Mangrove ecology: application of fatty acid biomarkers as trophic tracers in the ecosystem

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1 Kaiho Super Science High School Mangrove ecology: application of fatty acid biomarkers as trophic tracers in the ecosystem Prosper Mfilinge Advisor: Prof. Makoto Tsuchiya JSPS

2 Self Introduction

3 What are mangroves? Tropical & sub-tropical trees restricted to intertidal and adjacent communities and adapted to live in salt water Tropical: Intertidal: Adjacent:

4 Mangal Mangal community (all animals and plants) that contains mangrove plants

5 Mangrove ecology The study of interrelationships between living organisms and their environment Ecosystem ecological system made up of various communities of organisms interacting with one another and influenced by the environment Interrelationships: Interact with:

6 Ecological significance of mangroves Provide food, shelter, protection and nurseries for many species of fish Sustain high biodiversity Support aquatic and terrestrial food webs e.g. Birds, insects, reptiles which come to feed in mangroves Terrestrial: Source: ISME 1995

7 Ecological significance of mangroves Mangrove assimilated energy and nutrients exported to surrounding coral reefs through export and food chain Assimilate nutrient

8 Mangrove trophic ecology Food chain/ food web (Energy transferred by feeding and nutrient recycled)

9 Examples of mangrove food chains in Okinawa mangroves

10 Example of food pyramid in Okinawa mangroves 3 rd trophic level (Carnivores) 2 nd trophic level (Herbivores) 1 st trophic level (Primary producers) Bacteria

11 In order to study trophic (food chains) relationships we need biomarkers Trophic:

12 What are biomarkers Chemical compound/compounds characteristic of an organism, which can be used to indicate the presence of the organism/plant in the environment and to estimate its biomass e.g. bacteria, diatoms, dinoflagellates, mangrove, algae Estimate

13 Characteristic of a useful biomarker Must be synthesized at low trophic levels e.g. mangrove and remain unchanged when transferred to higher trophic levels 3 rd trophic level (Carnivores) 2 nd trophic level (Herbivores) 1 st trophic level (Primary producers)

14 Why do we use biomarkers? Because it is difficulty to identify types of food in stomachs once ingested Or identify types of organic material in sediments after decay To find links between ecosystems Ingest: decay Mangroves Sea-grasses Coral reefs

15 Examples of biomarkers used in marine ecology Stable isotopes e.g. carbon ( 13 C), nitrogen ( 15 N), sulfur etc Lipids e.g. fatty acids fatty acids carbon: nitrogen Classes of lipids Phospholipids (Polar lipids), Sterols, Triacylglycerols, Free fatty acids, Waxes, Wax esters

16 What are fatty acids? Major components of lipids Essential components of all living cells and cell membrane lipids Source of metabolic energy Plant cell From Animal cell Fatty acids cell membrane Metabolic:

17 General features of a fatty acid structure A long hydrocarbon chain ( 4-32 carbons), is most common A carboxylic acid group COOH Hydrocarbon chain Carboxylic acid group

18 Classification of fatty acids Based on the number of double bonds 0 double bonds: Saturated fatty acids e.g. 16:0 (Palmitic acid) CH 3 (CH 2 ) 14 COOH One double bond: Monounsaturated fatty acids e.g. 16:1ω7 (Palmitoleic acid) CH 3 (CH 2 ) 5 CH=CH(CH 2 ) 7 COOH Palmitic acid Monounsaturated

19 Classification of fatty acids 2 or more double bonds: Polyunsaturated fatty acids e.g. 18:2ω6 (linoleic acid) CH 3 (CH 2 ) 4 CH=CHCH 2 CH=CH(CH 2 ) 7 COOH Branched fatty acids (odd and even number) e.g. 15:0 iso CH 3 CH 2 (CH 2 ) 11 COOH H 3 C 15:anteiso Polyunsaturated CH 3 CH 2 CH(CH 2 ) 10 COOH H 3 C Branched

20 Nomenclature of Fatty acids There are 4 common naming systems 1) Trivial 2) IUPAC 3)Carboxylic reference 4) ω-reference Palmitic acid Hexadecanoic acid 16:0 16:0 Oleic acid 9-Octadecenoic acid 18:1 9 18:1ω9 Linoleic acid 9,12-Octadecenoic acid 18:2 9,12 18:2ω6 Palmitic acid

