Alternative Complement Activation Research Articles

Contribution of adipose-derived factor D/adipsin to alternative pathway complement activation: Lessons from lipodystrophy

Contribution of adipose-derived factor D/adipsin to alternative pathway complement activation: Lessons from lipodystrophy

The potential role of complements in cocaine-induced thrombotic microangiopathy.

Thrombotic microangiopathy (TMA) is a rare disorder characterized by microvascular injury and occlusion resulting in tissue ischemia and dysfunction. TMA occurs in a variety of settings including cocaine use. Although cocaine is widely used in the United States, cocaine-associated TMA is only rarely reported. Therefore, other factors may predispose cocaine users to the development of TMA. Emerging evidence indicates that cocaine activates complements. Therefore, complement activation may contribute to the development of cocaine-induced TMA. Here, we report a cocaine user who presented with renal failure. Renal biopsy demonstrated TMA. Laboratory tests revealed reduced serum complement C3 and normal complement C4 levels indicative of alternative complement activation. We postulate that complement activation is involved in the pathogenesis of cocaine-induced TMA.

Complement-Mediated Regulation of Apolipoprotein E in Cultured Human RPE Cells.

Purpose. Complement activation is implicated in the pathogenesis of age-related macular degeneration (AMD). Apolipoprotein E (ApoE) and complement activation products such as membrane attack complex (MAC) are present in eyes of individuals with AMD. Herein, we investigated the effect of complement activation on induction of ApoE accumulation in human retinal pigment epithelial (RPE) cells.Methods. Cultured human RPE cells were primed with a complement-fixing antibody followed by treatment with C1q-depleted (C1q-Dep) human serum to elicit alternative pathway complement activation. Controls included anti-C5 antibody-treated serum and heat-inactivated C1q-Dep. Total protein was determined on RPE cell extracts, conditioned media, and extracellular matrix (ECM) by Western blot. ApoE and MAC colocalization was assessed on cultured RPE cells and human eyes by immunofluorescent stain. ApoE mRNA expression was evaluated by quantitative PCR (qPCR).Results. Complement challenge upregulated cell-associated ApoE, but not apolipoprotein A1. ApoE accumulation was blocked by anti-C5 antibody and enhanced by repetitive complement challenge. ApoE mRNA levels were not affected by complement challenge. ApoE was frequently colocalized with MAC in complement-treated cells and drusen from human eyes. ApoE was released into complement-treated conditioned media after a single complement challenge and accumulated on ECM after repetitive complement challenge.Conclusions. Complement challenge induces time-dependent ApoE accumulation in RPE cells. An understanding of the mechanisms by which complement affects RPE ApoE accumulation may help to better explain drusen composition, and provide insights into potential therapeutic targets.

Nutritional evaluation of Gracilaria pulvinata as partial substitute with fish meal in practical diets of barramundi (Lates calcarifer)

A 40-day feeding trial was carried out to investigate the potential of red algal (Gracilaria pulvinata) meal as a protein source in formulated diets for barramundi (Lates calcarifer) (initial mean body weight of 28.5 ± 0.5 g). Five practical diets were prepared using a fishmeal-based diet used as positive control (PC) and blends of soybean meal and fishmeal used as a negative control (NC), in which NC was supplemented with 3 (GL3%), 6 (GL6%), and 9% (GL9%) G. pulvinata. Each dietary treatment was replicated in triplicate. Results indicated that growth and feeding performance including specific growth rate, feed conversion, protein efficiency ratios, and feed intake in all treatments were not significantly different (P > 0.05). Crude protein, lipid, and ash contents of fish carcass were not markedly altered in all dietary treatments. Serum lysozyme, alternative complement activities, serum immunoglobulin, and total protein content significantly decreased in fish fed GL9 diets. Increasing dietary supplementation of G. pulvinata decreased the serum triglycerides and cholesterol when compared to the NC. Gracilaria inclusion levels did not affect intestinal total protease and amylase activities (P < 0.05); however, intestine lipase activity in fish fed GL6 diets was significantly higher than other groups (P < 0.05). The results of the present study recommend the inclusion of Gracilaria meal up to 3%, without significant negative effects on the growth performance, body composition, and health parameters of L. calcarifer.

