Humoral Innate Immune Parameters Research Articles

Immunomodulatory effect of different dietary ARA/EPA/DHA ratios in gilthead sea bream ( Sparus aurata ) juveniles after infection with Photobacterium damselae subsp. piscicida

The immunomodulatory effect of different dietary arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) ratios was evaluated in gilthead sea bream juveniles after intraperitoneal injection (i.p.) with formalin-killed Photobacterium damselae subsp. piscicida (Phdp). Fish were fed with four isoproteic (47% crude protein) and isolipidic (18% crude lipids) diets with the following long-chain polyunsaturated fatty acids (LC-PUFA) ratios: Diet A (2.0% ARA: 0.2% EPA: 0.1% DHA); Diet B (1.0% ARA: 0.4% EPA: 0.4% DHA); Diet C (0% ARA: 0.6% EPA: 0.6% DHA); Diet D (0% ARA: 0.3% EPA: 1.5% DHA). After being fed for 56 days with the experimental diets, fish from each dietary treatment were randomly selected and i.p. injected with 0.2 ml of killed Phdp (1 × 108 CFU ml−1) or phosphate-buffered saline (control group). Four and twenty-four hours (i.e., twice) after the i.p., the haematological profile, plasma humoral innate immune parameters, and the intestine expression of immune-related genes were assessed. Results showed that fish fed diet D (high DHA content) had increased complement activity and higher intestinal expression of cyclooxygenase 2 (cox2) and interleukin 1β (il-1β) at 4 and 24 hours after i.p., as well as increased intestinal tumour necrosis factor α (tnfα) expression at 4 hours. On the other hand, fish fed with diet A (high ARA content) showed higher plasma bactericidal activity against Phdp than fish fed with diet D. Overall, the results of this study suggest a positive modulatory effect of DHA in response to the pathogenic agent.

Effects of pre-treatment with waterborne selenium on redox homeostasis and humoral innate immune parameters in African catfish, Clarias gariepinus (Burchell, 1822), experimentally challenged with Serratia marcescens.

Bacterial infections have been associated with immune dysfunction and oxidative stress in cultured fish species while essential elements could boost immunity and exhibit antioxidant properties in fish. This study was therefore aimed at determining the effects of pre-treatment with waterborne selenium on humoral immunity and redox status of Clarias gariepinus experimentally challenged with Serratia marcescens. Juveniles C. gariepinus were pre-treated with 50µg/L selenium for 14days after which they were challenged with 5 × 103CFU/mL of S. marcescens via oral gavage for 24 or 48h. The control fish were not pre-treated with selenium and not challenged with bacteria. Thereafter, fish were sacrificed, blood collected into EDTA bottles for the determination of plasma nitric oxide levels and respiratory burst, and the liver excised for the determination of reduced glutathione, lipid peroxidation, and activities of catalase, superoxide dismutase, and glutathione peroxidase. Fish that were pre-treated with selenium prior to bacterial challenge (Sel + Bact) had decreased levels of nitric oxide and lipid peroxidation but a significant increase in the levels of reduced glutathione (at 48-h post-infection period only) compared to the fish challenged with bacteria without prior selenium pre-treatment (Bact). The respiratory burst and catalase activity decreased significantly in the Sel + Bact group especially at 48-h post-infection period while the activity of glutathione peroxidase increased significantly in the Sel + Bact group (at 24-h post-infection period only) compared to the Bact group. The results from this study showed that infection with S. marcescens is capable of disrupting the immune system and redox homeostasis in C. gariepinus, while pre-treatment with selenium has the ability to improve the physiological status of fish that were challenged with bacteria probably through its antioxidant properties. HIGHLIGHT: The pre-treatment of Clarias gariepinus to waterborne selenium for 14days improved the redox homeostasis and innate immunity of fish that were experimentally challenged with the bacterium, Serratia marcescens.

Effect of different dietary arachidonic, eicosapentaenoic, and docosahexaenoic acid content on selected immune parameters in gilthead sea bream (Sparus aurata)

The increasing replacement of fish oil (FO) by vegetable oils (VO) in aquafeeds for marine carnivorous species may lead to modifications of the dietary arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) levels and ratios that may compromise the fish immune status. Therefore, this study aimed to assess how different dietary ratios of ARA, EPA, and DHA modulate the immune response of gilthead sea bream juveniles. For that purpose, four isoproteic and isolipidic plant-based diets were formulated to include different dietary Long-Chain-Polyunsaturated Fatty Acids (LC-PUFA) ratios (%DM): Diet A (2.0% ARA: 0.2% EPA: 0.1% DHA); Diet B (1.0% ARA: 0.4% EPA: 0.4% DHA); Diet C (0% ARA: 0.6% EPA: 0.6% DHA); Diet D (0% ARA: 0.3% EPA: 1.5% DHA). Fish were fed the experimental diets for 56 days and then the haematological profile, humoral innate immune parameters, and expression of immune-related genes in the gut were determined. Our results showed that fish fed diet B exhibited an increase in the monocyte numbers, alternative complement activity, and bactericidal activity against Photobacterium damselae compared to the values found in fish fed other diets. Contrarily, peroxidase, antiproteases, and proteases activity and nitric oxide levels were not affected by the dietary treatments. Immune-related gene expression in the distal intestine did not show differences between dietary treatments. Overall, a diet with a balanced n-6 (ARA) and n-3 (EPA+DHA) LC-PUFA ratio seemed to better modulate the fish innate immune response, thus better preparing the fish innate immune system against potential immunological insults.

