Integrative Approach for the Reliable Detection and Specific Identification of the Microsporidium Loma morhua in Atlantic Cod (Gadus morhua).

Microsporidia are fungal parasites that infect diverse invertebrate and vertebrate hosts. Finfish aquaculture supports epizootics due to high host density and the high biotic potential of these parasites. Reliable methods for parasite detection and identification are a necessary precursor to empirical assessment of strategies to mitigate the effects of these pathogens during aquaculture. We developed an integrative approach to detect and identify Loma morhua infecting Atlantic cod. We show that the spleen is more reliable than the commonly presumed gills as best organ for parasite detection in spite of substantial morphological plasticity in xenoma complexes. We developed rDNA primers with 100% sensitivity in detecting L. morhua and with utility in distinguishing some congeneric Loma species. ITS sequencing is necessary to distinguish L. morhua from other congeneric microsporidia due to intraspecific nucleotide variation. 64% of L. morhua ITS variants from Atlantic cod have a 9-nucleotide motif that distinguishes it from Loma spp. infecting non-Gadus hosts. The remaining 36% of ITS variants from Atlantic cod are distinguished from currently represented Loma spp., particularly those infecting Gadus hosts, based on a 14-nucleotide motif. This research approach is amenable to developing templates in support of reliable detection and identification of other microsporidian parasites in fishes.

Description of five new Loma (Microsporidia) species in pacific fishes with redesignation of the type species Loma morhua Morrison & Sprague, 1981, based on morphological and molecular species-boundaries tests.

Five new species of Loma were described from five Pacific fishes using light-microscopic and ultrastructural features along with phylogenetic analysis of the gene sequences of ribosomal RNA (rRNA) and elongation factor 1-alpha. Morphological data revealed both qualitative and quantitative differences in developmental stages and timing, vesicles, xenoma features, and spore sizes with statistical support that differentiated Loma pacificodae n. sp. in Pacific cod, Loma wallae n. sp. in walleye pollock, Loma kenti n. sp. in Pacific tomcod, Loma lingcodae n. sp. in lingcod, and Loma richardi n. sp. in sablefish from each other and other species in the genus. Phylogenetic analyses combined with monophyly tests supported species designations, but with low resolution in two cases perhaps due to rRNA paralogs or recent speciation. Loma branchialis in haddock was shown to be separate from Loma morhua in Atlantic cod, thereby making L. morhua, and not L. branchialis, the type species. A species from brook trout was shown to be a separate species from Loma salmonae, not a variant strain selected in the laboratory. By comparison with gadid host phylogeny, these Loma species appear to have coevolved with their hosts, first colonizing the Pacific basin about 12 million years ago.

Morphological and genetical description of Loma psittaca sp. n. isolated from the Amazonian fish species Colomesus psittacus.

A previously unrecognised fish-infecting microsporidia (Loma psittaca n. sp.), found adherent to the intestinal mucosa of the freshwater puffer fish Colomesus psittacus (Teleostei, Tetraodontidae) from lower Amazon River, was described based on light and transmission electron microscope and phylogenetic analysis. The whitish xenoma was completely filled by numerous spores, including several developmental stages of the parasite. In all of these stages, the nuclei were monokaryotic. The merogonial plasmodium divided by binary fission and the sporont gave rise to disporoblastic ovoid spores measuring 4.2 +/- 0.4 x 2.8 +/- 0.4 microm. In mature spores, the polar filament was arranged in 10-11 (rarely 12) coils in one row in turn of posterior vacuole. The polaroplast had two distinct regions around the manubrium. The polyribosomes were organised in coiled tapes. The small subunit rRNA gene was sequenced and maximum parsimony analysis placed the microsporidian described here in the clade that includes the genera Ichthyosporidium, Loma and Pseudoloma. Based on differences from previously described microsporidians, such as ultrastructural characteristics of the xenoma, developmental stages including the spore and phylogenetic analysis supported the recognition of a new species, herein named L. psittaca n. sp.