To document the first dictyostelid report in San Fernando City, La Union, Philippines, soils from upland and coastal areas were randomly collected and processed in the laboratory using Hay Infusion Agar. After several weeks of incubation, the spore touch technique was employed to isolate the six determinable morphospecies of dictyostelids from 38% of positive plates: Heterostelium pallidum, Dictyostelium discoideum, Dictyostelium purpureum, Dictyostelium rosarium, Dictyostelium sp.1 (the most abundant), and Raperostelium minutum. This is the first attempt to isolate dictyostelids using the coastal soils in the tropical part of the world.

To quickly increase the cells number of the cultivated bacterivorous protist Nuclearia thermophila and minimize the diversity of bacteria in culture, for the convenience of bioinformatic processing of the sequencing results, the cultivation of N. thermophila was switched to feeding on monoculture of the bacteria Escherichia coli.Experimental selection of the medium composition showed that the PJ+WG medium with the addition of 2.5% of the LB medium ensures the propagation of E. coli and is suitable for rapid reproduction of many N. thermophila cells.A cell density of 200 cells/mm 2 was reached by the end of the first week and was about 600 cells/mm 2 by the end of the third week, with stabilization at the end of the first month at the level of ca.700 cells/ mm 2 .The approximate duration of the cell cycle in the first week was about 24 hours, the rate of increase in the number of cells was 1.9 times a day.This enhancement is convenient for transcriptomic study of differential gene expression in the cell cycle.It is highly likely that in such cultivation conditions, many of the early culture cells will be at different stages of cell development.

a remarkable Russian protistologist and geneticist, passed away in Moscow after serious illness at the age of 71. We lost a wonderful scientist -an enthusiastic experimental researcher who was keen on her work and able of analyzing deeply various aspects of protistan biology (Fig.

Among all ciliates of the Paramecium genus described in the literature, not a single entirely marine species has ever been recorded.Several paramecia belonging to the Cypriostomum and the Helianter subgenera could be regarded as euryhaline species, but these are, in fact, mainly brackish-water inhabitants that, in rare cases, could be found in fresh water bodies as well.More than a quarter of a century ago (in 1997) at Tbingen University, Germany, a culture of the paramecia perfectly adapted to marine salinity (33) was found by the author.Morphological investigation showed that the ciliate was a new, probably, initially marine species.This suggestion was further supported by the acclimation experiment results.Unfortunately, the molecular investigation aimed at clarifying the position of the newly discovered Paramecium sp.among other congeners was not successful, and after several years (in 2003) the culture was lost.The origin of the ciliate is uncertain as it was transferred as a teaching material between two institutions in Europe after it had been received from an unknown sender and place in the USA.Nevertheless, this case implies that the genus Paramecium incudes at least one strictly marine species describe here as P. marinum sp.nov.

The search for new therapeutics and strategies to suppress microsporidiosis of domesticated insects requires cultivation of honeybee and silkworm parasites in available insect cell cultures as well as reliable methods of their detection. In this study, we infected the commonly used lepidopteran Sf9 cell line with the Vairimorpha ceranae and Nosema bombycis spores to evaluate molecular methods for microsporidia growth assay. The silkworm parasite N. bombycis effectively develops in lepidopteran cells and, according to literature data, its growth can be detected by qPCR analysis of -tubulin gene copies in infected Sf9 cultures. Here, we used Western blotting with antibodies against N. bombycis -tubulin to analyze Sf9 cultures infected with the parasite spores and demonstrated the prospects of immunochemical methods to assay its intracellular growth. Analysis of five genes of N. bombycis spore wall and polar tube proteins in infected cultures by reverse transcription (RT) PCR showed that expression of the polar tube protein PTP2 may serve as a specific marker of the parasite growth because only its transcripts were not detected in freshly inoculated Sf9 cells. The honeybee parasite V. ceranae infects Sf9 cells less efficiently. To find a sensitive and specific marker of the growth of this parasite, we analyzed the transcripts of its 13 genes in infected cultures using the same RT-PCR method. The spore wall protein SWP32 gene demonstrated the highest expression at the 4 th -day post infection with V. ceranae spores, alongside with specificity of PCR-amplification, and the absence of transcripts in freshly inoculated cultures. Thus, quantitative PCR analysis of its expression may help to assay the V. ceranae intracellular growth in the Sf9 cell line.

We have sequenced and described the mitochondrial genome of Thecamoeba quadrilineata (Amoebozoa, Discosea, Thecamoebida).This is the first sequenced mitochondrial genome for amoebae of Thecamoebida lineage.The circular mitochondrial DNA of this species has 50942 bp in length and contains 23 proteincoding genes, 2 ribosomal RNAs, 18 transfer RNAs, and 26 open reading frames.In contrast with the shorter mitochondrial genomes of vannellid amoebae, it shows no evidence of RNA editing.This finding supports the hypothesis on the multiple origins of editing in different phylogenetic lineages of Amoebozoa.