Trade practices are main factors involved in the transmission of viral haemorrhagic septicaemia

Viral haemorrhagic septicaemia (VHS), caused by the novirhabdovirus viral haemorrhagic septicaemia virus (VHSV), causes significant economic problems to European rainbow trout, Oncorhynchus mykiss (Walbaum), production. The virus isolates can be divided into four distinct genotypes with additional subgroups. The main source of outbreaks in European rainbow trout farming is sublineage Ia isolates. Recently, this group of isolates has been further subdivided in to two subclades of which the Ia‐2 consists of isolates occurring mainly in Continental Europe outside of Denmark. In this study, we sequenced the full‐length G‐gene sequences of 24 VHSV isolates that caused VHS outbreaks in Polish trout farms between 2005 and 2009. All these isolates were identified as genotype Ia‐2; they divided however into two genetically distinct subgroups, that we name Pol I and Pol II. The Pol I isolates mainly caused outbreaks in the southern part of Poland, while Pol II isolates predominantly were sampled in the north of Poland, although it seems that they have been transmitted to other parts of the country. Molecular epidemiology was used for characterization of transmission pathways. This study shows that a main cause of virus transmission appears to be movement of fish. At least in Polish circumstances trading practices appear to have significant impact on spreading of VHSV infection.     

Isolation of viral haemorrhagic septicaemia virus from mummichog, stickleback, striped bass and brown trout in eastern Canada

Viral haemorrhagic septicaemia virus (VHSV) was isolated from mortalities occurring in populations of mummichog, Fundulus heteroclitus, stickleback, Gasterosteus aculeatus aculeatus, brown trout, Salmo trutta, and striped bass, Morone saxatilis, in New Brunswick and Nova Scotia, Canada. The isolated viral strains produced a cytopathic effect on the epithelioma papillosum cyprini cell line. Serum neutralization indicated the virus was VHSV and sequencing identified the rhabdovirus isolates as the North American strain of VHSV. Phylogenetic analysis indicated that the isolates are closely related and form a distinguishable subgroup of North American type VHSV. To our knowledge, this is the first report of VHSV in mummichog and striped bass.     

Cholesterolaemia and triacylglycerolaemia in farmed rainbow trout,Oncorhynchus mykiss

Physiological (reference) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for blood plasma cholesterol (CHOL) and triacylglycerol (TGL) in farmed rainbow trout, Oncorhynchus mykiss, in raceway culture. The fish were given dry pelleted diets that contained 37–47% crude protein and 7–14% crude fat and were kept at a stocking density of 50 kg m^?3 in tanks provided with running freshwater at an ambient temperature of 3–16°C and dissolved oxygen, 8.4–11.5 mg L^?1. Blood was sampled between September and November at a photoperiod of 9:00–13:00 hours: 11:00–15:00 hours (light:dark). Cholesterol levels were significantly (P = 0.0001) greater in males (4.7–12.1 mmol L^?1, n = 34, mean weight 406 ± 138 g) than immature females (3.2–9.7 mmol L^?1, n = 386, mean weight 416 ± 147 g). Physiological range for TGL in immature females and males was 2.4–14.4 mmol L^?1 (n = 249, mean weight 418 ± 149 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for CHOL in males and in females, but not for TGL, in which a sinistral asymmetry was found. Correlation and regression analyses indicated significant (P = 0.0000) dependence between the CHOL and total protein (r² = 76.2%), CHOL and Fulton's condition factor (r² = 43.3%) and CHOL and absolute weight of liver (r² = 45.5%). Fluctuation in cholesterolaemia and triacylglycerolaemia, depending on nutrition and the aquaculture method is discussed below.     

Transfer of toxaphene and chlordane into farmed rainbow trout, Oncorhynchus mykiss (Walbaum) via feed

The study was carried out to quantitate the transfer of toxaphene and chlordane compounds from commercial fish feed into the edible part of rainbow trout, Oncorhynchus mykiss (Walbaum) under normal rearing conditions. Trout were fed with unspiked high energy feed for salmon (fat content 26–30%) over a period of 19 months. The average weight of trout increased from 10 g to more than 2092 g, reaching sizes of 51 cm length. Considerable amounts of toxaphene and chlordane residues were transferred from fish feed into trout muscle. Toxaphene concentrations increased up to 8.6 µg (Σ toxaphene indicator compounds 1–3) kg^?1 wet weight (w.w.) and chlordane reached 5.3 µg Σoxy‐, trans‐, cis‐chlordane + t‐nonachlor kg^?1 w.w. Results are also discussed on the contaminant levels based on the fat content and the effect of sexual maturation is considered in this study. The data allow the establishment of transfer rates for toxaphene and chlordane congeners from high energy diet into the edible part of farmed rainbow trout.     

Feeding behaviour of fast- and slow-growing strains of rainbow trout, Oncorhynchus mykiss (Walbaum), during first feeding

The development of feeding behaviour in two rainbow trout strains and their hybrid was compared at the transition from endogenous nutrition to exogenous feeding. After 60 min of acclimatization, the swimming pattern, spatial distribution, agonistic behaviour and snapping of fish were quantified by making 5-min observations on groups of five fish. Behavioural patterns were recorded in four replicate groups per treatment, before and after the introduction of dry trout feed or Artemia nauplii (actometric tests) or extracts of these (olfactometric tests). Fish of the slow growing strain were less active than those of the other two strains. Greater swimming activity observed in the fish of the fast growing strain was associated with more rapid feeding and a higher number of snapping responses. Such behavioural precocity is of interest because it could provide the basis for the growth differences between the fast and slow growing strains.     

