Alternative Complement Activation Research Articles

Betaine and the probiotic, Lactobacillus rhamnosus in the diet of the Common carp, Cyprinus carpio: Effects on growth, digestive enzyme activities, antioxidant system, humoral and mucosal immunity and resistance to Streptococcus iniae

The probiotic, Lactobacillus rhamnosus (LAC-R) and Betaine (BT) alone and combined were used in the diet of the common carp, Cyprinus carpio to evaluate their potentials in improving growth, immunity and disease resistance. Seven experimental treatments (LAC-R: 1 × 108 CFU/g LAC-R; BT1: 0.5 % BT; BT2: 1 % BT; BT3: 2 % BT; LAC-R + BT1; LAC-R + BT2; LAC-P + BT3) and one control group (control: non-supplemented fish) were designed in three replicates and fish (13.3 ± 1.5 g) fed with the test diets for two months. After the feeding experiment, fish were exposed to Streptococcus iniae for 14 days to evaluate the fish disease resistance. The fish supplemented with BT and LAC-R alone or combined exhibited better growth compared to control (P<0.01), however, fish of LAC-R + BT had the highest growth performance (P<0.01). Digestive enzyme activities increased in response to LAC-R, BT and their combinations, in which the highest activities were for LAC-R + BT2 and LAC-R + BT3 (P<0.01). The lactic acid bacteria (LAB) populations in intestine increased after the probiotic supplementation (P<0.01), while the results were non-significant for total bacteria count (P>0.01). The total antioxidant capacity (T-AOC), antioxidant enzymes [serum catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx)] and immune components [lysozyme, total immunoglobulin (Ig), myeloperoxidase (MPO), alternative complement activity (ACH50), Nitroblue tetrazolium (NBT) and total protein in blood and total Ig, lysozyme, alkaline phosphatase (ALP) and protease in mucus] were mostly enhanced by LAC-R, BT2, BT3 and their combinations (P<0.01). The oxidative stress was ameliorated by LAC-R, BT and their combinations, as the levels of malondialdehyde (MDA) were lower in these treatments (P<0.01). Fish supplemented with LAC-R, BT and LAC-R + BT had lower mortality rate compared to control after challenge with S. Iniae (P<0.01). In conclusion, dietary LAC-R and BT improved growth and immune performance in the fish. Also, the best performance was found for fish supplemented with a combination of LAC-R and 1–2 % BT.

Probiotic, fructooligosaccharide and yeast extract mixture improves gut health in rainbow trout, Oncorhynchus mykiss.

The present study investigated the effects of a feed additive (FA) containing a probiotic consortium, fructooligosaccharide and yeast extract on growth performance, humoral immune responses, hepatic antioxidant parametersand intestine digestive enzymes, morphology and transcripts in rainbow trout, Oncorhynchus mykiss. The fish were reared for 8 weeks, feeding on diets containing 0 (CNT), 0.3 (0.3FA), 1 (1FA)and 2 (2FA) g/kg FA. The results showed that fish growth parameters were significantly and quadratically related to FA levelsand FA treatments had better growth performance than CNT treatment. Intestinal amylase activity significantly increased in 2FA, whereas, intestinal protease activity increased in all FA treatments. Intestinal villus length and muscular layer thickness significantly increased in 0.3FA treatment. Blood leucocyte and lymphocyte counts, plasma lysozyme activityand hepatic glutathione content significantly increased in 0.3FA and 1FA treatments; whereas hepatic malondialdehyde significantly decreased in these treatments. Blood neutrophil and monocyte counts significantly increased in 0.3FA treatment, while plasma alternative complement activity significantly increased in 1FA treatments. Plasma bactericidal activities against Aeromonas hydrophila, Yersinia ruckeriand Streptococcus iniae, and intestinal expression of heat shock protein 70 and beta-defensin significantly increased in all FA treatments. The abundance of A. hydrophila, Y. ruckeriand S. iniae in fish gut significantly decreased in 0.3FA treatment; these bacteria were absent in the intestines of 1FA and 2FA treatments. The present results suggest that dietary 0.3-1 g/kg of FA can significantly improve growth performance, immune response, intestinal healthand hepatic antioxidant capacity in rainbow trout.

