taxonID	type	description	language	source
03F7B7434E5C8A0875D6FD7F67A266AA.taxon	description	Data based on Chatton – Lwoff silver impregnated cells. Macro- and micronuclei were measured from Feulgen-stained ciliates. Measurements in micrometers x arithmetic mean, SD standard deviation, Min minimum, Max maximum, CV coefficient of variation in percentage, n number of cells investigated 2.9 μm on average) was situated close to the cell’ s equator (Table 2, Figs. 1, 9, 10). A single ellipsoidal micronucleus (MI) had sizes of 2.5 – 3.2 × 5.5 – 6.0 μm (2.9 ± 0.9 × 5.8 ± 1.8 μm on average) and was usually located close to the MA or in its depression (Table 2, Figs. 1, 9, 10, 12). This single MI is of the compact morphological type, but the chromatin of the nucleus always looked fine and homogenous without any achromatin cape (Figs. 9, 10). The general cell form was intermediate, between the classical cigar shape and those of P. woodroffi, which is posteriorly rounded and to some extent dorsoventrally compressed (Figs. 1 – 5). The width-to-length ratio was 0.28. When undisturbed, the cells were slowly moving or gliding along substrate using typical cilia beating. During the normal swimming procedure, the cells rotated both clockwise and counter-clockwise (Table 2, Figs. 1 – 8, 9 – 16, and 17 – 22). Using artificial treatment of the culture with KCl, acriflavine and Ca-poor conditions (Miyake 1968), conjugation could be induced (Fig. 17). In the prophase of the first meiotic division, the MI does not manifest a clear parachote stage; however, this species apparently has no crescent MI stage as well (Fig. 18). Exchange of pronuclei started before the old MA break down. The old MA became fragmented only after the first synkaryon division. The number of old MA fragments was always quite small (15 – 25) and of diverse dimensions and forms (10 – 25 μm; Figs. 20 – 22). The three consecutive synkaryon divisions resulted in four MA anlagen and four further MI (Figs. 19 – 22), but only one of the generative nuclei survived in progeny. Despite many attempts, we did not succeeded in establishing stable monoclonal cultures, but the ciliate could be maintained as a multi-clonal population on rice grain medium. Other food sources such as lettuce medium inoculated with E. aerogenes or medium with the freshwater micro-alga Chlorogoniom sp. never served as a proper medium for P. boetschlii sp. nov. Unfortunately, the stock culture died off after 2 years of maintenance under laboratory conditions.	en	Krenek, Sascha, Berendonk, Thomas U., Fokin, Sergei I. (2015): New Paramecium (Ciliophora, Oligohymenophorea) congeners shape our view on its biodiversity. Organisms Diversity & Evolution (New York, N. Y.) 15 (2): 215-233, DOI: 10.1007/s13127-015-0207-9, URL: http://dx.doi.org/10.1007/s13127-015-0207-9
03F7B7434E5D8A0A766EF92D60CE6488.taxon	description	Characteristics Statistics B Eocandidatos P. germanicum ^ B Eocandidatos P. brazilianum ^ B Eocandidatos P. hungarianum ^ Body, length x 190.45 SD 13.4 Min 185.0 Max 210.0 CV 7.0 n 20 Body, width x 60.65 SD 4.0 Min 49.0 Max 69 CV 6.6 n 20 Buccal cavity, length x 37.8 SD 1.5 Min 34.0 Max 40.0 CV 4.0 n 20 Macronucleus, length x 55.45 SD 5.4 Min 45 Max 68 CV 9.8 n 20 Macronucleus, width x 42.6 SD 4.9 Min 35 Max 50 CV 11.4 n 20 Somatic ciliary rows, number x 86.8 SD 4.6 Min 80 Max 95 CV 5.3 n 16 Excretory pores, number x 1 SD 0 Min 1 Max 1 CV 0 n 16 Micronucleus, number x 2.7 SD 0.4 Min 2 Max 4 CV 21.9 n 15 Micronucleus, size x 1.8 202.1 84.9 12.6 6.6 182.5 72 225.0 98 6.2 7.8 25 20 57.6 37.1 5.2 3.4 45.1 30 64.8 43 9.0 9.2 25 20 39.6 18.5 3.5 1.4 35.0 16 42.0 21 9.3 7.3 20 15 48.1 23.0 a 23.4 2.0 2.5 a 3.5 4512 a 15 5225 a 28 3.6 12.6 a 15.0 2050 a 15 30.2 17.2 2.7 1.1 24 14 35 18 9.1 6.1 20 15 93.0 56.9 5.5 4.8 80 44 100 65 5.9 8.4 15 20 1.1 1 0.3 0 1 1 2 1 27.3 0 20 20 1.48 