Recent data on the occurrence of species of the Paramecium aurelia complex in Europe

Among 15 species of the Paramecium aurelia complex known world-wide, 10 have been found in Europe, namely: P. primaurelia, P. biaurelia, P. triaurelia, P. tetraurelia, P. pentaurelia, P. sexaurelia, P. septaurelia, P. novaurelia, P. dodecaurelia, and P. tredecaurelia. Recent data on the frequency of occurrence of the species in Europe are given in the paper.     

Paramecium aurelia species complex in the catchment area of the River Raba mountain course (the Carpathians)

In the water bodies of the Beskid Wyspowy Mts three species of the Paramecium aurelia complex have been found, i.e. P. biaurelia, P. tetraurelia and P. novaurelia. P. tetraurelia, which in Europe is a very rare species, was for the first time recorded in this region of the Carpathians.     

Species of the Paramecium aurelia complex in the middle Sudeten of Czecho-Slovakia.

On the investigated territory three species of the complex were identified, namely Paramecium biaurelia, P. triaurelia, and P. novaurelia. P. novaurelia dominated over the other species with regard to the number of clones established from nature as well as to habitats.     

Distribution of the Paramecium aurelia Species Complex (Protozoa,Ciliophora) in the Carpathian Chain of Poland*

Four species of the Paramecium aurelia complex were found in the Carpathian chain: P. primaurelia, P. biaurelia, P. tetraurelia and P. novaurelia. However, variation in the frequency of settlement of these species in the region was observed; P. biaurelia and P. novaurelia appeared with the greatest frequency, P. Primaurelia occurred less frequently. while P. tetraurelia was rather rare being found only in the Tatras.     

Species of the Paramecium aurelia complex in Russia (western region of European Russia) with molecular characteristics of Paramecium novaurelia

The presence of P. primaurelia, P. biaurelia, P. triaurelia, and P. novaurelia of the P. aurelia complex was revealed in the studied region of Russia. RAPD-PCR fingerprints (band patterns) of newly identified P. novaurelia strains from Russia were compared to those characteristic for the other chosen European strains of the species. The strains revealed intraspecific polymorphism as several groups of genotypes confirming the existence of polymorphism within P. novaurelia.     

Relationships of species of the Paramecium aurelia complex (Protozoa, Ph. Ciliophora, Cl. Oligohymenophorea) based on sequences of the histone H4 gene fragment

A fragment ofhistone H4 gene (160 bp long) was sequenced in the standard strains of P. primaurelia (DQ067620), P. biaurelia (DQ067621), P. tetraurelia (DQ067622), P. pentaurelia (DQ067623), P. septaurelia (DQ067624), P. octaurelia (DQ067625), P. decaurelia (DQ067626), P. undecaurelia (DQ067627), P. dodecaurelia (DQ067628), P. tredecaurelia (DQ067629), and P. quadecaurelia (DQ067630). The tree constructed according to the Kimura model presents two main species clusters, one comprising P. undecaurelia, P. octaurelia, P. septaurelia, and the second cluster with P. pentaurelia, P. tredecaurelia, P. quadecaurelia, P. tetraurelia, P. decaurelia, P. primaurelia, P. biaurelia. P. dodecaurelia was recovered as a separate branch. The tree constructed on the basis of the maximum likelihood method also presents two species clusters, one with P. undecaurelia, P. octaurelia, P. septaurelia, and the second with P. primaurelia, P. decaurelia, P. pentaurelia, P. tredecaurelia, P. quadecaurelia, P. tetraurelia. P. biaurelia forms a basal clade to the latter cluster, and P. dodecaurelia was recovered as a clearly distinct branch from the clusters.     

First European record of Paramecium septaurelia and the discovery of new European habitats of P. pentaurelia and P. sexaurelia in Russia (Astrakhan and Volgograd regions)

The presence of several species of the P. aurelia complex was revealed in the studied regions. In the Volgograd region P. primaurelia, P. biaurelia, P. triaurelia, and P. novaurelia were recorded. In the Astrakhan Nature Reserve P. primaurelia, P. pentaurelia, P. sexaurelia, and P. septaurelia were identified. Among these species, P. septaurelia was recorded for the first time in Europe, known before only from the territory of the USA, P. pentaurelia and P. sexaurelia are species rare in Europe. The studied regions are very rich in species of the P. aurelia complex and worthy of future studies.     

