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Acartia Tonsa Dana and Acute Testing with Copper on Different Life Cycle Stages

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Acartia Tonsa Dana and Acute Testing with Copper on Different Life Cycle Stages Faculty of Bioscience Engineering Academic Year 2014 – 2015 Culturing Acartia tonsa Dana and Acute Testing with Copper on Different Life Cycle Stages Jonathan Baetens Promotor: Prof. Dr. Eng. Karel De Schamphelaere Tutor: Msc. Olivier Berteloot Thesis Prepared for the Degree of Master of Science in Biochemical Engineering Technology Faculty of Bioscience Engineering Academic Year 2014 – 2015 Culturing Acartia tonsa Dana and Acute Testing with Copper on Different Life Cycle Stages Jonathan Baetens Promotor: Prof. Dr. Eng. Karel De Schamphelaere Tutor: Msc. Olivier Berteloot Thesis Prepared for the Degree of Master of Science in Biochemical Engineering Technology Copyright The author, the tutor and the promoter give the permission to use this thesis for consultation and to copy parts of it for personal use. Every other use is subject to the copyright laws, more specifically the source must be extensively specified when using the results from this thesis. De auteur en de promotor geven de toelating deze scriptie voor consultatie beschikbaar te stellen en delen van de scriptie te kopiëren voor persoonlijk gebruik. Elk ander gebruik valt onder de beperkingen van het auteursrecht, in het bijzonder met betrekking tot de verplichting de bron uitdrukkelijk te vermelden bij het aanhalen van resultaten uit deze scriptie. Written in Dendermonde and Ghent on 21/08/2015 Geschreven in Dendermonde en Gent op 21/08/2015 Jonathan Baetens Prof. Dr. Eng. Karel De Schamphelaere Msc. Olivier Berteloot Acknowledgements When it was time to choose a thesis subject, I was certain of two things. One, I wanted a subject that had an environmental ‘ring’ to it. Two, I wanted to work in a laboratory. My first choice was a different subject but when it was time to choose, someone else had already taken it. I wanted to work at GhEnToxLab so I asked my guidance counsellor Prof. Dr. Eng. Leen De Gelder to check if there was another topic available and there was, namely “something with copepods” under the guidance of Prof. Dr. Eng. Karel De Schamphelaere and Msc. Olivier Berteloot. I was immediately drawn to the topic and was happy to get started at GhEnToxLab. So it was decided to culture copepods and use them in classic ecotoxicological tests. Acartia tonsa was one of the prime candidates because the species was easily and cheaply procurable. The process of culturing and maintaining the copepods was arduous and a lot of mistakes were made. But because of these mistakes, I learned a lot of new techniques and for that I am grateful. I thank my promotor Prof. Dr. Eng. Karel De Schamphelaere for giving me the opportunity to work at GhEnToxLab. I thank my tutor Msc. Olivier Berteloot for guiding me despite his busy agenda and for sacrificing his vacation to ensure that everything was ready and finished. I would personally like to thank each staff member of GhEnToxLab. I thank Gisèle Bockstael for educating me on algae and helping me whenever I needed it. I would like to thank Nancy De Saeyer and Jolien Depecker for supplying the materials and answering my questions. I would also like to thank Marianne Van den Hove for taking care of the administrative side of things. I thank Emmy Pequeur for performing several analyses that I couldn’t do myself and I would like to thank Marc Van Der Borght for the technical assistance and brainstorming. It was a pleasant and fun team to work with. My only regret is that it was too short Before I knew it, it was over. I went through a difficult time during this thesis research but everyone was always there for me. Therefore, I would sincerely like to thank everyone for the words of comfort and support everyone has given me. I would also like to thank Joey De Bruyn for redrawing and designing several figures and schematics. Written in Dendermonde and Ghent on 21/08/2015 Jonathan Baetens IX Table of Contents 1 General Introduction ............................................................................................1 2 Literature Study.....................................................................................................3 2.1 Ecotoxicology ..........................................................................................................3 2.1.1 Toxicity Tests ...........................................................................................................3 2.1.2 The Dose-Response Curve ...................................................................................4 2.1.3 From Organism to Ecosystem ..............................................................................6 2.2 Copepods as a Test Organism in Ecotoxicology ...............................................6 2.2.1 General Characteristics of Copepods ..................................................................7 2.2.2 Acartia tonsa (Dana).............................................................................................10 2.2.3 Copepods in Aquaculture ....................................................................................12 2.3 Algae for feeding purposes .................................................................................13 2.3.1 Commercial production ........................................................................................14 2.3.2 Tetraselmis suecica ..............................................................................................15 2.3.3 Isochrysis galbana ................................................................................................16 2.4 Toxicity of Heavy Metals ......................................................................................17 2.4.1 Copper ....................................................................................................................19 3 Materials and Methods ......................................................................................20 3.1 Starting and Maintaining an Acartia tonsa Culture ..........................................20 3.1.1 Principle ..................................................................................................................20 -1 -1 3.1.2 Preparation ASW 30.0 g L ± 1.0 g L ..............................................................20 3.1.3 Preparation Walne Medium .................................................................................21 3.1.4 Algae Culture .........................................................................................................22 3.1.5 Concentrating and Counting Algae ....................................................................24 3.1.6 Acartia tonsa culture .............................................................................................24 3.2 Acute Testing .........................................................................................................25 3.2.1 Principle ..................................................................................................................25 X 3.2.2 Copper Solutions...................................................................................................26 3.2.3 Acute Tests ............................................................................................................27 3.3 Data analysis .........................................................................................................29 4 Results ...................................................................................................................32 4.1 30.0 PSU ................................................................................................................33 4.2 15.0 PSU ................................................................................................................51 4.3 Summary ................................................................................................................62 4.4 Experimental and practical findings ...................................................................65 5 Discussion ............................................................................................................66 5.1 Nauplius, copepodite, and copepod at 30 PSU ...............................................66 5.2 Nauplius, copepodite, and copepod at 15 PSU ...............................................67 5.3 Comparing results of 30 PSU and 15 PSU .......................................................68 5.4 Overcoming practical difficulties .........................................................................68 6 Conclusion ...........................................................................................................72 7 References ............................................................................................................73 1 Appendix A: SOP 001.01 Acartia tonsa Dana Culture ................................1 1.1 Principle ....................................................................................................................2 1.2 Materials ...................................................................................................................3 1.2.1 Algal culture for feeding purposes ........................................................................3 1.2.2 Preparing ASW 30.0 g L-1 ± 1.0 g L-1 ...................................................................1 1.2.3 Filtering ASW 30.0 g L-1 ± 1.0 g L-1 ......................................................................1 1.2.4 Start-up Acartia tonsa culture ...............................................................................1 1.2.5 Maintaining Acartia tonsa culture .........................................................................1
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