Effect of Feed Additives as Alternatives to In-feed Antimicrobials on Production Performance and Intestinal Clostridium perfringens Counts in Broiler Chickens.

Numerous non-antibiotic feed additives (alternatives to antibiotics, ATAs) have been marketed, but few have been evaluated under uniform testing conditions modelling commercial flocks. We compared 24 ATA treatments and the ionophorous coccidiostat narasin against a diet without any feed additives. Feed conversion ratio and body weight gain were registered from day 0 to 28 in Ross 308 chickens housed on litter floor. The chickens were challenged with Eimeria spp., and cecal Clostridium perfringens (CP) counts were investigated. Active components from all ATA classes had a positive impact on intestinal health or production performance. Whereas narasin had a strong CP-reducing effect in combination with performance-promoting impact, only two ATA treatments achieved significantly beneficial effects on CP counts as well as feed conversion during the time span following Eimeria challenge. Active components present in these two treatments include a Bacillus subtilis probiotic strain, short- and medium-chain fatty acids and Saccharomyces cerevisiae components. Different ATA classes had beneficial impact during distinct rearing phases and on specific performance targets, suggesting that optimizing combinations and use of active components can make ATAs even more useful tools in broiler rearing without the use of in-feed antimicrobials. Further studies of promising ATAs and ATA combinations are required.

Estimating risk factors for the daily risk of developing clinical cardiomyopathy syndrome (CMS) on a fishgroup level.

Cardiomyopathy syndrome (CMS) is a viral disease, causing significant mortality and decreased welfare in farmed salmon in the North Atlantic Ocean. In Norway, it has become the most important disease in animal husbandry, affecting more than 100 farms each year. Control of CMS is based on mitigation of risk factors, since no treatment or vaccine is available. However, little is known about how the disease spreads and develops, thus rendering disease control difficult for farmers and competent authorities. The objective of the present study was to identify risk factors leading to the development of clinical CMS, using data provided from the salmon producers. Daily production data from individual fishgroups in more than 120 salmon farms along the coast of Norway from fish put to sea in 2012-2014 was collected. The data included cause-specific mortalities, which was used to identify outbreaks of CMS and risk factors for disease. A model for describing the daily probability of outbreak of CMS in each fishgroup was developed. The model was run to find the most likely value for each of the parameters, given the observed outbreak data. From the data, we found that fish in the southern region of Norway have a much higher risk of developing CMS than fish in mid and west (parameter estimates (PE) 4.43 (CI: 2.54-7.04) vs. 3.27 and 2.58 (CI: 2.45-4.37 and 2.01-3.57). Further, across all regions, fish put to sea in the late fall develop CMS twice as often as fish put to sea in the early spring (PE 2.18-2.59; CI:1.54-4.6). Previous outbreaks of pancreas disease increased the risk of getting CMS with 3.36 (CI:2.97-3.78) in the west and 1.41 (CI: 1.24-1.63) in the mid regions and decreased the risk with 0.519 (CI: 0.456-0.611) in the south. Previous outbreaks of heart- and skeletal muscle inflammation increased the risk of CMS with 1.56-1.73 (CI:1.34-2.11) in the mid and south regions, and had no effect in the west. In addition, we found that fish groups originating from certain hatcheries had a higher risk of CMS than other fishgroups, independent on which farm they were farmed on. The risk of developing CMS also increased with the number of days at sea. The use of production data in the study gave the possibility to study disease development on a fish group level, and on a daily basis. Thus, the identification of risk factors provides new possibilities for control of disease.

A Practical ELISA for Azaspiracids in Shellfish via Development of a New Plate-Coating Antigen.

Azaspiracids (AZAs) are a group of biotoxins that appear periodically in shellfish and can cause food poisoning in humans. Current methods for quantifying the regulated AZAs are restricted to LC-MS but are not well suited to detecting novel and unregulated AZAs. An ELISA method for total AZAs in shellfish was reported recently, but unfortunately, it used relatively large amounts of the AZA-1-containing plate-coating conjugate, consuming significant amounts of pure AZA-1 per assay. Therefore, a new plate-coater, OVA-cdiAZA1 was produced, resulting in an ELISA with a working range of 0.30-4.1 ng/mL and a limit of quantification of 37 µg/kg for AZA-1 in shellfish. This ELISA was nearly twice as sensitive as the previous ELISA while using 5-fold less plate-coater. The new ELISA displayed broad cross-reactivity toward AZAs, detecting all available quantitative AZA reference materials as well as the precursors to AZA-3 and AZA-6, and results from shellfish analyzed with the new ELISA showed excellent correlation ( R2 = 0.99) with total AZA-1-10 by LC-MS. The results suggest that the new ELISA is suitable for screening samples for total AZAs, even in cases where novel AZAs are present and regulated AZAs are absent, such as was reported recently from Puget Sound and the Bay of Naples.

