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HRZZ: Appearance and interaction of biologically important organic molecules and micronutrient metals in marine ecosystem under environmental stress (AMBIOMERES)

HRZZ: Appearance and interaction of biologically important organic molecules and micronutrient metals in marine ecosystem under environmental stress (AMBIOMERES)
Projekti Hrvatske zaklade za znanost
Start date
Oct 1st 2014
End date
Jan 31st 2019

Principal investigator

Recently documented oligotrophication of the Adriatic Sea caused by global changes opened questions how this reflects on the quality and quantity of organic matter (OM) produced by the phytoplankton and on the consequent influence on the micronutrient metals availability and cycling. Within the project, a progressive (model and in-situ) approach will be accomplished to reveal crucial alterations in the marine ecosystem. Initial investigations will imply model experiments on the interaction of selected OM (polysaccharides, lipids, polymeric organic matter containing N catalytic atoms) with micronutrient metals (Fe, Co, Cu, Zn and Cd). Additionally, phytoplankton monoculture incubation under various growth conditions (nutrient concentrations and temperature variations) will be included in order to comprehend multiple impacts on the phytoplankton OM production and micronutrients metals distribution. Model experiments will be followed by in-situ investigations at the selected oligotrophic and eutrophic areas of the Adriatic Sea.

This will give holistic informations on the natural marine biochemical cycles and linkage between OM and micronutrient metals. In-situ investigations will be focused on the seasonal variations of the OM and micronutrient appearance and concentrations in combination with physico-chemical parameters (temperature, salinity, pH, O2), and dissolved and particulate organic carbon content, surface-active organic substances and macronutrients (PO4, NO3, NO2, NH4). These investigations will be conducted by intercomparable and multimethodological approaches using various chromatographic to electrochemical techniques. Finally, taking into account all obtained results an important part of the natural marine ecosystem processes will be characterized in the prospective of climate changes.  

Simplified schematic of assumed interactions between micronutrient metals (M=Fe, Cu, Co, Zn, Cd) and organic molecules (lipids, acidic polysaccharides and N-POM) released from phytoplankton in conditions of (A) oligotrophy and (B) eutrophy. Biological processes drive production of OM and speciation of micronutrient metals:

(A) Micronutrient metals (hydrated ionic form, M, or inorganic complexes, MX) interact with released OM forming complexes from which cells easily take micronutrient metal by ligand exchange reactions. This is mechanism through which OM enhance bioavailability of micronutrient metals in deficiency.

(B) In eutrophy phytoplankton produce altered organics that form complexes with micronutrient metals which do not enable micronutrient metal uptake by cell. This is mechanism through which OM is reducing toxicity of micronutrient metals when they are in surplus. There are enough micronutrients in inorganic form which are preferable for the cellular uptake.


6.10.2014 - Kickoff meeting

19.09.2016 workshop held at the Ruđer Bošković Institute. The program of the workshop is given in the enclosed brochure.

Final Workshop of the AMBIOMERES project. Workshop program is in the attached brochure.



