Current view on phylogeny within the genus Flabellula Schaeffer, 1926 (Amoebozoa: Leptomyxida).

The molecular phylogeny of Flabellula Schaeffer, 1926 has been updated by analysing 18S rRNA and actin gene sequences of 19 new strains collected and characterised by the authors over the past ten years. The genus Flabellula Schaeffer, 1926 (Amoebozoa: Leptomyxida) is a taxon in which species delineation based on morphological data by themselves is insufficient or even misleading. The description of two novel species, F. schaefferi n. sp. and F. sawyeri n. sp., is justified by the congruence of morphological data with 18S rRNA and actin gene sequence phylogenies, in-silico secondary structure prediction of the V2 region in the 18S rRNA, and by recognition of species-specific sequential motifs within this region.

A New Heterolobosean Amoeboflagellate, Tetramitus dokdoensis n. sp., Isolated from a Freshwater Pond on Dokdo Island in the East Sea, Korea.

The genus Tetramitus is a representative amoeboflagellate group within the Heterolobosea, and currently contains over a dozen species. Here, a new heterolobosean amoeboflagellate was isolated from a freshwater pond on Dokdo Island, Korea. The amoebae have eruptive pseudopodia, no uroidal filament, and a nucleus with a central nucleolus. The length and width of the amoebae are 15.5-28.0 µm and 5.4-12.6 µm, respectively. The flagellates are conical, with 4 flagella of equal length (~10 µm). There is a discrete rostrum in the subapical region of the flagellate form. The cyst has thin endo- and ectocyst layers and no cyst pores. The amoeba shows slow movement at 37 °C, but does not move at 42 °C under a light microscope. Phylogenies of the 18S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 sequence show that the strain belongs to a subclade of Tetramitus that includes Tetramitus rostratus, Tetramitus waccamawensis and Tetramitus entericus, amongst others. Nonetheless, the strain is distinct from other species in both molecular phylogenetic trees. Thus the strain isolated from the Dokdo Island is proposed as a novel species, Tetramitus dokdoensis n. sp.

Heterogeneity in the genus Allovahlkampfia and the description of the new genus Parafumarolamoeba (Vahlkampfiidae; Heterolobosea).

Heterolobosean amoebae are common and diverse members of soil protist communities. In this study, we isolated seven strains of amoebae from soil samples taken in Tibet (at high altitude), Sardinia and the Netherlands, all resembling to belong to a similar heterolobosean morphospecies. However, sequences of the small subunit (SSU) rDNA and internal transcribed spacers, including the 5.8S rDNA, revealed a high heterogeneity in the genus Allovahlkampfia to which six of the isolates belong. Some unnamed strains, of which the sequences had been published before, are also included within the genus Allovahlkampfia. One Allovahlkampfia isolated in the Netherlands harbors a twin-ribozyme, containing a His-Cys box, similar to the one found in strain BA of Allovahlkampfia. The other SSU rDNA sequence grouped in phylogenetic analyses with sequences obtained in environmental sequencing studies as sister to the genus Fumarolamoeba. This phylogenetic placement was supported by analyses of the 5.8S rDNA leading us to describe it as a new genus Parafumarolamoeba.

Potentially pathogenic free-living amoebae isolated from hospital wards with immunodeficient patients in Tehran, Iran.

This study investigated the occurrence of free-living amoebae (FLA) in immunodeficiency wards of hospitals in Tehran, Iran. A total of 70 dust and biofilm samples from wards serving transplant, pediatric (malignancies), HIV, leukemia and oncology patients of five university hospitals were collected and examined for the presence of FLA using culturing and molecular approaches. Based on the morphology of the amoebae in plate cultures, primer sets were applied for molecular identification of Acanthamoeba, vahlkampfiid amoebae and Hartmannella. Out of 70 samples, 37 (52.9%) were positive for FLA. Acanthamoeba belonged to the T4 genotype was the most prevalent isolate. Presence of the T4 genotype on medical instruments, including an oxygen mask in an isolation room of an immunodeficiency pediatric ward, should be of concern for health authorities. Acanthamoeba T5 genotypes, Hartmannella vermiformis, and Vahlkampfia avara were also present. These results highlight a clear need for greater attention to improved disinfection, especially where susceptible patients, such as those who are immune-suppressed, are served. To our knowledge, this is the first report of these FLA in immunodeficiency wards in Iran, and also the first to identify Acanthamoeba T5, Hartmannella, and Vahlkampfia in moist habitats, such as biofilms, in this country.

First report of a mixed infection due to Acanthamoeba genotype T3 and Vahlkampfia in a cosmetic soft contact lens wearer in Iran.