Effects of sodium selenite and L-selenomethionine on feed intake, clinically relevant blood parameters and selenium species in plasma, colostrum and milk from high-yielding sows.

A field study in periparturient sows fed different dietary concentrations of either sodium selenite or L-selenomethionine (SeMet) was conducted to evaluate feed intake, haematological and biochemical parameters as well as to describe some key selenium (Se) species, namely selenoprotein P (SelP), selenoalbumin (SeAlb) and selenomethionine (SeMet) as well as total Se in plasma, colostrum and milk. Thirty-two sows were allotted to four treatments from 30 days (d) prepartum throughout on average a 32 d lactation period. Sodium selenite supplemented diets contained 0.40 and 0.60 mg Se/kg feed, while SeMet supplemented feed contained 0.26 and 0.43 mg Se/kg feed. Concentrations of sodium selenite and SeMet in complete feed exceeded the upper limits for total dietary Se and added organic Se, respectively, according to the European Union legislation. Blood samples were collected at initiation of the study, at farrowing and at weaning. Colostrum samples were collected at farrowing and milk samples at weaning. Se species were subjected to liquid chromatography, and total Se and Se species were determined using inductively coupled plasma mass spectrometry. The SeMet supplemented diets resulted in higher feed intake and in higher levels of total Se, SelP, SeAlb and SeMet in colostrum compared with sows fed sodium selenite. Similar results were obtained for levels of total Se and SeMet in milk at weaning. The higher dietary sodium selenite concentration in sows' feed did not increase the Se transfer into colostrum or milk when compared with those receiving the lower level of sodium selenite. However, the increase in serum-Zn from initiation until farrowing, observed in sows fed SeMet as well as the higher glutamate dehydrogenase activity in sodium selenite supplemented sows in this period might indicate a higher requirement of antioxidant defence in sodium selenite-supplemented sows. To our knowledge, the present data on Se species in plasma, colostrum and milk of sows represent the most complete investigation of Se in sows conducted to date. A higher amount of the above-mentioned Se species in the colostrum of sows supplemented with SeMet might strengthen the piglets' antioxidative system and passive immunity as well as improve their average daily weight gain. The higher feed intake in sows fed diets supplemented with SeMet is an interesting finding that warrants further investigation.

Monitoring infection with Piscine myocarditis virus and development of cardiomyopathy syndrome in farmed Atlantic salmon (Salmo salar L.) in Norway.

An epidemiological study was carried out in Norway in 2015-2018, investigating the development of infection with Piscine myocarditis virus (PMCV) and development of cardiomyopathy syndrome (CMS) in farmed Atlantic salmon. Cohorts from 12 sites were followed and sampled every month or every other month from sea transfer to slaughter. PMCV was detected at all sites and in all sampled cages, and fish in six sites developed clinical CMS. The initial infection happened between 1 and 7 months post-sea transfer, and the median time from infection with PMCV until outbreak of CMS was 6.5 months. Generally, fish from sites with CMS had higher viral titre and a higher prevalence of PMCV, compared to sites that did not develop clinical CMS. The virus persisted until the point of slaughter at most (11 out of 12) of the sites. The detection of PMCV in all sites suggests that PMCV is more widespread than previously known. Screening for PMCV as a tool to monitor impending outbreaks of CMS must be supported by observations of the health status of the fish and risk factors for development of disease.

Infectious Salmon Anemia and Farm-Level Culling Strategies.

