Effects of dietary salidroside on intestinal health, immune parameters and intestinal microbiota in largemouth bass (Micropterus salmoides).

The largemouth bass has become one of the economically fish in China, according to the latest China Fishery Statistical Yearbook. The farming scale is constantly increasing. Salidroside has been found in past studies to have oxidative stress reducing and immune boosting properties. In this study, the addition of six different levels of salidroside supplements were 0、40、80、120、160 and 200 mg kg-1. A 56-day feeding trial was conducted to investigate the effects of salidroside on the intestinal health, immune parameters and intestinal microbiota composition of largemouth bass. Dietary addition of salidroside significantly affected the Keap-1ß/Nrf-2 pathway as well as significantly increased antioxidant enzyme activities resulting in a significant increase in antioxidant capacity of largemouth bass. Dietary SLR significantly reduced feed coefficients. The genes related to tight junction proteins (Occludin, ZO-1, Claudin-4, Claudin-5) were found to be significantly upregulated in the diet supplemented with salidroside, indicating that salidroside can improve the intestinal barrier function (p < 0.05). The dietary administration of salidroside was found to significantly reduce the transcription levels of intestinal tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) (p < 0.05). Furthermore, salidroside was observed to reduce the transcription levels of intestinal apoptosis factor Bcl-2 associated death promoter (BAD) and recombinant Tumor Protein p53 (P53) (p < 0.05). Concomitantly, the beneficial bacteria, Fusobacteriota and Cetobacterium, was significantly increased in the SLR12 group, while that of pathogenic bacteria, Proteobacteria, was significantly decreased (p < 0.05). In conclusion, the medium-sized largemouth bass optimal dosage of salidroside in the diet is 120mg/kg-1.

Immune cell phenotype and function patterns across the life course in individuals from rural Uganda.

Background: To determine the pattern of immune cell subsets across the life span in rural sub-Saharan Africa (SSA), and to set a reference standard for cell subsets amongst Africans, we characterised the major immune cell subsets in peripheral blood including T cells, B cells, monocytes, NK cells, neutrophils and eosinophils, in individuals aged 3 to 89 years from Uganda. Methods: Immune phenotypes were measured using both conventional flow cytometry in 72 individuals, and full spectrum flow cytometry in 80 individuals. Epstein-Barr virus (EBV) IFN-γ T cell responses were quantified in 332 individuals using an ELISpot assay. Full blood counts of all study participants were also obtained. Results: The percentages of central memory (TCM) and senescent CD4+ and CD8+ T cell subsets, effector memory (TEM) CD8+ T cells and neutrophils increased with increasing age. On the other hand, the percentages of naïve T (TN) and B (BN) cells, atypical B cells (BA), total lymphocytes, eosinophils and basophils decreased with increasing age. There was no change in CD4+ or CD8+ T effector memory RA (TEMRA) cells, exhausted T cells, NK cells and monocytes with age. Higher eosinophil and basophil percentages were observed in males compared to females. T cell function as measured by IFN-γ responses to EBV increased with increasing age, peaking at 31-55 years. Conclusion: The percentages of cell subsets differ between individuals from SSA compared to those elsewhere, perhaps reflecting a different antigenic milieu. These results serve as a reference for normal values in this population.

Comparison between inhalational anesthetics in terms of DNA damage and immunological markers.

This study compared genetic damage and immunological markers between surgical patients who underwent inhalational anesthesia with isoflurane or sevoflurane. Blood samples were collected from surgical patients (n = 18 in the isoflurane group and n = 17 in the sevoflurane group) at baseline (before the anesthesia procedure) and the day after anesthesia. DNA damage was detected using an alkaline comet assay; proinflammatory interleukin (IL)-6 was detected by flow cytometry, and white blood cells were detected via an automatic hematology analyzer. The characteristics of both groups were similar, and neither of the two anesthetics induced DNA damage. Similarly, mild neutrophilia was observed after anesthesia in both groups. Increased IL-6 levels were observed 1 day after anesthesia regardless of the type of anesthetic, but this increase was greater in the isoflurane group. Our study suggested that isoflurane and sevoflurane administration may contribute to changes in the immune parameters measured, though no genotoxic hazard was identified, in healthy adult patients who undergo low-stress surgery.