3 0.5 0.7 1 2 2 4 33.8 23.3 20 20 3.4 1.7 Data based on Chatton – Lwoff silver impregnated cells. Macro- and micronuclei were measured from Feulgen-stained ciliates. Measurements in micrometers x arithmetic mean, SD standard deviation, Min minimum, Max maximum, CV coefficient of variation in percentage, n number of cells investigated a Morphometry of the macronuclear fragments was 80 – 95 (87.0 ± 4.6 on average). The BC size was 37.8 ± 1.5 μm (ratio of BC size to the cell length, 0.20). Location of the BO was close to the cell’ s equator (Fig. 24). Two contractile vacuoles had always 1 PCV each and a number of collecting canals, 7 – 10 (most often 9). The cytoproct was situated in the middle between BO and the posterior end of the ciliate. The cortex contained numerous typical trichocysts. The ellipsoidal MA (42.6 ± 4.9 × 55.45 ± 5.4 μm on average) was situated close to the cell’ s equator. A number of rounded MI (2 – 4) with sizes of 1.6 – 2.0 μm (1.8 ± 0.2 μm on average) were usually located around and close to the MA (Table 3, Fig. 25) and belong to the compact morphological type (Figs. 25, 26). The general cell form had the classical cigar shape; however, cultured cells usually were rather fat, with a width-to-length ratio of 0.39 (Fig. 23). During the normal swimming procedure, the ciliate always rotated counter-clockwise (left spiral swimmer) (Table 3, Figs. 23 – 26).	en	Krenek, Sascha, Berendonk, Thomas U., Fokin, Sergei I. (2015): New Paramecium (Ciliophora, Oligohymenophorea) congeners shape our view on its biodiversity. Organisms Diversity & Evolution (New York, N. Y.) 15 (2): 215-233, DOI: 10.1007/s13127-015-0207-9, URL: http://dx.doi.org/10.1007/s13127-015-0207-9
03F7B7434E508A0775D6FE8866F266CC.taxon	description	Multivariate morphometric analysis Comparisons of mean values of morphometric data and some morphobiological features are represented as a dendrogram and topogram according to UPGMA and MDS methods (Fig. 35 a, b). For these analyses, the same set of characters obtained for other species previously investigated (12 morphospecies of the Parameciom genus) were used together with the data derived from the herein described new Parameciom species. The addition of the four new representatives does not change the previous tree topology much and fits with major morphological characters of the herein described paramecia (Fig. 35 a). P. boetschlii sp. nov. clusters within the woodroffi subgroup (subgenus Cypriostomom) but is separated by a quite large Euclidian distance from the rest of the species. Here, P. woodroffi was the nearest congener to P. boetschlii sp. nov. The same topology was indicated by the MDS analysis (Fig. 35 b). According to their morphometric and morphobiological features, the other three investigated Parameciom species, namely B Eocandidatos Paramecium brazilianum ^, B Eocandidatos Paramecium germanicum ^ and B Eocandidatos Paramecium hungarianum ^, can be associated to Parameciom moltimicronocleatom, Parameciom caodatom, and Parameciom polycaryom, respectively (Fig. 35 a, b). While these three Parameciom species show also rather high Euclidian distances to their nearest relatives, they cannot be clearly separated from congeners based on general morphology. For example, while B Eocandidatos P. germanicum ^ is closer related to P. caodatom and P. B aorelia ^ according to MDS and UPGMA analyses, it rather resembles P. moltimicronocleatom based on its general morphology. However, it could only be distinguished from P. moltimicronocleatom by the morphological type of MI (compact vs vesicolar, respectively), which is not an easy character to perceive because of its small size and the requirement of high quality staining techniques. Sequence comparison and molecular phylogeny of new Parameciom spp. Comparative sequence analysis and phylogenetic reconstructions