Infectious salmon anemia (ISA) is an infectious disease, and outbreaks must be handled to avoid spread between salmon sea farms. Intensive culling at infected farms is an important biosecurity measure to avoid further spread but is also a costly intervention that farmers try to avoid. A lack of action, however, may lead to new outbreaks in nearby salmon sea farms, with severe impacts on both economy and animal welfare. Here, we aim to explore how a time delay between a detected outbreak and the culling of both infected cages and entire farms affects the further spread of the disease. We use a previously published model to calculate how many salmon sea farms were directly infected in each outbreak. To investigate the effect of culling on the further spread of disease, we use the number of months elapsed from the detected outbreak to (a) the first cage being depopulated, and (b) to the entire salmon sea farm being depopulated as predictors of how many new farms the virus was transmitted to, after controlling for contact between the farms. We show that the lapse in time before the first cage is depopulated correlates positively with how many new salmon sea farms are infected, indicating that infected cages should be culled with as little time delay as possible. The model does not have sufficient power to separate between culling of only cages assumed to be infected and the entire farm, and, consequently, provides no direct empirical evidence for the latter. Lack of evidence is not evidence, however, and we argue that a high probability of spread between cages in infected salmon sea farms still supports the depopulation of entire farms as the safest option.

Risk Factors Associated With Outbreaks of Infectious Salmon Anemia (ISA) With Unknown Source of Infection in Norway.

The occurrence of infectious salmon anemia (ISA) outbreaks in marine farmed Atlantic salmon constitutes a recurring challenge in Norway. Here, we aim to identify risk factors associated with ISA outbreaks with an unknown source of infection (referred to as primary ISA outbreaks). Primary ISA outbreaks are here defined by an earlier published transmission model. We explored a wide range of possible risk factors with logistic regression analysis, trying to explain occurrence of primary ISA with available data from all Norwegian farm sites from 2004 to June 2017. Explanatory variables included site latitude and a range of production and disease data. The mean annual risk of having a primary outbreak of ISA in Norway was 0.7% during this study period. We identified the occurrence of infectious pancreatic necrosis (IPN), having a stocking period longer than 2 months, having the site located at high latitude and high fish density (biomass per cage volume) in the first six months after transfer to sea site as significant risk factors (p < 0.05). We have identified factors related to management routines, other disease problems, and latitude that may help to understand the hitherto unidentified drivers behind the emergence of primary ISA outbreaks. Based on our findings, we also provide management advice that may reduce the incidence of primary ISA outbreaks.

Molecular tracing confirms that infection with infectious pancreatic necrosis virus follows the smolt from hatchery to grow-out farm.

Infectious pancreatic necrosis (IPN) is an important restraint to production of salmonids in aquaculture globally. In order to implement efficacious mitigation strategies for control of this disease, it is important to understand infection routes under current production systems. IPN virus has been shown to be transmitted vertically in Rainbow trout, from broodstock to fingerlings in hatcheries, and there is circumstantial evidence suggesting that vertical transmission can also occur in Atlantic salmon, in addition to horizontal transmission between grow-out fish in farms. In this study, we show that the smolt carries infection with IPN from hatchery to the marine farm. We do this by comparing sequences from fish groups taken both in hatcheries and on corresponding marine grow-out farms. We use statistical analysis to prove that sequences obtained from the same fish group in both hatchery and marine farm are more similar than sequences obtained from random fish groups on hatcheries and marine farms.

Effects of dietary sodium selenite and organic selenium sources on immune and inflammatory responses and selenium deposition in growing pigs.

The study was conducted to compare effects of different dietary Se sources (sodium selenite [NaSe], Se-enriched yeast [Se yeast] or L-selenomethionine [SeMet]) and one Se-deficient control diet on the expression of selected genes, hematological and clinical biochemical parameters, and muscle morphology in two parallel trials with finisher pigs. Se concentrations in blood plasma and tissues were also monitored. From the pigs in one of the parallel groups, muscle samples obtained from Musculus longissimus dorsi (LD) before and during the trial were examined. The pigs in the other parallel group were challenged once with lipopolysaccharide (LPS) intravenously. Transcriptional analyses of LD showed that selenogenes SelenoW and H were higher expressed in pigs fed Se-supplemented diets compared with control. Furthermore, the expression of interferon gamma and cyclooxygenase 2 was lower in the Se-supplemented pigs versus control. In whole blood samples prior to LPS, SelenoN, SelenoS and thioredoxin reductase 1 were higher expressed in pigs fed NaSe supplemented feed compared with the other groups, possibly indicating a higher level of oxidative stress. After LPS exposure glutathione peroxidase 1 and SelenoN were more reduced in pigs fed NaSe compared with pigs fed organic Se. Products of most above-mentioned genes are intertwined with the oxidant-antioxidant system. No significant effects of Se-source were found on hematologic parameters or microscopic anatomy. The Se-concentrations in various skeletal muscles and heart muscle were significantly different between the groups, with highest concentrations in pigs fed SeMet, followed by those fed Se yeast, NaSe, and control diet. Consistent with previous reports our results indicate that dietary Se at adequate levels can support the body's antioxidant system. Our results indicate that muscle fibers of pigs fed organic Se are less vulnerable to oxidative stress compared with the other groups.

