Bioaccumulation of lanthanum in marbled crayfish (Procambarus sp.) Master Thesis Eyerusalem Goitom Wageningen, August 2011 Bioaccumulation of lanthanum in marbled crayfish (Procambarus sp.) Eyerusalem Goitom Wageningen, July 2011 M.Sc. Thesis Aquatic Ecology and Water Quality Management Group Report no. 007/2011 WAGENINGEN UNIVERSITY Supervisors: Dr.Ir. Miquel Lurling Drs. Frank Van Oosterhout Dr. Ir. Ivo Roessink DATA FOR INTERNAL USE ONLY EXTERNAL USE OF DATA IS ONLY ALLOWED WITH PERMISSION OF THE PROJECT LEADER Table of content Abstract ............................................................................................................................ i 1. Introduction .................................................................................................................. 1 1.1. Eutrophication ................................................................................................................... 1 1.2. The Cyano team research on Phoslock® .............................................................................. 2 1.3. Characteristics of Phoslock® ............................................................................................... 2 1.4. Occurrence and Distribution of Rare Earth Elements ........................................................... 3 1.5. Chemical Properties of Lanthanides ................................................................................... 3 1.6. Physiochemical property of Lanthanum .............................................................................. 3 1.7.Toxicity of Lanthanum ........................................................................................................ 4 1.8. Effect of Rare Earth Elements (REEs) on Aquatic organism .................................................. 4 1.9. Mechanism of Metal Uptake by Aquatic Invertebrate ....................................................... 5 1.9.1. Trace metals ......................................................................................................................................... 5 1.9.2. Heavy metals ........................................................................................................................................ 6 2. Research objectives, questions and hypothesis .............................................................. 7 2.1 Objectives .......................................................................................................................... 7 2.2. Research questions ............................................................................................................ 7 2.3. Hypothesis ........................................................................................................................ 7 3. Materials and methods ................................................................................................. 8 3.1. Test organism .................................................................................................................... 8 3.2. Crayfish anatomy ............................................................................................................... 8 3.2. Experimental settings ........................................................................................................ 9 3.3. Principle of the experiment .............................................................................................. 10 3.3. Bioaccumulation Procedure ............................................................................................. 10 3.2.1. Preparation ....................................................................................................................................... 10 3.2.2. Stratification and Randomisation Procedure ..................................................................................... 11 3.2.3. Exclusion Criteria................................................................................................................................ 11 3.2.4. Phoslock® dosing and replenishing .................................................................................................... 11 3.2.5. Water quality variables ..................................................................................................................... 12 3.2.6. Sampling of experimental unit .......................................................................................................... 12 3.3. Analytical methods .......................................................................................................... 13 3.4. Data analysis ................................................................................................................... 15 4. Results ........................................................................................................................ 17 4.1. Water quality variables .................................................................................................... 17 4.2. Lanthanum in water and crayfish tissue samples .............................................................. 18 4.2.1. Lanthanum in water samples ............................................................................................................. 18 4.2.2. Lanthanum in Crayfish tissue samples ............................................................................................... 20 5. Discussion ................................................................................................................... 22 5.1. Water quality variables .................................................................................................... 22 5.2. Lanthanum in crayfish tissues .......................................................................................... 22 6. Conclusion and Recommendation ............................................................................... 24 6.1. Conclusions ..................................................................................................................... 24 6.2. Recommendations ........................................................................................................... 24 Acknowledgement .......................................................................................................... 25 References...................................................................................................................... 26 8. Appendices ................................................................................................................. 32 Appendix 1. Descriptive and statistical test analysis for Water quality variables ....................... 32 Appendix 2. Total body weight and total body length of the crayfish at start and end of the experiment ............................................................................................................................ 37 Appendix 3. Normality, descriptive and statistical test results for lanthanum in tissue samples for control and Phoslock® groups ............................................................................................ 38 Appendix 4. Multiple comparison of Crayfish tissues ............................................................... 43 Table 11. Descriptive and Wilcoxon Signed Ranks test results for control and Phoslock® groups at day 0, after 14 and 28 days of exposure .............................................................................. 43 Appendix 6. Laboratory Protocol ............................................................................................ 47 Lists of figures Figure 1: External and internal anatomy of the crayfish .....................................................................................................9 Figure 2: Experimental units in water bath compartment with aeration tubes ....................................................................10 Figure 3: Turbidity after Phoslock® application ..................................................................................................................11 Figure 4: Principle of the experiment and sampling scheme of the test organism...............................................................12 Figure 5: Dissection of crayfish...........................................................................................................................................13 Figure 6: Tissue samples in a dry Freezer............................................................................................................................14 Figure 7: Crushed tissues for micro destruction ...............................................................................................................14 Figure 8: Micro destruction block containing crayfish tissue samples................................................................................15 Figure 9: Average turbidity value in controlled and Phoslock® groups during experiment..................................................18 Figure 10: Average ammonia concentrations in controlled and Phoslock® groups...............................................................18 Figure 11: Average filterable lanthanum concentrations (µg l-1) in CFW, Controlled and Phoslock® experimental units during the exposure period..................................................................................................................19 Figure 12: Average filterable lanthnaum concentrations after 4 days gut cleaning period ....................................................19 Figure 13: Average