Project description:The transcriptional profile of intracellular P. salmonis was determined using a whole-genome microarray. Our microarray contained 5,309 different probes targeting 2,850. In our experimental design, the gene expression profile obtained from intracellular P. salmonis was compared with the transcriptome of control bacteria grown in culture medium. RNA from P. salmonis and SHK-1 cells were extracted simultaneously to avoid changes in transcriptomic profile after the bacterial purification process and eukaryotic RNA was depleted using the MicrobEnrich Kit.
Project description:The aquaculture industry has confronted severe economic losses due to infectious diseases in the last years. Piscirickettsiosis or Salmonid Rickettsial Septicaemia (SRS) is the bacterial disease caused by Piscirickettsia salmonis. This Gram-negative, non-motile, cellular pathogen has the ability to infect, survive, replicate, and propagate in salmonid monocytes/macrophages generating a systemic infection characterized by the colonization of several organs including kidney, liver, spleen, intestine, brain, ovary and gills. In this study, we attempted to determine whether global gene expression differences can be detected in different genetic groups of Atlantic salmon as a result of Piscirickettsia salmonis infection. Moreover, we sought to characterize the fish transcriptional response in order to reveal the mechanisms that might confer resistance in Atlantic salmon to an infection with Piscirickettsia salmonis. In doing so, after challenging with Piscirickettsia salmonis, we selected the families with the highest (HS) and the lowest (LS) recorded susceptibility for gene expression analysis using 32K cGRASP microarrays. Our results revealed in LS families expression changes are linked to iron depletion, as well as, low contents of iron in kidney cells and low bacterial load, indicated that the iron-withholding strategy of innate immunity is part of the mechanism of resistance against Piscirickettsia salmonis. This information contributes to elucidate the underlying mechanisms of resistance to Piscirickettsia salmonis infection in Atlantic salmon and to identify new candidate genes for selective breeding programmes.
Project description:The aquaculture industry has confronted severe economic losses due to infectious diseases in the last years. Piscirickettsiosis or Salmonid Rickettsial Septicaemia (SRS) is the bacterial disease caused by Piscirickettsia salmonis. This Gram-negative, non-motile, cellular pathogen has the ability to infect, survive, replicate, and propagate in salmonid monocytes/macrophages generating a systemic infection characterized by the colonization of several organs including kidney, liver, spleen, intestine, brain, ovary and gills. In this study, we attempted to determine whether global gene expression differences can be detected in different genetic groups of Atlantic salmon as a result of Piscirickettsia salmonis infection. Moreover, we sought to characterize the fish transcriptional response in order to reveal the mechanisms that might confer resistance in Atlantic salmon to an infection with Piscirickettsia salmonis. In doing so, after challenging with Piscirickettsia salmonis, we selected the families with the highest (HS) and the lowest (LS) recorded susceptibility for gene expression analysis using 32K cGRASP microarrays. Our results revealed in LS families expression changes are linked to iron depletion, as well as, low contents of iron in kidney cells and low bacterial load, indicated that the iron-withholding strategy of innate immunity is part of the mechanism of resistance against Piscirickettsia salmonis. This information contributes to elucidate the underlying mechanisms of resistance to Piscirickettsia salmonis infection in Atlantic salmon and to identify new candidate genes for selective breeding programmes. Forty full-sibling families of Atlantic salmon (Salmo salar) were infected by intraperitoneal injection with 0.2 mL Piscirickettsia salmonis (PS889, isolated from Oncorhynchus kisutch, 1M-CM-^W104 PFU/mL). After forty days, the fishes were harvested and the cumulative mortality (dead fish / total fish) for each family was calculated. For the second challenge, the six families with the highest cumulative mortality levels were considered of relatively high susceptibility (HS) and the six families with the lowest cumulative mortality levels were considered of relatively low susceptibility (LS) to the infection. Five control and five infected fish from three HS and three LS families were analyzed. For each HS and LS family, pools of RNA from control and infected fish were prepared separately and were reverse transcribed. Four slides were for each used family hybridized including two dye-swaped slides. Labeled samples were hybridized on a 32K cDNA microarray, developed at the Consortium for Genomics Research on All Salmonids Project (cGRASP), GEO accession number: GPL8904.