Intraspecies Variability in the Esterases and Acid Phosphatases of Four Species of the Paramecium aurelia Complex1

ABSTRACT. One hundred eighty-eight stocks of Paramecium primaurelia. P. biaurelia, P. tetraurelia. and P. octaurelia were grown axenically and screened for variation in four different esterases and acid phosphatase using starch gel electrophoresis. Major observations: frequency of intraspecies variation for these enzymes is much lower in these four species than in other organisms; hypervariability for two esterases occurs in P. biaurelia both in isolates from worldwide locales and in a restricted locale; clustering of variations occurs in a high proportion of variant stocks in all four species; frequency of intraspecies variation is highest in Central and South America for all four species; and geographical differentiation is lacking between stocks in the same species both for common as well as variant phenotypes despite the cosmopolitan distribution of these species. These results are not correlated with adaptations that favor inbreeding over outbreeding. nor is the possession of bacterial endosymbionts strongly correlated with enzyme variation. When the riequency of intraspecies variation was examined for the aurelia complex of species as a whole for 13 enzymes, mitochondrial DNA, and ribosomal DNA, differences between enzymes in frequency of variation could be seen, ranging from less than 2% for seven enzymes to 12.4% for glucosephosphate isomerase, a value similar to that observed for malic dehydrogenase, mitochondrial DNA, and ribosomal DNA in P. tetraurelia. The percentage of polymorphic enzyme loci in the complex as a whole was found to be much lower than that observed for other organisms. For the species more intensely studied in this paper the level of genetic polymorphism was also much lower, although P. biaurelia showed greater variability for two of the enzymes.     

Factors Determining the Frequency of the Killer Trait within Populations of the Paramecium aurelia Complex

The factors maintaining the cytoplasmically inherited killer trait in populations of Paramecium tetraurelia and Paramecium biaurelia were examined using, in part, computer simulation. Frequency of the K and k alleles, infection and loss of the endosymbionts, recombination during conjugation and autogamy, cytoplasmic exchange and natural selection were incorporated in a model. Infection during cytoplasmic exchange at conjugation and natural selection were factors that would increase the proportion of killers in a population. Conversely, k alleles reduced the proportion of killers in a population, acting through conjugation and autogamy. Field studies indicate that the odd mating type is prevalent in P. tetraurelia isolated from nature. Conjugation and therefore transmission by cytoplasmic transfer would be rare. Competition studies indicate a strong selective disadvantage for sensitives at concentrations found in nature. Natural selection must therefore be the factor maintaining the killer trait in P. tetraurelia.     

New, world-wide data on the distribution of species of the Paramecium aurelia complex (Ciliophora, Protozoa)

This is the first report on the presence of P. biaurelia in Tasmania, an island that has probably never been investigated before for the occurrence of the P. aurelia species. P. tetraurelia was recorded in Brazil, another very poorly investigated country in terms of this species complex. New stands of P. biaurelia and P. tetraurelia were also recorded in Japan. We present data concerning the occurrence and distribution of the P. aurelia species on different continents as a background for the newly described stands of P. aurelia spp.     

Interspecies Relationships in the Paramecium aurelia Complex: Acid Phosphatase Variation1

ABSTRACT. Up to five zones of acid phosphatase activity appear in gels after electrophoresis of detergent-treated extracts from 13 of the 14 species of the Paramecium aurelia complex. The overall pattern is somewhat similar for all species: differences in intensity and mobility of individual zones permit the grouping of these sibling species into eight groups. All 14 species can be identified using the procedure of enzyme electrophoresis, although two of them are more similar than is usually the case. Problems of misclassification are discussed in terms of the nature and frequency of variants. With the judicious choice of enzymes used to screen new stocks, these problems can be circumvented. Species relationships are updated using 11 enzymes. A dendrogram constructed from the matrix of genetic distances shows four clusters of species: (i) P. biaurelia, P. triaurelia; (ii) P. primaurelia, P. pentaurelia, P. sexaurelia, P. novaurelia; (iii) P. septaurelia, P. undecaurelia, P. tredecaurelia, P. quadecaurelia; and (iv) P. tetraurelia, P. octaurelia, P. decaurelia, P. dodecaurelia. Distances between the species are large, on the order of the differences between Drosophila species. The species are characterized by an extraordinary lack of geographical differentiation and great morphological similarity, which contrasts strongly with the molecular differentiation.     

