littoraria scabra </li></ul><ul><li>collected at intertidal area </li></ul><ul><li>15 animals per treatment </li></ul><ul><li>Shell length of 17.46 – 20.16 mm </li></ul><ul><li>TBT </li></ul><ul><li>Pure TBT + tween 80 </li></ul>a. Accumulation via TBT-contaminated water TBT : 1, 2 and 3 ppb + control (treatment) Seawater : vol. 250 ml, 22.6 ppt, 28-30 o C, pH 7.50-7.70 b. Accumulation via TBT-contaminated sediment and food <ul><li>TBT : 1, 2 and 3 ppb + control (treatment) </li></ul><ul><li>Seawater : vol. 250 ml, 22.6 ppt, 28-30 o C, pH 7.50-7.70 </li></ul><ul><li>Daily observation was carried out on each treatment along 15 days </li></ul>
  7. 7. MATERIALS AND METHODS 2. Intersex Occurrence <ul><li>Measurement on Intersex Stage has been done by calculating </li></ul><ul><li>Intersex Index (ISI) as the mean value of the Intersex Stage in a </li></ul><ul><li>population </li></ul>ISI > 1.0 indicate that at least some of the females in the sample are sterilized due to intersex development ISI = Intersex Stage values of all females sampled / number of females <ul><li>Two different locations were chosen: </li></ul><ul><li>Amurang Bay : indicated by small scale/traditional harbour </li></ul><ul><li>Bitung Coastal Waters : indicated by large scale harbour </li></ul><ul><li>(this location then is divided by 5 sub-locations (A, B, C, D and E) to see the gradient of TBT contamination) </li></ul>(continued)
  8. 8. MATERIALS AND METHODS <ul><li>Intersex Occurrence </li></ul>Littoraria scabra <ul><li>Shell length : 17.46 – 20.16 mm </li></ul><ul><li>Sampled during low-tide at the area </li></ul><ul><li>Anesthetic in 7% MgCl 2 for maximum relaxation </li></ul><ul><li>Observation under microscope </li></ul>Tin-layer Chromatography technique was applied to confirmed residues of TBT accumulated in the body of animals (continued)
  9. 9. RESULTS Visual Observation on L. scabra contaminated to TBT at different pathways yes (before 9 days) yes, very rough produce mucus Behaviour : yes (before 9days) Water condition: very rough Other : produce mucus 3 ppb TBT yes (before 9 days) yes, rough produce mucus Behaviour : yes (before 9days) Water condition: yes, rough Other : produce mucus 2 ppb TBT no (after 9 days) no -- Behaviour : no (after 9 days) Water condition: no Other : -- 1 ppb TBT no no -- Behaviour : no Water condition: no Other : -- Control TBT-contaminated sediment & food TBT-contaminated water Treatments Pathways
  10. 10. RESULTS Qualitative measurement of TBT residue accumulated in the body (soft part) of L. scabra (continued) no : undetected residues yes : detected residues yes yes no TBT-contaminated sediment & food yes yes no TBT-contaminated water 3 2 1 ways TBT Concentration (ppb) Path-
  11. 11. Intersex Determination in Littoraria scabra RESULTS (continued) ISI > 1.0 indicate that at least some of the females in the sample are sterilized due to intersex development 0.27 9 29 E 0.50 17 28 D 0.93 22 31 C 0.76 13 30 B 0.15 5 26 A Bitung Coastal Water (BCW) 0.08 5 36 Amurang Bay ISI Intersex (%) Number of Samples (Female) Locations
  12. 12. Locations ISI Values Intersex Intensity Comparison Qualitative residue of TBT in body of L. scabra RESULTS (continued) no : undetected residues, yes : detected residues no yes yes yes no no E D C B A Bay Bitung Coastal Waters Amurang
  13. 13. DISCUSSION Organotin (TBT) Availability <ul><li>Water </li></ul><ul><li>Sediment & food </li></ul>Body system of Littoraria scabra Effects Accumulation <ul><li>Uptake </li></ul><ul><li>diffusions </li></ul><ul><li>Sublethal: </li></ul><ul><li>Spend energy </li></ul><ul><li>Intersex </li></ul><ul><li>behavioral </li></ul><ul><li>Lethal: </li></ul><ul><li>mortality </li></ul>Depuration If Accumulation > depuration = Effects Contamination
  14. 14. CONCLUSION <ul><li>TBT compound was accumulated in both pathways (via TBT- </li></ul><ul><li>contaminated water and TBT-contaminated Sediment & food) by </li></ul><ul><li>the marine snail Littoraria scabra , start from 2 ppb of </li></ul><ul><li>experimental concentration </li></ul><ul><li>Bitung Coastal Waters (BCW) is more pollute by TBT than </li></ul><ul><li>Amurang Bay (AB), although it is not in dangerous stage of intersex </li></ul><ul><li>Intersex is found as a sensitive indicator for TBT contamination in </li></ul><ul><li>low concentration </li></ul><ul><li>Intersex occurrence in marine snail Littoraria scabra can be used in </li></ul><ul><li>Control & Monitoring Program for TBT contamination </li></ul>
  15. 15. ACKNOWLEDGEMENT I am greatly indebted to the DANIDA sponsored Tropical Marine Mollusc Programme (TMMP) for the possibility to present this paper at the 11 th TMMP Conference/Workshop at Kodaikanal, Tamilnadu, India, to Dr. Jorgen Hylleberg for his kindly help and advice. I am particularly grateful to the Organizing Committee, Dr. P. Edward and Dr. A. Murugan for their warm welcome and hospitality during the congress. “ Thank you”

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