Effects of Greek juniper (Juniperus excelsa) extract on immune responses and disease resistance against Yersinia ruckeri in rainbow trout (Oncorhynchus mykiss).

This study investigated the effects of Greek juniper extract on immune responses of rainbow trout. In this experiment, 4 doses [0 (Control), 1 (J1), 4 (J4) and 8 (J8) mg/kg] of the extract were administered orally using an oral gavage twice a day for 14 days. Immune responses were measured on 7th and 14th days. On 14th day, Yersinia ruckeri was injected intraperitoneally to all fish of all groups. On 14th day, ORP in fish of J1 group increased significantly. Lysozyme activity (LA) was increased in J8 group on 7th day (p < .05). On 14th day, a significant decrease was determined in J1 and J4 treatments in LA. Myeloperoxidase activity was significantly decreased in all groups irrespective of sampling times (p < .05). Interleukin (IL)-1ß was significantly elevated in fish of J8 group on 7th day. IL-8 increased in fish of J8 and J4 groups on 7th day of the study. IL-12 gene expression was significantly up-regulated in J8 fish group on 7th day, and in J4 fish group on 14th day. Survival rate was higher in J8 treatment compared to the control and other treatments (p < .05). The results suggest that Juniperus excelsa provides protection against Y. ruckeri in rainbow trout.

Synbiotic feed supplementation significantly improves lipid utilization and shows discrete effects on disease resistance in rainbow trout (Oncorhynchus mykiss).

Enteric redmouth disease caused by the bacterial pathogen Yersinia ruckeri is the main reason for antimicrobial prescription, and a cause of substantial economic losses and decreased animal welfare in aquaculture. Given the importance of the intestinal microbiota in digestion and disease, our aim was to investigate whether synbiotic feed supplementation strategies could improve feed performance and disease resistance. Four experimental synbiotic feeds formulated with pre- and probiotics were tested against a commercially available probiotic control feed. Each experimental feed was evaluated for feed performance, effects on gross as well as intestinal morphometrics, and finally their effect on resistance against a waterborne experimental infection with Yersinia ruckeri serotype O1, biotype 2. While co-supplementing Pediococcus acidilactici with citrus flavonoids or bacterial paraprobiotics significantly improved utilization of feed lipid content relative to the control group, a decrease in lipid utilization was observed for feeds that combined P. acidilactici with yeast paraprobiotics. No significant improvements on disease resistance were observed. Still, synbiotic formulations including P. acidilactici led to reduced risks relative to that of the control group, while an increased relative risk was observed for a Bacillus-based formulation. In conclusion, two of the synbiotic supplements significantly improved lipid utilization and contributed to minor increases in disease resistance.

Effect of a multi-citrus extract-based feed additive on the survival of rainbow trout (Oncorhynchus mykiss) following challenge with Lactococcus garvieae.

Growing global concerns about antibiotic resistance have generated a considerable interest in the search for alternative environmental-friendly approaches. This study was aimed to assess the antimicrobial activity of a multi-citrus extract-based feed additive (Biocitro®) against some fish pathogens, as well as evaluate its capacity to protect rainbow trout (Oncorhynchus mykiss) to lactococcosis. A broth dilution method was used to determine the minimum inhibitory concentration (MIC) of Biocitro®, and the results showed a strong antibacterial activity against Aeromonas salmonicida, Lactococcus garvieae and Yersinia ruckeri with MIC values of 2.0 µg/mL. Afterwards, rainbow trout juveniles were fed a Biocitro®-enriched diet (750 mg/kg feed) at a daily rate of 1.5% body weight for 4 weeks, then they were challenged with L. garvieae by the cohabitation method. At the end of the experimental period, fish treated with Biocitro® showed significantly (P < 0.001) improved protection against L. garvieae compared to control fish. Although further studies are needed to understand how Biocitro® increases rainbow trout resistance to L. garvieae, this feed additive could be considered as a useful alternative to chemotherapeutic treatment in aquaculture.

Effects of dietary chitosan and nano-chitosan loaded clinoptilolite on growth and immune responses of rainbow trout (Oncorhynchus mykiss).