Immuno-protective response of Asian seabass (Lates calcarifer) to inactivated vaccines against Streptococcus iniae and Vibrio harveyi.

BACKGROUND: In this study, the protective immunity and immunogenicity of the monovalent and bivalent Streptococcus iniae and Vibrio harveyi vaccine were evaluated in Asian seabass. To analyze immune responses, 1200 Asian seabass with an average weight of 132.6 ± 25.4 g were divided into eight treatments in triplicates (50 fish per tank) as follows: S. iniae immunized by injection (SI), V. harveyi immunized by injection (VI), bivalent S. iniae and V. harveyi (SVI) immunized by injection, S. iniae immunized by immersion (SIM), V. harveyi (VIM) immunized by immersion, bivalent S. iniae and V. harvei (SVIM) immunized by immersion, phosphate-buffered saline (PBS) by injection (PBSI) and control group without vaccine administration (CTRL). Blood and serum samples were taken at the end of the 30th and 60th days. Then the vaccinated groups were challenged with two bacteria (S. iniae) and (V. harveyi) separately and mortality was recorded for 14 days. RESULTS: This study reveals that there is no significant difference in the hematological parameters on the 30th and 60th days of the experiment in the vaccine-immunized groups compared to the CTRL group (P > 0.05). Meanwhile, there was no significant difference in the amount of serum albumin level, respiratory burst activity, and serum bactericidal activity in the vaccine-immunized groups compared to the CTRL group on the 30th and 60th days of the experiment (P > 0.05). Total protein on the 60th day (in the VI and SVI groups), globulin on the 30th day (in the VI and SVI groups) and the 60th day (in the VI group) compared to the CTRL and PBSI groups had a significant increase (P < 0.05). Complement activity (in the VI and SVI groups) and lysozyme (in the SI and SVI groups) increased significantly compared to the control group (P < 0.05). Serum antibody titer against S. iniae had a significant increase in the SI, VI, SVI and SVIM groups compared to the CTRL and PBSI groups (P < 0.05). Serum antibody titer against V. harveyi had a significant increase in the groups immunized with the vaccine compared to the CTRL and PBSI groups (P < 0.05). A significant increase in the relative percentage survival (RPS) following challenge with S. iniae in the SVI (86.6%), SI (83.3%,) and VI (73.3%) groups were observed compared to the CTRL (43.3%) and PBSI (40%) groups (P < 0.05). Also, a significant increase in the RPS after challenge with V. harveyi in the SVI group, VI 86.6%, SVI 83.3%, VIM 80% and SVIM 76.6% were observed compared to the CTRL (46.6%) and PBSI (50%) groups (P < 0.05). CONCLUSION: Overall, the results demonstrated that the bivalent vaccine of S. iniae and V. harveywas able to produce significant immunogenicity and RPS in Asian seabass.

Effect of feeding fermented distiller's grains diets on immune status and metabolomics of spleen and mesenteric lymph nodes in finishing cattle.