Elevated factor H–related protein 1 and factor H pathogenic variants decrease complement regulation in IgA nephropathy

IgA nephropathy (IgAN), a frequent cause of chronic kidney disease worldwide, is characterized by mesangial deposition of galactose-deficient IgA1-containing immune complexes. Complement involvement in IgAN pathogenesis is suggested by the glomerular deposition of complement components and the strong protection from IgAN development conferred by the deletion of the CFHR3 and CFHR1 genes (ΔCFHR3-CFHR1). Here we searched for correlations between clinical progression and levels of factor H (FH) and FH-related protein 1 (FHR-1) using well-characterized patient cohorts consisting of 112 patients with IgAN, 46 with non-complement-related autosomal dominant polycystic kidney disease (ADPKD), and 76 control individuals. Patients with either IgAN or ADPKD presented normal FH but abnormally elevated FHR-1 levels and FHR-1/FH ratios compared to control individuals. Highest FHR-1 levels and FHR-1/FH ratios are found in patients with IgAN with disease progression and in patients with ADPKD who have reached chronic kidney disease, suggesting that renal function impairment elevates the FHR-1/FH ratio, which may increase FHR-1/FH competition for activated C3 fragments. Interestingly, ΔCFHR3-CFHR1 homozygotes are protected from IgAN, but not from ADPKD, and we found five IgAN patients with low FH carrying CFH or CFI pathogenic variants. These data support a decreased FH activity in IgAN due to increased FHR-1/FH competition or pathogenic CFH variants. They also suggest that alternative pathway complement activation in patients with IgAN, initially triggered by galactose-deficient IgA1-containing immune complexes, may exacerbate in a vicious circle as renal function deterioration increase FHR-1 levels. Thus, a role of FHR-1 in IgAN pathogenesis is to compete with complement regulation by FH.

Apple cider vinegar boosted immunomodulatory and health promoting effects of Lactobacillus casei in common carp (Cyprinus carpio)

The present study was performed to investigate the immunomodulatory and health promoting effects of combined or singular administration of apple cider vinegar (ACV) and Lactobacillus casei in common carp (Cyprinus carpio) diet. An 8-week feeding trial was designed with following treatments: Control (basal diet), Pro (contains 107 CFU g−1L. casei), LACV (contains 1% ACV), HACV (contains 2% ACV), Pro + LACV (contains 107 CFU g−1L. casei plus 1% ACV) and Pro + HACV (contains 107 CFU g−1L. casei plus 2% ACV). Evaluation of skin mucus revealed notable increase of total Ig level and lysozyme activity in Pro + LACV and Pro + HACV treatments compared other groups (P < 0.05). Similarly, serum total Ig and lysozyme activity in HACV, Pro + LACV and Pro + HACV fed carps was remarkably higher than other groups (P < 0.05). However, regarding serum alternative complement (ACH50) activity significant difference was observed just between Pro + HACV and control treatment (P < 0.05). The highest expression of immune related (LYZ, TNF-alpha, IL1b, IL8) and antioxidant enzymes genes (GSR, GST) were observed in carps fed Pro + HACV and Pro + LACV. The expression of GH gene expression in Pro, LACV and HACV treatments was significantly higher than those in control group (P < 0.05). The highest expression level of GH and IGF1 was observed in fish fed combined Pro and ACV (P < 0.05). These results indicated that co-administration of ACV boosted immunomodulatory and health promoting effects of L. casei and can be considered as a promising immunostimulants in early stage of common carp culture.

The effects of dietary inulin and Jerusalem artichoke (Helianthus tuberosus) tuber on the growth performance, haematological, blood chemical and immune parameters of Nile tilapia (Oreochromis niloticus) fingerlings