In this study, rainbow trout Oncorhynchus mykiss weighing 27.75 ± 0.34 g were orally subjected to eight experimental diets each in three replicates containing varying amounts of chitosan and nano-chitosan (0.05, 0.5 and 5 g kg-1) loaded in clinoptilolite (14.28 g kg-1) for 70 days; and the growth and immune responses were evaluated. Results showed that growth parameters in fish fed diets chit + clin2, chit + clin3, nchit + clin1, nchit + clin2 and nchit + clin3 were significantly higher than in fish fed the control diet. All feeds, except chit + clin3, and nchit + clin3, significantly increased the total protein level. Feeds containing chit + clin2, nchit + clin1, and nchit + clin2 significantly elevated serum lysozyme activity compared with the control group. All treatments, except chit + clin3, and nchit + clin3 exhibited higher serum immunoglobulin (Ig) level than control one. In contrast, diet nchit + clin1 significantly unregulated the expression of Ig M gene in fish head-kidney compared to other groups. Additionally, all feeds, except clinoptilolite, and nchit + clin3, significantly improved the serum complement activity. Diets chit + clin2, nchit + clin1, and nchit + clin2 also significantly elevated antibacterial activity against Yersinia ruckeri compared with the control diet. Expression of inducible nitric oxide synthase (iNOS) gene in fish fed diets clinoptilolite, chit + clin1, chit + clin3, nchit + clin1, nchit + clin2, and nchit + clin3 was significantly higher than the control diet. All diets, except clinoptilolite, increased IL-1ß gene expression compared to the control group. Present results suggest that diets supplemented with nchit + clin, especially at 0.05 g kg-1 nano-chitosan inclusion, could improve growth performance and immune parameters of rainbow trout.

Extract of common mallow (Malvae sylvestris) enhances growth, immunity, and resistance of rainbow trout (Oncorhynchus mykiss) fingerlings against Yersinia ruckeri infection.

The dietary effects of a native medicinal plant from Iran, common mallow (Malvae sylvestris), was evaluated on growth performance, innate immune parameters, mucosal immune parameters, and resistance of rainbow trout (Oncorhynchus mykiss) against Yersinia ruckeri. Therefore, 360 fish (initial weight 10.42 ± 0.09 g) were randomly distributed into 12 fiberglass tanks. Experimental diets supplemented with 0 (as control- C), 1% (M1), 3% (M2) and 5% (M3) levels of M. sylvestris flowers extract were fed to the fish based on 3% of body weight for 8 weeks. At the terminal sampling, growth performance, liver and digestive enzymes activities, blood and mucosal immune responses were determined. Results showed that M2 and M3 had greater final weight, weight gain, SGR, survival rate and lower FCR; higher levels of total protein, albumin, globulin, and lower cortisol levels in comparison to control; 5% extract also lowered cholesterol and glucose levels as well as Lactate Dehydrogenase (LDH) activity. We reported higher values of hematocrit, hemoglobin, Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Volume (MCV), White Blood Cell (WBC), Red Blood Cell (RBC) and lymphocytes for treated groups. Innate immune responses (Alternative complement activity (ACH50) in M2 and M3 group, total Immunoglobulin (Ig) and lysozyme in M3), mucosal immune parameters (ACH50, total Ig for M2 and M3 group and lysozyme in all treated groups) were enhanced. Activities of digestive enzymes (protease in all treated groups, amylase for 3 and 5%, while lipase only for 5%) and lower activity of liver ALT enzyme in individuals treated with highest dose was observed. Overall results indicated that the extract can positively affect growth performance and immune responses of rainbow trout.

Growth performance, haematological changes, immune response, antioxidant activity and disease resistance in rainbow trout (Oncorhynchus mykiss) fed diet supplemented with ellagic acid.

The present study was performed to investigate the effects of various levels of dietary ellagic acid (EA) on growth performance, haematological values, immune response, protection against Yersinia ruckeri infection, and oxidant/antioxidant status in rainbow trout, Oncorhynchus mykiss. Fish were fed with the control diet and three different experimental diets containing three graded levels of EA (50, 100 and 200 mg kg-1 diet) for 8 weeks. At the end of the experiment, the growth performance [weight gain (WG), specific growth rate (SGR) and feed conversion ratio (FCR)], haematological values [the red blood cell (RBC) count, haemoglobin (Hb) concentration, haematocrit (Ht) level and erythrocyte indices: mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC)], immune response [white blood cell (WBC) count, oxidative radical production (nitroblue tetrazolium (NBT) assay), phagocytic activity (PA) and phagocytic index (PI), total protein (TP) and immunoglobulin M (IgM) levels, serum bactericidal activity (BA), lysozyme (LYZ) and myeloperoxidase (MPO) activities] and oxidant/antioxidant status [tissue malondialdehyde (MDA) levels and superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities] were analysed. In addition, fish were challenged by Y. ruckeri and survival rate was recorded for 14 days. In the groups fed with EA the growth parameters such as WG, SGR, and FCR did not vary significantly. The RBC count, Hb concentration, and Ht level increased in the groups fed with EA when compared with the control group. However, there were no significant differences in the MCV, MCH and MCHC values among the groups. The results demonstrated enhancement in all the immunological parameters in the groups fed with EA compared to the control group. The results obtained from challenge with Y. ruckeri revealed reduction in the mortalities in the groups fed with EA. The dietary EA stimulated the SOD, CAT and GSH-Px activities in liver, head kidney and spleen as compared to the control group; however, a reverse trend was observed in the MDA levels of tissues. The present study suggest that EA can effectively enhance the haematological values, immune response, antioxidant capacity, and disease resistance in rainbow trout.