To explore the effect of feeding fermented distiller's grains (FDG) diets on spleen and mesenteric lymph nodes (MLN) immune status and metabolomics in finishing cattle, eighteen Guanling crossbred cattle (18 months old, 250.0 ± 25 kg) were randomly divided into 3 groups: a basal diet (Control) group, an FDG-15% group, and an FDG-30% group (containing 0%, 15% and 30% FDG to partially replace the concentrates, respectively). After 75 days, the spleens and MLN were collected for detection of relative spleen weight, immune parameters, and metabolomic analysis. Compared with the Control group, FDG-30% group significantly increased (P<0.05) the relative spleen weight. In addition, the level of IL-17A in the spleen of the FDG-30% group was significantly higher than that of the FDG-15% group. Metabolomic analysis showed that differential metabolites (VIP＞1, P＜0.05) of spleen and MLN in FDG-15% and FDG-30% groups are mostly lipids and lipid molecules. KEGG analysis illustrated that choline metabolism in cancer, glycerophospholipid metabolism, biosynthesis of unsaturated fatty acids and insulin resistance were metabolic pathways in spleen shared by FDG-15% group vs.Control group and FDG-30% group vs.Control group, and choline metabolism in cancer was a metabolic pathway in MLN shared by FDG-15% group vs.Control group and FDG-30% group vs.Control group. These results suggest that feeding FDG may promote spleen development by regulating choline metabolism in cancer, glycerophospholipid metabolism, biosynthesis of unsaturated fatty acids and insulin resistance. Additionally, it may affect MLN development by regulating choline metabolism in cancer. SIGNIFICANCE: Fermented distiller's grains (FDG) is a high quality alternative to feed because it is rich in beneficial microorganisms and nutrients. The spleen and mesenteric lymph nodes (MLN) are important peripheral immune organs in animals, whose status reflects the health of the animal. However, there are few reports on the effect of feeding FDG diets on spleen and MLN immune status and metabolomics in domestic animals. In this study, we found that feeding FDG may promote spleen development by regulating choline metabolism in cancer, glycerophospholipid metabolism, biosynthesis of unsaturated fatty acids and insulin resistance metabolic pathways, and may affect MLN development by regulating choline metabolism in cancer. This study extends our understanding of the metabolomics of the spleen and MLN in FDG and helps to further understand of the immunomodulatory effects of the FDG diet.

Synergism of fermented feed and ginseng polysaccharide on growth performance, intestinal development, and immunity of Xuefeng black-bone chickens.

Microbial fermented feed (MF) is considered a valuable strategy to bring advantages to livestock and is widely practiced. Oral supplementation of Ginseng polysaccharide (Gps) eliminated weight loss in chickens following vaccination. This study investigated the effects of the combined use of Gps and MF on growth performance and immune indices in Xuefeng black-bone chickens. A total of 400 Xuefeng black-bone chickens at the age of 1 day were randomly assigned to four groups. Normal feed group (Control group), ginseng polysaccharide (200 mg/kg) group (Gps group), microbially fermented feed (completely replace the normal feed) group (MF group), and microbially fermented feed and add ginseng polysaccharide just before use (MF + Gps group). Each group contained 5 pens per treatment and 20 birds per pen. The body weight and average daily gain in the Gps, MF, and MF + Gps groups increased significantly (P < 0.01), while the feed conversion ratio decreased significantly (P < 0.01). The combined use of MF and Gps showed a synergistic effect. There was no significant difference in villus height (cecal) between the experimental group and the Con group. The crypt depth of the three experimental groups exhibited a significantly lower value compared to the Control group (P < 0.05). The V/C ratio of the Gps group and MF + Gps was significantly increased (P < 0.05), but there was no significant difference in the MF group. Moreover, the diarrhea rate of the Gps and the MF + Gps groups was lower than that of the Con group, while that of the MF + Gps group decreased the mortality rate (P < 0.05). The serum tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6) levels in the MF, Gps, and MF + Gps groups decreased significantly (P < 0.01), the serum immunoglobulin G (IgG) levels increased significantly (P < 0.01), while the combination of MF and Gps had a synergistic effect. The combined use of Gps and MF not only further improved growth performance and immune parameters, but also reduced the diarrhea rate and mortality.

Impacts of florfenicol on immunity, antioxidant activity, and histopathology of Oreochromis niloticus: a potential protective effect of dietary spirulina platensis.