This study investigated the effects of dietary inulin or Jerusalem artichoke (JA) on the growth performance, haematological, blood chemical and immune parameters of Nile tilapia fingerlings. Five treatment diets were designed to incorporate inulin at 0 (basal diet), 2.5 and 5.0 g/kg and JA at 5.0 and 10.0 g/kg. Two basal diets including fish meal and formulated experimental feed were used for fry and fingerling growing periods, respectively. During the fry growing period, larvae were fed treatment diets for 4 weeks. There were no significant differences in growth performance or survival rate. Fingerlings were then nursed with the formulated experimental diets from weeks 5 to 12. Fingerlings fed on inulin at 5.0 g/kg or JA at either level had better growth performance and survival rate than that fed on the basal diets. There were no significant differences in body composition. Dietary prebiotic inulin and JA increased red blood cell number (p < .05). Among the five blood chemistry parameters examined, both inulin at 5.0 g kg and JA (5.0 and 10.0 g/kg) increased blood protein (p < .05). Dietary inulin at 5.0 g/kg and JA at 5.0 and 10.0 g/kg increased total immunoglobulin and lysozyme activity (p < .05). Both inulin and JA inclusion diets increased alternative complement activity (p < .05). Taken together, dietary inulin at 5 g/kg and JA at 5.0 and 10.0 g/kg had beneficial effects on the growth performance, survival rate and immune of Nile tilapia fingerlings.

A comparison between marine and terrestrial invertebrate meals for mirror carp (Cyprinus carpio ) diets: Impact on growth, haematology and health

Invertebrate meals (e.g. polychaetes and insects) present novel and sustainable high-quality nutrient sources for use in fish feed formulations. To test this innovative source, an eleven-week feeding trial was conducted evaluating the effects of replacing the fishmeal (FM) component as an example of a superior protein source (FM CTRL) with ragworm meal (RW, Nereis virens) and/or silkworm pupae (SWP, Bombyx mori) in mirror carp (Cyprinus carpio) diets. Three experimental diets with partial replacement of FM (diets: RW + FM, SWP + FM and RW + SWP + FM) were formulated. All diets were formulated to be iso-nitrogenous, iso-lipidic and iso-energetic. Growth performance and feed utilization indices were assessed, and the feeding trial concluded with the analysis of haematological parameters to provide an indication of carp physiological and health status. Mean weight gain was greatest in mirror carp fed RW + FM (60.83 fish−1 day−1; P < 0.05 vs. all other diets) followed by SWP + FM (40.62 g fish−1 day−1; P < 0.05 vs. all other diets). The least weight gain was achieved in fish fed FM + SWP + RW+ and FM CTRL (34.34 and 33.96 g fish−1 day−1, respectively; not significantly different from each other). Fish fed on RW + FM diet had significantly lower plasma ammonia concentrations than any other dietary groups (P = 0.04). Mirror carp fed on SWP + FM diet (111.52 units mL−1) were observed to have a marked enhancement in alternative complement activity than FM CTRL (79.21 units mL−1, P = 0.041). Both ragworm and silkworm pupae meal present attractive sustainable functional feed component in carp diets, with benefits on enhancing growth performance and specific physiological parameters.

Glutathione inhibits antibody and complement-mediated immunologic cell injury via multiple mechanisms

Antioxidant glutathione (GSH) plays an important role in the regulation of immunity. However, little is known about its effects on humoral immunity, especially its action on effector molecules like antibody and complement. Given that these molecules contain abundant disulfide bonds, we speculated that GSH might influence the action of these proteins via its thiol function. Using a model of a glomerular mesangial cell (MC) lysis induced by antibodies plus complement, we addressed this hypothesis. Exposure of rat MCs to anti-Thy-1 antibody plus complement or anti-MC rabbit serum caused a complement-dependent cell lysis, which was completely blocked by GSH. Moreover, GSH potently prevented the antibody-mediated agglutination of red blood cells and aggregation of antibody-sensitized microspheres. Further analysis revealed that GSH inhibited antibody binding to antigens and promoted the conversion of the antibodies to its reduced forms. GSH also potently inhibited the formation and deposition of C5b-9 in MCs and suppressed both the classic and alternative complement activation pathway. Lastly, GSH attenuated P38 activation, an oxidative sensitive kinase that partially mediated the antibody- and complement-dependent MC lysis. Depletion of GSH via inhibiting gamma-glutamylcysteine synthetase or xCT transporter augmented P38 activation and sensitized MCs to the cell lysis. Collectively, our results indicate that GSH protects cells from immunological cell damage via mechanisms involving inhibition of antibody binding to the antigens, suppression of complement activation and augmentation of cellular defense mechanism. Our study provides novel mechanistic insights into the actions of GSH in the regulation of immune responses and suggests that GSH might be used to treat certain immune disorders.

