Microsporidia Alfvenia sibirica sp. n. and Agglomerata cladocera (Pfeiffer) 1895, from Siberian microcrustaceans and phylogenetic relationships within the "Aquatic outgroup" lineage of fresh water microsporidia

Abstract

Here we report on two microsporidia from freshwater crustaceans collected during the ongoing survey for microsporidia in the river Karasuk and adjacent waterbodies (Novosibirsk region, Western Siberia). The first species parasitized hypoderm and fat body of a cyclopid Cyclops sp. (Maxillopoda, Copedoda) and produced oval spores, measured 2.0×3.6μm (fixed) enclosed individually or in small groups in fragile sporophorous vesicles (SVs). We describe it here as Alfvenia sibirica sp. n. The second species infected the same tissues of a cladoceran Daphnia magna (Branchiopoda, Phyllopoda). Its spores were pyriform, 2.3×4.0μm (fixed), and resided in relatively persistent SVs in groups of 8-16. This species was identified as a Siberian isolate (Si) of Agglomerata cladocera (Pfeifer) because ultrastructurally it was hardly distinguishable from A. cladocera recorded from England from the same host (Larsson et al., 1996). A. cladocera (Si) shared 99% SSU rDNA sequence similarity to Binucleata daphniae from D. magna collected in Belgium (Refardt et al., 2008). The major outcome of our work is that we present molecular (SSUrDNA) characterization coupled with EM description, for representatives of two genera, Alfvenia (encompasses 3 described so far species) and Agglomerata (7 species), which allowed us to place these two genera on the phylogenetic tree. We also summarized the literature data on Alfvenia and Agglomerata spp., and provided their comparative morphological analysis. These two genera belong to so called "Aquatic outgroup" (Vossbrinck et al., 2004), a poorly resolved lineage, a sister to Amblyosporidae. This lineage probably includes majority of fresh water forms of microsporidia, most of which remain undescribed. SSUrDNA-based phylogenetic analysis and analysis of hosts suggest that diversification within the "Aquatic outgroup" could have been connected with the host switch from dipterans or copepods to cladocerans that had occurred in some ancestral Amblyospora-related lineage(s).

Keywords: Cladocera; Copepoda; Cyclopidae; Daphnia; Microsporidiosis; Molecular phylogeny; Ultrastructure.