A trypsin-like protease with apparent dual function in early Lepeophtheirus salmonis (Krøyer) development

<p>Abstract</p> <p>Background</p> <p>Trypsin-like serine proteases are involved in a large number of processes including digestive degradation, regulation of developmental processes, yolk degradation and yolk degradome activation. Trypsin like peptidases considered to be involved in digestion have been characterized in <it>Lepeophtheirus salmonis</it>. During these studies a trypsin-like peptidase which differed in a number of traits were identified.</p> <p>Results</p> <p>An intronless trypsin-like serine peptidase (<it>LsTryp10</it>) from <it>L., salmonis </it>was identified and characterized. <it>LsTryp10 </it>mRNA is evenly distributed in the ovaries and oocytes, but is located along the ova periphery. LsTryp10 protein is deposited in the oocytes and all embryonic cells. <it>LsTryp10 </it>mRNA translation and concurrent degradation after fertilization was found in the embryos demonstrating that LsTryp10 protein is produced both by the embryo and maternally. The results furthermore indicate that LsTryp10 protein of maternal origin has a distribution pattern different to that of embryonic origin.</p> <p>Conclusion</p> <p>Based on present data and previous studies of peptidases in oocytes and embryos, we hypothesize that maternally deposited LsTryp10 protein is involved in regulation of the yolk degradome. The function of LsTryp10 produced by the embryonic cells remains unknown. To our knowledge a similar expression pattern has not previously been reported for any protease.</p

No environmental effect on vaccine-induced reduced growth in Atlantic salmon Salmo salar smolts

Oil-adjuvanted vaccines reduce long-term growth in farmed Atlantic salmon Salmo salar, possibly via an increase in metabolic rate due to the energetic demands of the immune system. We tested this hypothesis by comparing sham-vaccinated to vaccinated smolts (total n = 2096, ca. 80 g) under different scenarios of water temperature (12 vs. 17°C, n = 1048 per temperature) and oxygen (O2) saturation (60, 70, 80, and 100%, n = 524 per O2 saturation level) in order to manipulate metabolic rate and O2 availability. We expected a more severe vaccination effect under conditions of high water temperature and low O2 saturation. Groups were kept in duplicate tanks under controlled temperature and hypoxia conditions for 7 wk post-vaccination before being transferred to uncontrolled common-garden natural conditions for 5 mo in a sea-cage. Body mass and length were recorded at the initiation and end of the controlled and uncontrolled environmental conditions. Vaccination and low O2 saturation at 17°C significantly reduced body mass (13 and 3% through vaccination and 9 and 20% through 60% O2 saturation at the end of the tank and sea-cage periods, respectively). However, there was no interaction between vaccination, temperature, and O2 saturation at the end of the tank or sea-cage period, lending no support to our hypothesis. A secondary observation was that emaciated ‘loser’ fish were mainly associated with the 17°C and low (mainly 60%) O2 saturation treatment. In conclusion, although vaccination led to a reduction in body mass, this effect was not influenced by environmental conditions expected to alter metabolic rate.publishedVersio

Phenotypic and genetic characterization of Piscirickettsia salmonis from Chilean and Canadian salmonids

Background. The study presents the phenotypic and genetic characterization of selected P. salmonis isolates from Atlantic salmon and rainbow trout suffering from SRS (salmonid rickettsial septicemia) in Chile and in Canada. The phenotypic characterization of the P. salmonis isolates were based on growth on different agar media (including a newly developed medium), different growth temperatures, antibiotics susceptibility and biochemical tests. Results. This is the first study differentiating Chilean P. salmonis isolates into two separate genetic groups. Genotyping, based on 16S rRNA-ITS and concatenated housekeeping genes grouped the selected isolates into two clades, constituted by the Chilean strains, while the Canadian isolates form a branch in the phylogenetic tree. The latter consisted of two isolates that were different in both genetic and phenotypic characteristics. The phylogenies and the MLST do not reflect the origin of the isolates with respect to host species. The isolates included were heterogeneous in phenotypic tests. Conclusions. The genotyping methods developed in this study provided a tool for separation of P. salmonis isolates into distinct clades. The SRS outbreaks in Chile are caused by minimum two different genetic groups of P. salmonis. This heterogeneity should be considered in future development of vaccines against this bacterium in Chile. Two different strains of P. salmonis, in regards to genetic and phenotypic characteristics, can occur in the same contemporary outbreak of SRS.publishedVersio

Complete sequence of RNA1 and subgenomic RNA3 of Atlantic halibut nodavirus (AHNV)