Occurrence of Paramecium species in Western Siberia, Russia

Paramecium strains collected in Central Russia, Western Siberia (the West Siberian Lowland and the Altai Mountains in the south) were studied. The presence of P. caudatum, P. bursaria, P. multimicronucleatum, P. polycaryum, and four species of the P. aurelia complex, i.e. P. primaurelia (in Omsk), P. biaurelia (in Krasnoyarsk and the Altai Mountains), P. triaurelia (in Krasnoyarsk), and P. pentaurelia (in Novosibirsk, Altai Foreland, and Altai Mountains) was revealed. P. triaurelia and P. pentaurelia were found for the first time in Asia.     

Phylogenetic relationships of the genus Paramecium inferred from small subunit rRNA gene sequences

The genus Paramecium includes species that are well known and very common in freshwater environments. Species of Paramecium are morphologically divided into two distinct groups: the "bursaria" subgroup (foot-shaped) and the "aurelia" subgroup (cigar-shaped). Their placement within the class Oligohymenophorea has been supported by the analysis of the small subunit rRNA gene sequence of P. tetraurelia. To confirm the stability of this placement and to resolve relationships within the genus, small subunit rRNA gene sequences of P. bursaria, P. calkinsi, P. duboscqui, P. jenningsi, P. nephridiatum, P. primaurelia, and P. polycaryum were determined and aligned. Trees constructed using distance-matrix, maximum-likelihood, and maximum-parsimony methods all depicted the genus as a monophyletic group, clustering with the other oligohymenophorean taxa. Within the Paramecium clade, P. bursaria branches basal to the other species, although the remaining species of the morphologically defined "bursaria" subgroup do not group with P. bursaria, nor do they form a monophyletic subgroup. However, the species of the "aurelia" subgroup are closely related and strongly supported as a monophyletic group.     

First American stand of Paramecium novaurelia and intra-specific differentiation of the species

A stand of Paramecium novaurelia was found in Boston Massachusetts, USA, the first on this continent. Molecular studies (RAPD and sequencing ofrRNA [3'SSUrRNA-ITS1, 5' LSU rRNA] and COI mtDNA fragments) of P. novaurelia strains show intra-specific polymorphism within the species as strain clusters characterized by variable relationships.     

Influence of extremely low frequency electromagnetic fields on the swimming behavior of ciliates

Different species of ciliates (Paramecium biaurelia, Loxodes striatus, Tetrahymena thermophila) have been taken as model systems to study the effects of extremely low-frequency electromagnetic fields (50 Hz, 0.5–2.0 mT) on the cellular level. A dose-dependent increase in the mean swimming velocity and a decrease in the linearity of cell tracks were observed in all wild-type cells. In contrast, field-exposure did not increase the number of directional turns of the Paramecium tetraurelia pawn mutant (d4–500r), which is characterized by defective Ca²⁺-channels. The described changes indicate a direct effect of low frequency electromagnetic fields on the transport mechanisms of the cell membrane for ions controlling the motile activity of cilia. Bioelectromagnetics 18:491–498, 1997. © 1997 Wiley-Liss, Inc.     

Esterase Variants in Four Species of the Paramecium aurelia Complex1

One hundred eighty-eight stocks of Paramecium primaurelia, P. biaurelia, P. tetraurelia, and P. octaurelia were grown axenically and tested for five esterases, visualized by starch gel electrophoresis, in a search for variant stocks. The five esterases can be distinguished on the bases of their substrate specificity, sensitivity to an inhibitor, and response to different growth conditions. This paper addresses the nature of the electrophoretic change in mobility of the variant stocks in order that species relationships can be more accurately assessed. Crosses carried out in all four species show that single genes determine the differences in mobility between variant and common subtypes. Extracts of variant stocks that gave similar patterns were run against each other, tested for their sensitivity to the inhibitor, and the pattern was compared to that found in extracts of stocks with variant and common subtypes in other species. The majority of the variants in P. primaurelia, P. tetraurelia, and P. octaurelia show an electrophoretic mobility characteristic of a common subtype, or a variant, in another species. The same proportion of variant subtypes as common subtypes have mobilities similar to esterase subtypes found in other species. Of the four species examined in this paper, P. tetraurelia and P. octaurelia appear to be most closely related on the basis of shared esterase subtypes. In P. biaurelia the mobilities of most of the variants are unique, as are the common esterase subtypes in this species. P. biaurelia stands out as having the greatest number of esterase subtypes, with very few of them homologous to subtypes found in other species. This observation supports the idea of greater diversification of stocks within P. biaurelia than for the other three species.     

