Paramecium bursaria (EHRENBERG 1831), a freshwater ciliate, typically harbors hundreds of green algal symbionts inside the cell. The aim of present study was the molecular identification of newly analyzed P. bursaria symbionts. The second aspect of the present survey was testing a hypothesis whether endosymbionts prefer the specified syngen of the host, and the specified geographical distribution. Ten strains of endosymbionts isolated from strains of P. bursaria originating from different geographical locations were studied. We analyzed for the first time, both the fragment of plastid genome containing 3'rpl36-5'infA genes and a fragment of a nuclear gene encoding large subunit ribosomal RNA (LSU rDNA). The analysis of the LSU rDNA sequences showed the existence of 3 haplotypes and the haplotype diversity of 0.733, and 8 haplotypes for the 3'rpl36-5'infA gene fragment and haplotype diversity of 0.956. The endosymbionts isolated from P. bursaria strains were identified as Chlorella vulgaris, Ch. variabilis and Micractinium conductrix. There was no correlation between the syngen of P. bursaria and the species of endosymbiont.

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Paramecium bursaria is composed of five syngens that are morphologically indistinguishable but sexually isolated. The aim of the present study was to confirm by molecular methods (analyses of mitochondrial COI) the identification of P. bursaria syngens originating from different geographical locations. Phylograms constructed using both the neighbor-joining and maximum-likelihood methods based on a comparison of 34 sequences of P. bursaria strains and P. multimicronucleatum, P. caudatum and P.calkinsi strains used as outgroups revealed five clusters which correspond to results obtained previously by mating reaction. Our analysis shows the existence of 24 haplotypes for the COI gene sequence in the studied strains. The interspecies haplotype diversity was Hd = 0.967. We confirmed genetic differentiation between strains of P. bursaria and the occurrence of a correlation between geographical distribution and the correspondent syngen.

Paramecium bursaria is an unicellular organism, widely distributed in the freshwater environment, where heavy metals are common contaminants. The ciliates, also including Paramecium bursaria, are a very abundant group in aquatic ecosystems, what makes them effective biological indicators of water pollutants. Paramecium bursaria is the only Paramecium which has evolved a mutualistic relationship with algae and it harbors these endosymbionts in its own cytoplasm. The algae are also very effective bioindicators of some pollutants because of their ability to biosorption and bioaccumulation of heavy metals. The aim of this study was to determine the acute toxicity of two metals’ compounds: nickel chloride (NiCl2) and zinc chloride (ZnCl2) to Paramecium bursaria and its endosymbionts. The ciliates were incubated in solutions with 5x10-8 to 5x10-2g/dm3 of NiCl2 and with 5x10-8 to 5x10-2g/dm3 of ZnCl2, at the temperature of 180C, in the light/dark conditions (12L/12D). Microscopic observations of cell divisions rate, cell shape changes as well as the swimming behavior, were conducted after 24, 48, 72 and 120 hours of incubation in the tested solutions and were compared to the control sample. Microscopic observations revealed the lethal doses for both compounds, for nickel chloride 5x10-5g/dm3 and for zinc chloride 5x10-3. These observations also revealed that in lesser concentrations than the lethal one, the slowdown and characteristic movements occur after metal addition. The PEA measurements of Fv/Fm parameter were carried out within 4 days, the first one after 24 hours of incubations. The results of this investigation has given us a view of a fluorescence efficiency by revealing that both compounds solutions can have the stimulating effect on Photosystem II, because the lowest fluorescence efficiency was measured in control samples.

Keywords: Albumins, brain, acrylamide, oxidative stress