C5 inhibition prevents renal failure in a mouse model of lethal C3 glomerulopathy

C3 glomerulopathy is a potentially life-threatening disease of the kidney caused by dysregulated alternative pathway complement activation. The specific complement mediator(s) responsible for kidney injury in C3 glomerulopathy are yet to be defined and no specific therapy is currently available. We previously developed a mouse model of lethal C3 glomerulopathy with factor H and properdin gene double mutations. Therefore, we used this model to examine the role of C5 and C5a receptor (C5aR) in the pathogenesis of the disease. Disease severity in these factor H/properdin double-mutant mice was found to be correlated with plasma C5 levels, and prophylactic anti-C5 mAb therapy was effective in preventing lethal C3 glomerulopathy. When given to these double-mutant mice that had already developed active disease with severe proteinuria, anti-C5 mAb treatment also prevented death in half of the mice. Deficiency of C5aR significantly reduced disease severity, suggesting that C5aR-mediated inflammation contributed to C3 glomerulopathy. Thus, C5 and C5aR have a critical role in C3 glomerulopathy. Hence, early intervention targeting these pathways may be an effective therapeutic strategy for patients with C3 glomerulopathy.

A microplate assay to measure classical and alternative complement activity.

We developed and validated a kinetic microplate hemolytic assay (HA) to quantify classical and alternative complement activity in a single dilution of human plasma or serum. The assay is based on monitoring hemolysis of sensitized sheep (or uncoated rabbit) red blood cells by means of a 96-well microplate reader. The activity of the calibrator was evaluated by reference to 200 healthy adults. The conversion of 50% hemolysis time into a percentage of activity was obtained using a calibration curve plotted daily. The linearity of the assay as well as interference (by hemolysis, bilrubinemia and lipemia) was assessed for classical pathway (CP). The within-day and the between-day precision was satisfactory regarding the performance of commercially available liposome immunoassay (LIA) and ELISA. Patients with hereditary or acquired complement deficiencies were detected (activity was measured <30%). We also provided a reference range obtained from 200 blood donors. The agreement of CP evaluated on samples from 48 patients was 94% with LIA and 87.5% with ELISA. The sensitivity of our assay was better than that of LIA, and the cost was lower than either LIA or ELISA. In addition, this assay was less time consuming than previously reported HAs. This assay allows the simultaneous measurement of 36 samples in duplicate per run of a 96-well plate. The use of a daily calibration curve allows standardization of the method and leads to good reproducibility. The same technique was also adapted for the quantification of alternative pathway (AP) activity.

The effects of dietary kefir and low molecular weight sodium alginate on serum immune parameters, resistance against Streptococcus agalactiae and growth performance in Nile tilapia (Oreochromis niloticus)

The present study evaluates the effects of dietary kefir and low molecular weight sodium alginate (LWMSA) (singular or combined) on non-specific immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Fish with average weight of 18.60 ± 0.04 g were supplied and randomly stocked in sixteen glass tanks (150 L) at density of 20 fish per tank. Fish were fed experimental diets as follows: 0 g kg−1 LMWSA (Control, Diet 1), 10 g kg−1 LMWSA (Diet 2), 40 g kg−1 kefir (Diet 3), and 10 g kg−1 LMWSA + 40 g kg−1 kefir (Diet 4) for 50 days. At the end of the feeding trial, serum lysozyme (SL), phagocytosis (PI), respiratory burst (RB), and alternative complement (ACH50) activities as well as growth performance were measured. Singular and combined administration of kefir and low molecular weight sodium alginate (LMWSA) significantly increased serum SL, PI, RB, and ACH50 activities compared control group (P < 0.05); the highest innate immune responses were observed in fish fed combinational diet (kefir + LMWSA) (P < 0.05). The results of experimental challenge revealed significantly higher resistance against Streptococcus agalactiae in fish fed supplemented diets and the highest post challenge survival rate was observed in synbiotic diet (P < 0.05). Similar results obtained in case of growth parameters. Feeding on supplemented diet significantly improved SGR and FCR and the highest growth parameters was observed in fish fed synbiotic diet (P < 0.05). These finding revealed that combined administration of dietary kefir and LMWSA can be considered for improving immune response, disease resistance and growth performance of Nile tilapia.

