On June 3, 2024, PhD student Tina Bonačić participated in the Scientific Gathering in Split, organized by the Croatian Society of Biochemistry and Molecular Biology. During the event, Tina presented the main objectives and the accomplished results of the PSEUDOTOX project.
The 15th EDM took place from May 7 th – 9 th , 2024 in the town of Ohrid (North Macedonia). Tina Bonačić and Jasna Arapov presented their posters on the second day of the conference. The presented results included: 1. molecular data based on three markers: ITS, LSU, and rbcL, accompanied by phylogeny analyses and morphological data of established Pseudo-nitzschia cultures (T. Bonačić); 2. the phytoplankton community and Pseudo-nitzschia genus diversity during a two-year field study (J.
The results of the morphological analyses were presented at the 2nd Croatian Microscopy Symposium in Split (November 24-25, 2023). In this study, the application of STEM (scanning transmission electron microscopy) in phytoplankton taxonomy is presented. STEM combines some of the advantages of SEM and TEM methods for easier observation of ultrafine structural features of phytoplankton frustules. In this study, STEM is used to image and morphologically identify the marine diatom genus Pseudo-nitzschia from the central and southern Adriatic Sea using the Tescan FE-SEM /STEM MIRA3 microscope at nanoscale resolution.
The first results were presented at the 8th European Phycological Congress in Brest, France (August 20-26, 2023). On the third day of the congress, Tina Bonačić and Jasna Arapov presented their posters with the preliminary results. During our stay, we visited the Roscoff Biological Station, one of the oldest marine biology laboratories in the world, founded in 1872.
The first results of a comprehensive study on the diversity of Pseudo-nitzschia, combining morphological and molecular analyses, were published in the journal JMSE. The integrated approach resulted in the confirmation of P. brasiliana, P. hasleana, P. galaxiae and P. linea. The findings of P. brasiliana and P. hasleana represent the first confirmation for the Adriatic Sea, while P. galaxiae and P. linea have not yet been previously reported along the eastern Adriatic coast.
On the 26th and 27th of January 2023 PhD. student Tina Bonačić was attending personalized (individual) workshop in BIOCentre about optimising PCR reaction based on products, analysing gel electoforesis, discusing about electrophotograms and data editing, measuring DNA concentration on NanoDrop sprectrophotometer, and also about design of DNA primers. The workshop included both, lectures and practical work on own samples. Here is a link.
Project objectives and some preliminary results were presented in oral presentation at the scientific conference „Croatian Naturalist 31- contribution of naturalists to the development of science in the Split-Dalmatia Country“ in Split on 25-26th November 2022. Here is a link.
On the 26th of October we participated the presentation and the demonstration of new SEM (FEG-SEM JEL JSM 7610F Plus) and the new TEM (JOEL JEM 1400 Flash) at the University of Split.
On October 14, the doctoral student, Tina Tomašević, gave a lecture to second and third grade students about the ecology and toxicity of species of the genus Pseudo-nitzschia in the coastal waters of the central and southern Adriatic. After the introduction about the HRZZ PSEUDOTOX project, the students had the opportunity, with our help, to determine the species of the genus Pseudo-nitzchia cells photographed on a scanning electron microscope (SEM).
Our Ph.D. students, Mia Pejković and Tina Pavelin, were hosted from 7th to 18th February 2022 in Laboratory of Marine Ecology, Campus Enrico Mattei Urbino by project consultant prof.ssa Antonella Penna. During their stay, they performed molecular analysis on Pseudo-nitzschia spp. cultured strains. The methods used were DNA isolation, quantification, PCR amplification, DNA electrophoresis, and sequence alignment. Also, the Ph.D. students learned more about protocols for culturing phytoplankton species, cell isolation from seawater samples and cell enumeration by light microscopy.
The FE-SEM Mira 3 Tescan microscope has been upgraded with a new HADF R- STEM detector, fully funded by Croatia Science Foundation, project PSEUDOTOX. After the successful training held at IOF, the first photos of cultured Pseudo-nitzschia cells are made :).
The preliminary clonal Pseudo-nitzschia cultures from field samples collected in the Central and Southern Adriatic Sea has been established. The cultures are further used for species identification by molecular methods in our newly equipped molecular laboratory. After adjustment of DNA isolationmethods and PCR conditions, by shifting polymerase chain reactions and polymerases, we successfully amplificated the gene that encodes for the large ribosomal unit (LSU) in Pseudo-nitzschia. The analyses are going on, and optimization of PCR reaction for ITS region is in progress……
PSEUDOTOX project was presented in the IOF-virtual tour within the Festival of Science 2021. Here is a link.
The kick-off meeting took place on March 11th 2021, at the Institute of Oceanography and Fisheries. The working plan and activities for the first year were discussed.
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