Risk factors for outbreaks of infectious pancreatic necrosis (IPN) and associated mortality in Norwegian salmonid farming.

Infectious pancreatic necrosis (IPN) has for many years been considered one of the most important restraints to the production of salmonids in European aquaculture. In Norway, the disease is responsible for high losses in post-smolts in the first few weeks after sea transfer. Despite the importance of IPN, there are few epidemiological studies on risk factors and mitigation strategies. In this paper, we present analyses of data from all cohorts put to sea in 2009 to 2012 on Norwegian marine salmonid farms. The data used were obtained from national registers on salmonid production and disease outbreaks. The results showed that the risk of IPN outbreak was higher for spring versus autumn cohorts, Atlantic salmon versus rainbow trout and for cohorts on farms with previous history of IPN. The risk increased with increasing cohort size and infection pressure, whereas increasing temperature and weight at sea transfer decreased the risk. Estimations from a model of cumulative mortality within the first 6 mo after sea transfer showed that mortality in cohorts with IPN increased to approximately 7.2% as compared to a 'baseline' cohort with a mortality of 3.4%. If the cohort had both IPN and pancreas disease (PD), the estimated mortality increased to 12.9%, and cohorts with both IPN, PD and heart and skeletal muscle inflammation (HSMI) had an estimated mortality of 16.6%, when all other significant factors were kept constant (these were cohort type, year, temperature at sea transfer and weight at sea transfer). Our results provide valuable inputs for mitigation strategies and for economic modelling of consequences of disease.

Large scale modelling of salmon lice (Lepeophtheirus salmonis) infection pressure based on lice monitoring data from Norwegian salmonid farms.

Infection by parasitic sea lice is a substantial problem in industrial scale salmon farming. To control the problem, Norwegian salmonid farms are not permitted to exceed a threshold level of infection on their fish, and farms are required to monitor and report lice levels on a weekly basis to ensure compliance with the regulation. In the present study, we combine the monitoring data with a deterministic model for salmon lice population dynamics to estimate farm production of infectious lice stages. Furthermore, we use an empirical estimate of the relative risk of salmon lice transmission between farms, that depend on inter-farm distances, to estimate the external infection pressure at a farm site, i.e. the infection pressure from infective salmon lice of neighbouring farm origin. Finally, we test whether our estimates of infection pressure from neighbouring farms as well as internal within farm infection pressure, predicts subsequent development of infection in cohorts of farmed salmonids in their initial phase of marine production. We find that estimated external infection pressure is a main predictor of salmon lice population dynamics in newly stocked cohorts of salmonids. Our results emphasize the importance of keeping the production of infectious lice stages at low levels within local networks of salmon farms. Our model can easily be implemented for real time estimation of infection pressure at the national scale, utilizing the masses of data generated through the compulsory lice monitoring in salmon farms. The implementation of such a system should give the salmon industry greater predictability with respect to salmon lice infection levels, and aid the decision making process when the development of new farm sites are planned.

Climate and environmental change drives Ixodes ricinus geographical expansion at the northern range margin.

BACKGROUND: Global environmental change is causing spatial and temporal shifts in the distribution of species and the associated diseases of humans, domesticated animals and wildlife. In the on-going debate on the influence of climate change on vectors and vector-borne diseases, there is a lack of a comprehensive interdisciplinary multi-factorial approach utilizing high quality spatial and temporal data. METHODS: We explored biotic and abiotic factors associated with the latitudinal and altitudinal shifts in the distribution of Ixodes ricinus observed during the last three decades in Norway using antibodies against Anaplasma phagocytophilum in sheep as indicators for tick presence. Samples obtained from 2963 sheep from 90 farms in 3 ecologically different districts during 1978 - 2008 were analysed. We modelled the presence of antibodies against A. phagocytophilum to climatic-, environmental and demographic variables, and abundance of wild cervids and domestic animals, using mixed effect logistic regressions. RESULTS: Significant predictors were large diurnal fluctuations in ground surface temperature, spring precipitation, duration of snow cover, abundance of red deer and farm animals and bush encroachment/ecotones. The length of the growth season, mean temperature and the abundance of roe deer were not significant in the model. CONCLUSIONS: Our results highlight the need to consider climatic variables year-round to disentangle important seasonal variation, climatic threshold changes, climate variability and to consider the broader environmental change, including abiotic and biotic factors. The results offer novel insight in how tick and tick-borne disease distribution might be modified by future climate and environmental change.