Heat-killed Bacillus sp. SJ-10 probiotic acts as a growth and humoral innate immunity response enhancer in olive flounder (Paralichthys olivaceus)

Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) Bacillus sp. SJ-10 (BSJ-10) probiotic (1 × 108 CFU g−1) on the growth and immunity of olive flounder (Paralichthys olivaceus). Flounder (averagely 9.64 g) were divided into two groups, and fed control and HK BSJ-10 (HKBSJ-10)-inoculated diets for 8 weeks. Investigations were carried out on growth and feed utilizations, innate immunity, serum biochemical parameters, microvilli length, and pro- and anti-inflammatory cytokine gene (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, and IL-10) transcriptions. Compared to control, HKBSJ-10 diet significantly (P < 0.05) enhanced weight gain and protein efficiency ratio, 1.17 and 1.11 folds respectively. Humoral innate immune parameters, lysozyme and superoxide dismutase in treatment group were also elevated by 1.34 and 1.16 folds. Similarly, an increased (P < 0.05) relative expressions of TNF-α, IL-1β, IL-6 were recorded in liver (2.71, 3.38, and 4.12 folds respectively), and gill (2.08, 1.98, and 1.81 folds respectively) than that of controls. Moreover, after challenge with Streptococcus iniae (1 × 108 CFU mL−1), the HKBSJ-10-fed group exhibited significantly higher protection (P < 0.05) against streptococcosis compared to controls, validating the observed changes in immune parameters and induction on the cytokine-encoding genes. Therefore, HKBSJ-10 increases growth, modulates innate immune parameters, and protects olive flounders against streptococcosis.

Increased stocking density causes changes in expression of selected stress- and immune-related genes, humoral innate immune parameters and stress responses of rainbow trout (Oncorhynchus mykiss).

The present study investigated the effects of various stocking densities on the health status (stress and immune responses) of rainbow trout (Onchorhynchus mykiss). Juvenile rainbow trout were acclimated, placed in circular tanks under stocking densities of 10, 40 and 80kgm(-3) and reared for 30 days. The relative expression of genes involved in stress and immunity such as HSP70, LyzII, TNF-1α, IL-1β, IL-8 and IFN-γ1 in the head kidney was determined. Serum cortisol, ACTH, total antioxidant capacity, osmolality and lactate were measured after 30 days of culture at different stocking densities (D1:10kgm(-3), D2: 40kgm(-3) and D3: 80kgm(-3)) as indices of stress responses. In addition, the effects of stocking densities on serum complement, bactericidal activity, agglutinating antibody titers, serum IgM, anti-protease activity, serum total protein and alkaline phosphatase of the fish were measured. HSP70 gene expression was significantly density-dependent upregulated in D2 and D3 densities compared to D1 (P<0.05). Also, there was significant downregulation in expression of LyzII, TNF-1α, IL-1β, IL-8 and IFN-γ1 in fish reared at density of either D2 or D3 (P<0.05). In terms of stress responses, serum ACTH, cortisol and lactate level showed significant density-dependent increase (P<0.05) while serum osmolality and total antioxidant capacity showed significant decline (P<0.05) in fish reared at higher densities (D2 and D3) compared to fish reared at lower density (D1) (P<0.05). Concordant with the expression of the immune-related genes, the serum complement and bactericidal activity as well as specific antibody titer against Aeromonas hydrophila, IgM and anti-protease activity decreased along with elevation of stocking density from D1 to D3 (P<0.05). However, different stocking densities had no significant effect on serum total protein level and alkaline phosphatase activity. These results suggested that elevation of stocking densities and crowding resulted in the increase in HSP70 gene expression and the levels of selected stress responses in the serum. However, there was down-regulation of immune genes expression and decreased innate immune responses in the fish. The mRNA expression of the genes and immune parameters that were measured in this study could be helpful in monitoring the health status and welfare of the fish in aquaculture systems particularly in relation to increased stocking densities.

Effects of dietary β-glucan on the growth and innate immune response of juvenile Persian sturgeon, Acipenser persicus.

