Charlotte Axén

In 2023, all but one Swedish restocking farm rearing Baltic salmon and sea trout fry experienced massive mortality caused by lipoid liver disease (LLD). The struck farms used the same feed, and this was suspected to be the cause of the outbreak. We performed two parallel feed trials in three salmon populations as well as fatty acid and antioxidant analyses of feed and fish to investigate the causal relationship between the feed and the LLD mortality. Significant mortality was induced in one population and LLD histopathology was induced in all three populations in groups fed the suspected feed (test feed) but not in the control groups, linking the test feed to the outbreak in 2023. Fatty acid analysis revealed a difference in bound fatty acid composition between the suspected feed and the control feeds, including higher amounts of erucic acid. Erucic acid has negative health effects for fish by altering lipid metabolism. It is present in for example, capelin, a main ingredient in this feed. The test feed also contained lower levels of several antioxidants, especially vitamin A. Antioxidative status of the fish could not be fully analysed but results indicated a difference in antioxidative status between the populations.

During routine sampling of northern pike, a male Esox lucius with circular blue-metallic granular spots mainly located on the head and back was identified. Histological investigations presented multifocally thickened epidermis rich in basophilic large structures with a granulated rim and a dense, non-granulated center. Other organs showed no signs of infection. Ultrastructural analysis of the skin revealed three different types of herpes-like structures predominantly located within enlarged vacuoles. PCR analysis and NGS of dissected skin tissue verified the presence of EsHV1 DNA. In this study, we describe the first identification of EsHV1 in mainland Europe. In addition, for the first time, full sequences of both the DNA polymerase and terminase of the virus is available, thus allowing for an improved phylogenetic placement of EsHV1 within the Alloherpesviridae family. In addition to the EsHV1 infected pike, we also observed that 11.1% of the pike were affected by lymphosarcoma, a hyperplasia-disease caused by retroviruses. In conclusion, viral infections in pike are relatively common and likely have consequences for the local population dynamics.

In November 2016, infectious pancreatic necrosis virus (IPNV) was isolated from a broodstock female of landlocked sea trout (Salmo trutta) in Lake Vänern in Sweden. VP2 gene sequencing placed the IPNV isolate in genogroup 6, for which pathogenicity is largely unknown. Lake Vänern hosts landlocked sea trout and salmon populations that are endangered, and thus the introduction of new pathogens poses a major threat. In this study we characterized the novel isolate by conducting an infection trial on three salmonid species present in Lake Vänern, whole genome sequencing of the isolate, and prevalence studies in the wild sea trout and salmon in Lake Vänern. During the infection trial, the pathogenicity of the Swedish isolate was compared to that of a pathogenic genogroup 5 isolate. Dead or moribund fish were collected, pooled, and analyzed by cell culture to identify infected individuals. In the trial, the Swedish isolate was detected in fewer sample pools in all three species compared to the genogroup 5 isolate. In addition, the prevalence studies showed a low prevalence (0.2–0.5%) of the virus in the feral salmonids in Lake Vänern. Together the data suggest that the novel Swedish IPNV genogroup 6 isolate is only mildly pathogenic to salmonids.

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum (Rs), can be transmitted both horizontally and vertically and there is no available cure or prophylaxis. The control of BKD requires continuous surveillance, which is challenging in aquaculture as well as in programs for conservation and restoration of salmonid fish strains. BKD is a notifiable disease in Sweden and is monitored through the mandatory health control program using a polyclonal ELISA for detection of the Rs p57 protein in kidney. Fish must be killed for sampling, an obvious disadvantage especially regarding valuable broodfish. The present study shows that gill-/cloacal swabs collected in vivo for real-time PCR (qPCR(gc)), allow a sensitive and specific detection of Rs. The sensitivity of qPCR(gc) was estimated to 97.8% (credible interval (ci) 93.8%-100%) compared to 98.3% (ci 92.7%-100%) and 48.8% (ci 38.8%-58.8%) of kidney samples for qPCR (qPCR(k)) and ELISA (ELISA(k)) respectively, by use of the Bayesian Latent Class Analysis (BLCA). Since the goal of the program is eradication of BKD the most sensitive test is preferrable. Using qPCR(gc) instead of ELISA(k) will result in a lower false negative rate and can be useful for surveillance in aquaculture and in breeding programs with valuable fish. However, a higher false positive rate warrants confirmatory lethal testing before a previously Rs negative farm is subject to restrictions.

Enteritis is the major cause of mortality in neonatal calves. Diagnostic work to detect the pathogens involved and identification of risk factors for such infections are imperative to improve calf health. Bovine norovirus (BNoV) and nebovirus prevalence was investigated using RT-PCR in 50 dairy herds in five geographic regions of Sweden. BNoV and nebovirus were found to be common infections in young dairy calves. BNoV was detected in 20% of 250 calves in 48% of the herds. Nebovirus was detected in 5% of the calves in 16% of the herds. Both infections were present in three herds (6%). BNoV-infected calves were younger (median age 7 days) than nebovirus-infected calves (median age 21 days). These infections were not detected in adult cows and young stock over 6 months of age tested in selected herds. Risk factors identified for BNoV infection were a high number of preweaned calves present at the time of sampling, colostrum uptake by suckling the dam only, calf kept in a single pen, and geographic location of herd, while no risk factors for nebovirus infection were identified in the multivariable analysis. Univariable analysis suggested risk factors for nebovirus infection to be geographic location and early separation from the dam.