The misuse of antibiotics enhances the development of resistant microorganisms and decreases the efficacy of treatments. Florfenicol (FF) is one of the antibiotics approved for use in aquaculture in Egypt. Because of its extensive usage, potential negative impacts on aquatic creatures are a major concern. This motivates us to search for an appropriate neoadjuvant to work synergistically with FF and reduce adverse effects. Results from this study will contribute towards improving the understanding of the impacts of FF on Oreochromis niloticus and the possible amelioratory effects of Spirulina platensis algae (SP). O. niloticus (n = 240; 40 ± 2.5 g) were fed on two diets supplemented with or without SP for 4 weeks, then divided into four treatments each in three replicates (n = 60/treatment). G1; was fed a control diet, and the other groups were fed diets supplemented with FF (10 mg /kg of BW, G2), SP (2 g/kg of diet, G3), or FF + SP (G4) for 10 days. Among the four groups, the SP group (G3) had the best immunostimulatory effects as observed by a significant (p < 0.05) elevation in phagocytic activity, phagocytic index, IL6, and TNF-α. The treatment with FF had significantly impacted hepatic and renal tissues, as the values of liver enzymes and creatinine demonstrated tissue deterioration and also resulted in oxidative stress, which was expressed by an increase of GPx, CAT, and SOD in (G2). Additionally, the combined FF + SP improved the hematological parameters and decreased the oxidative damage induced by FF (G4). Thus, it was clear that FF has harmful effects on O. niloticus and that SP can modulate such impacts. These data recommend the use of SP as an effective immunostimulant and a probable adjuvant to FF in O. niloticus diets to attain maximum disease resistance.

Dietary Inclusion of Halobacterium salinarum Modulates Growth Performances and Immune Responses in Farmed Gilthead Seabream (Sparus aurata L.).

The use of natural immunostimulants is considered the most promising alternative to promote fish health, productive performance and quality, increasing the aquaculture profitability, sustainability and social acceptance. The purpose of this study was to evaluate the effect of the integration of a potential probiotic strain, Halobacterium salinarum, belonging to the Archaea domain, in the formulated diets of farmed gilthead seabream (Sparus aurata L.) in terms of growth performances and immunity responses. The experiment was set up to test two different levels of inclusion of the bacteria in the diet: 0.05% (D1) and 0.1% (D2). The effects on fish growth performances; humoral (peroxidase, protease, antiprotease and IgM levels) and cellular immunity parameters (phagocytosis, respiratory burst and myeloperoxidase), along with bactericidal activity, were evaluated after 15 and 30 days of experimental feeding. The obtained results showed that the inclusion of H. salinarum at the highest concentration (D2 0.1%) improved growth performances, bactericidal activity against Vibrio anguillarum and some parameters related both to the humoral and cellular immune response, suggesting exploring other aspects of welfare in view of future supplementations of this probiotic strain in the diet of S. aurata.

Responses of broilers challenged by Eimeria maxima fed with different levels of dietary balanced protein.

This study aimed to evaluate the effects of different dietary balanced protein (BP) levels on the gut health, amino acid apparent ileal digestibility (AID), footpad dermatitis lesions, and litter quality in broiler chicks infected with Eimeria maxima. A total of 2400 male 14-day-old Cobb500 broilers were randomly allotted into 10 treatments with six replications containing 40 birds each in a factorial design of 5 × 2. The treatments consisted of five levels of BP (6.66%, 13.32%, 19.98%, 26.64%, and 33.3%), and broilers unchallenged (NCH) or challenged (CH). Broilers in the CH group received 1 mL of Eimeria maxima inoculum (7 × 103 sporulated oocysts/mL). Oocyst count in excreta, visual intestinal modifications score, morphology, and morphometrics of the ileum were used to determine gut health status. Additionally, amino acids and CP AID, litter quality, and footpad dermatitis were evaluated. An ANOVA and Kruskal-Wallis tests followed by post-hoc tests were performed. The oocyst count in the CH group increased with an increase in dieatary BP (P = 0.08). The incidence of intestinal modifications was higher in the CH group (P < 0.05) and increased with increasing dietary BP (P < 0.05). Morphometrics were impaired by the challenge (P < 0.05), and by the two highest BP levels (P < 0.05). Amino acids AID (methionine, methionine + cystine, arginine, and serine) were reduced by E. maxima challenge. An increase in dietary BP resulted in poor litter quality and high prevalence of of footpad dermatitis (P < 0.05). The E. maxima challenge and increased BP decreased gut health, litter quality, and cause a high incidence of footpad dermatitis.