The present study evaluated the effects of different levels of dietary β-glucan (MacroGard(®)) on growth performance and innate immune parameters in juvenile Persian sturgeon (Acipenser persicus). Fish (20.1 ± 0.8 g) were allocated into 12 tanks (15 fish per tank) and triplicate groups were fed a control diet or diets containing 0.1%, 0.2% and 0.3% β-glucan. After six weeks of daily feeding (3.0% body weight day(-1)), humoral innate immune parameters (serum immunoglobulin (Ig) levels, lysozyme activity and alternative complement activity (ACH50)) and growth parameters (final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR)) were examined. Innate immune responses (lysozyme activity and ACH50) were significantly higher in 0.2% and 0.3% β-glucan fed fish (P < 0.05). Similarly, elevated growth performance (final weight, SGR and FCR) was observed in fish fed 1%, 2% and 3% β-glucan compared to the control group (P < 0.05). However, dietary β-glucan had no significant effect on survival rate of fish (P > 0.05). These results indicate that β-glucan can be considered as a beneficial dietary supplement for improving the immune response and growth performance of Persian sturgeon.

Enhancement of secondary metabolites from Bacillus Licheniformis XY-52 on immune response and expression of some immune-related genes in common carp, Cyprinus carpio

This study was undertaken to isolate active secondary metabolites from immunostimulatory Bacillus Licheniformis XY-52 and evaluate their activities at 0%, 0.5%, 1.0%, and 2.0% doses supplementation with feed on immune response in common carp at weeks 1, 2, and 3. By applying chromatography techniques and successive recrystallization, two purified metabolites were obtained and identified by spectral data (mass spectrometry and nuclear magnetic resonance) as: Cyclo-(Phe–Tyr) and Cyclo-(Phe–Gly). The results revealed that humoral innate immune parameters (lysozyme activity, phagocytic activity and bactericidal activity) were significantly (P < 0.05) increased after feeding on the two active compounds-supplemented diet. Furthermore, administration of the two active compounds significantly (P < 0.05) up regulated IL-1β, Type 1 IFN, IFN g2b, IL10 and TNF-α gene expression. Heat shock protein 70 (HSP70) gene expression was significantly (P < 0.05) lower as compared to control group at the end of trial. Common carp fed with the two compounds had higher survival rates (69.3%) compared to the controls (32.0%) after challenged with the pathogen, Aeromonas hydrophila. The present study indicates that the two isolated active compounds could positively influence immune response and enhance disease resistance of common carp against A. hydrophila infection.

Evaluation of different stocking densities in a Senegalese sole (Solea senegalensis) farm: Implications for growth, humoral immune parameters and oxidative status

Abstract Fish are usually exposed to farm operations that can activate the stress system, which in turn diverts energy from normal metabolic processes (i.e. growth or immune defence) to deliver it into the physiological systems activated to cope with stress. The main goal of this study was to assess the effect of high stocking densities on growth performance of Senegalese sole (Solea senegalensis) juveniles reared under intensive farming conditions. The impact of high densities on plasma cortisol and metabolites, humoral immune parameters and hepatic oxidative status was evaluated. Senegalese sole juveniles (45 ± 5 g average body weight) were stocked at three different initial densities (7, 17 and 24 kg m− 2) throughout a 60 days rearing period. Final stocking densities were 13, 31 and 40 kg m− 2 with no differences in terms of growth performance. Similar levels of plasma cortisol, circulating leucocytes and metabolites and humoral innate immune parameters suggest that stocking densities do not affect the hematological and biochemical status of Senegalese sole juveniles, at least under the current experimental conditions. Overall, this study presents the possibility for Senegalese sole on-growing at high stocking densities, strengthening the needs for its intensive commercial farming.

The effects of Ficus carica polysaccharide on immune response and expression of some immune-related genes in grass carp, Ctenopharyngodon idella

The present study investigated the effect of Ficus carica polysaccharide (FCP), isolated from the fruit of F. carica L., at 0%, 0.1%, 0.5% and 1.0% doses supplementation with feed on genes Interleukin 1-β (IL-1β), Tumor Necrosis Factor α (TNF-α) and heat shock protein 70 (HSP70) gene expression in blood, humoral innate immune parameters and resistant to Flavobacterium columnare of grass carp at weeks 1, 2 and 3. The results revealed that administration of FCP significantly (P < 0.05) up regulated IL-1β and TNF-α gene expression. HSP70 gene expression was significantly (P < 0.05) lower in FCP-fed fish at the end of trial. The serum total protein, albumin and globulin did not significantly increased in any diet on the first week whereas it was significantly enhanced in 0.5% and 1.0% supplementation diets on weeks 2 and 3 when compared to control. The serum complement C3 was significantly (P < 0.05) increased on weeks 1 and 2 when compared to control, however, no significant difference was found in this activity after 3 weeks of treatment. All diets significantly enhanced the serum lysozyme activity, bactericidal activity from weeks 1–2 as compared to control. Grass carp fed with FCP showed remarkably higher resistance against F. columnare (60% survival) compared to the control group (30% survival). These results confirm that FCP can up regulate immune related genes expression, stimulates immune response that per se enhances disease resistance in grass carp.