Bacterial kidney disease (BKD) can be a devastating bacterial infection in salmonids, and it is present in aquaculture throughout the world. BKD is caused by the Gram‐positive facultative intracellular bacterium Renibacterium salmoninarum (R. salmoninarum) that is spread both horizontally and vertically. Disease signs include external ulcerations and blisters and internal signs such as organ swelling, granulomas, petechiae and ascites. In Sweden, BKD accounts for a significant income loss in aquacultures due to expensive decontamination of the facility and increased disease susceptibility for the immunocompromised fish leading to higher mortality rates. In addition, uncontrolled spread in aquaculture may threaten the survival of wild fish populations. The aim of our study was to investigate the prevalence of R. salmoninarum in wild salmonids caught in Swedish waters where net pen farms with a recent history of BKD are present. Four rivers with at least one BKD‐positive or recently BKD‐positive farm were selected. In addition, we evaluated the use of environmental DNA (eDNA) for surveillance and monitoring of ongoing infections at these locations. In total, 1058 fish were sampled from four different river systems, and of them 52 (4.9%) were positive for R. salmoninarum by antigen ELISA. Surprisingly, these fish were not evenly distributed between the four river systems, but 50 were caught in the same river (Ljungan). This accounts for an alarmingly high rate of 17% R. salmoninarum‐positive samples in wild salmonids in this area. This number is far above what was expected and clearly shows the risk with an open farming system as well as the importance of effective health monitoring programmes to avoid an uncontrolled spread of the disease. The use of eDNA for monitoring BKD is somewhat difficult to evaluate. Few of the water samples analysed were PCR positive for R. salmoninarum (2 of 38) and those were collected where no ELISA positive fish were identified. In addition to water, sediment samples were collected under a net pen farm that had recently slaughtered all fish due to ongoing R. salmoninarum infections. Sediment samples are more promising than water as 4 of 5 samples at one farming facility where positive for R. salmoninarum. Thus, sediment samples may be valuable for monitoring potential ongoing BKD in farms, without the need to sacrifice valuable fish.

Infectious disease is a major challenge in aquaculture and poses a constraint for the development of farming of new species. In 2017, Siberian sturgeon (Acipenser baerii) juveniles were imported from Italy to a Swedish farm. Transport conditions were suboptimal. Thirty percent died during transport and within the first days after arrival. Ten days after arrival, mortalities started to occur again, which prompted initiation of an investigation into the mortalities. Diseased fish were transported live to the National Veterinary Institute (SVA) for necropsy and further analysis. Pathological and histopathological investigation was conducted. Virology was performed on gills and internal organs by cell culture isolation and using specific PCR protocols against nervous necrosis virus (NNV) and Acipenser iridovirus European (AcIV-E). The juveniles displayed neurological signs such as lethargy, inability to maintain an upright position, and erratic swimming. Body condition was below normal, and gills were pale. One fish had petechial hemorrhages on the abdomen and the snout. Two specimens had intestinal hyperemia. Ventricles were air-filled, and swim bladders were deflated. Viral cell cultures gave negative results, but PCR analysis of gills and internal organs detected the presence of AcIV-E. We conclude that AcIV-E was associated with disease and high mortality in the sturgeon juveniles, and stress probably aggravated the course of the infection.

Background: Infectious disease is a major challenge in aquaculture and poses a constraint for development of farming of new species. In 2017, Siberian sturgeon (Acipenser baerii) juveniles were imported from Italy to a Swedish farm. Due to stressful conditions, 30% died during transport and in the first days after arrival. Ten days after arrival, mortalities started to occur again. Within two months, only 5% of the juveniles were still alive. Methods: Diseased fish were transported live to the National Veterinary Institute (SVA) for necropsy and further analysis. Pathological and histopathological investigation was conducted. Virology was performed on gills and internal organs by cell culture isolation and specific PCR protocols. Results: The juveniles displayed neurological signs such as lethargy, inability to maintain upright position and erratic swimming. Body condition was low. Gills were pale. One fish had petechial hemorrhage on the abdomen and the snout. The ventricles were air-filled with, but swim bladders were deflated. One specimen had intestinal hemorrhage. Viral cell cultures were negative, but PCR of gills and internal organs detected the presence of Acipenser Iridovirus European (AcIV-E). Conclusions: AcIV-E was associated with disease and high mortality in the sturgeon juveniles. Stress probably aggravated the course of the infection.

We report the first description of a new Rhabdoviridae tentatively named eelpout rhabdovirus (EpRV genus Perhabdovirus). This virus was associated with mass mortalities in eelpout (Zoarces viviparous, Linnaeus) along the Swedish Baltic Sea coast line in 2014. Diseased fish showed signs of central nervous system infection, and brain lesions were confirmed by histology. A cytopathogenic effect was observed in cell culture, but ELISAs for the epizootic piscine viral haemorrhagic septicaemia virus (VHSV), infectious pancreas necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and spring viraemia of carp virus (SVCV) were negative. Further investigations by chloroform inactivation, indirect fluorescence antibody test and electron microscopy indicated the presence of a rhabdovirus. By deep sequencing of original tissue suspension and infected cell culture supernatant, the full viral genome was assembled and we confirmed the presence of a rhabdovirus with 59.5% nucleotide similarity to the closest relative Siniperca chuatsi rhabdovirus. The full‐genome sequence of this new virus, eelpout rhabdovirus (EpRV), has been deposited in GenBank under accession number KR612230. An RT‐PCR based on the L‐gene sequence confirmed the presence of EpRV in sick/dead eelpout, but the virus was not found in control fish. Additional investigations to characterize the pathogenicity of EpRV are planned.