Harnessing paraprobiotics and postbiotics for enhanced immune function in Asian seabass (Lates calcarifer): Insights into pattern recognition receptor signaling

The Asian seabass, Lates calcarifer, is a key species in Asian aquaculture due to its nutritional value and adaptability. However, disease outbreaks, particularly viral and bacterial infections, pose significant challenges to its production. Immunostimulants offer promising solutions but raise safety concerns. Paraprobiotics and postbiotics (CPP) emerge as safer alternatives, exerting health benefits without live microorganisms. This study investigated the potential of probiotic paraprobiotic and postbiotic supplements derived from Bacillus subtilis to enhance the immune response and antioxidant capacity of Asian seabass and improve their resistance to Streptococcus iniae infection. Analysis of antioxidant activity and lipid peroxidation revealed significant improvements in fish supplemented with CPP, indicating their effectiveness in mitigating oxidative stress. Immunological assays demonstrated enhanced growth performance and serum immunity, including increased alternative complement activity, immunoglobulin levels, and phagocytic activity, in supplemented fish. Furthermore, upregulated expression of proinflammatory cytokines (TNF-α, IL-6, IL-1β) and pattern recognition receptors (NLRC3, TLR22, MDA5) in immune tissues. Fish supplemented with CPP exhibited higher resistance and survival rates against S. iniae infection challenge compared to control groups. The study elucidates the mechanisms underlying the immunomodulatory effects of CPP, shedding light on their potential applications in aquaculture.

Targeted complement inhibition using bispecific antibodies that bind local antigens and endogenous complement regulators.

Complement activation protects against infection but also contributes to pathological mechanisms in a range of clinical conditions such as autoimmune diseases and transplant rejection. Complement-inhibitory drugs, either approved or in development, usually act systemically, thereby increasing the risk for infections. We therefore envisioned a novel class of bispecific antibodies (bsAbs) which are capable of site-directed complement inhibition by bringing endogenous complement regulators in the vicinity of defined cell surface antigens. Here, we analyzed a comprehensive set of obligate bsAbs designed to crosslink a specific target with either complement regulator factor H (FH) or C4b-binding protein (C4BP). The bsAbs were assessed for their capacity to inhibit complement activation and cell lysis in an antigen-targeted manner. We observed that the bsAbs inhibited classical, lectin, and alternative pathway complement activation in which sufficient endogenous serum FH and C4BP could be recruited to achieve local inhibition. Importantly, the bsAbs effectively protected antigen-positive liposomes, erythrocytes, and human leukocytes from complement-mediated lysis. In conclusion, localized complement inhibition by bsAbs capable of recruiting endogenous human complement regulators (such as FH or C4BP) to cell surfaces potentially provides a novel therapeutic approach for the targeted treatment of complement-mediated diseases.

Dietary lysophospholipid improves growth performance, antioxidant capacity and immunity response of red tilapia (Oreochromis niloticus × O. mossambicus)

Lysophospholipids (LPL) are usually used as emulsifiers in fish nutrition because of their ability to ameliorate nutrient digestibility and lipid metabolism. The present study aims to assess the effects of a commercial dietary lysophospholipid on growth performance, feed utilization, antioxidant activity, innate immunity and biochemical indices of red tilapia (Oreochromis niloticus x O. mosambicus) fingerlings. Five isonitrogenous (35.8% CP) and isocaloric (18.7 MJ kg−1) diets were prepared as follows: two control diets coded as C1 and C2, containing 2 and 1% soybean oil, respectively. The other three diets were coded as LPL1, LPL2 and LPL3, and were formulated by supplementing the C2 diet with 0.1, 0.2 and 0.3% lysophospholipid. The fingerlings of red tilapia (2.13 ± 0.42 g fish−1) were fed the diets for 70 days. The highest significant (p < 0.05) final weight and weight gain were recorded in LPL2. The best values of feed efficiency and protein efficiency (PER)were in all fish groups fed LPL in their diets (p < 0.05). Protein productive value (PPV) showed the highest result in LPL3 (p < 0.05). Lysozyme activity showed the best result in LPL3 (p < 0.05). The highest values of alternative complement activity (ACH50) and total immunoglobulin (Ig) were in LPL3 and LPL2 (p < 0.05). The best results of superoxide dismutase (SOD) and catalase activity were recorded in LPL3 (p < 0.05). The lowest values of triglycerides (TG) and total cholesterol (CHO) were detected in LPL1 and LPL2 (p < 0.05). Thus, the inclusion of 0.2% of the commercial lysophospholipid in red tilapia diets enhanced the growth performance, feed utilization, innate immunity, antioxidant activity and lipase activity, and reduced CHO and TG of red tilapia fingerlings.

CFH Haploinsufficiency and Complement Alterations in Early-Onset Macular Degeneration.