Transfer of PCDDs and PCDFs into the edible parts of farmed rainbow trout, Oncorhynchus mykiss (Walbaum), via feed

This study was carried out to quantify the transfer of polychlorinated dibenzo‐p‐dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) from commercial fish feed into the edible part of rainbow trout (Oncorhynchus mykiss, Walbaum) under normal rearing conditions. Trout were fed with high‐energy feed for salmon (fat content 26–30%) over a period of 19 months. The average weight of the fish increased from 10 g to more than 2092 g, reaching a size of up to 51‐cm length (Tl). Considerable amounts of PCDDs and PCDFs were transferred from the fish feed into the tissue of the trout. Dioxin concentrations increased during the time of feeding from 0.054 up to 0.914 ng WHO‐PCDD/F‐TEQs kg^?1 wet weight and from 4.991 to 15.815 ng WHO‐PCDD/F‐TEQs kg^?1 fat. A correlation was found between the dioxin concentration of the feed and the resulting concentration in the fat of the muscle tissue. The data allow the establishment of transfer rates from a high‐energy diet to farmed rainbow trout.     

Identification and pathogenicity to rainbow trout, Oncorhynchus mykiss (Walbaum), of some aeromonads

Twelve strains of fish pathogenic aeromonads were identified by 16S rRNA sequencing as Aeromonas bestiarum , A. hydrophila , A. hydrophila subsp. dhakensis , A. salmonicida subsp. salmonicida , A. sobria biovar sobria and A. veronii biovar sobria. Following intramuscular injection, A. hydrophila subsp. dhakensis caused dark liquefying, raised furuncle-like lesions in rainbow trout within 48 h. Extracellular products of all cultures contained gelatinase and lecithinase, and most revealed lipase. Congo red absorption and siderophore production was recorded, but not so the suicide phenomenon or slime production. Sodium dodecyl sulphate polyacrylamide gel electrophoresis profile of the outer membrane proteins (OMP) revealed 10–25 bands, of which major bands were seen in the region of 32.5–47.5 and 62–83 kDa. Marked heterogenicity of the OMP and whole cell protein (WCP) profiles within and among the species was observed. Polypeptides of 83–173 kDa were detected in the WCP profile of the cultures, but they were not expressed in OMP fractions.     

Pancreas disease in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in Norway

The present paper describes, for the first time, clinical signs and pathological findings of pancreas disease (PD) in farmed Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), in sea water in Norway. Similarities and differences with reports of PD from Ireland and Scotland are discussed. Samples of 68 rainbow trout from disease outbreaks on 14 farms and from 155 Atlantic salmon from outbreaks on 20 farms collected from 1996 to 2004 were included in the present study. The histopathological findings of PD in Atlantic salmon and rainbow trout in sea water were similar. Acute PD, characterized by acute necrosis of exocrine pancreatic tissues, was detected in nine Atlantic salmon and three rainbow trout. Salmonid alphavirus (SAV) was identified in acute pancreatic necroses by immunohistochemistry. Most fish showed severe loss of exocrine pancreatic tissue combined with chronic myositis. Myocarditis was often but not consistently found. Kidneys from 40% and 64% of the rainbow trout and Atlantic salmon, respectively, had cells along the sinusoids that were packed with cytoplasmic eosinophilic granules. These cells resembled hypertrophied endothelial cells or elongated mast cell analogues. Histochemical staining properties and electron microscopy of these cells are presented. SAV was identified by RT-PCR and neutralizing antibodies against SAV were detected in blood samples.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

Immunization of rainbow trout, Oncorhynchus mykiss (Walbaum), with a low molecular mass fraction isolated from Flavobacterium psychrophilum

Flavobacterium psychrophilum, the causative agent of rainbow trout fry syndrome has become a widespread fish pathogen in freshwater aquaculture worldwide. In this study, a low molecular mass fraction (P25-33), with an approximate weight of 25-33 kDa, was identified among F. psychrophilum strains in an immunoblotting analysis with anti-F. psychrophilum sera. The immunogenic efficacy of the isolated and extracted P25-33 was investigated in two intraperitoneal immunization trials with rainbow trout, Oncorhynchus mykiss (Walbaum). The first trial included immunizations using P25-33 with Freund's complete adjuvant (FCA) and the second trial included immunizations using P25-33, formalin-inactivated whole and sonicated F. psychrophilum cell preparations without FCA. In both trials, antibody titres against F. psychrophilum were analysed with an enzyme-linked immunosorbent assay and the efficacy of the immunizations was determined by a challenge with F. psychrophilum. The P25-33 was shown to give rise to a protective immune response in rainbow trout after immunization with FCA, but not without FCA when a low concentration of P25-33 was used. Instead formalin-inactivated whole and sonicated cells of F. psychrophilum were able to protect the immunized fish more effectively when immunized without FCA. The results suggest that whole or sonicated F. psychrophilum cells could be better candidates for a cost-effective water-based injection vaccine than the immunogenic fraction.