Dietary effects of Coriandrum sativum extract on growth performance, physiological and innate immune responses and resistance of rainbow trout (Oncorhynchus mykiss) against Yersinia ruckeri.

This investigation was aimed to determine the efficacy of coriander seed extract (Coriandrum sativum) on physiological responses, immunity and disease resistance of rainbow trout, Oncorhynchus mykiss for eight weeks. A total number of six hundred rainbow trout (62 ±â€¯0.81 g) were divided into four feeding groups including 0 (control), 0.5%, 1% and 2% of coriander seed extract (CSE). In the present study, rainbow trout fed with 2% of CSE showed significantly higher values of specific growth rate (SGR), final weight (FW) and condition factor (CF) in comparison with control group after eight weeks (P < 0.05). Regarding hematological indices results, the 2% dosage of CSE showed the highest amount of hematocrit and hemoglobin compared to control group (P < 0.05). In addition, significant improvement of lysozyme and alternative complement activity, were observed in 2% of CSE treatment (P < 0.05). After eight weeks post-feeding, 30 fish from each treatment were challenged with Yersinia ruckeri for 14 days. The findings presented that fish fed with CES, especially 2% of CSE inclusion, improved survival rate of rainbow trout against Y. ruckeri; however, there were no significant differences among the fish in control and treatment groups at the end of the eight weeks feeding with coriander seed extract. The present study demonstrated, dietary incorporation of coriander extract can improve growth factors, immunological indices and resistance of rainbow trout (O. mykiss) against Y. ruckeri infection.

Possible effect of hala extract (Pandanus tectorius) on immune status, anti-tumour and resistance to Yersinia ruckeri infection in rainbow trout (Oncorhynchus mykiss).

The possible effect of dietary administration of hala extract (Pandanus tectorius) on rainbow trout (Oncorhynchus mykiss) immune status as well as its effect as an anti-tumour agent was studied. Fish were divided into 4 groups before feeding with commercial diet (0%, control; 0.5%, 1% and 2% of hala extract) for 2 weeks. The effect of diet on the humoral immune parameters, ie total protein, myeloperoxidase content, antiproteases, lysozyme and bactericidal activities were studied. Also, the effect of the diets on the expression of some immune-related genes in rainbow trout head-kidney (TNF, LYZ2, IL-8 and CD-4) as well as tumour suppressor gene (WT-1a) was investigated. At the end of the feeding trial fish groups were challenged with Yersinia ruckeri. The results demonstrated enhancement in all the immune parameters in fish fed hala extract diets compared to control fish especially with the highest dose (2%) which recorded the highest significant increase (p < 0.05) in some parameters (total protein, myeloperoxidase content, antiproteases, and bactericidal activities) compared to the control. The results obtained from challenge with Y. ruckeri revealed reduction in the mortalities in fish groups fed with 1% and 2% doses of hala extract. Feeding with hala extract provoked upregulation in all immune- related genes. Again, the highest dose of hala extract showed a significant upregulation in WT1a expression (p < 0.05). The current study suggest that the hala extract, especially the highest dose, could be considered a good food additive to improve the immune status, resist tumour formation and to resist or control infectious diseases of rainbow trout.

Dietary olive leaf (Olea europea L.) extract alters some immune gene expression levels and disease resistance to Yersinia ruckeri infection in rainbow trout Oncorhynchus mykiss.