Complement dysregulation is a key component in the pathogenesis of age-related macular degeneration (AMD) and related diseases such as early-onset macular drusen (EOMD). Although genetic variants of complement factor H (CFH) are associated with AMD risk, the impact of CFH and factor H-like protein 1 (FHL-1) expression on local complement activity in human retinal pigment epithelium (RPE) remains unclear. We identified a novel CFH variant in a family with EOMD and generated patient induced pluripotent stem cell (iPSC)-derived RPE cells. We assessed CFH and FHL-1 co-factor activity through C3b breakdown assays and measured complement activation by immunostaining for membrane attack complex (MAC) formation. Expression of CFH, FHL-1, local alternative pathway (AP) components, and regulators of complement activation (RCA) in EOMD RPE cells was determined by quantitative PCR, western blot, and immunostaining. Isogenic EOMD (cEOMD) RPE was generated using CRISPR/Cas9 gene editing. The CFH variant (c.351-2A>G) resulted in loss of CFH and FHL-1 expression and significantly reduced CFH and FHL-1 protein expression (∼50%) in EOMD iPSC RPE cells. These cells exhibited increased MAC deposition upon exposure to normal human serum. Under inflammatory or oxidative stress conditions, CFH and FHL-1 expression in EOMD RPE cells paralleled that of controls, whereas RCA expression, including MAC formation inhibitors, was elevated. CRISPR/Cas9 correction restored CFH/FHL-1 expression and mitigated alternative pathway complement activity in cEOMD RPE cells. Identification of a novel CFH variant in patients with EOMD resulting in reduced CFH and FHL-1 and increased local complement activity in EOMD iPSC RPE supports the involvement of CFH haploinsufficiency in EOMD pathogenesis.

C3 Hypocomplementemia Predicts the Progression of CKD towards End-Stage Kidney Disease in IgA Nephropathy, Irrespective of Histological Evidence of Thrombotic Microangiopathy.

Background: IgA nephropathy (IgAN) is the most common primary glomerulonephritis worldwide. IgAN causes end-stage kidney disease (ESKD) in 30-40% of all cases. The activation of the complement system by pathological circulating IgAs, which is often associated with low serum C3 levels (LowC3), seems to play a crucial role. Previous studies have shown an association between histological evidence of TMA, which is the result of alternative complement activation, and poor outcomes. However, it is not known to what extent the decrease in serum C3 levels reflects ongoing TMA injury. Our study aimed at assessing the association between LowC3 and ESKD and whether this association reflects ongoing TMA. Methods: We enrolled all patients with biopsy-proven IgAN and followed-up patients until their last visit, ESKD, or death. Results: Of the 56 patients included in the study, 12 (21%) presented low serum C3 (LowC3) at the time of renal biopsy. TMA was significantly more frequent in the LowC3 group [7/12 (58%) vs. 9/44 (20%), p = 0.02]. After adjusting for potential confounders, LowC3 was strongly associated with an increased hazard of ESKD (hazard ratio [HR]: 5.84 [95%CI: 1.69, 20.15; p = 0.005). The association was not affected by adjusting for TMA. The estimated overall proportion of the relation between C3 and ESKD mediated by TMA was low and not statistically significant. Conclusions: Our study provides evidence that C3 hypocomplementemia is associated with an increased risk of ESKD through mechanisms that are largely independent from TMA.

Nano-selenium (nano-Se) removes the detrimental impacts of plant-based diets on the production performance and well-being of common carp (Cyprinus carpio)

Fishmeal (FM) is not a sustainable source of nutrients due to the over-exploitation of fisheries resources and increasing aquafeed costs. Plant protein (PP) sources have been utilized as FM alternatives in diets. A 60-day nutritional experiment was conducted to study the effects of the complete replacement of FM with PP ingredients in diets supplemented with nano-Se on growth performance, plasma physiological and immunological parameters, and liver antioxidant capacity of common carp (Cyprinus carpio). Five experimental diets were prepared, where FM-Control contained 35 % FM, PP-Control contained complete PP ingredients, and NSe1, NSe2, and Nse3 contained complete PP ingredients supplemented with 1, 2, and 3 mg/kg nano-Se, respectively. The growth performance of fish fed the FM-Control and NSe3 diets was higher than in the PP-Control group (P < 0.05). The best feed conversion and protein efficiency ratios were detected in the NSe3 group but were not significantly different from the FM-Control group (P > 0.05). While the highest levels of antioxidant parameters, including reduced glutathione, superoxide dismutase, catalase, and glutathione peroxidase, were detected in the liver of fish fed with the FM-Control diet, the lowest malondialdehyde concentration was measured in the NSe3 group (P < 0.05). Plasma levels of lysozyme activity, alternative complement activity, and total immunoglobulin of the FM-Control and NSe3 groups were significantly higher than in other groups (P < 0.05). The best plasma physiological parameters including the lowest levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, cholesterol, triglycerides, and low-density lipoprotein, and the highest levels of total protein, triiodothyronine (T3), and high-density lipoprotein were observed in fish fed with the FM-Control and NSe3 diets (P < 0.05). These results suggest that FM can be entirely substituted by PP in diets containing nano-Se at 3 mg/kg without harmful impacts on the performance and immunity of common carp.