Effects of ribonucleotide supplementation in modulating the growth of probiotic Bacillus subtilis and the synergistic benefits for improving the health performance of Asian seabass (Lates calcarifer).

In aquaculture, due to the requirements for high-density culture, the diseases caused by bacterial pathogens have become a serious issue. To solve this problem, we performed synbiotic application of RNA and Bacillus subtilis as a sustainable and eco-friendly approach to improve the health and immunity of Asian seabass (Lates calcarifer) during cultivation without using any harmful antibiotics or chemicals. Among various forms of nucleic acids, such as mononucleotides and DNA, RNA was found to be most effective in promoting the growth performance of probiotic B. subtilis in all the tested minimal medium conditions. Accordingly, we used the synbiotic combination of B. subtilis and RNA for Asian seabass cultivation. After feed supplementation for fourteen days, the fish that received the combination treatment exhibited a significant increase in innate cellular and humoral immune parameters, including phagocytic activity, phagocytic index, respiratory burst, serum lysozyme and bactericidal activities, as well as upregulated expression of immune-related genes, including HEPC1, A2M, C3, CC, CLEC, LYS, HSP70, and HSP90. Furthermore, significant increases were observed in the ileal villus height and goblet cell numbers in the intestinal villi in all fish treatment groups. The combination treatment did not cause histopathological abnormalities in the intestine and liver, suggesting that the synbiotic treatment is safe for use in fish. The treated Asian seabass also exhibited a significantly increased survival rate after Aeromonas hydrophila challenge. These results indicate that the synbiotic mixture of B. subtilis and RNA can be considered a beneficial feed additive and immunostimulant for Asian seabass cultivation.

Exopolysaccharides-Mediated ZnO Nanoparticles for the Treatment of Aquatic Diseases in Freshwater Fish Oreochromis mossambicus.

Bacterial fish disease outbreaks are a key concern for aquaculture. Complementary feed additives such as immunostimulants can serve as an ideal solution for disease prevention. Herein, we scrutinized the efficacy of exopolysaccharides (EPSs) from probiotic Bacillus licheniformis and EPS-mediated zinc oxide nanoparticles (EPS-ZnO NPs) for a diet to evaluate growth parameters, antioxidant enzyme activities, and immune stimulation together with disease resistance against Aeromonas hydrophila and Vibrio parahaemolyticus in Mozambique tilapia Oreochromis mossambicus. Fish were separated into seven groups, with six experimental groups fed with EPS and EPS-ZnO NPs at 2, 5, and 10 mg/g and a control fed a basal diet. The fish ingesting feed supplemented with EPS and EPS-ZnO NPs at 10 mg/g showed improved growth performance. Cellular and humoral-immunological parameters were tested in serum and mucus after 15 and 30 days of feeding. These parameters were substantially enriched with a 10 mg/g diet (p < 0.05) of EPS and EPS-ZnO NPs in comparison with the control. Furthermore, the EPS and EPS-ZnO NP supplemental diet actively enhanced the antioxidant response (glutathione peroxidase, superoxide dismutase, and catalase). In addition, the supplemental diet of EPS and EPS-ZnO NPs lowered the death rate and improved the disease resistance of O. mossambicus following assessment with A. hydrophila and V. parahaemolyticus at 50 µL. Hence, the overall results suggest that the supplemental diet of EPS and EPS-ZnO NPs might be used to ensure aquaculture feed additives.

Effects of ß-1,3/1,6 glucan dietary supplements on some immunological and hematological health markersin Siberian sturgeon (Acipenser baerii) infected with Aeromonas hydrophila.