  1. Gašparović, B.*, Vrana, I., Frka, S., Marić Pfannkuchen, D., Vlašiček, I., Djakovac, T., Ivančić, I., Smodlaka Tanković, M., Milinković, A., Flanjak, L., Chaux, F., Novak, T., Medić, N., Godrijan, J. 2023. Paradox of relatively more phospholipids in phytoplankton in phosphorus limited sea. Limnol. Oceanogr. 68, 2800-2813.
  2. Godrijan, J., Marić Pfannkuchen, D., Djakovac, T., Frka, S., Gašparović, B.* 2023. Lipids of different phytoplankton groups differ in sensitivity to degradation: implications for carbon export. Global Change Biology. 29, 5075-5086.
  3. Gašparović, B.*, Penezić, A., Lampitt, R.S., Sudasinghe, N., Schaub, T. 2023. Molecular-level evidence of early lipid transformations throughout oceanic depths. Geochim. Cosmochim. Acta 343, 49-63.
  4. Vrana, I.*, Gašparović, B.*, Geček, S., Godrijan, J., Novak, T., Kazazić, S.P., Mlakar, M., Kužat, N., Pfannkuchen, M., Marić Pfannkuchen, D. 2023. Successful acclimation of marine diatoms Chaetoceros curvisetus/pseudocurvisetus to climate change. Limnol. Oceanogr. 68, S158-S173.
  5. Penezić, A., Drozdowska, V., Novak, T., Gašparović, B.* 2022. Distribution and characterization of organic matter within the sea surface microlayer in the Gulf of Gdansk. Oceanologia, 64, 631-650.
  6. Flanjak, L., Vrana, I., Cvitešić Kušan, A., Godrijan, Novak, T., Penezić, A., Gašparović, B.* 2022. The effects of high temperatures and nitrogen availability on the growth and composition of the marine diatom Chaetoceros pseudocurvisetus. J. Exp. Bot. 73, 4250-4265.
  7. Vrana, I., Bakija Alempijević, S., Novosel, N., Ivošević DeNardis, N., Žigon, D, Ogrinc, N., Gašparović, B. 2022. Hyposalinity induces significant polar lipid remodeling in the marine microalga Dunaliella tertiolecta (Chlorophyceae). J. Appl. Phycol. 34, 1457-1470.
  8. Kužat, N., Smodlaka Tanković, M., Baričević, A., Ivančić, I., Vrana, I., Gašparović, B., Marić Pfannkuchen, D., Pfannkuchen, M. 2022. Morpho-physiological adaptations of Leptocylindrus aporus and L. hargravesii to phosphate limitation in the northern Adriatic. Sci. Rep. 12, 2687.
  9. Penezić, A., Gašparović, B.*, Cuculić, V., Strmečki, S., Djakovac, T., Mlakar, M. 2022. Dissolved trace metals and organic matter distribution in the northern Adriatic, an increasingly oligotrophic shallow sea. Water 14, 349.
  10. Strmečki, S., Pereža, L. 2021. Electrochemistry of chitosan amino-glycan and BSA protein mixture under seawater conditions. J.  Electroanal. Chem. 898, 115630.
  11. Hao, Y-Y, Zhu, Z-Y, Fang, F-T., Novak, T., Čanković, M., Hrustić, E., Ljubešić, Z., Li, M., Du, J-Z., Zhang, R-F., Gašparović, B. 2021. Tracing Nutrients and Organic Matter Changes in Eutrophic Wenchang (China) and Oligotrophic Krka (Croatia) Estuaries: A Comparative Study. Front. Mar. Sci. 8, 663601.
  12. Vrana Špoljarić, I., Novak, N., Gašparović, B.*, Kazazić, S.P., Čanković, M., Ljubešić, Z., Hrustić, E., Mlakar, M., Du, J., Zhang, R., Zhu, Z. 2021. Impact of environmental conditions on phospholipid fatty acid composition: Implications from two contrasting estuaries. Aquat. Ecol. 5, 1-20.
  13. Cabanes, J.E.D., Norman, L., Bowie, R.A., Strmečki, S., Hassler, S.C. 2020. Electrochemical evaluation of iron-binding ligands along the Australian GEOTRACES southwestern Pacific section (GP13). Mar. Chem. 219, 103736.
  14. Mucko, M., Padisák, J., Gligora Udovič, M., Pálmai, T., Novak, T., Medić, N., Gašparović, B., Peharec Štefanić, P., Orlić, S., Ljubešić, Z. 2020. Characterization of a lipid-producing thermotolerant marine photosynthetic pico-alga in the genus Picochlorum (Trebouxiophyceae), Eur. J. Phycol. 55, 384-399.
  15. Stolle, C., Ribas-Ribas, M., Badewien, T.H., Barnes, J., Carpenter, L.J., Chance, R., Riis Damgaard, L., Durán Quesada, A.M., Engel, A., Frka, S., Galgani, L., Gašparović, B., Gerriets, M., Hamizah Mustaffa, N.I., Herrmann, H., Kallajoki, L., Pereira, R., Radach, F., Revsbech, N.P., Rickard, P., Saint, A., Salter, M., Striebel, M., Triesch, N., Uher, G., Upstill-Goddard, R.C., van Pinxteren, M., Zäncker, B., Zieger, P., Wurl, O. 2019. The MILAN campaign: Studying diel light effects on the air-sea interface. B. Am. Meteorol. Soc. 101, 146-166.