PolySialic Acid Nanoparticles Actuate Complement-Factor-H-Mediated Inhibition of the Alternative Complement Pathway: A Safer Potential Therapy for Age-Related Macular Degeneration.

The alternative pathway of the complement system is implicated in the etiology of age-related macular degeneration (AMD). Complement depletion with pegcetacoplan and avacincaptad pegol are FDA-approved treatments for geographic atrophy in AMD that, while effective, have clinically observed risks of choroidal neovascular (CNV) conversion, optic neuritis, and retinal vasculitis, leaving room for other equally efficacious but safer therapeutics, including Poly Sialic acid (PSA) nanoparticle (PolySia-NP)-actuated complement factor H (CFH) alternative pathway inhibition. Our previous paper demonstrated that PolySia-NP inhibits pro-inflammatory polarization and cytokine release. Here, we extend these findings by investigating the therapeutic potential of PolySia-NP to attenuate the alternative complement pathway. First, we show that PolySia-NP binds CFH and enhances affinity to C3b. Next, we demonstrate that PolySia-NP treatment of human serum suppresses alternative pathway hemolytic activity and C3b deposition. Further, we show that treating human macrophages with PolySia-NP is non-toxic and reduces markers of complement activity. Finally, we describe PolySia-NP-treatment-induced decreases in neovascularization and inflammatory response in a laser-induced CNV mouse model of neovascular AMD. In conclusion, PolySia-NP suppresses alternative pathway complement activity in human serum, human macrophage, and mouse CNV without increasing neovascularization.

Influences of makiang (Syzygium nervosum) seed powder on growth performance, immunological response, antioxidant and immune related gene expression in juvenile Nile tilapia (Oreochromis niloticus)

A makiang seed powder (MSP) feeding trial was performed on Nile tilapia (Oreochromis niloticus) for eight weeks to evaluate the effects on growth performance, immune response, and gene expression. Three hundred fingerlings (13.20 ± 0.04 g) were fed five diets with various amounts of MSP at 0, 5, 10, 20 and 40 g kg−1. After four and eight weeks of the feeding trial, growth rates and immune responses were tested, while relative immune and antioxidant gene expression were performed after eight weeks of MSP feeding. Accordingly, the growth indices (SGR, WG) and immune activity (skin mucus and serum lysozyme, skin mucus and serum peroxidase, alternative complement, phagocytosis and respiratory burst activities) in Nile tilapia were significantly increased in the treated diets compared with the control diet (P < 0.05). The maximum (P ≤ 0.05) growth and immune activity values were noted in fish fed 5 g kg−1 MSP. IL1, IL8, LBP, GPX, GSTa and GSR gene expressions were also shown to be upregulated in fish on MSP diets, with the highest value reported in fish fed 5 g kg−1 MSP. Finally, MSP has the potential to be employed as a feed supplement in the biofloc system for the cultivation of Nile tilapia.

Terminalia arjuna Bark Powder as a Potential Immunomodulator in Labeo rohita: Enhanced Hematological, Adaptive, and Humoral Responses against Bacterial Pathogens and Concordant Liver Histomorphology.

This study investigated the dietary immunomodulatory effects of Terminalia arjuna bark powder (TABP) in Labeo rohita, a freshwater fish model. Four iso-nitrogenous and iso-caloric diets containing graded levels of TABP (0, 1, 10, and 15 g/kg were fed to fish for 90 days, followed by a 10 day challenge with pathogenic bacteria Aeromonas hydrophila and Edwardsiella tarda. An integrated biomarker response (IBR) approach assessed the impact of TABP on hematological, adaptive, and humoral immune parameters, along with liver histomorphology. Dietary TABP at 10 g/kg significantly enhanced (p < 0.05) hematological indices (hemoglobin, red blood cell count, hematocrit), specific immune parameters (lysosomal enzyme activity, phagocytosis, respiratory burst), and non-specific immune parameters (serum lysozyme, alternative complement activity), and exhibited improvements in liver architecture consistent with the enhanced immune response. Broken line regression analysis showed 11.5 g/kg to be an optimum dose. However, at 15 g/kg, a compromised trend was observed in some parameters. These findings suggest an optimal dosage range for TABP's immunomodulatory effects. The study highlights the potential of TABP as a natural immunomodulator in fish aquaculture. The improved immune response and concomitant liver health observed in Labeo rohita opens avenues for further research on TABP's applicability in animal health, using fish as a model organism. Additionally, the IBR approach proved effective in evaluating TABP's immunomodulatory properties, paving the way for similar studies on other natural products in aquaculture.