This experimental study aimed to investigate some hematological and immunological changes as a result of Aeromanas hydrophila infection in Siberian sturgeon (Acipenser baerii Brandt, 1869). Their feeds were supplemented with ß-1,3/1,6 glucan at different ratios, 250 mg/kg (ßG250); 500 mg/kg (ßG500) and 750 mg/kg (ßG750). To create an experimental infection, 4×106 cfu/ml Aeromonas hydrophila inoculum was intraperitoneally injected to fish. 0.1 ml intraperitoneal bacteria injection was given to the fish in 12 of 15 tanks, each consisting of 10 fish (the fish in the control group were not given bacteria). Considering the ßG500 and ßG750 group fish as positive (C+) and negative control (C-) groups in terms of hematological parameters, it was found that RBC (erythrocyte) and Hb (hemoglobin) values, as well as RBC indices (MCV, MCHC, MCH) significantly increased. The immunological parameters, including WBC (leukocyte), leukocyte cell percentages (lymphocyte, monocyte, neutrophil, eosinophil), as well as cytokines, including IL-1ß, IL-6, IL-8, and TNF-α values showed similar increases in the ßG500 and ßG750 groups. It was found that the addition of 500 and 750 mg/kg doses of ß-1,3/1,6 glucan to the feed stimulated non-specific immunity of fish against bacterial agents and/or septicemic diseases and beta glucan at this dosage range was determined to be ideal for fish health and that it may be a herbal immunostimulant that can be an alternative to many medicaments.

Efficacy of polyvalent vaccine on immune response and disease resistance against streptococcosis/lactococcosis and yersiniosis in rainbow trout (Oncorhynchus mykiss).

Diseases are the most significant challenge in the development and stability of aquaculture. In this study, the immunogenic efficiency of polyvalent streptococcosis/lactococcosis and yersiniosis vaccines was evaluated by injection and immersion methods in rainbow trout.. The 450 fish with an average weight of 50 ± 5 g were divided into three treatments and three replications as follows: injection vaccine treatment, immersion vaccine treatment and control group without vaccine administration. Fish were kept for 74 days and sampling was done on days 20, 40 and 60. Then, from the 60th to the 74th day, the immunized groups were challenged with three bacteria Streptococcus iniae (S. iniae), Lactococcus garvieae (L. garvieae) and Yersinia ruckeri (Y. ruckeri) separately. A significant difference was observed in the weight gained (WG) in the immunized groups compared to the control group (P < 0.05). The relative survival percentage (RPS) after 14 days of challenge with S. iniae, L. garvieae and Y. ruckeri in the injection group compared to the control group increased respectively (60%, 60% and 70%), (P < 0.05). Also, RPS in the immersion group had an increase respectively (30%, 40% and 50%) after the challenge with S. iniae, L.garvieae and Y. ruckeri compared to the control group. Immune indicators such as antibody titer, complement and lysozyme activity significantly increased in comparison to the control group (P < 0.05). In general, it can be concluded that applying three vaccines by injection and immersion method has significant effects on immune protection and survival rate. However, the injection method is more effective and more suitable than the immersion method.

The antibacterial activity and immunomodulatory effect of naturally synthesized chitosan and silver nanoparticles against Pseudomonas fluorescence infection in Nile tilapia (Oreochromis niloticus): An in vivo study.

A sixty-day feeding trial was conducted to assess the effects of silver nanoparticles (AgNPs) and chitosan nanoparticles (CNPs) on the growth and immunity of Nile tilapia (Oreochromis niloticus), compared with the control group. CNPs and AgNPs were green synthesized and added to a control diet (30% crude protein) at levels of 2.0 g CNPs/kg diet and 1.0 mg AgNPs/kg diet. One hundred and eighty fish (101 ± 3.98 g) were randomly distributed into nine fiberglass tanks (200 cm × 200 cm x 100 cm, twenty fish each) to represent three equal groups (60 fish per group). After one and two months of the feeding trial, parameters of water quality, growth indices, hematology, and liver and kidney biomarkers were evaluated. At the end of the experiment, 10 fish from each group were challenged experimentally via the intraperitoneal injection with Pseudomonas fluorescence and fish mortality was observed for further ten days. Then, specimens from the liver, kidney, spleen, and anterior intestine were examined to assess the histopathological alterations. Incorporating a 2.0 g CNPs/kg diet was a promising growth enhancer; however, a 1.0 mg AgNPs/kg diet had no effects on tilapia performance. Furthermore, AgNPs appeared to reduce water pollution, leading to water filtration via decreasing both total dissolved solids (TDS) and electrical conductivity (EC). A significant role of AgNPs in improving tilapia's erythrogram (RBCs number and Hb concentration) was evident. Compared with the control group, both groups of CNPs and AgNPs improved non-specific immune parameters and showed defense effects against P. fluorescence. The fish mortality after P. fluorescence infection in CNPs and AgNPs-fed fish groups revealed significant decreases (P < 0.05) of 10% and 25%; respectively, while the control group exhibited a mortality rate of 40%. The current investigation evoked that using dietary CNPs (2.0 g/kg feed) as an antibacterial agent against P. fluorescence infection in Nile tilapia culture was better than dietary AgNPs (1.0 mg/kg diet) which, induced cells inflammation causing tissues necrosis.