Risk factors for cardiomyopathy syndrome (CMS) in Norwegian salmon farming.

Cardiomyopathy syndrome (CMS) has been an economically important disease in Norwegian aquaculture since the 1990s. In this study, data on monthly production characteristics and case registrations were combined in a cohort study and supplemented with a questionnaire-based case-control survey on management factors in order to identify risk factors for CMS. The cohort study included cases and controls from 2005 to 2012. From this dataset differences between all cases and controls were analyzed by a mixed effect multivariate logistic regression. From this we found that the probability of CMS increased with increasing time in the sea, infection pressure, and cohort size, and that cohorts which had previously been diagnosed with heart and skeletal muscle inflammation or which were in farms with a history of CMS in previous cohorts had double the odds of developing CMS. The model was then used to calculate the predicted value for each cohort from which additional data were obtained via the questionnaire-based survey and used as offset for calculating the probability of CMS in a semi-univariate analysis of additional risk factors. Finally, the model was used to calculate the probability of developing CMS in 100 different scenarios in which the cohorts were subject to increasingly worse conditions with regards to the risk factors from the dataset. We believe that this exercise is a good way of communicating the findings to farmers, so they can make informed decisions when trying to avoid CMS in their fish cohorts.

Ultra-deep pyrosequencing of partial surface protein genes from infectious Salmon Anaemia virus (ISAV) suggest novel mechanisms involved in transition to virulence.

Uncultivable HPR0 strains of infectious salmon anaemia viruses (ISAVs) infecting gills are non-virulent putative precursors of virulent ISAVs (vISAVs) causing systemic disease in farmed Atlantic salmon (Salmo salar). The transition to virulence involves two molecular events, a deletion in the highly polymorphic region (HPR) of the hemagglutinin-esterase (HE) gene and a Q266→L266 substitution or insertion next to the putative cleavage site (R267) in the fusion protein (F). We have performed ultra-deep pyrosequencing (UDPS) of these gene regions from healthy fish positive for HPR0 virus carrying full-length HPR sampled in a screening program, and a vISAV strain from an ISA outbreak at the same farming site three weeks later, and compared the mutant spectra. As the UDPS data shows the presence of both HE genotypes at both sampling times, and the outbreak strain was unlikely to be directly related to the HPR0 strain, this is the first report of a double infection with HPR0s and vISAVs. For F amplicon reads, mutation frequencies generating L266 codons in screening samples and Q266 codons in outbreak samples were not higher than at any random site. We suggest quasispecies heterogeneity as well as RNA structural properties are linked to transition to virulence. More specifically, a mechanism where selected single point mutations in the full-length HPR alter the RNA structure facilitating single- or sequential deletions in this region is proposed. The data provides stronger support for the deletion hypothesis, as opposed to recombination, as the responsible mechanism for generating the sequence deletions in HE.

Molecular and epidemiological characterization of avian influenza viruses from gulls and dabbling ducks in Norway.