Enhancement of Skin Mucus Immunity, Carotenoid Content, Sexual Parameters, and Growth Response in Guppy Fish (Poecilia reticulata) Fed with Green Algae (Chaetomorpha aerea) Diets

The research aimed to analyze the influences of adding marine green algae Chaetomorpha aerea to the diet of guppy fish (Poecillia reticulate) on growth, immunological responses in skin mucus, total carotenoid content, and sexual characteristics. A total of 450 fish, with a mean body weight of 0.19 ± 0.1 g and 30 fish per tank (triplicate), were randomly fed into 15 experimental tanks, each containing 50 L. Five different diets with 0, 1, 2, 4, 8, and 10% of C. aerea g/kg diets were fed to P. reticulate for 30 days. After 30 days, growth, immunological responses in skin mucus, total carotenoid content, and sexual characteristics were investigated. The results observed that the feed conversion rate and fry output were significantly (p &gt; 0.05) decreased in experimental groups compared to the control group. The results revealed that the dietary inclusion of C. aerea algal significantly increased (p &lt; 0.05) in mucosal immunological parameters containing lysozyme activity, myeloperoxidase activity, total immunoglobulins, and alternative complement activity, which were the highest in the group with 4% of C. aerea g/kg. Additionally, lateral skin and the caudal fin of fish had higher total carotenoid levels from the dietary C. aerea algae diet than the control group, which were the highest in the groups with 4%. Among them, 4 and 8% of C. aerea g/kg diet resulted in better growth performance and feed conversion ratio. Thus, the study suggested that 4% of C. aerea g/kg diet has enrichment of immunity, total carotenoid concentrations, and skin mucus immunity of P. reticulate.

Efficacy of nutritional selenium nanoparticles on growth performance, immune response, antioxidant capacity, expression of growth and immune-related genes, and post-stress recovery in juvenile Sobaity seabream (Sparidentex hasta)

This study evaluated the impacts of nano-Se on the growth, immunity, antioxidant capacity, physiological parameters, gene expression, and stress resistance of fingerling Sobaity seabream (Sparidentex hasta). The fish with an average weight of 21.5 ± 0.1 g were divided into four treatment groups in triplicates that received one of the test diets supplemented with varying levels of nano-Se: 0 (control), 0.5 (Se-0.5), 1 (Se-1), and 2 (Se-2) mg/Kg for 60 days. The results showed that final weight, weight gain rate, specific growth rate, feed intake, and feed conversion ratio improved with significant linear and quadratic trends (P < 0.05) in response to nano-Se-supplemented diets, and the best values were measured in the Se-2 group. Superoxide dismutase activity level remained unaffected among the four groups (P > 0.05). Catalase activity increased in nano-Se-supplemented groups, with the highest level measured in fish fed the Se-0.5 diet. Glutathione peroxidase activity levels were not significantly different between the control and nano-Se groups, but the lowest malondialdehyde concentration was detected in the Se-2 group. Nano-Se had no marked effect on total plasma Ig levels; however, the highest lysozyme activity and alternative complement activity (ACH50) were observed in the Se-0.5 and Se-2 groups, respectively. No significant differences (P > 0.05) were observed in plasma total protein, albumin, globulin, triglyceride, and thyroid hormone (T3 and T4) contents among the groups. However, the lowest cholesterol and low-density lipoprotein values and the highest high-density lipoprotein concentration were measured in the Se-2 group. The Se-0.5 and Se-1 groups exhibited significantly lower levels of aspartate aminotransferase activity, and the lowest alkaline phosphatase activity level was detected in the Se-1 group. The expression level of insulin-like growth factor I gene in all nano-Se-fed groups was significantly higher than the control. Also, the expression of interleukin-1β and lysozyme genes was significantly upregulated in nano-Se-supplemented groups, with the highest values in the Se-2 group. Following acute crowding stress, plasma cortisol and lactate levels at all post-stress time intervals were not significantly different among the experimental groups. Fish fed the Se-0.5 and Se-2 diets tended to have lower plasma glucose concentrations than other groups. In conclusion, dietary nano-Se at 2 mg/kg is recommended to promote growth performance and enhance antioxidant and immune parameters in Sobaity juveniles.