The Nodaviridae are divided into the alphanodavirus genus, which infects insects, and the betanodavirus genus, which infects fishes. Betanodaviruses are the causative agent of viral encephalopathy and retinopathy (VER) in a number of cultivated marine fish species. The Nodaviridae are small non-enveloped RNA viruses that contain a genome consisting of 2 single-stranded positive-sense RNA segments: RNA1 (3.1 kb), which encodes the viral part of the RNA-dependent RNA polymerase (RdRp); and RNA2 (1.4 kb), which encodes the capsid protein. In addition to RNA I and RNA2, a subgenomic transcript of RNA1, RNA3, is present in infected cells. We have cloned and sequenced RNAI from the Atlantic halibut Hippoglossus hippoglossus nodavirus (AHNV), and for the first time, the sequence of a betanodaviral subgenomic RNA3 has been determined. AHNV RNAI was 3100 nucleotides in length and contained a main open reading frame encoding a polypeptide of 981 amino acids. Conservative motifs for RdRp were found in the deduced amino acid sequence. RNA3 was 371 nucleotides in length, and contained an open reading frame encoding a peptide of 75 amino acids corresponding to a hypothetical B2 protein, although sequence alignments with the alphdnodavirus B2 proteins showed only marginal similarities. AHNV RNA replication in the fish cell-line SSN-1 (derived from striped snakehead) was analysed by Northern blot analysis, which indicated that RNA3 was synthesised in large amounts (compared to RNA1) at an early point in time post-infection

Consequences of Haemorrhagic Smolt Syndrome (HSS) for the Immune Status of Atlantic salmon (Salmo salar L.) (Case Study)

Haemorrhagic smolt syndrome (HSS) is a disorder of unknown aetiology causing losses in the fresh water phase of Atlantic salmon farming. Normally, the mortality is limited and symptoms disappear upon seawater exposure. In this case study, classical HSS pathology with internal organ haemorrhages and nephrocalcinosis was diagnosed, and the losses were substantial. Microarray analyses of head kidney revealed association between HSS and enhanced expression of stress genes and proteins reducing bioavailability of iron, heme, and retinol. In parallel, suppression of multiple metabolic pathways was observed. Up-regulation of genes encoding acute phase proteins, complement, and lectins indicated mild inflammation but without characteristic features of viral or bacterial infections. Microarray analyses highlighted several members of tumor necrosis factor receptor superfamily that may control development of B-cell immunity. Examination of IgM at the mRNA and protein levels showed the impact of HSS on vaccine responses. In fish without HSS symptoms (non-HSS), titres of vaccine specific antibodies to A-layer of Aeromonas salmonicida subsp. salmonicida and Moritella viscosa and antibodies binding to DNP-keyhole limpet hemocyanin (DNP-KLH), which are presumably polyreactive, were respectively four- and 14-fold higher than in HSS-diseased fish. Parallel sequencing of variable regions of immunoglobulin Mrevealed a larger size of most abundant clonotypes shared by multiple individuals in the non-HSS group. The results of the current case study indicated that, in addition to direct damage, HSS suppresses humoral immune responses including the production of specific and polyreactive antibodies.Consequences of Haemorrhagic Smolt Syndrome (HSS) for the Immune Status of Atlantic salmon (Salmo salar L.) (Case Study)publishedVersio

Consequences of Haemorrhagic Smolt Syndrome (HSS) for the Immune Status of Atlantic salmon (Salmo salar L.) (Case Study)

Haemorrhagic smolt syndrome (HSS) is a disorder of unknown aetiology causing losses in the fresh water phase of Atlantic salmon farming. Normally, the mortality is limited and symptoms disappear upon seawater exposure. In this case study, classical HSS pathology with internal organ haemorrhages and nephrocalcinosis was diagnosed, and the losses were substantial. Microarray analyses of head kidney revealed association between HSS and enhanced expression of stress genes and proteins reducing bioavailability of iron, heme, and retinol. In parallel, suppression of multiple metabolic pathways was observed. Up-regulation of genes encoding acute phase proteins, complement, and lectins indicated mild inflammation but without characteristic features of viral or bacterial infections. Microarray analyses highlighted several members of tumor necrosis factor receptor superfamily that may control development of B-cell immunity. Examination of IgM at the mRNA and protein levels showed the impact of HSS on vaccine responses. In fish without HSS symptoms (non-HSS), titres of vaccine specific antibodies to A-layer of Aeromonas salmonicida subsp. salmonicida and Moritella viscosa and antibodies binding to DNP-keyhole limpet hemocyanin (DNP-KLH), which are presumably polyreactive, were respectively four- and 14-fold higher than in HSS-diseased fish. Parallel sequencing of variable regions of immunoglobulin Mrevealed a larger size of most abundant clonotypes shared by multiple individuals in the non-HSS group. The results of the current case study indicated that, in addition to direct damage, HSS suppresses humoral immune responses including the production of specific and polyreactive antibodies