21 ω & Designation ω designated as X:YωZ Where X= number of carbon atoms Y= number of double bonds Z= Position of double bond from the terminal methyl group e.g. 16:0 and 18:2ω6 CH 3 (CH 2 ) 14 COOH 16:0 CH 3 (CH 2 ) 4 CH=CHCH 2 CH=CH(CH 2 ) 7 COOH 18:2ω6 (ωdesignated) 18:2 9,12 ( designated) double bonds methyl CH 3

22 Why two common designations? The ω -designation is useful because it allows nutritionists and ecologists to track families of fatty acids or link diet with tissue fatty acid composition The -designation is useful to describe the biochemical reactions

23 Fatty acid metabolism Synthesis in plants..where? in chloroplast Synthesis: Chloroplast From

24 Fatty acid metabolism Synthesis in animals Where? Phospholipids of smooth endoplasmic reticullum (ER) Smooth ER endoplasmic reticullum Rough ER From

25 Fatty acid metabolism Fatty acids are synthesized from acetyl-coa First products of biosynthesis 16:0 Subsequent metabolism and modification occur in endoplasmic reticulum From Subsequent: Modification: Endoplasmic: Reticulum

26 Why do we use fatty acids as biomarkers High structural diversity High taxonomic specificity e.g. Terebralia palustris Terebralia sulcata Specificity

27 FA distributions in green and yellow leaves Green leaves % of total FAs 17:0 20:0 22:0 23:0 24:0 26:0 28:0 14:0 18:0 16:1ω7 16:0 18:1ω9 18:2ω6 18:3ω3 Yellow leaves % of total FAs 14:0 17:0 20:0 22:0 23:0 24:0 26:0 28:0 30:0 16:1ω7 18:0 18:1ω9 18:2ω6 18:3ω3 16:0 Even-Long chain FAs

28 FA distributions in decayed leaves and mangrove sediment After 132 days 14:0 16:1ω7 17:0 18:0 18:1ω9/7 18:2ω6 18:3ω3 20:0 20:3ω6 22:0 23:0 22:6ω3 24:0 26:0 28:0 30:0 16:0 Mangrove sediments % of total FAs % of total FAs 14:0 17:0 16:1ω7 18:1ω9/7 18:0 20:0 20:3ω6 22:0 23:0 24:0 26:0 28:0 30:0 16:0 Sediment Even-Long chain FAs

29 FA should be transferred unchanged to high trophic level 20:5 3 18:2 6 18:3 6 18:3 3 26:0 28:0 30:0 32:0 15:0 iso; 15:0 anteiso; 17:0 iso 17:0 anteiso; 18:1 7 Bacteria 18:4 3 22:6 3

30 Diet composition of gastropods High 12 26:0 28:0 30:0 32:0 Mangrove Macroalgae Diatoms 26:0 28:0 30:0 32:0 26:0 28:0 30:0 32:0 18:2 6 18:3 6 18:3 3 20:5 3 18:4 3 22:6 3 15:0 iso 15:0 anteiso 17:0 iso 17:0 anteiso; 18:1 7 % 8 4 Low High 0 6 Oura Okukubi Nagura Komi Urauchi Dinoflagellates Bacteria 18:1ω7 Algae 18:2 6 18:3 6 18:3 3 Diatoms (20:5 3) % Low Oura Okukubi Nagura Komi Urauchi Bacteria 15:0 iso; 15:0 anteiso; 17:0 iso17:0 anteiso; 18:1 7 Dinoflaggelates18:4 3 22:6 3

31 Fatty acid as indicators of sources & fate of organic matter Mfilinge et al., (2005) Estuarine Coastal and Shelf Science, Vol. 63, Mud flat mangrove Sand flat Sand flat

32 Remarks and conclusions Vascular plants (mangrove and terrestrial plants) share common fatty acid markers (even long chain FAs). Therefore it is impossible to distinguish between mangrove and terrestrial plants organic matter inputs. Ongoing research: Find new fatty acid biomarkers that can distinguish between mangrove and terrestrial plants. Vascular plants: Fungi:

33 Acknowledgements Prof. Makoto Tsuchiya For his kind advice throughout my studies in Japan Japan Society for the Promotion of Science (JSPS) For the post-doctoral fellowship THANK YOU

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