  16. Novak, T., Godrijan, J., Marić Pfannkuchen, D., Djakovac, T., Medić, N., Ivančić, I., Mlakar, M., Gašparović, B. 2019. Global warming and oligotrophication lead to increased lipid production in marine phytoplankton. Sci. Total Environ. 668, 171–183.
  17. Rickard, P.C., Uhera, G., Upstill-Goddarda, R.C., Frka, S., Hamizah Mustaffa, N.I., Banko-Kubis, H.M., Cvitešić Kušan, A., Gašparović, B., Stolle, C., Wurl, O., Ribas-Ribas, M. 2019. Reconsideration of seawater surfactant activity analysis based on an inter-laboratory comparison study. Marine Chemistry, 208, 103-111.
  18. Gašparović, B.*, Penezić, A., Lampitt, R.S., Sudasinghe, N., Schaub, T. 2018. Phospholipids as a component of the oceanic phosphorus cycle. Mar. Chem. 250, 70-80.
  19. Smodlaka Tanković, M., Baričević, A., Ivančić I., Kužat, N., Medić, N., Pustijanac, E., Novak, T., Gašparović, B., Marić, D. 2018. Insights into the life strategy of the common marine diatom Chaetoceros peruvianus Brightwell, PLoS ONE, 13(9): e0203634.
  20. Strmečki, S., Paleček, E. 2018. Adsorption/desorption of biomacromolecules involved in catalytic hydrogen evolution. Bioelectrochem. 120, 87-93.
  21. Gašparović, B.*, Penezić, A., Frka, S., Kazazić, S., Lampitt, R.S., Holguin, F.O., Sudasinghe, N., Schaub, T. 2018. Particulate sulfur-containing lipids: production and cycling from the epipelagic to the abyssopelagic zones. Deep-Sea Res. I, 134, 12–22.
  22. Novak, T., Godrijan, G., Marić Pfannkuchen, D., Djakovac, T., Mlakar, M., Baricevic, A., Smodlaka Tanković, M., Gašparović, B.* 2018. Enhanced dissolved lipid production as a response to the sea surface warming. J. Mar. Sys. 180, 289–298.
  23. Mlakar, M., Cuculić, V., Frka, S., Gašparović, B. 2018. Copper - phospholipid interaction at cell membrane model hydrophobic surfaces. Bioelectrochem. 120, 10–17.
  24. Gašparović, B.*, Penezić, A., Lampitt, R.S., Sudasinghe, N., Schaub, T. 2017. Depth-related cycling of suspended nitrogen-containing lipids in the northeast Atlantic. Org. Geochem. 113, 55-66.
  25. Mlakar, M., Pavković, S., Tumir, L-M., Vukosav, P., Gašparović, B. 2017. Electrochemical characterization of biochemically active Cu(II) mixed ligand complex with histidine and cysteine. Electroanalysis 29, 392-397.
  26. Gašparović, B., Kazazić, S.P., Cvitešić, A., Penezić, A., Frka, S. 2017. Corrigendum to “Improved separation and analysis of glycolipids by Iatroscan thin-layer chromatography–flame ionization detection” [J. Chromatogr. A 1409 (2015) 259–267]. J. Chromtogr. A 1521, 168–169.
  27. Gašparović, B.*, Penezić, A., Lampitt, R.S., Sudasinghe, N., Schaub, T. 2016. Free fatty acids, tri-, di- and monoacylglycerol production and depth-related cycling in the Northeast Atlantic. Mar. Chem. 186, 101–109.
  28. Vukosav, P., Mlakara, M., Selmani, A., Bregović, N., Gašparović, B. 2016. Electroanalytical characterization of Zn (II) complexes with D-mannosamine and Glycine in aqueous solutions. Electrochim. Acta 188, 671–678.
  29. Plavšić, M., Strmečki, S. 2016. Carbohydrate polymers as constituents of exopolymer substances in seawater, their complexing properties towards copper ions, surface and catalytic activity determined by electrochemical methods. Carbohydrate polymers, 135 48-56
  30. Gašparović, B.*, Kazazić, S.P., Cvitešić, A., Penezić, A., Frka, S. 2015. Improved separation and analysis of glycolipids by Iatroscan thin–layer chromatography–flame ionization detection. J. Chromtogr. A 1409, 259–267.
  31. Mlakar, M., Fiket, Ž., Geček, S., Cukrov, N., Cuculić, V. 2015. Marine lake as in situ laboratory for studies of organic matter influence on speciation and distribution of trace metals. Cont. Shelf Rec. 103, 1-11.


