The phylum Chlamydiae contains obligate intracellular bacteria, several of which cause disease in their hosts. Morphological studies have suggested that this group of bacteria may be pathogens of fish, causing cysts in epithelial tissue - epitheliocystis. Recently, the first genetic evidence of a chlamydial aetiology of this disease in seawater reared Atlantic salmon from Norway and Ireland was presented, and the agent was given the name 'Candidatus Piscichlamydia salmonis'. In this article we present molecular evidence for the existence of a novel Chlamydiae that also may cause epitheliocystis in Norwegian salmonids. This novel Chlamydiae has been found in salmonid fish from freshwater, and based on its partial 16S rRNA gene, it may constitute a third genus in the family Chlamydiaceae, or a closely related sister family. By using whole-mount RNA-RNA hybridization we demonstrate how infected cells are distributed in a patchy manner on a gill arch. The morphology of the novel Chlamydiae includes the characteristic head-and-tail cells that have been described earlier from salmonid fish suffering from epitheliocystis. We propose the name 'Candidatus Clavochlamydia salmonicola' for this agent of epitheliocystis in freshwater salmonids.

In 2004, a new disease was detected in cod (Gadus morhua) in western Norway. Affected cod had white granulomas in the visceral organs and skin. A species of Francisella was isolated on blood agar plates from moribund cod. The bacterium could be grown at temperatures ranging from 6 to 22 degrees C, but did not grow at 37 degrees C. Challenge experiments showed that Francisella sp. was the cause for the new disease. The 16S rDNA gene sequence from Francisella sp. showed 99.17% similarity to F. philomiragia, and the 16S-23S ribosomal RNA intergenic spacer (249 nt), shows a similarity with that from Francisella isolated from tilapia and F. tularensis of 96.8 and 35.9%, respectively. The 23S sequence is more similar to F. tularensis, 97.7% (2,862 nt), compared to the tilapia isolate 96.8% (2,131 nt). The partial putative outer membrane protein (FopA) sequence (781 nt) from Francisella sp. shows a similarity with that from F. tularensis and F. philomiragia of 77.3 and 98.2%, respectively. Based on sequence data, culturing temperatures and pathogenicity for cod, it is suggested that this Francisella sp. from cod could be a new species of Francisella, Family Francisellaceae.

Paranucleospora theridion n. gen, n. sp., infecting both Atlantic salmon (Salmo salar) and its copepod parasite Lepeophtheirus salmonis is described. The microsporidian exhibits nuclei in diplokaryotic arrangement during all known life-cycle stages in salmon, but only in the merogonal stages and early sporogonal stage in salmon lice. All developmental stages of P. theridion are in direct contact with the host cell cytoplasm or nucleoplasm. In salmon, two developmental cycles were observed, producing spores in the cytoplasm of phagocytes or epidermal cells (Cycle-I) and in the nuclei of epidermal cells (Cycle-II), respectively. Cycle-I spores are small and thin walled with a short polar tube, and are believed to be autoinfective. The larger oval intranuclear Cycle-II spores have a thick endospore and a longer polar tube, and are probably responsible for transmission from salmon to L. salmonis. Parasite development in the salmon louse occurs in several different cell types that may be extremely hypertrophied due to P. theridion proliferation. Diplokaryotic merogony precedes monokaryotic sporogony. The rounded spores produced are comparable to the intranuclear spores in the salmon in most aspects, and likely transmit the infection to salmon. Phylogenetic analysis of P. theridion partial rDNA sequences place the parasite in a position between Nucleospora salmonis and Enterocytozoon bieneusi. Based on characteristics of the morphology, unique development involving a vertebrate fish as well as a crustacean ectoparasite host, and the results of the phylogenetic analyses it is suggested that P. theridion should be given status as a new species in a new genus.