Pregnancy-associated atypical hemolytic uremic syndrome: A case report and literature review

Atypical hemolytic uremic syndrome (aHUS) is a rare disease of uncontrolled alternative pathway complement activation and pregnancy-associated aHUS (P-aHUS) is a more rare but life-threatening disorder. Besides gene screening, ADAMTS-13 detection and renal biopsy is helpful to establish diagnosis and prognosis assessment when kidney is involved. Combined liver-kidney transplant and eculizumab therapy has generally been the initial approach to this disease management besides therapeutic plasma exchange and plasma infusion. Though transplant and eculizumab therapy may be more effective than traditional therapy, it is much more expensive and not easily available. Herein we report a case of P-aHUS and our case indicates that plasma exchange and hemodialysis may be effective to P-aHUS for those not suitable/available for eculizumab treatment or transplant.

Dietary phytoimmunostimulant Persian hogweed (Heracleum persicum) has more remarkable impacts on skin mucus than on serum in common carp (Cyprinus carpio)

Immunostimulation through the use of sustainable and eco-friendly dietary additives is one of the current prophylactic strategies in fish husbandry. Plant-based immunostimulants are highly considered for this intent, both for their scientific and practical advantages. Persian hogweed (Heracleum persicum) is a flower-bearing herb that possesses interesting pharmacological importance due to its bioactive compounds. It is commonly used as spice, food additive, dietary supplement and traditional remedy. The present study evaluated the potential of H. persicum as a dietary phytoimmunostimulant in common carp (Cyprinus carpio). The powder form of H. persicum was incorporated in the basal diet at three different inclusion levels: 2.5, 5 and 10 g kg−1. The basal diet (0 g kg−1 of H. persicum) served as control. Experimental diets were administered to the fish for a period of 8 weeks. At the termination of the feeding experiment, impacts on fish immunity and performance were evaluated.Inclusion of H. persicum in the diet significantly elevated several immunological factors such as immunoglobulins, lysozyme, protease and alternative complement activities in carp. Interestingly, the changes were more pronounced in the skin mucus than in the serum. Performance was significantly improved in the fish groups that received the candidate phytoimmunostimulant. Specifically, final weight, weight gain, specific growth rate and feed conversion ratio were significantly improved in the fish that received dietary H. persicum at inclusion levels 5 g kg−1 and higher. This study demonstrated the potential of Persian hogweed as a candidate dietary phytoimmunostimulant in carp, impacting mainly the skin mucosal defenses. The study supports the current trend in the exploration of sustainable plant-based dietary supplements that are capable of boosting the immunological defenses of farmed fish.

Combined administration of low molecular weight sodium alginate boosted immunomodulatory, disease resistance and growth enhancing effects of Lactobacillus plantarum in Nile tilapia (Oreochromis niloticus)

The present study investigates the effects of combined or singular administration of low molecular weight sodium alginate (LWMSA) and Lactobacillus plantarum on innate immune response, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Three hundred and twenty fish were supplied and randomly stocked in sixteen glass tanks (150 L) assigned to four treatments as follows: 0 g kg−1 LMWSA (Control, Diet 1), 10 g kg−1 LMWSA (Diet 2), 108 CFU g−1L. plantarum (Diet 3), and 10 g kg−1 LMWSA + 108 CFU g−1L. plantarum (Diet 4). Following 30 and 60 days of the feeding trial, serum lysozyme, phagocytosis, respiratory burst and alternative complement activities as well as growth performance parameters (specific growth rate, feed conversion ratio) were measured. Serum lysozyme, phagocytosis, respiratory burst, and alternative complement activities of fish were significantly stimulated by both LMWSA and L. plantarum diets, however, the highest innate immune response were observed in fish fed synbiotic diet. At the end of the experiment, eight fish per replication were randomly selected for a challenge test against Streptococcus agalactiae. The survival rate of the fish fed supplemented diets was significantly greater than the control treatment and the highest post challenge survival rate was observed in synbiotic diet. Furthermore, SGR and FCR were significantly improved in fish fed supplemented diets after 60 days and the highest growth performance was observed in fish fed synbiotic diet. These results suggest combined LMWSA and L. plantarum can be considered as a promising immunostimulant and growth enhancer in Nile tilapia diet.