Effects of Dietary Phytol Supplementation on Growth Performance, Immunological Parameters, Intestinal Bacteria, and Prevention of Oxidative Stress Following Transportation of Nile Tilapia, Oreochromis niloticus

Nile tilapia, Oreochromis niloticus, (2.00 ± 0.02 g) were reared in 16 70-L tanks (40 individual/tank) and fed diets (approx. 345 g/kg protein, approx. 87 g/kg crude fat) containing 0 (CTL), 75 (PH-75), 150 (PH-150), and 300 (PH-300) mg/kg phytol (n = 4). After 60 days of feeding (4% daily), growth performance, humoral immune parameters, and gut bacteria were analyzed. Also, hepatic antioxidant parameters were determined before and after the fish were transported in plastic bags for 6 hr. The results showed that PH-75 exhibited the highest final weight (P&lt;0.001), weight gain (P&lt;0.001), feed intake (P&lt;0.001), feed efficiency (P=0.015), plasma lysozyme activity (P=0.004), and intestinal Lactobacillus sp. population (P=0.017), among the treatments. The highest plasma alternative complement activity (P=0.006) and the lowest intestinal total viable bacteria (P=0.027) were observed in PH-75 and PH-150. The highest plasma alkaline phosphatase activities were observed in PH-75 and PH-300 (P=0.014). The highest blood leukocyte (P=0.008), monocyte (P=0.010), and eosinophil (P&lt;0.001) were observed in PH-300, while the highest blood neutrophil was observed in all phytol treatments (P&lt;0.001). The highest hepatic lipid peroxidation was observed in PH-300, whereas PH-75 and PH-150 showed the lowest values (P&lt;0.001). The highest hepatic reduced glutathione was observed in PH-75, also PH-150 exhibited significant elevation in this parameter, compared to CTL (P&lt;0.001). Transportation led to significant elevations in the hepatic antioxidant enzymes’ activities in CTL, PH-75, and PH-150; the highest activities were related to PH-75 and PH-150 treatments, which had also the highest post-transportation survivals (P&lt;0.001). In conclusion, phytol is a suitable feed supplement for Nile tilapia, improving growth performance and welfare, particularly at 75 mg/kg.

Effects of Background Color on Growth, Stress, Biochemical, Hematological, and Immunological Responses, and Expression of Growth-Related Genes in Oscar Fish (Astronotus ocellatus)

The aim of the current study was to assess the impacts of tank color on the growth, stress, biochemical, hematological, and immunological responses, and expression of growth-related genes in juvenile Oscar (Astronotus ocellatus). Therefore, a total of 120 fish (9.14 ± 0.34 g) were distributed into 12 aquariums (60 × 50 × 35 cm) and divided into four treatments (aquariums with blue, white, yellow, and red colors) in three replicates (10 fish per aquarium). The fish were daily hand-fed ad libitum in three meals at 7:00, 12:00, and 17:00 hr for 56 days. Results showed that Oscar fish cultured in the red aquariums had higher final weight and weight gain and lower feed conversion ratio. Plasma cortisol, glucose, and lactate values of fish were significantly higher, and the amount of white blood cells was lower in the blue aquariums compared to other tank colors. Plasma triglyceride values were significantly higher in the white and yellow groups than the blue aquariums. Moreover, fish cultured in yellow and red aquariums had significantly higher melatonin levels than the blue aquariums. Plasma cholesterol, total protein, albumin, complement 3, and total immunoglobulin values were highest in Oscar fish cultured in the red aquariums. Also, lysozyme and alternative complement (ACH50) activities and complement 4 values of fish cultured in yellow and red aquariums were meaningfully higher compared to blue and white groups. Growth hormone relative gene expression levels were meaningfully higher in Oscar fish cultured in white, yellow, and red aquariums than the blue aquariums. Furthermore, insulin-like growth factor-1 (IGF-1) relative gene expression levels were significantly higher in fish cultured in yellow and red aquariums compared to those cultured in blue and white tank colors. Thus, the most suitable tank color for rearing juvenile A. ocellatus is red, while blue tank color is unsuitable.

Complement System Deficiencies in Elite Athletes.