based on 18 S-rRNA and mitochondrial COI gene sequences of the herein described new Parameciom spp. gave us trees with different topology (Figs. 36, 37). According to the 18 S-rDNA analyses, P. boetschlii sp. nov. represents a sister taxon to Parameciom potrinom with a sequence similarity of 95.6 % (Fig. 36, Table S 1). The average genetic distance to P. doboscqoi, another member of the Helianter subgenus like P. potrinom, is comparably low (0.048 vs 0.051, cf. Table S 1) indicating that P. boetschlii sp. nov. belongs to the Helianter subgenus. However, as shown in Fig. 36 the Helianter subgenus does not form a monophyletic clade within our analyses. While the relationship of P. boetschlii sp. nov. to P. potrinom is fairly supported by the tree reconstruction methods used, the two more basal branch nodes are not supporter by either method. The other new cryptic species, namely B Eocandidatos P. brazilianum ^ and B Eocandidatos P. germanicum ^ belong to the Parameciom subgenus, while BEocandidatos P. hungarianum ^ appears to be a member of the Cypriostomom subgenus (Fig. 36). Here, B Eocandidatos P. germanicum ^ is inferred to be a sister species to P. caodatom with a sequence similarity of 97.8 %. B Eocandidatos P. brazilianum ^, on the other hand, clusters within one of the two P. moltimicronocleatom clades. However, it should be stressed that both P. moltimicronocleatom clusters exhibit a comparatively large averaged genetic distance of 0.021 with an averaged sequence similarity of only 98.0 %. B Eocandidatos P. hungarianum ^, however, seems to be a sister species to P. woodroffi and P. nephridiatom with a comparatively high sequence similarity of 99.5 % each. Using the mitochondrial COI gene as a molecular marker, the position of P. boetschlii sp. nov. is the most basal in the Parameciom genus with representatives of P. potrinom and P. doboscqoi as closest relatives (Fig. 37). Interestingly, the inclusion of P. boetschlii sp. nov. and the other new cryptic Parameciom species into the COI dataset dramatically changed the relationships between the different Parameciom subgenera, resulting in Chloroparameciom appearing more closely related to Parameciom than to Helianter (data not shown, but cf. Fig. 4 in Boscaro et al. 2012). According to the COI tree topology, B Eocandidatos P. germanicum ^ is placed at a very basal position in the Parameciom subgenus (subgroup B caodatom ^) with three P. moltimicronocleatom strains as apparently closest relatives (averaged genetic analyses, respectively, while asterisks denote full support (1.00 / 100) from both tree reconstruction methods. The scale bar corresponds to 0.05 nucleotide substitutions per site. Sqoared brackets to the right of the tree designate the five different subgenera Chloroparameciom, Cypriostomom, Helianter, Parameciom, and Viridoparameciom as proposed previously (Fokin et al. 2004; Kreutz et al. 2012) distance 0.2), which cluster outside of all other so far haplotyped P. moltimicronocleatom strains (see Fig. 37). The position of B Eocandidatos P. hungarianum ^, based on its COI gene sequence, is similar to its 18 S-rDNA phylogenetic position within the Cypriostomom subgenus with a close relationship to P. woodroffi (strain BB- 5, sequence similarity 89.2 %). However, as shown in Fig. 37, P. calkinsi does not form a monophyletic clade and the closest relative to B Eocandidatos P. hungarianum ^ is actually P. calkinsi strain BOB 130 - 7 from Olkhon Island in Lake Baikal (Przybos et al. 2013) with a sequence similarity of 94.8 %.	en	Krenek, Sascha, Berendonk, Thomas U., Fokin, Sergei I. (2015): New Paramecium (Ciliophora, Oligohymenophorea) congeners shape our view on its biodiversity. Organisms Diversity & Evolution (New York, N. Y.) 15 (2): 215-233, DOI: 10.1007/s13127-015-0207-9, URL: http://dx.doi.org/10.1007/s13127-015-0207-9