Tihana Novak, Snježana Kazazić, Jelena Godrijan, Daniela Marić, Tamara Djakovac, Marina Mlakar, Blaženka Gašparović, Effects of temperature on lipid synthesis of diatom Chaetoceros pseudocurvisetusand the northern Adriatic plankton community. Lipids in the ocean,structure, function, ecological role and applications, Brest, Francuska, 20-22.11.2018.

Blaženka Gašparović, Tihana Novak, Jelena Godrijan, Marina Mlakar, Daniela Marić, Tamara Djakovac, „Dissolved lipid production in the Northern Adriatic (Mediterranean) in response to sea surface warming“. „European Geosciences Union General Assembly 2017“, April 23–28, 2017, Vienna, Austria (

Tihana Novak, Blaženka Gašparović, Jelena Godrijan, Daniela Marić, Tamara Djakovac i Marina Mlakar,"Effects of temperature on growth and lipid synthesis of diatom Chaetoceros curvisetus and the Northern Adriatic plankton community“. „European Geosciences Union General Assembly 2017“, April 23–28, 2017, Vienna, Austria. (

Blaženka Gašparović, Tihana Sesar, Milan Čanković, Zrinka Ljubešić, Enis Hrustić, Zhouyi Zhu, Ruifeng Zhang, Jinzhou Du: How Phytoplankton Membranes Cope With Steep Ionic Strength (Salinity) Gradient? Ocean Sciences Meeting 2016, New Orleans, USA, 21-26 02. 2016.

Marina Mlakar, Željka Fiket, Vlado Cuculić, Neven Cukrov, Sunčana Geček: Speciation and Distribution of Trace Metals and Organic Matter in Marine Lake as In Situ Laboratory. Ocean Sciences Meeting 2016, New Orleans, USA, 21-26 02. 2016.

Tihana Sesar: Phytoplankton cellular lipid response on replete and deplete phosphorous conditions, COST Action ES1408: 1st EUALGAE workshop of algae bioproducts for early stage researchers, Valladolid, Spain, 04. 04. 2016.

Marta Plavšić, Slađana Strmečki: The characterization of polysaccharides in seawater accessed by electrochemical methods. Book of abstracts ( ISSBN 978-84-608-9990-7), 04-07. 09. 2016, Interfaces Against Pollution Conference ( IAP 2016), Llieda, Spain.



Blaženka Gašparović, Abra Penezić, Richard S. Lampitt, Nilusha Sudasinghe, Tanner Schaub: Phospholipid Characterization as a Component of the Oceanic Phosphorus and Carbon Cycle. 2018 Ocean science meeting, Portland, Oregon, USA,

Marina Mlakar, Vlado Cuculić, Sanja Frka Milosavljević, Blaženka Gašparović, Sanja Frka:Copper - Phospholipid Interaction at Cell Membrane Model Hydrophobic Surfaces. 2018 Ocean science meeting, Portland, Oregon, USA,

Marina Mlakar, Biogeochemistry and speciation of trace elements in karst marine lakes. Zurich, Switzerland, 16-20. 07.2017.

