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Notes: Preliminary experiments are described which aimed to identify compounds that could inhibit the attachment of Flavobacterium branchiophilum strains LAB4a and ATCC 35035 to the gills of rainbow trout. Total inhibition was never achieved, regardless of the compound tested. Formalin-killed or acetone-killed F. branchiophilum cells retained at least some of their adherent nature, relative to untreated (live) cells. Adherence was reduced by 22–33% following immersion of fish in one litre of water containing 0.21 mg of a homologous crude fimbrial extract. When fish were immersed in water containing hyperimmune rainbow trout antiLAB4a serum, a dose-dependent decrease in attachment (a reduction of 15% to 63%) of LAB4a to the gills was observed. Rainbow trout anti-LAB4a serum also reduced the attachment of ATCC 35035 to the gills, but this reduction was not significant. Adherence of LAB4a was not inhibited following exposure of fish to group 1 carbohydrates (arabinose, mannose and xylose), group 2 carbohydrates (dextrose, galactose and lactose), group 3 carbohydrates (galactosamine, glucosamine and fucose) or group 4 carbohydrates (N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, N-acetyl-neuraminic acid and the globoceramide glycolipid from human erythrocytes). In contrast, when rainbow trout erythrocytes were incubated with the bacteria prior to bath challenge, this resulted in an 87% and 53% reduction in gill-associated LAB4a and ATCC 35035 antigen, respectively, following immersion of rainbow trout in this suspension.

Notes: A severe necrotic myositis characterized by large intramuscular bullae was identified in 100–500 g rainbow trout, Oncorhynchus mykiss (Walbaum), maintained in freshwater netpens. Mortality increased as the temperature decreased, with mortality rates peaking at 10% per month. Flexibacter psychrophilus was recovered from the muscle lesions and kidneys of affected fish. The gross and histological description of the disease, and the phenotypic characteristics of the causative agent are described. The necrotic myositis isolate was utilized to experimentally reproduce the disease.

Notes: Abstract A skin disease of intensively reared salmonids in Ontario hatcheries, known to the farmers as‘no-mucus skin disease’, is reported for the first time. It was characterized by erosive and ulcerative lesions found mainly on the flanks of fingerlings, which resulted in exposure of the tips of the scales. Associated with these lesions were colonies of bacteria seen in the SEM to be clustered round the mucous cell pores and under-running the margins of epithelial cells. The cause of this condition is unknown, although the response of fish to formalin treatment, and the presence of bacilli seen in skin scrapings and in the SEM, suggest that bacteria are responsible.

Notes: Abstract. Recent shipments of goldfish into Canada have suffered major mortality within a short time of arrival. The pathology and bacteriology of several of these diseased fish are described. Lesions were either acute and necrotizing with haemorrhage, or they were granulomatous. In either case, the spleen was a prime target, although in severely affected animals other organs were similarly involved; these included anterior and posterior kidney (plus endocrine elements), liver, intestine, mesentery and brain. Associated with these lesions were intracellular bacteria, which were often arranged in a Chinese letters-like configuration, and which are presumed to be the cause of the disease. The Gram-negative bacterium was isolated from all fish, but its characteristics were such that it was not easily classified and its taxonomy therefore remains uncertain.

Notes: A crude extracellular preparation (CEP) from a strain of Flavobacteriumpsychrophilum recovered from a case of necrotic myositis affecting rainbow trout was capable of causing severe muscle necrosis in rainbow trout following intramuscular injection. Cell wall-associated preparations, however, were unable to produce similar lesions in experimentally injected fish. The CEP degraded gelatin and type II collagen but not type I or type IV collagen. Furthermore, the CEP did not degrade 2-furanacryloyl- l-leucylglycyl- l-prolyl-alanine (FALGPA), chondroitin sulphates A, B or C, heparan sulphate, keratan sulphate, hyaluronic acid, elastin or rainbow trout erythrocytes. The addition of the protease inhibitors 1,10-phenanthroline, ethylenediamine-tetraacetic acid (EDTA) and EGTA to the CEP halted its ability to degrade gelatin in vitro and to produce muscle necrosis in rainbow trout in vivo. In vitro and in vivo activity was restored following the addition of 1 m m zinc chloride to the protease inhibitor-treated CEP, suggesting that this strain of F. psychrophilum secretes a protein complex with zinc metalloprotease-like activity. This protein complex, therefore, appears to be involved in the pathogenesis of necrotic myositis in rainbow trout.

Notes: Recent reports of the isolation of infectious salmon anaemia virus (ISAV) from Atlantic salmon, Salmo salar L., affected by haemorrhagic kidney syndrome (HKS) suggest that ISAV can cause severe renal haemorrhage and necrosis in addition to well-known pathognomonic hepatocellular necrosis and haemorrhage. The prevalence of ISAV-induced pathognomonic renal HKS lesions and their correlation to pathognomonic hepatic lesions of infectious salmon anaemia (ISA) is not known. The present experimental infection of Atlantic salmon with a Canadian isolate of ISAV found that pathognomonic hepatic ISA lesions were present in 90.6% and pathognomonic renal HKS lesions in 78.1% of fish which died after the experimental challenge. Both pathognomonic hepatic ISA lesions and pathognomonic renal HKS lesions were found together in 65.6% of fish which died after ISAV challenge. The present study clearly demonstrates that ISAV can cause a very high prevalence of both HKS and ISA pathognomonic lesions.