A two-locus molecular characterization of Paramecium calkinsi

Paramecium calkinsi (Ciliophora, Protozoa) is a euryhaline species which was first identified in freshwater habitats, but subsequently several strains were also collected from brackish water. It is characterized by clockwise spiral swimming movement and the general morphology of the "bursaria type." The present paper is the first molecular characterization of P. calkinsi strains recently collected in distant regions in Russia using ITS1-5.8S- ITS2-5'LSU rDNA (1100bp) and COI (620bp) mtDNA sequenced gene fragments. For comparison, our molecular analysis includes P. bursaria, exhibiting a similar "bursaria morphotype" as well as species representing the "aurelia type," i.e., P. caudatum, P. multimicronucleatum, P. jenningsi, and P. schewiakoffi, and some species of the P. aurelia species complex (P. primaurelia, P. tetraurelia, P. sexaurelia, and P. tredecaurelia). We also use data from GenBank concerning other species in the genus Paramecium and Tetrahymena (which used as an outgroup). The division of the genus Paramecium into four subgenera (proposed by Fokin et al. 2004) is clearly presented by the trees. There is a clear separation between P. calkinsi strains collected from different regions (races). Consequently, given the molecular distances between them, it seems that these races may represent different syngens within the species.     

Telomeres inhibit end to end fusion and enhance maintenance of linear DNA molecules injected into the Paramecium primaurelia macronucleus.

Direct injection into the macronucleus of Paramecium tetraurelia of DNA molecules coding for the A-antigen leads to expression of the gene and autonomous replication. When injected into Paramecium primaurelia DNA from probably any origin, procaryote or eucaryote, can replicate as linear telomerized molecules and the number of copies maintained can be very high (up to 20000 copies). We present here evidence that if the injected linear DNA molecules harbour preexisting telomeres at both extremities they are protected from degradation, the number of DNA molecules maintained being 15- to 30-fold higher than if the molecules are injected without telomeres. Some of the injected molecules replicate as multimers, but, only when the fused ends are devoid of preexisting telomeric repeats.     

Variety of serotypes of Paramecium primaurelia: single epitopes are responsible for immunological differentiation

Paramecium primaurelia expresses three major types of surface antigens. We report here the identification of the gene for serotype S, which completes the sequence data of expressed serotypes of P. primaurelia. The complete open reading frame of surface antigen S was identified using a novel technique, based upon the presence of conservative regions in the non-coding areas of the multigene family. We were able to isolate the 7194-bp-long open reading frame from the macronuclear DNA for Serotype 156S. The corresponding mRNA was detected in the two serotype S-expressing stocks, 60 and 156, of P. primaurelia, which clarifies that both stocks are using the same S allele. Comparisons of the nucleic acid and the deduced amino-acid sequence showed high identity to surface antigen 51B of P. tetraurelia, sufficient to cause an immunological cross-reaction in vivo. Immunologically relevant epitopes in vivo were identified in the central regions of the genes, constructed of nearly perfect tandem repeats.     

Paramecium GPI proteins: variability of expression and localization

In Paramecium primaurelia, the two major classes of cell surface proteins, the surface antigen (SAg) and the surface GPI proteins (SGPs), are linked to the plasma membrane through a glycosylphosphatidylinositol (GPI) anchor. In the present study, we have characterized the expression of the SGPs in several geographical strains of P. primaurelia and P. tetraurelia at different temperatures, 23 degrees C and 32 degrees C. The identification of the expressed SGPs was performed on purified cilia, by establishing the SGP SDS-PAGE profiles under four different conditions: with or without their anchoring lipid, cleaved with a Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PI-PLC), and either in a reduced or in an unreduced state. This screening revealed the existence of specific sets of ciliary SGPs, as a function of temperature and the geographical origin of the strains. The SGPs the most abundant at 23 degrees C and 32 degrees C displayed a rapid turnover. We also looked for the presence of PI-PLC releasable proteins in purified cortices. In addition to the SAg and SGPs, the cortical fraction was shown to contain other PI-PLC releasable proteins, not found in the ciliary fraction, thus localized exclusively in the interciliary region.