Acanthamoeba keratitis cases have emerged in the recent years in Iran. In this case, an amoebic keratitis due to a mixed infection with Acanthamoeba and Vahlkampfia species is reported. Corneal scrapes, contact lenses and contact lens cases obtained from the patient were analysed and were positive for cysts of Acanthamoeba and Vahlkampfia genera. Genus-specific PCR was carried out for both genera, confirming the microscopic observations. To our knowledge, this is the first reported case of a possible mixed amoebic infection due to Acanthamoeba and Vahlkampfia and raises awareness within contact lens wearers in Iran.

Tetramitus thermacidophilus n. sp., an amoeboflagellate from acidic hot springs.

Tetramitus thermacidophilus n. sp. is a novel thermophilic and acidophilic amoeboflagellate isolated from acidic hot springs in the Caldera Uzon (Kamchatka, Russia) and in Pisciarelli Solfatara (Naples, Italy). We describe it based on physiological, morphological, and sequence data. It was grown in monoxenic culture on the archaeon Acidianus brierleyi as food. Tetramitus thermacidophilus multiplies in a pH range from 1.2 to 5 and in a temperature range from 28 °C to 54 °C. The shortest doubling time was 4.5 h at pH 3 at 45 °C. Its spindle-shaped biflagellated stage was only rarely found in culture. The amoeboid stage shows the typical locomotive form of vahlkampfiid amoebae. Sequence comparisons of the internal transcribed spacer sequences and the small subunit rRNA genes confirm that T. thermacidophilus is a novel species within the genus Tetramitus and that both isolates belong to that species.

[Parasitic amoebae and amoebo-flagellates from the Lobosea and heterolobosea classes]. / Parazitichekie ameby i ameboflagelliaty klassov Lobosea i Heterolobosea.

Different cited evidences on parasitic amoebae and amoebo-flagellates belonging to the Lobosea and Heterolobosea classes (Pages, 1987) have been reviewed. Special attention is paid to various degree of their adaptation to parasitic mode of life, which ranges from a parasitism on a border with commensalism to true parasitism (both facultative and obligatory ones). Besides the coprophilous and commensal species, the number of true parasites among the Lobosea and Heterolobosea classes is comparatively small. In many cases, both facultative and obligatory parasites cause the death of their hosts. Apparently this strongly pronounced pathogenicity of parasitic amoebae and amoebo-flagellates suggest a recent origin of such parasite-host systems. Pathogens of amoebic dysentry, primary amoebic meningoencephalitis and granulomatous amoebic encephalitis are specially considered. In the Russian text-books on a parasitology the information on most of them, except Entamoeba hystolytica, is either totally absent or very scare and out of date.

On the identity of the amoeboflagellates Didascalus thorntoni and Adelphamoeba galeacystis.

Didascalus thorntoni, Singh 1952 has been classified alternately as a separate genus or as a species of Naegleria. In the 18th edition of the American Type Culture Collection catalogue it is classified as Naegleria thorntoni. To resolve the question of its identity we have used riboprinting and sequencing of the small subunit ribosomal DNA. The results indicate that D. thorntoni does not belong to the genus Naegleria. The sequence of the small subunit ribosomal DNA differs only in 20 nucleotides (1%) from that of Paratetramitus jugosus. The difference is much smaller than between some species of Nageleria. Therefore, it is not clear whether D. thorntoni should be considered as a species of Paratetramitus or as a separate genus. The strain used in different laboratories as the type strain of Adelphamoeba galeacystis has been identified as a Naegleria strain. We believe that the type strain of A. galeacystis was mislabeled prior to submission to the American Type Culture Collection and to the Culture Collection of Algae and Protozoa. A recent isolate, which on the basis of morphology was identified as a strain of A. galeacystis, has the identical small subunit ribosomal DNA sequence as D. thorntoni. Our results prove Page was right when he stated that Adelphamoeba might be a synonym of Didascalus.

The evolution of the Vahlkampfiidae as deduced from 16S-like ribosomal RNA analysis.

The amoebae, a phenotypically diverse, paraphyletic group of protists, have been largely neglected by molecular phylogeneticists. To better understand the evolution of amoebae, we sequenced and analyzed the 16S-like ribosomal RNA genes of three vahlkampfiid amoebae: Paratetramitus jugosus, Tetramitus rostratus and Vahlkampfia lobospinosa. The Vahlkampfiidae lineage is monophyletic, branches early along the eukaryotic line of descent, and is not a close relative of the multicellular amoebae that also reversibly transform from amoebae to flagellates.