Proliferative gill disease (PGD) is an emerging problem in Norwegian culture of Atlantic salmon (Salmo salar). Parasites (Ichthyobodo spp.) and bacteria (Flexibacter/Flavobacterium) may cause PGD, but for most cases of PGD in farmed salmon in Norway, no specific pathogen has been identified as the causative agent. However, Neoparamoeba sp. and several bacteria and viruses have been associated with this disease. In the spring of 2006, a new poxvirus, salmon gill poxvirus (SGPV), was discovered on the gills of salmon suffering from PGD in fresh water in northern Norway. Later the same year, this virus was also found on gills of salmon at two marine sites in western Norway. All farms suffered high losses associated with the presence of this virus. In this study, we describe the entry and morphogenesis of the SGP virus in epithelial gill cells from Atlantic salmon. Intracellular mature virions (IMVs) are the only infective particles that seem to be produced. These are spread by cell lysis and by "budding" of virus packages, containing more that 100 IMVs, from the apical surface of infected cells. Entry of the IMVs appears to occur by attachment to microridges on the cell surface and fusion of the viral and cell membranes, delivering the cores into the cytoplasm. The morphogenesis starts with the emergence of crescents in viroplasm foci in perinuclear areas of infected cells. These crescents consist of two tightly apposed unit membranes (each 5 nm thick) that seem to be derived from membranes of the endoplasmic reticulum. The crescents develop into spheres, immature virions (IVs), that are 350 nm in diameter and surrounded by two unit membranes. The maturation of the IVs occurs by condensation of the core material and a change from spherical to boat-shaped particles, intracellular mature virions (IMVs), that are about 300 nm long. Hence, the IMVs from the SGP virus have a different morphology compared to other vertebrate poxviruses that are members of the subfamily Chordopoxvirinae, and they are more similar to members of subfamily Entomopoxvirinae, genus Alphaentomopoxvirus. However, it is premature to make a taxonomic assignment until the genome of the SGP virus has been sequenced, but morphogenesis clearly shows that this virus is a member of family Poxviridae.

The microsporidian Paranucleospora theridion was discovered in Atlantic salmon Salmo salar suffering from proliferative gill disease in a marine farm in western Norway in 2008. The parasite develops in cells of the reticuloendothelial system, cells important for normal immune function. The aim of this study was to see if P. theridion could play a part in some of the diseases with unclear causes in salmon production in Norway, i.e. proliferative gill disease (PGI), pancreas disease (PD), heart and skeletal muscle inflammation (HSMI) and cardiomyopathy syndrome (CMS). P. theridion was present in all areas with salmon farming in Norway, but high prevalence and densities of the parasite in salmon and salmon lice were only seen in southern Norway. This region is also the main area for PGI and PD in Norway. Quantification of pathogens associated with PGI, PD, HSMI and CMS diagnoses showed that P. theridion levels are high in southern Norway, and may therefore play a role in susceptibility and disease development. However, among the different diagnoses, fish with PGI are particularly heavily infected with P. theridion. Therefore, P. theridion appears as a possible primary agent in cases with high mortality in connection with PGI in western Norway. KEY WORDS: Paranucleospora theridion · Atlantic salmon · Microsporidia · Proliferative gill disease · PGI · Pancreas disease · PD Resale or republication not permitted without written consent of the publisherDis Aquat Org 94: [41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56][57] 2011 Clavochlamydia salmonicola , Atlantic salmon paramyxovirus (Kvellestad et al. 2005), and salmon gill poxvirus . These agents may all contribute to the gill pathology, hypertrophia, hyperplasia, inflammation and necrosis of gill tissues, but it is not known whether they are primary pathogens or secondary invaders.It is well documented that 'pancreas disease' is associated with the presence of different strains of SAV (Nelson et al. 1995, McLoughlin et al. 1996, Villoing et al. 2000, Hodneland et al. 2005, Fringuelli et al. 2008). However, the observed disease and mortality in salmon and rainbow trout farms are different from what can be seen in challenge experiments using the different SAV strains . The explanations for this could be presence of different pathogens or other stress factors that may trigger the disease or be the primary cause of disease, reducing SAV to a secondary cause of the observed mortality in farms. Lack of comparable mortality and to a certain extent also pathology has also been observed when comparing challenge experiments using CMS and HSMI homogenates with the observed situation in salmon farms affected by these 2 diseases (Kongtorp et al. 2004a, Fritsvold et al. 2009. Hence, the trigger or primary causes for these diseases could be environmental factors or other pathogens.Paranucleospora theridion was first discovered in farmed Atlantic salmon suffering from gill disease in western Norway in (A. Nylund et al. 2009a,b, S. Nylund et al. 2009. Th...