Occurrence of viral haemorrhagic septicaemia virus (VHSV) in wild marine fish species in the coastal regions of Norway

This study was aimed at determining the occurrence of viral haemorrhagic septicaemia virus (VHSV) in selected stocks of wild fish species in the coastal regions of Norway. Six cruises were undertaken covering areas which included the coastal regions of northern Norway, the Norwegian Sea, the Barents Sea, and the Skagerrak area down to the Danish border. Collected, pooled samples of internal organs (kidney, spleen and heart) from 8395 fish were examined for the presence of VHSV by inoculation on BF-2 cells. Identification of virus was performed by a standard ELISA procedure with monoclonal antibody IP5B11, which is specific for the VHSV nucleocapsid protein (N-protein). VHSV was isolated from blue whiting and Norway pout in Skagerrak. No positive samples were detected in the northern coastal regions of Norway, the Norwegian Sea, or the Barents Sea. The findings indicate a very low occurrence of VHSV in the coastal regions of Norway. Geographically, the only positive samples were obtained from fish collected in areas where VHSV has previously been found in different species of fish.     

Screening for Viral Hemorrhagic Septicemia Virus in Marine Fish along the Norwegian Coastal Line

Viral hemorrhagic septicemia virus (VHSV) infects a wide range of marine fish species. To study the occurrence of VHSV in wild marine fish populations in Norwegian coastal waters and fjord systems a total of 1927 fish from 39 different species were sampled through 5 research cruises conducted in 2009 to 2011. In total, VHSV was detected by rRT-PCR in twelve samples originating from Atlantic herring (Clupea harengus), haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and silvery pout (Gadiculus argenteus). All fish tested positive in gills while four herring and one silvery pout also tested positive in internal organs. Successful virus isolation in cell culture was only obtained from one pooled Atlantic herring sample which shows that today''s PCR methodology have a much higher sensitivity than cell culture for detection of VHSV. Sequencing revealed that the positive samples belonged to VHSV genotype Ib and phylogenetic analysis shows that the isolate from Atlantic herring and silvery pout are closely related. All positive fish were sampled in the same area in the northern county of Finnmark. This is the first detection of VHSV in Atlantic herring this far north, and to our knowledge the first detection of VHSV in silvery pout. However, low prevalence of VHSV genotype Ib in Atlantic herring and other wild marine fish are well known in other parts of Europe. Earlier there have been a few reports of disease outbreaks in farmed rainbow trout with VHSV of genotype Ib, and our results show that there is a possibility of transfer of VHSV from wild to farmed fish along the Norwegian coast line. The impact of VHSV on wild fish is not well documented.     

Prevalence of viral haemorrhagic septicaemia virus in Danish marine fishes and its occurrence in new host species

In order to analyse the occurrence of viral haemorrhagic septicaemia virus (VHSV) in the marine waters around Denmark, staff from the Danish Institute for Food and Veterinary Research participated in 5 research cruises during 1998 to 2002 as a follow-up to 4 research cruises performed in 1996 to 1997. In total, 16,655 fish were examined virologically as 3569 samples. Forty fish species and 3 invertebrate species were represented. VHSV was isolated from 133 samples representing 8 species: herring Clupea harengus, sprat Sprattus sprattus, dab Limanda limanda, flounder Platichthys flesus, plaice Pleuronectes platessa, cod Gadus morhua, sand eel Ammodytes sp. and sand goby Pomatochistus minutus. Calculations showed that VHSV was more prevalent in the Baltic Sea in an area between Zealand and the island of Bornholm and the waters surrounding Bornholm than in the Kattegat, Skagerrak and along the North Sea coast of Denmark. This is the first report on the isolation of VHSV from dab, flounder and plaice and the first publication on VHSV from sand eel from Europe and sand goby.     

Observations on the food and diel feeding behaviour of pelagic O-group gadoids in the northern North Sea

The diets and diel feeding behaviour of six O-group gadoid species are described, the cod Gadus morhua (L.), the haddock Melanogramus aeglefinus (L.), whiting Merlangius merlangus (L.), saithe Pollachius virens (L.), Norway pout Trisopterus esmarkii (L.) and the blue whiting Micromesistius poutassou (R.). There were differences, in the main food groups selected by each species with respect to the type, size and number. The main prey consumed did not vary with depth. Cod, saithe and Norway pout had only one period of feeding activity whereas haddock and whiting apparently had two. The diets of haddock and whiting varied in association with these two periods. It is suggested that the differences between the diets were the result of each species having a different feeding behaviour which reflects that adopted in adult life.     

