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The invasion of the Laurentian Great Lakes by the fish-parasitic sea lamprey has led to catastrophic consequences, including the potential introduction of fish pathogens. Aeromonas salmonicida is a bacterial fish pathogen that causes devastating losses worldwide. Currently, there are five accepted subspecies of Aeromonas salmonicida: A. salmonicida subsp. salmonicida, masoucida, smithia, achromogenes, and pectinolytica. We discuss the discovery of an isolate of A. salmonicida that is pathogenic to rainbow trout (Oncorhynchus mykiss) and exhibits unique phenotypic and molecular characteristics. We examined 181 adult sea lamprey (Petromyzon marinus) from the Humber River (Lake Ontario watershed) and 162 adult sea lamprey from Duffins Creek (Lake Ontario watershed) during the spring seasons of 2005-11. Among those, 4/343 (1.2%) sea lamprey were culture positive for A. salmonicida, whereby biochemical and molecular studies identified three of the isolates as A. salmonicida subsp. salmonicida. The remaining isolate (As-SL1) recovered from Humber River sea lamprey was phenotypically more similar to A. salmonicida subsp. salmonicida than to the four other A. salmonicida subspecies. However, unlike A. salmonicida subsp. salmonicida, As-SL1 was sucrose positive, produced an acid-over-acid reaction on triple-sugar iron medium and did not amplify with A. salmonicida subsp. salmonicida specific primers. Phylogenetic analysis based on partial stretches of the 16S rRNA and DNA gyrase subunit B genes further confirmed that the As-SL1 isolate was not A. salmonicida subsp. masoucida, smithia, achromogenes, or pectinolytica. Based on our analyses, the As-SL1 isolate is either an unusual strain of A. salmonicida subsp. salmonicida or a novel A. salmonicida subspecies. The four A. salmonicida isolates that were recovered from sea lamprey were pathogenic to rainbow trout in experimental challenge studies. Our study also underscores the potential role of sea lamprey in the ecology of infectious fish diseases.

Since the 1990s, populations of the benthic amphipod Diporeia spp. (Diporeia) have sharply declined across much of the Laurentian Great Lakes. This study was undertaken to identify contemporary and historical community composition, structure, and dynamics of parasites infecting Diporeia collected from nine sites in the southern basin of Lake Michigan, where declines of the amphipod have been well documented over the past 20 years. An additional aim of this study was to assess whether infection dynamics and dreissenid densities could explain the declines in Diporeia densities that have occurred. We found that Diporeia were host to eight groups of uni- and multicellular pathogens. Of the 3082 amphipods analyzed, 1624 individuals (52.7%) were infected with at least one type of parasite. Ciliophora was the most prevalent parasite (50.08% prevalence of infection), followed by Gregarinasina (2.79%), Microsporidia (0.68%), Cestoda (0.45%), Acanthocephala (0.36%), Haplosporidia (0.23%), Yeast (0.32%), and filamentous Fungi (0.10%). Considerable spatial and temporal variability were observed in parasite prevalences, with prevalences frequently appearing to cycle between low and high values. Parasite species belonging to Microsporidia and Haplosporidia were associated with tissue alteration and host inflammatory response; however, parasite prevalences explained very little in terms of Diporeia density declines at assessed sites. Despite these findings, we do not discount the possibility that parasitic infections may have played a role in declining Diporeia densities in the Great Lakes, as the cyclical prevalences that were observed are possibly suggestive of parasitic outbreaks that are followed by die-offs at affected sites. This study suggests that if parasites have affected Diporeia densities in the Laurentian Great Lakes, then the relationship may be a complicated one.

Dictyocoela diporeiae n. sp. is described from Diporeia spp. (Amphipoda, Gammaridea) collected from Lake Superior (USA), and its morphology and taxonomic affiliation are discussed. In hematoxylin- and eosin-stained sections of infected amphipods, the microsporidian was observed to infect muscle tissue surrounding the ovaries. Melanized hemocytic encapsulations were often observed in or near masses of microsporidians. The microsporidians appeared as spores measuring 1.99 ± 0.09 μm long by 1.19 ± 0.05 μm wide. Each spore contained eight coils of isofilar polar filaments that were arranged in single ranks. Polar filaments measured 71 ± 3 nm in diameter. A prominent lamellar polaroplast composed of ordered concentric membranes was found at the apical end of the spore surrounding the polar filament. A distinct posterior vacuole was observed at the distal end of the spore. Phylogenetic analysis based on 16s RNA sequences showed that the microsporidian belongs to the genus Dictyocoela, and is most similar to D. berillonum, yet distinctly different. The species is new, based on its morphology, genetic sequence, host, and location within the host.

Background The phylum Haplosporidia contains coelozoic and histozoic, spore-forming, obligate protozoan endoparasites that infect a number of freshwater and marine invertebrates including bivalves, crustaceans, and polychaetes. In amphipods, haplosporidians cause systemic infection resulting in a range of pathologies. While amphipods belonging to the genus Diporeia (Gammarideae) have been shown to host haplosporidians, the taxonomic relationship of the Diporeia haplosporidian(s) is largely unknown due to the lack of phylogenetic and detailed ultrastructural studies. Methods The infection characteristics and taxonomic identity of a haplosporidian infecting Diporeia spp. (Diporeia) were based on microscopical investigation, electron microscopy, and Bayesian phylogenetic inference using haplosporidian 16S rRNA gene sequences. Results In stained sections, the haplosporidian was observed to cause systemic infections in Diporeia that were often accompanied with host tissue degeneration. The haplosporidian appeared as binucleate plasmodia and sporocysts containing different spore maturation stages in the coelom, connective tissue, digestive tissue, and muscle. All of the observed systemic infections progressed to sporogenesis. Transmission electron microscopy revealed that fixed mature spores were slightly ellipsoidal and had a mean spore length X width of 5.34 ± 0.17 × 4.09 ± 0.15 μm. A hinged opercular lid with a length of 3.1 ± 0.17 μm was observed for a number of developing spores. The average thickness of the cell wall was 90.0 ± 8.33 nm. Thin filaments (70 nm) composed of spore wall material were observed projecting from an abopercular thickening of the spore wall. Phylogenetic analysis showed that the haplosporidian is novel bearing some similarities with the oyster pathogen Haplosporidium nelsoni, yet distinctly different. Conclusions Based on its morphology, genetic sequence, and host, it became evident that the Diporeia haplosporidian is taxonomically novel and we propose its nomenclature as Haplosporidium diporeiae. This is the first report of a haplosporidian infecting Diporeia in Lake Superior.

The mode of viral hemorrhagic septicemia virus (VHSV) transmission in the Great Lakes basin is largely unknown. In order to assess the potential role of macroinvertebrates in VHSV transmission, Diporeia spp., a group of amphipods that are preyed upon by a number of susceptible Great Lakes fishes, were collected from seven locations in four of the Great Lakes and analyzed for the presence of VHSV. It was demonstrated that VHSV is present in some Diporeia spp. samples collected from lakes Ontario, Huron, and Michigan, but not from Lake Superior. Phylogenetic comparison of partial nucleoprotein (N) gene sequences (737 base pairs) of the five isolates to sequences of 13 other VHSV strains showed the clustering of Diporeia spp. isolates with the VHSV genotype IVb. This study reports the first incidence of a fish-pathogenic rhabdovirus being isolated from Diporeia, or any other crustacean and underscores the role macroinvertebrates may play in VHSV ecology.