The aim for the present study was to investigate the effects of olive leaf (Olea europea L.) extract (OLE) on the control of Yersinia ruckeri infection in rainbow (Oncorhynchus mykiss) trout and to assess the impact on the expression of immune-related genes in the spleen and serum biochemical parameters of rainbow trout. Five experimental diets were prepared by adding 0.0%, 0.1%, 0.25%, 0.50% and 1.0% of OLE. Each diet was fed to triplicate groups of fish (mean body weight 51.22 ±â€¯3.04 g) twice a day (at 09:00 and 17:00 h) for 60 days. The dietary supplementation of OLE did not affect growth performance and feed utilization (P > 0.05). Major changes due to graded levels of OLE in the diets were observed in blood biochemical parameters (P < 0.05). TNFα, IL1-ß and IL-8 gene expressions were significanlty up-regulated in OLE 0.1% group compared with others (P < 0.05). Also, diet supplemented with OLE reduced mortality in rainbow trout fed with OLE 0.1% added diet. Present study suggests that OLE especially at 0.1% added feed may effectivelly enhance the serum biochemical parameters, survival rate and immune gene expression in rainbow trout.

Trans-cinnamic acid application for rainbow trout (Oncorhynchus mykiss): I. Effects on haematological, serum biochemical, non-specific immune and head kidney gene expression responses.

The present study investigated the effects of dietary trans-cinnamic acid (CA) on pre- and post-challenge haematological, serum biochemical, non-specific immune and head kidney gene expression responses of rainbow trout, Oncorhynchus mykiss juveniles. In this regard, fish with an average weight of 17.01 ±â€¯0.05 g were divided into five groups, and fed daily with an additive free basal diet (control); 250, 500, 750 or 1500 mg kg-1 CA for a 60-day period. Fish were sampled every 20 days during the experiment. On days 20, 40 and 60 (the pre-challenge period), the dietary CA especially at 250 and/or 500 mg kg-1 significantly increased blood granulocyte percentage, and serum total protein, globulin, lysozyme and total immunoglobulin values. Furthermore, dietary CA increased activities of phagocytic activity, respiratory burst and potential killing, and increased the expression levels of immune related genes [serum amyloid A (SAA), interleukin 8 (IL-8), interleukin 1, beta (IL-1ß), transforming growth factor beta (TGF-ß), tumor necrosis factor (TNF-α), and immunoglobulin T (IgT)] in the head kidney of fish fed with 250 and/or 500 mg kg-1 CA. Following 60 days of feeding, fish were challenged with Yersinia ruckeri and mortality was recorded for 20 days. Highest percentage survival (%) rate was found in the 250 and/or 500 mg kg-1 CA-supplemented feeding groups. During the post-challenge period, red blood cell (RBC) count, hematocrit (%), respiratory burst activity, and total antiprotease activity increased in fish fed with feed containing 500 mg kg-1 content. Moreover, markedly up-regulated the expression of related genes (SAA, IL-8, IL-1ß, TGF-ß, TNF-α, IFN-γ and IgM) in fish fed 250, 500 and/or 750 mg kg-1 CA. Therefore, feeding O. mykiss for 60 days with dietary CA at 250-500 mg kg-1 CA incorporation levels can be suggested as optimal to enhance the immunity and disease resistance against Y. ruckeri.

Immunological responses and disease resistance of rainbow trout (Oncorhynchus mykiss) juveniles following dietary administration of stinging nettle (Urtica dioica).

The present study investigated the effects of dietary supplementation of stinging nettle (Urtica dioica) on growth performance, skin mucus, immune response and disease resistance of rainbow trout (Oncorhynchus mykiss) fed with diets supplemented with U. dioica at 0, 1, 2 and 3%. After 8 weeks of feeding, the addition of U. dioica at 3% level resulted in improved weight gain, specific growth rate and feed conversion ratio significantly when compared to the other groups (P < 0.05). Hematological responses including: hematocrit (Htc), hemoglobin (Hb), lymphocyte and neutrophil populations enhanced significantly in fish fed 3% of stinging nettle when measured after 4 weeks; while, total red blood cells, white blood, Htc, Hb, lymphocyte and neutrophil populations significantly increased after 8 weeks in the same group (P < 0.05). Total serum protein and glucose contents increased significantly in fish fed stinging nettle at 3% when compared to the other groups after 8 weeks; however, triglycerides decreased significantly in the same group on the 4th and 8th week (P < 0.05). Additionally, several immune parameters, namely, IgM, lysozyme, complement components C3 and C4, and respiratory burst of blood leukocytes significantly increased in the 3% fed group on the 4th week; while, after 8 weeks the immune responses enhanced in fish fed 2 and 3% diets (P < 0.05). At the end of the feeding trial, mucus samples obtained from the fish fed stinging nettle supplementation exhibited improved antagonistic activities against several bacterial pathogens (Streptococcus iniae, Yersinia ruckeri, Vibrio anguillarum and Lactococcus garviae), skin mucus enzymes activities (alkaline phosphatase, lysozyme, protease and esterase) and protein levels in 2 and 3% groups with the highest being in case of 3% group when compared to the other groups (P < 0.05). The cumulative mortality of rainbow trout subjected to Y. ruckeri infectious exhibited relatively low mortality levels in all supplemented groups with the lowest being in fish fed 3% stinging nettle. The present findings demonstrated that dietary administration of U. dioica enhanced growth and stimulated fish immunity; thus, enabling the fish to be more resistant against bacterial infections.