BACKGROUND: Wild aquatic birds constitute the natural reservoir for avian influenza viruses (AIVs). Separate Eurasian and American AIV gene pools exist. Here, the prevalence and diversity of AIVs in gulls and dabbling ducks in Norway were described. The influence of host species and temporal changes on AIV prevalence was examined. Five AIVs from Norway, including three from common gull (Larus canus), were analyzed along with 10 available AIV genomes from gulls in Eurasia to search for evidence of intracontinental and intercontinental reassortment of gene segments encoding the internal viral proteins. METHODS: Swabs collected from 2417 dabbling ducks and gulls in the south-west of Norway during five ordinary hunting seasons (August-December) in the period 2005-2010 were analyzed for presence of AIV. Multivariate linear regression was used to identify associations between AIV prevalence, host species and sampling time. Five AIVs from mallard (Anas platyrhynchos) (H3N8, H9N2) and common gull (H6N8, H13N2, H16N3) were full-length characterized and phylogenetically analyzed together with GenBank reference sequences. RESULTS: Low pathogenic AIVs were detected in 15.5% (CI: 14.1-17.0) of the samples. The overall AIV prevalence was lower in December compared to that found in August to November (p = 0.003). AIV was detected in 18.7% (CI: 16.8-20.6) of the dabbling ducks. A high AIV prevalence of 7.8% (CI; 5.9-10.0) was found in gulls. A similar temporal pattern in AIV prevalence was found in both bird groups. Thirteen hemagglutinin and eight neuraminidase subtypes were detected. No evidence of intercontinental reassortment was found. Eurasian avian (non H13 and H16) PB2 or PA genes were identified in five reference Eurasian gull (H13 and H16) AIV genomes from GenBank. The NA gene from the Norwegian H13N2 gull isolate was of Eurasian avian origin. CONCLUSIONS: The similar temporal pattern in AIV prevalence found in dabbling ducks and gulls, the relatively high virus prevalence detected in gulls and the evidence of intracontinental reassortment in AIVs from gulls indicate that gulls that interact with dabbling ducks are likely to be mixing vessels for AIVs from waterfowl and gulls. Our results support that intercontinental reassortment is rare in AIVs from gulls in Eurasia.

DNA-fragments are transcytosed across CaCo-2 cells by adsorptive endocytosis and vesicular mediated transport.

Dietary DNA is degraded into shorter DNA-fragments and single nucleosides in the gastrointestinal tract. Dietary DNA is mainly taken up as single nucleosides and bases, but even dietary DNA-fragments of up to a few hundred bp are able to cross the intestinal barrier and enter the blood stream. The molecular mechanisms behind transport of DNA-fragments across the intestine and the effects of this transport on the organism are currently unknown. Here we investigate the transport of DNA-fragments across the intestinal barrier, focusing on transport mechanisms and rates. The human intestinal epithelial cell line CaCo-2 was used as a model. As DNA material a PCR-fragment of 633 bp was used and quantitative real time PCR was used as detection method. DNA-fragments were found to be transported across polarized CaCo-2 cells in the apical to basolateral direction (AB). After 90 min the difference in directionality AB vs. BA was >10(3) fold. Even undegraded DNA-fragments of 633 bp could be detected in the basolateral receiver compartment at this time point. Transport of DNA-fragments was sensitive to low temperature and inhibition of endosomal acidification. DNA-transport across CaCo-2 cells was not competed out with oligodeoxynucleotides, fucoidan, heparin, heparan sulphate and dextrane sulphate, while linearized plasmid DNA, on the other hand, reduced transcytosis of DNA-fragments by a factor of approximately 2. Our findings therefore suggest that vesicular transport is mediating transcytosis of dietary DNA-fragments across intestinal cells and that DNA binding proteins are involved in this process. If we extrapolate our findings to in vivo conditions it could be hypothesized that this transport mechanism has a function in the immune system.

Risk mapping of heart and skeletal muscle inflammation in salmon farming.

Heart and skeletal muscle inflammation (HSMI) is an infectious disease causing losses to the Norwegian salmon farming industry due to increased mortality and high morbidity in infected salmon. The disease is listed as a notifiable disease on list 3 (national list) by the Norwegian Food Safety Authority. HSMI is believed to be a viral disease, but the association to the recently discovered Piscine reovirus (PRV) remains unclear. Undoubtedly, other factors interact to determine whether PRV-infected fish develop disease or not. In this study, logistic regression was used to model the risk of an outbreak of HSMI at the cohort level, by including spatio-temporal risk factors. The data consisted of fish cohorts grown on geo-referenced farms from 2002 to 2010. The risk factors included were: infection pressure, cohort size (maximum number of fish), cohort index (smolt characteristics), cohort lifespan (months in sea) and a geo-index calculated as the position along a local polynomial regression line based on the longitude and latitude of each farm included in the study. The results showed that the risk of developing HSMI increased with increasing cohort lifespan, increasing infection pressure and increasing cohort size, and was mostly low for cohorts grown on farms in Southern-Norway, high for farms in Mid-Norway and variable for farms in Northern-Norway (based on the geo-index). The final model was used to explore three different scenarios with regards to the risk of developing HSMI, and to calculate the probability for each cohort of developing HSMI, independent of their actual disease-status. The model suggested that the probability of developing HSMI was much higher in Mid-Norway than in the rest of the country. Even though PRV seems to be widely distributed in the environment, the finding that infection pressure has a large influence on the probability of developing HSMI, suggests that it might be possible to reduce the number of clinical outbreaks, if measures are taken to reduce infection pressure. However, the prospects of controlling the spread of HSMI and reducing clinical outbreaks might be difficult because of indications of large distance spread of the disease.