Susceptibility of juvenile Atlantic cod Gadus morhua to viral haemorrhagic septicaemia virus isolated from wild-caught Atlantic cod

A European strain of viral haemorrhagic septicaemia virus (VHSV) isolated from wild-caught cod Gadus morhua (H16/7/95) was shown to cause clinical disease and mortality in excess of 80% in juvenile Atlantic cod when administered by the intra-peritoneal (i.p.) route. No virus was recovered from cod cohabiting with experimentally infected fish at a ratio of 1:1, and no VHSV-associated mortality was demonstrated following immersion infection. External signs of disease in cod were the presence of exophthalmia and ascites. Virus was identified as VHSV by enzyme-linked immunosorbent assay (ELISA) and was recovered from both brain and organ pools (kidney, liver and spleen) of 100% of i.p. infected cod mortalities. Virus was also detected using an indirect immunofluorescence test on tissue imprints of kidney, liver, spleen and brain taken from moribund fish. The fact that cod were not susceptible to VHSV following waterborne exposure raises important questions surrounding the propagation, maintenance and impact of a naturally occurring reservoir of virus in the marine environment.     

The food of five gadoid species during the pelagic O-group phase in the northern North Sea

The diets of five gadoid species in the northern North Sea are described: the cod, Gadus morhua (L.); haddock, Melanogrammus aeglefinus (L.); whiting, Merlangius merlangus (L.); saithe, Pollachius virens (L.); and the Norway pout, Trisopterus esmarkii (L.). The diet of each species was shown to be a gradual progression to different prey species as the predators increased in length. There were differences in the food taken by each species with respect to numbers, species and size. These differences in diet suggest that the degree of competition between pelagic O-group gadoids is not great.     

New host and locality records for two protozoan (Myxosporea, Coccidia) parasites of marine fishes

This is the first report of the myxosporean Ortholinea orientalis from Atlantic herring Clupea harrengus. It infects the kidney tubules and previously was known from Pacific herring Clupea pallasii and navaga Eleginus navaga in the White Sea and North Pacific Ocean. This is also the first report of the coccidian Eimeria raibauti from Norway pout Trisopterus esmarkii. It infects the epithelium of the pyloric ceca and previously was known only from poorcod Trisopterus minutus in the Mediterranean Sea. The new records are both from the northern North Sea.     

Seasonal variations in the energy density of fishes in the North Sea

The energy density ( E _D, kJ g^-1 wet mass) of saithe Pollachius virens , haddock Melanogrammus aeglefinus , whiting Merlangius merlangus , Norway pout Trisopterus esmarki , herring Clupea harengus , sprat Sprattus sprattus , sandeel Ammodytes marinus and pearlsides Maurolicus Muelleri , from the North Sea, increased with total length, L _T. However, there was not always a significant ( P> 0·05) linear relationship between L _T and E _D. Seasonal differences in E _D were obvious in mature fish, while geographical differences were insignificant. For all species there was a highly significant correlation ( P< 0·0001) between the percent dry mass of the fish ( D S) and E _D. A general relationship was established for gadoids and sandeel E _D=–3·1492+0·3459 D _S and herring E _D=–4·6395+0·4170 D _S. Thus seasonal and size-specific data on E _D needed for bioenergetics and gastric evacuation models can be determined simply from D _S, which is considerably less costly and time consuming than calorimetry or proximate analysis.     

Susceptibility of juvenile sole Solea senegalensis to marine isolates of viral haemorrhagic septicaemia virus from wild and farmed fish

The susceptibility of sole Solea senegalensis to infection with 3 viral haemorrhagic septicaemia virus (VHSV) strains obtained from wild Greenland halibut Reinhardtius hippoglossoides and farmed turbot Psetta maxima was demonstrated. Fish were infected by an intraperitoneal (i.p.), immersion or cohabitational route, and maintained at 16 degrees C. Infection trials showed that VHSV isolates were pathogenic for sole fingerlings by i.p. injection and waterborne exposure causing moderate levels of mortality (10 to 55%). In addition, the mortality observed in fish cohabitating with i.p.-infected sole confirms horizontal transmission of the virus. However, the low rates of mortality registered in this challenge suggest that there is a low dissemination of virus by the i.p.-infected sole, which results in lower secondary challenge of the cohabitating fish. External signs of disease included haemorrhaging of the ventral area and ascitic fluid in the body cavity. Dead fish were tested for VHSV by both cell culture and RT-PCR assay, using pools of kidney and spleen from 10 individuals. Virus was recovered from most of the pools composed of dead fish. The results obtained in this study not only demonstrate the susceptibility of sole to the VHSV strains employed but also indicate that wild VHSV marine isolates represent a potential risk for sole aquaculture.     