Notes: An unusual form of bacterial gill disease (BGD) was identified which affected five species of cultured salmonids from Canada (i.e. rainbow trout, chinook salmon and Atlantic salmon), Norway (i.e. brown trout) and Chile (i.e. coho salmon). All outbreaks occurred at low water temperatures (〈 10 °C) and with clinical presentations distinct from classical BGD, which is caused by Flavobacterium branchiophilum. In contrast to classical BGD, fish did not show marked respiratory distress with flaring of the opercula, the animals did not orientate at the surface of the water column near inflow water or at the margins of the tanks, and the feed response of the fish was varied. While mortality was increased, it was not precipitous as in classical BGD. Eight outbreaks were examined in greater detail using histopathology, scanning electron microscopy, bacteriology and immunohistochemistry. Large numbers of small bacterial rods were seen adhering to the lamellar epithelium of affected gills from all outbreaks. Histologically, the lamellar epithelium appeared swollen, often with evidence of single cell degeneration and exfoliation. In more severe instances, the formation of lamellar synechiae was seen, usually associated with sequestration of bacteria between fused lamellae. By contrast with typical BGD, overt epithelial hyperplasia, lamellar fusion and filamental clubbing were not common sequelae to infection; instead, the end result was shortened and somewhat stubby lamellae covered with swollen epithelial cells. The predominant bacterium recovered from affected gills was a small, Gram-negative, motile, fluorescent pigment-producing rod that shared phenotypic characteristics with Pseudomonas fluorescens. Polyclonal antisera prepared against three representative isolates indicated a weak antigenic similarity among them. Immunohistochemistry corroborated this finding, in that the antisera reacted strongly with gill sections containing the homologous bacteria, but not against morphologically similar bacteria in heterologous sections. A Gram-negative, yellow pigmented bacterium (YPB), identified as Flavobacterium psychrophilum, was also recovered, but only from the gills in the Ontario outbreaks. Antiserum prepared against this YPB indicated an antigenic similarity among isolates recovered from the Ontario outbreaks, but immunohistochemistry failed to recognize antigenically related bacteria on the gills of fish from the other outbreaks. Based on the unusual clinical presentation and the histopathological appearance of the gills, in conjunction with the absence of filamentous bacteria associated with and recovered from affected gills, the present authors have called this condition ‘atypical bacterial gill disease’ or ABGD.

Notes: The early humoral responses of rainbow trout, Oncorhynchus mykiss (Walbaum), and brook trout, Salvelinus fontinalis (Mitchill), with sterile inflammation induced by intraperitoneal Lipogen Triple vaccination were compared to determine if genetic differences in susceptibility to furunculosis in salmonids correlated with different acute phase responses to vaccination. Similar severe acute sterile peritonitis occurred in response to Lipogen Triple in both species. Both species also had a rapid transient reduction in plasma iron concentration at 3 days. Moderate hypoferraemia persisted to day 14 in brook trout, but returned to normal by day 7 in rainbow trout. Plasma zinc decreased sharply 3 days after vaccination in rainbow trout and returned nearly to control levels by day 10; however, plasma zinc did not change in brook trout. Two-dimensional sodium dodecyl sulphate polyacrylamide electrophoresis of plasma proteins revealed that increased amounts of a 48-kDa protein group coincided with the hypoferraemic response in rainbow trout. In addition, a modest elevation in a 16-kDa protein group also occurred in rainbow trout. These studies demonstrated the rapid changes in plasma iron in both species and mild elevation of two putative acute phase plasma proteins associated with vaccine-induced inflammation in rainbow trout.

Notes: To further characterize the putative role of constitutive and inducible plasma proteins in innate resistance to furunculosis, the present authors compared the alterations in profiles of plasma proteins in resistant and susceptible salmonids, i.e. rainbow trout, Oncorhynchus mykiss (Walbaum), and brook trout, Salvelinus fontinalis (Mitchill), respectively. Rainbow trout were injected with prednisolone acetate and exposed to higher water temperature (18 °C versus 10 °C), or injected with purified lipopolysaccharide (LPS) from a virulent strain of Aeromonas salmonicida, and plasma components were examined by two-dimensional polyacrylamide electrophoresis. Two days after A. salmonicida LPS exposure, rainbow trout had a four- to five-fold increase in concentrations of plasma proteins composed of p48, p19 and p16 subunits, and a significant decrease in a 100-kDa protein group. Consistent elevation or depletion of proteins corresponding to previously reported rainbow trout A. salmonicida LPS-binding pentraxins and lectins in plasma were not observed. Brook trout exposed to A. salmonicida LPS did not have any consistent plasma protein changes. There were no significant alterations in major plasma proteins following temperature shock and prednisolone acetate administration in rainbow trout plasma. These studies demonstrate that rainbow trout with LPS-induced sterile inflammation have few alterations in major plasma proteins or LPS-binding proteins, and do not exhibit the spectrum of acute phase changes induced by inflammation in mammals.

Notes: The effect of hydrogen peroxide (H2O2) treatment on the clearance of formalin-killed Flavobacterium branchiophilum from the gills of rainbow trout was examined. In untreated control fish, clearance was characterized by a rapid initial phase, with 50% reduction in bacterial antigen in the first 12 h after exposure. The bacteria then cleared more gradually, with total clearance being achieved by 40 h. Treatment of fish with 100 p.p.m. H2O2 did not influence bacterial clearance compared to untreated controls. Exposure for 1 h to H2O2 concentrations ≤ 100 p.p.m. caused no detectable clinical signs or evidence of ultrastructural damage to the respiratory epithelium. However, levels in excess of this caused significant gill damage.