Slađana Strmečki Kos, Emil Paleček, Tensammetry of proteins and polysaccharides on mercury electrode under conditions of catalytic activity, 6th RSE-SEE Conference 2017, Balatonkenese, Hungary, 11-15.06.2017.

Daniela Marić Pfannkuchen, Ingrid Ivančić, Mirta Smodlaka Tanković, Ana Baričević, Tihana Sesar, Blaženka Gašparović, Martin Pfannkuchen, Life strategies of harmful planktonic algae (with respect to phosphate limitation) in marine environments, Book of abstracts, 2016, ICHA, Florianopolis, Brasil, 28-28.

Daniela Marić Pfannkuchen, Ana Baričević; Mirta Smodlaka Tanković, Ingrid Ivančić, Nataša Kužat, Emina Pustijanac, Grozdan Kušpilić, Tihana Novak, Blaženka Gašparović, Martin Pfannkuchen, Exploring the structure of a planktonic ecosystem. Book of abstracts, 2017, International Phycological Society, Szczecin, Poland, 126-126.

Nataša Kužat, Mirta Smodlaka Tanković, Ana Baričević, Ingrid Ivančić, Emina Pustijanac, Kušpilić Grozdan, Tihana Novak, Blaženka Gašparović, Martin Pfannkuchen, Daniela Marić Pfannkuchen, Interspecific variation in metabolic response to P limitation helps explaining sympatric congeneric species diversity (in the northern Adriatic), Book of abstracts, 2017, International Phycological Society, Szczecin, Poland, 111-111.

Master thesis

2018: Matea Bertanjoli (student PMF, Department of Chemistry): Lipid-composition changes of the diatom Chaetoceros pseudocurvisetus depending on the phosphate availability

2018: Petra Mlacović (student PMF, Department of Biology): Adaptation of selected marine diatoms to limited phosphorus resources by the lipid remodeling

Paper written for The Rector's Award:

2017: Nikola Medić (student PMF, Department of Biology): The abundance and composition of lipids in the southern Adriatic pikoeukariotaPicochlorum sp. in enriched and phosphorus-limiting media.

Field work

As part of the project, we sampled the sea water in the northern Adriatic, on the transit between Rovinj and Po River, at two stations 107 and 108 (Figures D6.1.1-3). The station selection is determined by the station's trophic status. Station 107 is oligotrophic (lack of nutrients), while the station is meso-eutrophic (the nutrient concentration is favorable to too high). We sampled at three depths of 0, 5 and 10 m. We sampled with the Rudjer Bošković Institute's "Villa Velebita" research vessel. Sampling was performed monthly, including dates: November 16, 2016; December 12, 2016; January 27, 2017; February 17, 2017; March 28, 2017; May 3, 2017; June 12, 2017; July 12, 2017; August 2017 and October 4, 2017.

Picture D6.1.1. Sampling stations in the northern Adriatic Sea.


Picture D6.1.2. Sampling on the research vessel „Vila Velebita“.


D6.1.2. Sample filtering on the research vessel „Vila Velebita“.

Within the project, sampling of seawater in the Krka River Estuary was carried out at two stations (Figure D51.1). Two stations, clean estuarine waters and waters under the influence of the unloading port in Sibenik were selected. Three depths were sampled: brackish water 0 m, halocline at about 2 m and saline layer at 20 m. Sampling was carried out in four seasons, in November 2017 and in January, April and July 2018.

Members of the project AMBIOMERES are participating in the following COST Actions:

1. 2016 COST Action CA15121 -Advancing marine conservation in the European and contiguous seas(MarCons)

2. 2015 COST Action ES1408 -The European network for algal-bioproducts(EUALGAE)

3. 2016 COST Action CA15217 - Ocean Governance for Sustainability - challenges, options and the role of science.


Purchased equipment:


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