The effects of increasing dietary levels of soy protein concentrate (SPC) on the immune responses and disease resistance (furunculosis) of vaccinated and non-vaccinated Atlantic salmon (Salmo salar L.) parr

Juvenile salmon, with an initial weight of 9 g, were fed three experimental diets, formulated to replace 35 (SPC35), 58 (SPC58) and 80 (SPC80) of high quality fishmeal (FM) with soy protein concentrate (SPC) in quadruplicate tanks. Higher dietary SPC inclusion was combined with increased supplementation of methionine, lysine, threonine and phosphorus. The experiment was carried out for 177 days. On day 92 salmon in each tank were bulk weighed. Post weighing eighty salmon from each tank were redistributed in two sets of 12 tanks. Salmon from the first set of tanks were vaccinated, while the second group was injected with phosphate buffer saline (PBS). Salmon were sampled on day 92 (pre-vaccination), day 94 (2 days post vaccination [dpv]/PBS injection [dpPBSinj]) and day 154 (62 dpv/dpPBSinj) of the trial for the assessment of their immune responses, prior to the performance of salmon bulk weights for each tank. On day 154, fish from each tank were again bulk weighed and then seventeen salmon per tank were redistributed in two sets of twelve tanks and intra-peritoneally infected with Aeromonas salmonicida. At Day 154, SPC80 demonstrated lower performance (weight gain, specific growth rate and thermal growth coefficient and feed conversion ratio) compared to SPC35 salmon. Reduced classical and total complement activities for salmon fed diets with over 58% of protein from SPC, were demonstrated prior to vaccination. Reduced alternative complement activity was detected for both SPC58 and SPC80 salmon at 2 dpv and for the SPC80 group at 62 dpv. Total and classical complement activities demonstrated no differences among the dietary groups after vaccination. Numerical increases in classical complement activity were apparent upon increased dietary SPC levels. Increased phagocytic activity (% phagocytosis and phagocytic index) was exhibited for the SPC58 group compared to SPC35 salmon at 62 dpPBSinj. No differences in serum lysozyme activity, total IgM, specific antibodies, protein, glucose and HKM respiratory burst were detected among the dietary groups at any timepoint or state. Mortalities as a result of the experimental infection only occurred in PBS-injected fish. No differences in mortality levels were demonstrated among the dietary groups. SPC58 diet supported both good growth and health in juvenile Atlantic salmon while SPC80 diet did not compromise salmon’ immunity or resistance to intraperitoneally inflicted furunculosis.

Influence of probiotic, Lactobacillus plantarum on serum biochemical and immune parameters in vaccinated rainbow trout (Oncorhynchus mykiss) against streptococcosis/lactococosis

This study evaluated the effects of probiotic, Lactobacillus plantarum on serum biochemical and some immune parameters of immunized rainbow trout weighting 29.6±1.84 g, with streptococcosis/lactococosis vaccine at 16±1.5°C, for 60 days. A commercial diet was used as the control. Fish in the first treatment were immunized with streptococcosis/lactococosis vaccine in bathing route for 1 min. In the second group, the vaccinated trout were also fed diet containing L. plantarum (10 8 CFU g -1 ). In the third treatment, fish were only fed the diet supplemented with L. plantarum (10 8 CFU g -1 ). The results showed that vaccinated trout with or without L. plantarum feeding diets significantly decreased heterophils. Meanwhile it enhances serum lysozyme, alternative complement activities, antibody titer, total leukocytes, lymphocytes, and serum biochemical parameters, including ALP, IgM, and total protein levels compared to control groups. Moreover, the highest levels of above mentioned parameters were found in vaccinated fish that fed L. plantarum . In addition, the vaccinated fish that fed L. plantarum showed significantly elevated cholesterol levels compared to the control group. The results showed that the dietary L. plantarum improved the immunity of immunized trout with streptococcosis/lactococosis vaccine.