Parental exposure to natural mixtures of POPs reduced embryo production and altered gene transcription in zebrafish embryos.

Determination of toxicity of complex mixtures has been proposed to be one of the most important challenges for modern toxicology. In this study we performed genome wide transcriptome profiling to assess potential toxicant induced changes in gene regulation in zebrafish embryos following parental exposure to two natural mixtures of persistent organic pollutants (POPs). The mixtures used were extracted from burbot (Lota lota) liver originating from two lakes (Lake Mjøsa and Lake Losna) belonging to the same freshwater system in Norway. The dominating groups of contaminants were polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane metabolites (DDTs). Because both mixtures used in the present study induced similar effects, it is likely that the same toxicants are involved. The Mjøsa mixture contains high levels of PBDEs while this group of pollutants is low in the Losna mixture. However, both mixtures contain substantial concentrations of PCB and DDT suggesting these contaminants as the predominant contributors to the toxicity observed. The observed effects included phenotypic traits, like embryo production and survival, and gene transcription changes corresponding with disease and biological functions such as cancer, reproductive system disease, cardiovascular disease, lipid and protein metabolism, small molecule biochemistry and cell cycle. The changes in gene transcription included genes regulated by HNF4A, insulin, LH, FSH and NF-κB which are known to be central regulators of endocrine signaling, metabolism, metabolic homeostasis, immune functions, cancer development and reproduction. The results suggest that relative low concentrations of the natural mixtures of POPs used in the present study might pose a threat to wild freshwater fish living in the lakes from which the POPs mixtures originated.

Cohort study of effect of vaccination on pancreas disease in Norwegian salmon aquaculture.

Pancreas disease (PD) is an economically important viral disease in Norwegian aquaculture, with 75 to 89 annual outbreaks from 2009 to 2011. To hinder further spread of disease from an initial endemic area on the west coast of Norway, measures for surveillance and control are in place, and the disease is notifiable on a national level. Since 2008, the Norwegian coastline has been divided into 2 administrative zones separated by a production-free area of 10 nautical miles at approximately 63°N. At the same time, a vaccination program involving most marine salmonid farms was initiated by the industry, using a vaccine against PD that was made commercially available in 2007. The effects of the vaccine in the field have been questioned, since the annual number of PD outbreaks has not decreased as expected. However, other production parameters can be used for evaluation of vaccine effect, and in this study the effects of vaccination on cumulative mortality, growth rate, feed conversion factor and number of discarded fish were analyzed using data collected from fish cohorts with and without PD put to sea between spring 2007 and spring 2009. The results show that vaccination against PD has a positive effect in reducing the number of outbreaks, and decreasing cumulative mortality and the number of fish discarded at slaughter.

Bovine lactoferrin digested with human gastrointestinal enzymes inhibits replication of human echovirus 5 in cell culture.

Many infant formulas are enriched with lactoferrin (Lf) because of its claimed beneficial effects on health. Native bovine Lf (bLf) is known to inhibit in vitro replication of human enteroviruses, a group of pathogenic viruses that replicate in the gut as their primary infection site. On the basis of a model digestion and human gastrointestinal enzymes, we hypothesized that bLf could retain its antiviral properties against enterovirus in the gastrointestinal tract, either as an intact protein or through bioactive peptide fragments released by digestive enzymes. To test our hypothesis, bLf was digested with human gastric juice and duodenal juice in a 2-step in vitro digestion model. Two gastric pH levels and reduction conditions were used to simulate physiological conditions in adults and infants. The antiviral activity of native bLf and of the digested fractions was studied on echovirus 5 in vitro, using various assay conditions, addressing several mechanisms for replication inhibition. Both native and digested bLf fractions revealed a significant inhibitory effect, when added before or simultaneously with the virus onto the cells. Furthermore, a significant stronger sustained antiviral effect was observed when bLf was fully digested in the gastric phase with fast pH reduction to 2.5, compared with native bLf, suggesting the release of antiviral peptides from bLf during the human digestion process. In conclusion, this study demonstrates that bLf may have a role in the prevention of human gastrointestinal virus infection under physiological conditions and that food containing bLf may protect against infection in vivo.