Blood chemistry and acid-base balance in rainbow trout Oncorhynchus mykiss with experimentally-induced acute bacterial gill disease

Rainbow trout were experimentally infected with the causative agent of bacterial gill disease (BGD) (Flavobacterium branchiophilum) via bath challenge. All fish were cannulated with dorsal aortic catheters, had nasogastric tubes sutured in place for feeding, and were maintained individually, in plexiglass boxes with a flow-through water system. Fish were either fed, or unfed during the trial. Acute changes in blood gas, serum biochemistry and clinical parameters were monitored. By 24h post-challenge, BGD-infected trout that had been fed had significant hypoxemia, hypercapnia, increased blood ammonia, hypoosmolality, hyponatremia, hypochloremia, and increased cough and respiratory rates when compared to control levels. Unfed BGD-infected trout had similar, but less severe blood gas and clinical changes, and no electrolyte disturbances. The BGD-induced hypoxemia is likely exacerbated by increased oxygen demands brought on by feeding. It is not known what association feeding has with the development of low serum ion levels in BGD-infected trout. This is the first study to report the use of fed fish, as opposed to unfed or starved trout, in obtaining blood chemistry values from indisturbed and cannulated animals.     

Atypical bacterial gill disease: a new form of bacterial gill disease affecting intensively reared salmonids

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.     

A field efficacy evaluation of emamectin benzoate for the control of sea lice on Atlantic salmon

This study evaluated the efficacy of emamectin benzoate, 0.2% aquaculture premix, against sea lice on Atlantic salmon in eastern Canada. Salmon pens received either emamectin benzoate, orally, in feed at 50 micrograms/kg body weight/day for 7 consecutive days, or the same diet with no added medication. The site veterinarian had the option of administering a bath treatment with azamethiphos to any pen in the trial. The mean number of lice per fish was lower (P < 0.05) in the experimental group when measured 1, 3, 4, and 6 weeks after the start of medication. Treatment efficacy was 70%, 88%, 95%, and 61%, respectively. Three azamethiphos bath treatments were applied to each control pen during the trial, while the treatment pens received no bath treatment. No gravid female parasites were observed on any fish in the treatment group, while these life stages were observed on fish in the control group. Orally administered emamectin benzoate was palatable and highly effective for control of sea lice on salmon.     

Samples sizes required to accurately quantify viral load and histologic lesion severity at the maternal–fetal interface of PRRSV-inoculated pregnant gilts

Porcine reproductive and respiratory syndrome virus (PRRSV) is transmitted vertically, causing fetal death in late gestation. Spatiotemporal distribution of virus at the maternal–fetal interface (MFI) is variable, and accurate assessment of viral concentration and lesions is thus subject to sampling error. Our objectives were: 1) to assess whether viral load and lesion severity in a single sample of endometrium (END) and placenta (PLC), collected near the base of the umbilical cord (the current standard), are representative of the entire organ; and 2) to compare sampling strategies and evaluate if spatial variation in viral load can be overcome by pooling of like-tissues. Spatially distinct pieces of END and PLC of 24 fetuses from PRRSV-2–infected dams were collected. PRRSV RNA quantified by RT-qPCR was compared in 5 individual pieces per fetus and in respective pools of tissue and extracted RNA. Three distinct pieces of MFI were assessed for histologic severity. Concordance correlation and kappa inter-rater agreement were used to characterize agreement among individual samples and pools. The viral load of individual samples and pools of END had greater concordance to a referent standard than did samples of PLC. Larger pool sizes had greater concordance than smaller pool sizes. Average viral load and lesion severity did not differ by location sampled, and no technical advantages of pooling tissues versus RNA extracts were found. We conclude that multiple pieces of MFI tissues must be evaluated to accurately assess lesion severity and viral load. Three pieces per fetus provided a reasonable balance of cost and logistic feasibility.     

Haemorrhagic kidney syndrome of Atlantic salmon, Salmo salar L.

This report describes a new syndrome affecting farmed Atlantic salmon on the Canadian east coast that has resulted in increased morbidity and mortality in affected stocks. The major pathological findings are apparent only microscopically and include renal interstitial haemorrhage and acute tubular necrosis and tubular casting. As a result, the disease has become known as haemorrhagic kidney syndrome (HKS). Affected fish are lethargic and anorectic, and lack external lesions. Clinically, HKS fish are anaemic, hypoproteinaemic and hyperosmolalic, with increased serum concentrations of sodium and chloride. At necropsy, internal changes ranged from apparently normal to include one or several of the following: swelling and/or patchy reddening of the kidney, pale gills, exophthalmos, serosanguinous ascites, darkening of the posterior intestine and splenomegaly. Ultrastructurally, viral inclusions were found in the cytoplasm of erythrocytes of HKS fish, and there were unusual electron‐dense inclusions within the tips of renal tubular microvilli of HKS fish. The significance and relevance of the ultrastructural findings to HKS are unknown. Virus isolation was attempted using CHSE, RTG‐2, FH‐10, BB and EPC cell lines; no virus was isolated. Bacteriological analysis failed to reveal significant pathogens. Analysis of tissues for heavy metals and pesticides was negative. Assays for clostridial toxins, lipopolysaccharide and verotoxins were negative. The aetiology of HKS remains unresolved.     

Inhibition of the attachment of Flavobacterium branchiophilum to the gills of rainbow trout, Oncorhynchus mykiss (Walbaum)

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.     

X-ray imaging of shock waves generated by high-pressure fuel sprays

Synchrotron x-radiography and a fast x-ray detector were used to record the time evolution of the transient fuel sprays from a high-pressure injector. A succession of 5.1-microsecond radiographs captured the propagation of the spray-induced shock waves in a gaseous medium and revealed the complex nature of the spray hydrodynamics. The monochromatic x-radiographs also allow quantitative analysis of the shock waves that has been difficult if not impossible with optical imaging. Under injection conditions similar to those found in operating engines, the fuel jets can exceed supersonic speeds and result in gaseous shock waves.