The effects of dietary Immunogen® on innate immune response, immune related genes expression and disease resistance of rainbow trout (Oncorhynchus mykiss)

This study investigates the effects of prebiotic Immunogen® on lysozyme, TNFα and HSP70 gene expression in head kidney, humoral innate immune parameters and resistant to Aeromonas hydrophila of rainbow trout. 120 healthy rainbow trout (81.65 ± 1.49 g) were distributed in six fiberglass tanks assigned to two groups fed control or diet supplemented with 2 g kg−1 Immunogen® for 45 days. The results revealed that administration of Immunogen® significantly (P < 0.05) up regulated lysozyme and TNFα gene expression. HSP70 gene expression was significantly (P < 0.05) lower in Immunogen® fed fish at the end of trial. Humoral innate immune parameters (lysozyme activity, ACH50 and bactericidal activity) were significantly (P < 0.05) increased whether 15 or 45 days after feeding on Immunogen® supplemented diet. However, significant (P < 0.05) increase in agglutination antibody titer observed just after 45 days feeding on Immunogen®. Rainbow trout fed with 2 g kg−1 Immunogen® showed remarkably higher resistance against A. hydrophila (64.44% survival) compared to the control group (24.44% survival). These results confirm that Immunogen® can up regulates immune related genes expression, stimulates immune response that per se enhances disease resistance in rainbow trout.

Use of probiotics intercropped with plant protein diets and their influence on the growth performance and immunological status of Senegalese sole (Solea senegalensis)

Use of probiotics intercropped with plant protein diets and their influence on the growth performance and immunological status of Senegalese sole (Solea senegalensis)

Dietary supplementation of fructooligosaccharide (FOS) improves the innate immune response, stress resistance, digestive enzyme activities and growth performance of Caspian roach ( Rutilus rutilus) fry

The present study investigated the effects of prebiotic fructooligosaccharide (FOS) on the innate immune response, stress resistance, digestive enzyme activities, growth factors and survival of Caspian Roach ( Rutilus rutilus) fry. After acclimation, fish (0.67 ± 0.03 g) were allocated into 12 tanks (50 fish per tank) and triplicate groups were fed a control diet or diets containing 1%, 2% or 3% FOS. At the end of the trial (7 weeks), humoral innate immune parameters (serum Ig levels, lysozyme activity and alternative complement activity (ACH50)), resistance to salinity stress (150 g L −1), digestive enzyme activities (amylase, lipase and protease) and growth factors (final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR), and condition factor) were assessed. At the end of the study the innate immune responses (Ig levels, lysozyme activity and ACH50) were significantly higher in 2% and 3% FOS fed fish ( P < 0.05), whereas, 1% dietary FOS only elevated serum lysozyme activity. All dietary FOS levels significantly increased resistance to a salinity stress challenge ( P < 0.05) and highest survival was observed in the 3% FOS group. Similarly, digestive enzyme activities were significantly elevated with increasing levels of dietary FOS ( P < 0.05). Subsequently, elevated growth performance (final weight, SGR and FCR) was observed in roach fed 2% and 3% FOS compared to the control group ( P < 0.05). These results indicate that FOS can be considered as a beneficial dietary supplement for improving the immune response, stress resistance, digestive enzyme activities and growth performance of Caspian roach fry.

Sharpsnout sea bream ( Diplodus puntazzo) humoral immune response against the parasite Enteromyxum leei (Myxozoa)

The humoral innate immune response of sharpsnout seabream Diplodus puntazzo against the myxozoan Enteromyxum leei was studied. Enteromyxosis was transmitted by cohabitation and a group of uninfected fish served as control. At 5, 12, 19, 26, 40 and 55 days post-exposure (p.e.), control and recipient fish were sampled to determine the prevalence of infection and some humoral innate immune parameters (antiprotease, antitumoral and peroxidase activities). Prevalence of infection was high from day 12 p.e. and reached 100% at days 40 and 55, when intensity of infection was medium to severe. The antiprotease activity was significantly increased in E. leei-exposed fish with respect to control fish at days 12 and 19 p.e. The serum antitumoral activity was slightly lower in recipient than in control fish at all sampling times, except at 40 days p.e., though no statistically significant differences were observed. Serum peroxidases were higher in all recipient fish than in control ones, with the highest stimulation index at 40 days p.e. Within recipient fish, no differences were detected between sampling times in any of the measured activities. The possible implication of these immune factors in the high susceptibility of D. puntazzo to this enteromyxosis is discussed.