Effect of feed supplementation with kynurenic acid on the morphology of the liver, kidney and gills in rainbow trout (Oncorhynchus mykiss Walbaum, 1792), healthy and experimentally infected with Yersinia ruckeri.

Kynurenic acid (KYNA) is an endogenous substance produced on the kynurenine pathway which is primarily known for its neuroactive properties. Recently, it has been proven that KYNA is a selective ligand for G protein-coupled receptor (GPR 35), presented on immunocompetent cells such as T lymphocytes. This opens up new possibilities of its application as an immunostimulating substance in aquaculture. Thus far, no histopathological investigations in fish have been completed to evaluate influence of KYNA supplementation in feed. This study has been undertaken to determine the effect of feed supplementation with KYNA (2.5, 25, 250 mg kg-1 of feed) for 28 days on the liver, gills and kidney in healthy fish and experimentally infected with Yersinia ruckeri. In a control group were observed a fatty liver, which is natural for this fish species in the autumn and winter season. As the dose of the supplement was increased, the fat liver changed, it decreased or completely disappeared. Additionally, inflammatory changes occurred in all the analysed organs, and their intensification was dose dependent. In the fish experimentally infected, KYNA caused aggravation of the signs in the liver, kidneys and gills, and the effect was dose dependent. The results implicate that KYNA may be a stressor for fish.

Genomic organization, expression and antimicrobial activity of a hepcidin from taimen (Hucho taimen, Pallas).

Hepcidin, an antimicrobial peptide, plays a crucial role in innate immune system of teleost fish. As a cysteine-rich peptide, hepcidin possesses a dual function including iron regulation and innate immunity. In the present study, a full-length hepcidin cDNA (HtHep) was cloned and characterized by RT-PCR and RACE techniques from taimen (Hucho taimen, Pallas), which is a type of rare, precious and cold-water fish species in China. The cDNA contains an open reading frame (ORF) of 267 bp encoding 88 amino acid (aa), with 170 bp located in the 5(') untranslated region (UTR) and 151 bp in the 3' UTR. The genomic sequences analysis showed that the HtHep gene consisted of three exons and two introns (with the length 94 and 251 bp, respectively). With a predicted molecular mass of 2881.4 Da and a theoretical pI of 8.53, the deduced amino acid encodes a signal peptide of 24 aa, prodomain of 39 aa and mature peptide of 25 aa. The signal peptidase (SA-VP) and the motif RX (K/R)R of propeptide convertase suggested the cleavage site of signal and mature peptide. Eight conserved cysteine residues were also identified and formed four disulfide bonds. Pair-wise alignments showed that HtHep clustered together with two fish species of Salmonidae family (Salmo salar and Oncorhynchus mykiss) in HAMP1 branch. Quantitative RT-PCR analysis indicated that the mRNA levels of HtHep were detected in a wide range of tissues and the highest level was detected in the liver. Its expression was also detected early during embryonic stage and could be up-regulated in the liver when challenged with pathogenic bacteria (Yersinia ruckeri). The recombinant HtHep (rHtHep) had antimicrobial activity against both gram-positive (Micrococcus lysodeikticus and Staphylococcus aureus) and gram-negative bacteria (Escherichia coli). Our results suggested that HtHep might be involved in the innate immune defense against bacterial pathogens in taimen.

Hemato - Immunological and biochemical parameters, skin antibacterial activity, and survival in rainbow trout (Oncorhynchus mykiss) following the diet supplemented with Mentha piperita against Yersinia ruckeri.