Project description:An effective and economical vaccine against the Piscirickettsia salmonis pathogen is needed for sustainable salmon farming and to reduce disease-related economic losses. Consequently, the aquaculture industry urgently needs to investigate efficient prophylactic measures. Three protein-based vaccine prototypes against Piscirickettsia salmonis were prepared from a highly pathogenic Chilean isolate. Only one vaccine effectively protected Atlantic salmon (Salmo salar), in correlation with the induction of Piscirickettsia-specific IgM antibodies and a high induction of transcripts encoding pro-inflammatory cytokines (i.e. Il-1β and TNF-α). In addition, we studied the proteome fraction protein of P. salmonis strain Austral-005 using multidimensional protein identification technology. The analyzes identified 87 proteins of different subcellular origins, such as the cytoplasmic and membrane compartment, where many of them have virulence functions. The other two prototypes activated only the innate immune responses, but did not protect Salmo salar against Piscirickettsia salmonis. These results suggest that the knowledge of the formulation of vaccines based on P. salmonis proteins is useful as an effective therapy, this demonstrates the importance of the different research tools to improve the study of the different immune responses, resistance to diseases in the Atlantic salmon. We suggest that this vaccine can help prevent widespread infection by P. salmonis, in addition to being able to be used as a booster after a primary vaccine to maintain high levels of circulating protective antibodies, greatly helping to reduce the economic losses caused by the pathogen.
Project description:Piscirickettsia salmonis, the biological agent of SRS (Salmon Rickettsial Syndrome), is a facultative intracellular bacterium that can be divided into two genogroups (LF-89 and EM-90) with different virulence levels and patterns. There are studies that have found co-infection of these genogroups in salmonid farms in Chile, but it is essential to assess whether this competitive interaction within the host is related to virulence and changes in pathogen dynamics. In this work, we studied one isolate from each genogroup, EM-90 and LF-89 . The aim was to evaluate how co-cultures could affect their growth performance and virulence factors expression at in vivo cultures in Atlantic salmon. During in vivo co-cultures, transcriptomic analysis revealed an upregulation of transposases, flagellum-related genes (fliI and flgK), transporters and permeases that could unveil novel virulence effectors used in the early infection process of P. salmonis. Thus, our work has shown that the cohabitation of the genogroups of P. salmonis can modulate their behavior and virulence effectors expression. These data can contribute to new strategies and approaches to improve current health treatments against this salmonid bacterium.
Project description:This study investigates transcriptomic responses of Atlantic salmon lice, Lepeophtheirus salmonis exposed to cypermethrin, a commonly used antiparasitic agent used in aquaculture. Copepodid L. salmonis were exposed to cypermethrin (Betamax®) at a concentration of 1.0ppb
Project description:This study investigates sex-biased gene expression between populations of Atlantic and Pacific salmon lice, Lepeophtheirus salmonis. Two Atlantic L. salmonis populations were previously used for an array study (GSE56024) while a third dataset using Pacific L. salmonis was novel. Using all three populations, a consensus-based, meta-analysis approach was used to identify sex-biased and sex-specific genes.
Project description:This SuperSeries is composed of the following subset Series: GSE26981: Responses to ectoparasite salmon louse (Lepeophtheirus salmonis) in skin of Atlantic salmon GSE26984: Responses to ectoparasite salmon louse (Lepeophtheirus salmonis) in spleen of Atlantic salmon Refer to individual Series
Project description:Salmon infected with an ectoparasitic marine copepod, the salmon louse Lepeophtheirus salmonis, incur a wide variety of consequences depending upon host sensitivity. Juvenile pink salmon (Oncorhynchus gorbuscha) migrate from natal freshwater systems to the ocean at a young age relative to other Pacific salmon, and require rapid development of appropriate defences against marine pathogens. We analyzed the early transcriptomic responses of naïve juvenile pink salmon of sizes 0.3g (no scales), 0.7g (mid-scale development) and 2.4g (scales fully developed) to a low-level laboratory exposure with early moult stage L. salmonis. All infected size groups exhibited unique transcriptional profiles. Inflammation and inhibition of cell proliferation was identified in the smallest size class (0.3g), while increased glucose absorption and retention was identified in the middle size class (0.7g). Tissue-remodelling genes were also up-regulated in both the 0.3g and 0.7g size groups. Profiles of the 2.4g size class indicated cell-mediated immunity and possibly parasite-induced growth augmentation. Understanding a size-based threshold of resistance to L. salmonis is important for fisheries management. This work characterizes molecular responses reflecting the gradual development of innate immunity to L. salmonis between the susceptible (0.3g) and refractory (2.4g) pink salmon size classes.