Comparative effects of Aloe vera gel and Freund’s adjuvant in vaccination of common carp (Cyprinus carpio L.) against Aeromonas hydrophila

Natural compounds of Aloe vera gel are well known for their immunostimulatory properties. In this study, adjuvant effects of Aloe vera in combination with formalin-killed A. hydrophila bacterin in immunization of common carp were evaluated and compared with Freund’s adjuvant. A total of 480 juvenile carp were randomly divided into four groups. Groups 1–4 were injected on days 0 and 14 with normal saline (as control), bacterin, bacterin + Freund’s adjuvant and bacterin + Aloe vera gel, respectively. Blood samples were taken on days 14, 28 and 42. Hematoimmunological parameters including white blood cell (WBC) count, alternative complement activity, serum lysozyme and bactericidal activity, respiratory burst activity (NBT) and antibody titer were evaluated and compared among different groups. At the end of experiment, all groups were challenged with live A. hydrophila and relative percentage survival (RPS) compared among groups. According to the results, administration of both adjuvants caused a positive change in fish immune response and most of the aforementioned parameters significantly improved in vaccinated groups compare with control group (P < 0.05). Moreover, WBC count, alternative complement activity, serum lysozyme activity and NBT activity were significantly higher in Aloe adjuvant group compared with Freund’s adjuvant (P < 0.05). Finally after challenge, groups 3 and 4 showed significantly lesser mortality than groups 1 and 2. Overall, our results indicated that combination of A. hydrophila bacterin with adjuvant can improve the vaccine efficacy and resistance against A. hydrophila infection, and in comparison with traditional adjuvants, Aloe vera gel could be used as a natural adjuvant with similar or even greater positive effects on vaccination of common carp.

Influence of dietary prebiotic mixture (α-mune) on growth performance, haematology and innate immunity of Beluga sturgeon (Huso huso) juvenile

The present study investigated the effects of prebiotic (α-mune), a mixture of mannan oligosaccharide, β-1,3 and β-1,6 glucan, on the growth performance, haematology and innate immunity of beluga sturgeon ( Huso huso ) juvenile. Fish (46±3 g) were allocated into 12 tanks (15 fish per tank) and triplicate groups were fed a control diet or diets containing 1.5, 3.0 and 4.5 g kg -1 prebiotic for 46 days. Fish fed 1.5 g kg -1 prebiotic displayed significantly higher final weight, specific growth rate and feed conversion ratio. WBC, RBC, MCV, MCH, haemoglobin, haematocrit and lymphocyte levels were also significantly higher in the fish fed 1.5 g kg -1 prebiotic. Furthermore, the highest haematocrit content and lymphocyte level were found in the fish fed a diet containing 1.5 g kg -1 prebiotic. Alternative complement activity (ACH50), lysozyme activity and Ig concentration were significantly higher in the fed 1.5 g kg -1 prebiotic. These results indicate that α-mune can be considered as a beneficial dietary supplement for improving the growth performance, haematological and immunological parameters of beluga sturgeon juvenile.

Hemato-Immunological Responses and Disease Resistance in Siberian Sturgeon Acipenser baerii Fed on a Supplemented Diet of Lactobacillus plantarum.

A feeding trial was conducted to investigate the effects of different levels of dietary Lactobacillus plantarum on hemato-immunological parameters and resistance against Streptococcus iniae infection in juvenile Siberian sturgeon Acipenser baerii. Fish (14.6±2.3g) were fed three experimental diets prepared by supplementing a basal diet with L. plantarum at different concentrations [1×107, 1×108 and 1×109 colony-forming units (cfu) g-1] and a control (non-supplemented basal) diet for 8weeks. Innate immune responses (immunoglobulin (Ig), alternative complement activity (ACH50) and lysozyme activity) were significantly higher in fish fed the 1×108 and 1×109 cfug-1 L. plantarum diet compared to the other groups (P<0.05). Furthermore, fish fed on various levels of L. plantarum significantly showed higher red blood cell (RBC), hemoglobin (Hb), white blood cell (WBC) and monocyte compared to those of the control group (P<0.05). At the end of the feeding experiment, some fish were challenged with S. iniae to quantify the level of disease resistance. The mortality after S. iniae challenge was decreased in fish fed a probiotic. These results indicated that dietary supplementation of L. plantarum improved immune response and disease resistance of Siberian sturgeon juvenile.