This study was aimed to assess the potential effects of Mentha piperita on the hemato - immunological and biochemical parameters, skin antibacterial activity and protection against Yersinia ruckeri infection in rainbow trout Oncorhynchus mykiss. Fish were divided into 4 groups before being fed diets supplemented with 0, 1, 2 and 3% of Mentha piperita (MP) plant extract for 8 weeks. Dose-dependent increases immune (both in skin mucus and blood serum) and hematological parameters (number of red and white cells, hematocrit and hemoglobin contents), as well as in respiratory burst activity, total protein, albumin, and neutrophil levels in fish fed supplemented diets compared to the control fish. Furthermore, dietary MP plant extract supplements have no significant effect on blood biochemical parameters and enzymatic activities of liver determined in serum of rainbow trout. After 8 weeks the cessation of feeding with MP plant extract, survival rates of 54.4%, 63.6% and 75.2% were recorded in groups which received 1, 2 and 3% of MP plant extract of feed, respectively, compared to 34.6% survivals in the control. This study underlying several positive effects of dietary administration of MP plant extract to farmed fish.

Green tea (Camellia sinensis) administration induces expression of immune relevant genes and biochemical parameters in rainbow trout (Oncorhynchus mykiss).

Present study elucidates the efficacy of green tea (Camellia sinensis) on growth performance, immune and antioxidant systems and cytokine gene expression in rainbow trout tissues. Green tea was supplemented at 20, 100, and 500 mg kg(-1) diet and fed to fish (average weight: 23.5 g) for 35 days. No remarkable changes in growth performance were observed among all test groups. Lower lipid peroxidation product and higher superoxide dismutase activity were noted in fish received the medium dose of green tea. Significant increase in serum bactericidal activity and total protein were recorded in all treatment groups. All doses of green tea up-regulated Interleukin-1ß transcription in the spleen, while Interleukin-1ß mRNA level decreased significantly in the kidney of low dose of green tea. Interleukin-6 mRNA level was up-regulated in the spleen of high dose of green tea and liver of middle and high doses of green tea. High dose and medium dose of green tea up-regulated the interleukin-8 transcription in the kidney and liver, respectively. Meanwhile, green tea inhibited the production of interleukin-10 in all treatment groups compared with control group. Medium dose of green tea up-regulated tumor necrosis factor-α transcription in all fish tissues, while high dose and low dose of green tea enhanced tumor necrosis factor-α mRNA levels in the kidney and spleen, respectively. Present study suggests that green tea especially at 100 mg kg(-1) feed may effectively enhance the antioxidant system and immune system in rainbow trout.

Sequencing of a second interleukin-10 gene in rainbow trout Oncorhynchus mykiss and comparative investigation of the expression and modulation of the paralogues in vitro and in vivo.

Interleukin-10 (IL-10) is a multifaceted cytokine that is produced by and effects a variety of cell populations, including macrophages, T, B and NK cells. The gene encoding for IL-10 has been isolated in mammals, birds, amphibians and recently in fish, with only single copy identified in each species. We report here a second IL-10 gene (tIL-10b) in rainbow trout that showed 92% identity in the coding region but only 50% identity in the 5'- and 3'-UTR to the known trout IL-10 paralogue, which we have now called tIL-10a. There is a short upstream open reading frame (uORF) within the 5'-untranslated region (UTR) of tIL-10a that may inhibit its translation, whilst in tIL-10b multiple mRNA instability motifs exist in the 3'-UTR, suggesting that the two IL-10 paralogues may have different mechanisms to regulate their expression post-transcriptionally. The expression of tIL-10a is generally higher than that of tIL-10b in most of the fourteen tissues examined and in the RTS-11, RTL and RTGill cell lines. However, the expression level of tIL-10b can exceed that of tIL-10a, as seen in vivo in the ovary of healthy fish and in the gills of Yersinia ruckeri challenged fish, and in vitro in head kidney (HK) leucocytes cultured for ≥ 8 h. The expression of the trout IL-10 paralogues can be up-regulated by LPS and polyIC in RTS-11 cells and by LPS, polyIC, PHA, PMA, calcium ionophore (CI) and IL-21 in head kidney leucocytes, as well as by Y. ruckeri infection, and can be modulated positively or negatively by IFN-γ. Synergistic effects on up-regulation of IL-10 expression were also seen between PHA and IL-21, as well as between PMA and CI. The expression kinetics of the IL-10 paralogues was also found to be different, suggesting that rainbow trout has evolved different pathways to regulate the expression of the two IL-10 paralogues at the transcriptional level.