The influence of feeding behaviour on growth of Atlantic cod (Gadus morhua,Linnaeus, 1758) in the North Sea

The objective of this study was to resolve key mechanisms driving individual growth patterns of Atlantic cod (Gadus morhua). Growth dynamics were analysed by linking growth patterns with stomach content composition and environmental temperature. Samples were collected in August/September of the years 2009, 2010 and 2011 in the north‐eastern part of the central North Sea. Prey selection was assessed by identification of individual prey items in the stomach content to species. Ten feeding groups were identified consisting of individuals with one prey type dominating their stomach contents (≥75% by mass), of which six were used for growth analyses: “Sandeel”, “Clupeids”, “Norway pout”, “Flatfishes”, “Crustaceans” and “Brittle stars”. For each group, growth patterns were estimated based on measures of otolith growth increments. The stomach contents showed that cod as a species are opportunistic in their prey selection, but at the same time indicated that the total, broad feeding niche width of the population is dominated by individual diet specialization and that many individuals temporally show a preference for a particular prey type. The contribution of invertebrates and particularly crustaceans decreased with increasing cod size, whereas that of fish and predominantly herring increased. Prey type had a significant effect on growth, while temperature had no effect. Slowest growth was observed in the cod group preying on sandeel, while cod preying on Norway pout showed the fastest growth. No significant difference was observed between groups preying on brittle stars, crustaceans, flatfishes and herring. Growth in the year before capture did however not differ between any of these groups. Across sampling years, growth chronology patterns were similar but not significantly influenced by temperature.     

Isolation and identification of Viral Haemorrhagic Septicaemia (VHS) viruses from cod Gadus morhua with the ulcus syndrome and from haddock Melanogrammus aeglefinus having skin haemorrhages in the North Sea

During fish disease surveys for marine rhabdoviruses in 1993 and 1995, the cod ulcus syndrome was seen widely in all ages of cod, especially the 2 to 5+ year classes. Viral Haemorrhagic Septicaemia Virus (VHSV) was isolated from a small proportion of the lesion-positive fish and these isolates were identified by immunofluorescence or ELISA. A serendipitous observation of dermal petechiae on haddock was made. VHSV was isolated from this lesion for the first time indicating a new host species for VHSV.     

Viral load of various tissues of rainbow trout challenged with viral haemorrhagic septicaemia virus at various stages of disease

Market-sized rainbow trout Oncorhynchus mykiss were challenged by waterborne exposure to viral haemorrhagic septicaemia virus (VHSV isolate of genogroup Ia). Fish were sampled at 4 stages of infection (before onset of clinical signs, clinically affected fish, mortalities and survivors) and the viral load determined in (1) internal organs, (2) muscle tissue and (3) brain and gill tissue. Virus levels were determined by virus titration and real-time RT-PCR. VHSV was detected by either method in the majority of fish before onset of clinical signs and in the survivor group as well as in all fish in the clinically affected fish and mortality groups. Mean virus amounts per mg of tissue determined by virus titration (TCID50) or real-time RT-PCR (copy number) were > 10(4) in preclinical fish, > 10(3.8) in clinically affected fish, > 10(3.9) in mortalities and > 10(1.2) in survivors. Virus levels tended to be highest in the internal organs of subclinical and clinically affected fish and in brain and gill tissue of survivors. The results demonstrate that significant levels of VHSV can be found in tissues of rainbow trout that may be marketed for human consumption, which may have relevance for the biosecurity of VHS-free areas.     

Effects of survey scale and water depth on the assessment of spatial distribution patterns of selected fish in the northern North Sea showing different levels of aggregation

Geostatistics was employed to investigate spatial structuring of herring, cod, dab, haddock and whiting at different spatial scales in the northern North Sea. Additionally, a structural analysis of the maximum water depth was carried out to assess habitat associations of fish. Linear, spherical, exponential and Gaussian models were fitted to the semivariograms, showing clear spatial autocorrelations. At the smaller scale, spatial structuring was weak for haddock, herring and dab, increasing at the greater spatial scale, with the exception of whiting. Mean catch rates, estimated classically and geostatistically, were in good agreement. Corresponding variances were clearly reduced at both spatial scales, when accounting for spatial distribution of the fish. At the greater survey scale a high level of habitat association was detected for haddock and whiting, while a poor habitat association was found for cod, dab and herring. The smaller scale seems to be the threshold at which spatial structuring of “cpue” could have marked influence on estimation error. Thus, survey scale is important when analysing spatial patterns and estimating mean biomass indices, and a sound analysis of relations in spatial structuring of fish and habitat conditions is essential to derive more precise estimates.     

