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Protistology ■ 69
species richness ofprokaryotes — associates has been registered in the culture Tulamoeba bucina. The research was performed in the Center of Shared Scientific Equipment «Persistence of microorganisms» of ICIS UB RAS and was supported by RFBR (16-44-560316, 14-04-01796).
CHARACTERIZATION OF MONOCLONAL ANTIBODIES FOR CATHEPSIN B AND CA-THEPSIN B-LIKE PROTEINS OF NAEGLERIA FOWLERI
Seong G.S.12, Sohn H.J.1-2, Kang H.K.12, Shin H.J.12
1 - DepartmentofMicrobiology, Ajou University School of Medicine
2 - Department of Biomedical Science, Graduate School of Ajou University, Suwon 443-721, Republic of Korea
Naegleria fowleri causes a fatal primary amoebic meningoencephalitis (PAM) in human and experimental animals. Cathepsin B (NfCPB) and cathepsin B-Like (NfCPBL) gene in N. fowleri, are consists of 1,038bp of DNA (345 amino acid) and 939bp of DNA (313 amino acid), and molecular weights of recombinant proteins are 38.4 and 34 kDa, respectively. In the previous study, rNfCPB and rNfCPB-L refolding protein may play important roles in host tissue invasion, immune evasion and nutrient uptake. In this study, we produced anti-NfCPB and anti-NfCPBL monoclonal antibody (rNfCPB-McAb and rNfCPBL-McAb) using the cell fusion technique and observed the immunological characteristics. Seven hybridoma cells secreting rNfCPB-McAb (2D6, 2D11, 2A7, 2B7, 2E2, 2E9, 2C9) and three hybridoma cells secreting rNfCPB-McAb (1E5, 1C8, 1D9) were produced. Among them, 2C9 (a cell line producing rNfCPB-McAb) and 1C8 (a cell line producing rNfCPBL-McAb) which showed high antibody titre, respectively, were selected. 2C9 monoclonal antibody was reacted with N. fowleri whole lysate or rNfCPB fusion protein by western blotting, as which showed about 28 kDa and 38.4 kDa ofband pattern, respectively. 1C9 monoclonal antibody was reacted with N. fowleri whole lysate or rNfCPBL fusion protein by western blotting, as which showed 24 kDa and 34 kDa ofband pattern. 2C9 and 1C8 monoclonal antibodies were not reacted with another amoebic lysates such as N. gruberi, Acanthamoeba castellanii, A. polyphagia in western blot analysis. In the results of the immunocytochemistry analysis, NfCPB and NfCPB-L protein were mainly detected in cytoplasm and cell membrane, especially pseudopodia, of N. fowleri trophozoites under a confocal microscope.
These results suggested that monoclonal antibodies against rNfCPB and rNfCPBL may be useful for the further immunological study.
BENTHIC PROTISTS (CILIOPHORA, GRO-MIIDA, FORAMINIFERA) IN THE BLACK SEA: THEIR CONTRIBUTION TO THE BOTTOM COMMUNITIES Sergeeva N.G.
Institute of Marine Biological Research of Russian Academy of Sciences, 2, Nakhimov ave, Sevastopol, Russia
Ciliophora, Gromiida and Foraminifera (hard- shell and soft-shell forms) are the significant components of the benthic communities of the Black Sea. These protozoans are of great role in the transfer of bacterial and algal production to the next trophic levels. They themselves are consumers of bottom bacteria, microalgae, pollen terrestrial vegetation and marine filamentous fungi; as predators, they might prey upon unicellular organisms. In addition, the representatives of above listed groups are an important food source for many bottom invertebrates in different habitats. The changes in taxa composition and abundance, and also the proportions of these protozoan representatives in the benthic communities along deep-water oxic/ anoxic interface (75-300m) in the Istanbul Strait's (Bosporus) outlet area of the Black Sea and NW shelf of Crimea Peninsula were studied. In the oxic zone of the Black Sea, benthic Protozoa and Metazoa were studied in area near the Kerch Strait's on NE shelf ofCrimean Peninsula. Our data allowed concluding about tolerance ofmany benthic protozoans to the hypoxic / anoxic conditions and sulfidic pollution of bottom sediments of the Black Sea and of their significant share in the deep-water benthic communities. Benthic Ciliophora, Gromiida and Foraminifera are numerous, specific and diverse in these conditions. The contribution of these protozoans in the meiobenthos communities had high values and extremely prone to variability. The spatial-bathymetrical distribution of these protists along with benthic multicellular organisms had uneven character in the Black Sea.
CHARACTERIZATION OF " CANDIDATUS GORTZIA SHAHRAZADIS", A NOVEL ENDO-SYMBIONT OF PARAMECIUM MULTIMI-CRONUCLEATUM FROM INDIA Serra V.1, Fokin S.I.1'2, Castelli M.1, Basuri C.K.3, Nitla V.M.3, Verni F.1, Sandeep B.V.34, Kalavathi C.3, Petroni G.1
1 - Department of Biology, University of Pisa, Pisa,