Comprehensive analysis of immune parameters, mRNA and miRNA profiles, and immune genes expression in the gill of rainbow trout infected with infectious hematopoietic necrosis virus (IHNV).

Rainbow trout (Oncorhynchus mykiss) is a species of cold-water fish with important economic values, widely cultivated worldwide. However, the outbreak of infectious hematopoietic necrosis virus (IHNV) caused the large-scale death of rainbow trout and seriously restricted the development of the trout farming industry. In this study, the changes of immune parameters in different periods (6-, 12-, 24-, 48-, 72-, 96-, 120-, and 144 h post-infection (hpi)), transcriptome profiles of 48 hpi (T48G) compared to control (C48G), and key immune-related genes expression patterns were measured in rainbow trout gill following IHNV challenge through biochemical methods, RNA sequencing (RNA-seq), and quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that alkaline phosphatase (AKP), acid phosphatase (ACP), total superoxide dismutase (T-SOD), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, as well as lysozyme (LZM) and malonaldehyde (MDA) content decreased and then increased during infection, and remained at a high level after 48 hpi (P < 0.05), whereas catalase (CAT) activity showed a significant peak at 48 hpi (P < 0.05). The mRNA and miRNA analysis identified 4343 differentially expressed genes (DEGs) and 11 differentially expressed miRNAs (DEMs), and numerous immune-related DEGs involved in the Toll-like receptor signaling pathway, apoptosis, DNA replication, p53 signaling, RIG-I-like receptor signaling pathway, and NOD-like receptor signaling pathway and expression were significantly up-regulated in T48Gm group, including tlr3, tlr7, tlr8, traf3, ifih1, trim25, dhx58, ddh58, hsp90a.1, nlrc3, nlrc5, socs3, irf3, irf7, casp7, mx1, and vig2. The integrated analysis identified several important miRNAs (ola-miR-27d-3p_R+5, gmo-miR-124-3-5p, ssa-miR-301a-5p_L+2, and ssa-miR-146d-3p) that targeted key immune-related DEGs. Expression analysis showed that tlr3, tlr7, traf3, ifih1, dhx58, hap90a.1, irf3, irf7, and mx1 genes increased and then decreased during infection, and peaked at 72 hpi (P < 0.05). However, trim25 expression peaked at 96 hpi (P < 0.05). This study contributes to understanding immune response of rainbow trout against IHNV infection, and provides new insights into the immune regulation mechanisms and disease resistance breeding studies.

Exposure of Cyprinus carpio var. larvae to PVC microplastics reveals significant immunological alterations and irreversible histological organ damage.