Although regular physical activity improves immune competency and reduces the prevalence of inflammatory diseases, strenuous training in elite athletes is associated with an increased susceptibility to infectious complications. Therefore, the objective of our study was to assess the routinely examined parameters of the complement system in elite athletes. The study was carried out in a cohort of elite athletes (n = 134) and healthy control subjects (n = 110). In all subjects, besides a routine laboratory check-up, serum concentrations of the C3 and C4 complement components, mannose-binding lectin (MBL), as well as activation of all three complement pathways were determined. Compared to healthy controls, lower C3 and C4 complement component concentrations were observed in elite athletes (0.96 ± 0.1 vs. 1.08 ± 0.2mg/L, and 0.18 ± 0.1 vs. 0.25 ± 0.1mg/L, respectively, p < 0.05); with much higher frequency rates of C3 and C4 deficiencies in athletes (31.3 vs. 14.5%, and 6 vs. 0%, p < 0.05). Simultaneously, athletes had much higher frequency rates of deficiencies of activation of classical and alternative complement pathways; while, deficiency of activation of the lectin pathway was similar in both cohorts. We confirmed a high frequency of defects in the complement system in elite athletes. Lower concentrations of C3 and C4 complement components, with high frequencies of deficiencies of the classical and alternative complement activation pathways were the most prevalent disorder of the complement system in elite athletes. Further studies are needed to uncover the functional impacts of these observations upon the susceptibility to infectious diseases.

Effects of dietary common juniper (Juniperus communis) essential oil on growth, immunity, antioxidant status, and disease resistance in rainbow trout, Oncorhynchus mykiss

The main aim of this study is to evaluate the effects of common juniper (Juniperus communis) essential oil (JCEO) on growth, digestive enzymes, biochemical profile, immune and antioxidant system, and liver enzymes in the rainbow trout, Oncorhynchus mykiss. 240 fish (10.63 ± 0.06 g) were fed for 60 days in 12 thanks (20 fish per tank) with experimental diets containing JCEO including: control or T1 (untreated), T2 (fed 0.5% JCEO), T3 (1% JCEO), T4 (2% JCEO). Results revealed an increase in final weight (FW) and weight gain (WG) in all treatments, with a significant difference with the control group (P < 0.05). Feed conversion ratio (FCR) decreased, after addition of JCEO compared to the control group (P < 0.05). Amylase, protease and lipase activity levels increased in all groups (T2, T3 and T4 groups), which were significantly different from the control (P < 0.05). The T2 group had the highest total protein (TP) level in serum compared to the control, and all experimental treatments exhibited a significant difference with the control (P < 0.05). The activity of antioxidant enzymes in serum, superoxide dismutase (SOD) and catalase (CAT) increased compared to the control group, with the highest levels in the T4 group. Inclusion of JCEO to the diet significantly reduced the levels of alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activity in blood compared to the control (P < 0.05). The highest level of serum lysozyme activity and total immunoglobulin (total Ig) were observed in T3 group, which displayed a significant difference with the control (P < 0.05). Serum alternative complement activity (ACH50) increased in the supplemented fish compared to the control (P < 0.05). The fish survival increased after challenge with Yersinia ruckeri in JCEO-supplemented fish. In conclusion, JCEO at dietary level of 1% could improve the nutrient digestion and absorption and provide better growth, as well as reinforce the immune system in rainbow trout.

Effects of exogenous bile acids (BAs) on growth, lipid profile, digestive enzymes, and immune responses of thinlip mullet, Liza ramada

An eight-week trial was designed to explore the dietary effects of commercially purchased exogenous bile acids (BAs) on growth, whole-body composition, lipid profile, intestinal digestive enzymes, liver function enzymes, oxidative stress biomarkers, and serum immunity of thinlip mullet, Liza ramada. Four triplicate groups (10.50 ± 0.05 g) were fed four soybean meal (SBM)-based diets supplied with several BAs levels at 0 (control), 50, 130, or 350 mg/kg feed. Results indicated that the growth was significantly increased in groups fed BAs-based diets, especially at 130 mg/kg feed. The body composition analysis showed that feeding fish on diets supplied with BAs up to 130 mg/kg decreased moisture (%) alongside increased crude protein (%). However, the body composition of fish fed a diet with 350 mg BAs/kg had the lowest moisture (%) and the highest crude protein (%). Moreover, there were significant increases in the intestinal (protease, α-amylase, and lipase) enzyme activities in the groups supplied with BAs up to 130 mg BAs/kg. Liver function enzymes (aspartate aminotransferase and alanine aminotransferase enzyme activities) were significantly decreased in BAs-supplemented groups compared to those fed the BAs-free group. On the other hand, the control group had higher total cholesterol, triglycerides, and low-density lipoprotein alongside the lower high-density lipoprotein than BAs-supplemented groups, especially at 350 mg BAs/kg feed. BAs significantly decreased hepatic malondialdehyde concentrations and increased the activity of hepatic catalase, superoxide dismutase, and total antioxidant capacity compared with those reared on the control diet. Serum lysozyme, respiratory burst, and alternative complement activities were significantly increased in BAs-supplied groups, particularly in the group supplied with 130 mg BAs/kg compared to those fed on the control diet. Accordingly, our findings recommend that including 130 mg BAs/kg in an SBM-based diet enhanced the growth, digestive enzyme activities, and liver functions, alleviated oxidative stress, boosted serum immunity, and lowered lipid metabolites in thinlip mullet. These findings will be beneficial for improving the quality of feed prepared for feeding mullets and an effective alternative strategy to support mullet farming.

