SummaryWe have cloned and sequenced the haemolysin gene locus from Edwardsiella tarda (ETH

ABSTRACT. Pathological findings associated with scuticociliatosis in farmed Japanese flounder in Japan are described. Ten moribund fishes, farmed in Tottori Prefectural Fisheries Experimental Station, showed cutaneous ulcers, darkened skin, fin and tail rot, exophthalmia and alterations in swimming behaviour. Histopathologically, severe epidermal degeneration and necrosis, hyperplasia of branchial epithelium, myositis, myelitis, encephalitis associated with heavy accumulation of scuticociliates in the periorbital cavity and optic nerve fiber were observed. Moreover, masses of ciliates were found to feed on the host tissues such as skeletal muscles, gills and brain, causing severe degenerative changes associated with abundant neutrophilic and lymphocytic infiltration. These findings suggest that the present scuticociliate, Miamiensis avidus, is a highly invasive and destructive pathogen infecting Japanese flounder and capable of developing systemic fatal infection.

A cDNA clone for coho salmon (Oncorhynchus kisutch) transferrin was isolated from a liver cDNA library. The total 2,511 nucleotides contained a 5' non-coding region, an open reading frame encoded with a transferrin polypeptide consisting of 687 amino acids and 3' non-coding region. The amino acid sequence alignment of the coho salmon transferrin had the duplicated structure, conserved anion-binding, iron-binding, and cysteine residues and two glycosylation sites observed in the previously cloned cDNA transferrin of vertebrates. The coho salmon transferrin is similar in size to the corresponding nucleotides from medaka and Atlantic salmon. The deduced amino acid sequence of the transferrin cDNA of coho salmon had 48%, 46%, 67% and 85% amino acid identities with that of human, Xenopus, medaka and Atlantic salmon, respectively. The coho salmon gene was recognized to be transcribed only in the liver by Northern blot hybridization.

ABSTRACT. Japanese flounder (Paralichthys olivaceus) were experimentally infected with the highly pathogenic scuticociliate Miamiensis avidus (syn. Philasterides dicentrarachi) using the immersion method to clarify/identify the possible neural routes of entry and possible ways of dissemination of the scuticociliate in the fish body. Scuticociliates were observed on the skin and gills right from day 0-1 postinfection, muscle tissue on day 2 post-infection, reached the brain, and spinal cord on day 3 post-infection, and systemic infection was prominent afterwards. Brain lesions were observed in most of the examined fish from days 3 and 4 post-infection and considered to be the cause of the sudden increase in mortality. Affected fish showed varying degrees of tissue damage including severe epidermal and dermal necrotic lesions, necrotic myositis, encephalitis and myelitis. Whereas, scuticociliates were frequently observed along the optic and/or olfactory nerve in the fish which were accompanied by severe brain lesions but by minimum lesions in the gills and skin, suggesting that in addition to skin and/or gills, neural routes including periorbital and nasal routes may play a role in scuticociliate invasion to the brain. Scuticociliates were also observed in the peripheral nerve fibers in the muscle tissue, cranial and spinal nerves, cranial cavity and in the vertebral canal, suggesting that nerve fibers and/or cerebrospinal fluid circulation may be involved in the spread of the scuticociliate throughout the body in addition to the blood circulation and connective tissue.

Genetic differentiation within the salmon louse Lepeophtheirus salmonis (Krøyer, 1837), was investigated by the sequencing of specific nucleotide regions. Partial sequences of the 18S ribosomal RNA gene and the ribosomal internal transcribed spacer (ITS-1) region from single sea lice were amplified by the polymerase chain reaction (PCR). Lice were collected from wild and farmed Atlantic salmon (Salmo salar L., 1758) from nine selected localities around the Scottish coastline. A 0.9kb fragment of the 18S ribosomal RNA gene was amplified and compared for several samples of lice which showed no observable differences between the lice from different collection sites confirming the absence of cryptic species. The 454 nucleotide ITS-1 sequence showed differences between derived sequences from 13 sea lice samples from 4 collection sites which included 2 farm sites and 2 sites where lice were taken from wild fish. Across all samples, there was a 92.14% similarity in the ITS-1 sequence. The percentage similarity in the ITS-1 sequence in samples office from two fish farms were 99.71% (site A) and 95.72% (site D) but only 86.90% (site B) and 86.03% (site C) similarity was shown in lice samples taken from sites where wild salmonids were caught. The greater similarity between the ITS-1 sequence within farm sites may be attributed to a restricted gene flow within lice populations in Atlantic salmon cage sites.