Consequences of Haemorrhagic Smolt Syndrome (HSS) for the Immune Status of Atlantic salmon (Salmo salar L.) (Case Study)

Haemorrhagic smolt syndrome (HSS) is a disorder of unknown aetiology causing losses in the fresh water phase of Atlantic salmon farming. Normally, the mortality is limited and symptoms disappear upon seawater exposure. In this case study, classical HSS pathology with internal organ haemorrhages and nephrocalcinosis was diagnosed, and the losses were substantial. Microarray analyses of head kidney revealed association between HSS and enhanced expression of stress genes and proteins reducing bioavailability of iron, heme, and retinol. In parallel, suppression of multiple metabolic pathways was observed. Up-regulation of genes encoding acute phase proteins, complement, and lectins indicated mild inflammation but without characteristic features of viral or bacterial infections. Microarray analyses highlighted several members of tumor necrosis factor receptor superfamily that may control development of B-cell immunity. Examination of IgM at the mRNA and protein levels showed the impact of HSS on vaccine responses. In fish without HSS symptoms (non-HSS), titres of vaccine specific antibodies to A-layer of Aeromonas salmonicida subsp. salmonicida and Moritella viscosa and antibodies binding to DNP-keyhole limpet hemocyanin (DNP-KLH), which are presumably polyreactive, were respectively four- and 14-fold higher than in HSS-diseased fish. Parallel sequencing of variable regions of immunoglobulin Mrevealed a larger size of most abundant clonotypes shared by multiple individuals in the non-HSS group. The results of the current case study indicated that, in addition to direct damage, HSS suppresses humoral immune responses including the production of specific and polyreactive antibodies

No environmental effect on vaccine-induced reduced growth in Atlantic salmon Salmo salar smolts

Oil-adjuvanted vaccines reduce long-term growth in farmed Atlantic salmon Salmo salar, possibly via an increase in metabolic rate due to the energetic demands of the immune system. We tested this hypothesis by comparing sham-vaccinated to vaccinated smolts (total n = 2096, ca. 80 g) under different scenarios of water temperature (12 vs. 17°C, n = 1048 per temperature) and oxygen (O2) saturation (60, 70, 80, and 100%, n = 524 per O2 saturation level) in order to manipulate metabolic rate and O2 availability. We expected a more severe vaccination effect under conditions of high water temperature and low O2 saturation. Groups were kept in duplicate tanks under controlled temperature and hypoxia conditions for 7 wk post-vaccination before being transferred to uncontrolled common-garden natural conditions for 5 mo in a sea-cage. Body mass and length were recorded at the initiation and end of the controlled and uncontrolled environmental conditions. Vaccination and low O2 saturation at 17°C significantly reduced body mass (13 and 3% through vaccination and 9 and 20% through 60% O2 saturation at the end of the tank and sea-cage periods, respectively). However, there was no interaction between vaccination, temperature, and O2 saturation at the end of the tank or sea-cage period, lending no support to our hypothesis. A secondary observation was that emaciated ‘loser’ fish were mainly associated with the 17°C and low (mainly 60%) O2 saturation treatment. In conclusion, although vaccination led to a reduction in body mass, this effect was not influenced by environmental conditions expected to alter metabolic rate

No environmental effect on vaccine-induced reduced growth in Atlantic salmon Salmo salar smolts

Oil-adjuvanted vaccines reduce long-term growth in farmed Atlantic salmon Salmo salar, possibly via an increase in metabolic rate due to the energetic demands of the immune system. We tested this hypothesis by comparing sham-vaccinated to vaccinated smolts (total n = 2096, ca. 80 g) under different scenarios of water temperature (12 vs. 17°C, n = 1048 per temperature) and oxygen (O2) saturation (60, 70, 80, and 100%, n = 524 per O2 saturation level) in order to manipulate metabolic rate and O2 availability. We expected a more severe vaccination effect under conditions of high water temperature and low O2 saturation. Groups were kept in duplicate tanks under controlled temperature and hypoxia conditions for 7 wk post-vaccination before being transferred to uncontrolled common-garden natural conditions for 5 mo in a sea-cage. Body mass and length were recorded at the initiation and end of the controlled and uncontrolled environmental conditions. Vaccination and low O2 saturation at 17°C significantly reduced body mass (13 and 3% through vaccination and 9 and 20% through 60% O2 saturation at the end of the tank and sea-cage periods, respectively). However, there was no interaction between vaccination, temperature, and O2 saturation at the end of the tank or sea-cage period, lending no support to our hypothesis. A secondary observation was that emaciated ‘loser’ fish were mainly associated with the 17°C and low (mainly 60%) O2 saturation treatment. In conclusion, although vaccination led to a reduction in body mass, this effect was not influenced by environmental conditions expected to alter metabolic rate