Initial investigation on the feasibility of porcine red blood cells from genetically modified pigs as an alternative to human red blood cells for transfusion.

The decline in blood donation rates and the ongoing shortage of blood products pose significant challenges to medical societies. One potential solution is to use porcine red blood cells (pRBCs) from genetically modified pigs as an alternative to human red blood cells (hRBCs). However, adverse immunological reactions remain a significant obstacle to their use. This study aimed to evaluate the compatibility of diverse genetically modified pRBCs with human serum. We acquired human complement-competent serum, complement 7 (C7)-deficient serum, and hRBCs from all ABO blood types. Additionally, we used leftover clinical samples from health checkups for further evaluation. pRBCs were collected from wild-type (WT) and genetically modified pigs: triple knockout (TKO), quadruple KO (QKO), and TKO/hCD55.hCD39 knockin (hCD55.hCD39KI). The extent of C3 deposition on RBCs was measured using flow cytometry after incubation in C7-deficient serum diluted in Ca++-enriched or Ca++-depleted and Mg++-enriched buffers. The binding of immunoglobulin (Ig) M/IgG antibody to RBCs after incubation in ABO-type human serum was evaluated using flow cytometry. Naïve human serum- or sensitized monkey serum-mediated hemolysis was also evaluated. Phagocytosis was assessed by incubating labeled RBCs with the human monocytic cell line THP-1 and measurement by flow cytometry. All three genetic modifications significantly improved the compatibility of pRBCs with human serum relative to that of WT pRBCs. The extent of IgM/IgG binding to genetically modified pRBCs was lower than that of WT pRBCs and similar to that of O-type hRBCs. Total and alternative pathway complement activation in all three genetically modified pRBCs was significantly weaker than that in WT pRBCs and did not differ from that in O-type hRBCs. The extent of serum-mediated hemolysis and phagocytosis of these genetically modified pRBCs was low and similar to that of O-type hRBCs. Sensitized monkey serum-mediated hemolysis in QKO and TKO/hCD55.hCD39KI pRBCs was higher than in O-type hRBCs but lower than in TKO pRBCs. The elimination of porcine carbohydrate antigens in genetically modified pigs significantly enhanced pRBC compatibility with naïve human sera, which was comparable to that of O-type hRBCs. These findings provide valuable insights into the development of pRBCs as potential alternatives to hRBCs.

Effect of dietary nutmeg (Myristica fragrans) on growth performance, antioxidant status, immune response, and gene expression of common carp (Cyprinus carpio)

The aim of this study was to assess the effects of dietary nutmeg (Myristica fragrans (MF)) on growth, innate immunity parameters of serum and mucus, antioxidant defence, and expression of immune genes in common carp (Cyprinus carpio). Three hundred fish (3.79 ± 0.02 g) in four groups were distributed in twelve glass aquariums with a capacity of 150-L. The groups were T1 or control (0% MF), T2 (0.5% MF), T3 (1% MF), and T4 (2% MF), and the fish were sampled after 42 days. According to the results, diet supplementation showed significantly higher weight gain (WG) and lower food conversion ratio (FCR). Final weight (FW) increased in groups fed 1% and 2% MF and specific growth rate (SGR) improved in those provided 1% MF (P < 0.05). Alternative complement activity (ACH50) and the total immunoglobulin (total Ig) level were found to be significantly higher in T4 and T3 groups, respectively than in the T1 (P < 0.05). However, lysozyme (LYZ) and total protein (TP) in all treatment groups showed no significant variation compared with the T1. Similarly, feeding fish with supplemented diets did not affect the antioxidant status and mucus immune parameters. Additionally, fish fed 1% MF showed a significantly higher intestinal catalase (CAT) and superoxide dismutase (SOD) gene expression (P < 0.05). However, LYZ expression significantly decreased in the fish fed MF (P < 0.05). The results of the present study revealed the addition of MF to the diet affects the expression of both antioxidant and immune genes in common carp.