Viral haemorrhagic septicaemia (VHS) outbreaks in Finnish rainbow trout farms

In Finland, viral haemorrhagic septicaemia virus (VHSV) was diagnosed for the first time in 2000 from 4 rainbow trout farms in brackish water. Since then the infection has spread and, by the end of 2004, VHSV had been isolated from 24 farms in 3 separate locations: 2 in the Baltic Sea and 1 in the Gulf of Finland. The pathogenicity of 3 of these isolates from 2 separate locations was analysed in infection experiments with rainbow trout fry. The cumulative mortalities induced by waterborne and intraperitoneal challenge were approximately 40 and 90 %, respectively. Pair-wise comparisons of the G and NV gene regions of Finnish VHSV isolates collected between 2000 and 2004 revealed that all isolates were closely related, with 99.3 to 100% nucleotide identity, which suggests the same origin of infection. Phylogenetic analysis revealed that they were closely related to the old freshwater isolates from rainbow trout in Denmark and to one old marine isolate from cod in the Baltic Sea, and that they were located close to the presumed ancestral source. As the Finnish isolates induce lower mortality than freshwater VHSV isolates in infection experiments, they could represent an intermediate stage of marine isolates evolving towards pathogenicity in rainbow trout.     

Detection of rainbow trout antibodies against viral haemorrhagic septicaemia virus (VHSV) by neutralisation test is highly dependent on the virus isolate used

Three serological tests, enzyme linked immunosorbent assay (ELISA), 50% plaque neutralisation test (50%PNT) and Western blotting (WB), were used to detect antibodies against viral haemorrhagic septicaemia virus (VHSV) in 50 rainbow trout broodstock from a rainbow trout farm endemically infected with VHS but with no clinical signs of infection. When the sera were examined by 50%PNT using the VHSV reference isolate DK-F1 or the heat attenuated DK-F25 mutant strain, no neutralizing antibodies were found. In contrast, when one of the virus isolates from the farm (homologous virus) was used in the 50%PNT, 90% of the fish were found to be positive. By examining a panel of different VHSV isolates in 50%PNT, it was demonstrated that the virus isolate used as test antigen could significantly affect the sensitivity and titre determination in 50%PNT for detection of rainbow trout antibodies against VHSV. When the sera were examined for the presence of VHSV antibodies by ELISA or WB, 61% were found to be positive. When conducting WB analysis, the viral glycoprotein was the protein most frequently recognized, followed by the viral nucleoprotein.     

Rapid detection of viral haemorrhagic septicaemia virus in fish by ELISA

An ELISA for rapid detection of viral haemorrhagic septicaemia virus (VHSV, Egtved virus) in fish tissue was developed and tested with regard to sensitivity. The test is a valuable addition to traditional isolation of VHSV in cell culture when a rapid diagnosis is needed.     

Experimental susceptibility of turbot Scophthalmus maximus to viral haemorrhagic septicaemia virus isolated from cultivated turbot

Juvenile pathogen-free turbot were infected with a viral haemorrhagic septicaemia virus (VHSV) isolate recovered from turbot cultivated on the island of Gigha, West Scotland. Mortality of 100% was recorded in fish infected via the intra-peritoneal (i.p.) route. Horizontal transmission of VHSV in sea water was demonstrated by cohabitation of naive fish with i.p. infected fish at a ratio of 1:1. The total cumulative average mortality in cohabiting fish was 60% by 60 d post-infection. Turbot infected via an immersion route exhibited a cumulative average mortality of 71% by the end of the experiment. VHSV identified by enzyme-linked immunosorbent assay (ELISA) was recovered from both organ (kidney and spleen) and brain samples of individual fish that died following infection by all experimental routes. These findings pose significant implications regarding the persistence of VHSV and its role in limiting natural populations of marine fish species. In addition, the establishment of infection models for the transmission of VHSV in sea water is of fundamental importance to the development of anti-VHSV vaccines in important commercial species such as turbot.     

Diet composition and biomass consumption of harbour seals in Telemark and Aust-Agder,Norwegian Skagerrak

ABSTRACT

To explore ecosystem dynamics and functions it is vital to obtain knowledge on predator–prey relations. Harbour seals are piscivorous predators that can come into conflict with fisheries. Recently, as the Skagerrak and Kattegat population of harbour seals has increased, claims have emerged that seals are depleting coastal cod populations. The diet of harbour seals in Norwegian Skagerrak was investigated based on otolith identification from scats. The overall seal diet included 20 different fish species/groups. The most important prey (combined index Q _i ) were haddock/pollack/saithe (32.7%), genus Trisopterus (Norway pout/poor cod/bib, 12.5%), plaice (12.4%) and herring (10.0%). Plaice also had the largest biomass (24.1%). Gadoids and pleuronectids comprised 88.6% of the diet (combined index Q _i ) and 87.1% in terms of biomass. Cod constituted 0.7% (combined index Q _i ) of the overall diet and 2.3% in terms of biomass. Fish length estimates showed that seals generally prefer small fish below minimum allowed landing size. Estimated total amount of fish consumed was 315 tons per year and was dominated by non-commercial species. Annual cod consumption was an estimated 7.1 tons, representing 5% of annual cod landings, suggesting that competition between local fisheries and harbour seals is limited.