Microplastics have become pervasive in ambient aquatic ecosystems over the last decade and are currently a serious global scale concern. To evaluate the potential toxic effects of PVC (polyvinyl chloride) microplastics on the immune functions of freshwater fish, this study undertook a conducted chronic 60-day dietary exposure experiment with Cyprinus carpio var. larvae. We exposed the fish to four microplastic treatments of different concentrations (food rationed diets): no-plastic (control), 10%, 20% and 30%. At the end of the experimental period the impacts of microplastics on the histology, biochemistry, ROS (reactive oxygen species) levels and gene transcription of immune organs were investigated. The results revealed that PVC microplastics induced cytoplasmic vacuolation in the liver, damaged villi in the intestine, inflammatory cell infiltration, hemosiderosis and vacuolar degeneration in the spleen, glomeruli tuft shrinkage and aggregation of melanin macrophage cells in the kidney. Moreover, following PVC microplastics exposure, ROS levels in the liver and protein levels of pro-inflammatory cytokines including IL-6, IL-8, and TNFα in the liver and serum were increased. Furthermore, modifications in the activities of non-specific immunoenzyme ACP (Acid phosphatase), AKP (alkaline phosphatase), LZM (lysozyme), and expression levels of a range of immune-related genes were observed. Using various techniques at the histological, biochemical and molecular levels, our findings demonstrated the effects of PVC microplastics on changes and imbalances in the immune status of carp. The results of this study provide basic toxicological data toward elucidating and quantifying the impacts of microplastics immunotoxicity on aquatic organisms.

Immunoprotective role of aloin and disease resistance in Labeo rohita, infected with bacterial fish pathogen, Aeromonas hydrophila.

The effect of aloin on mucosal immune response and disease resistance was elucidated in Labeo rohita infected with the bacterial fish pathogen, Aeromonas hydrophila. Fishes were divided into four groups: (i) control, (ii) vehicle control, (iii) bacteria infected and (iv) bacteria infected and aloin treated. Fish were intraperitoneally injected with A. hydrophila suspension at the dose of 2 × 106 CFU/fish at 0 day (d). Following bacterial injection at 0 d, fish were treated with aloin at a dose of 1 mg/kg body weight intraperitoneally at an interval of 24 h for 4 consecutive days. Mucus collected from fish of each group was analyzed at 2 d, 4 d, 6 d, 8 d and 10 d. In bacteria-infected fish, a significant decrease (P < 0.05) in the activity of certain enzymatic and non-enzymatic immune parameters was observed. The activity of these immune parameters showed a gradual recovery on administration of aloin in bacteria-infected fish. Cumulative mortality was also found to be low in the aloin-treated group as compared to that in the infected group. Thus, aloin could act as an immunostimulant and play a protective role against disease caused by bacteria.

Prognostic value of a nomogram based on peripheral blood immune parameters in unresectable hepatocellular carcinoma after intensity-modulated radiotherapy.

BACKGROUND: For patients with unresectable hepatocellular carcinoma (uHCC), intensity-modulated radiotherapy (IMRT) has become one of the options for clinical local treatment. Immune parameters, including platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR) and systemic immune inflammatory (SII), predict survival in various cancers. This study aimed to determine whether peripheral immune parameters can predict survival in patients with uHCC undergoing IMRT and establish a clinically useful prognostic nomogram for survival prediction. METHODS: The clinical data of 309 HCC patients were retrospectively analyzed and randomly divided into training (n = 216) and validation (n = 93) cohorts. PLR, NLR and SII were collected before and after IMRT. Univariate and multivariate Cox analyses were performed to identify independent prognostic factors affecting survival, which were used to generate a nomogram. RESULTS: The median survival was 16.3 months, and significant increases in PLR, NLR, and SII were observed after IMRT (P < 0.001). High levels of immune parameters were associated with poor prognosis (P < 0.001); enlarged spleen, Barcelona clinic liver cancer stage (B and C), post-SII, and delta-NLR were independent risk factors for survival and were included in the nomogram, which accurately predicted 3- and 5-year survival. The nomogram was well verified in the validation cohort. CONCLUSIONS: High levels of immune parameters are associated with poor prognosis in uHCC patients receiving IMRT. Our nomogram accurately predicts the survival of patients with uHCC receiving IMRT.

Transcriptome analysis reveals immune and metabolic regulation effects of Poria cocos polysaccharides on Bombyx mori larvae.

Poria cocos polysaccharides (PS) have been used as Chinese traditional medicine with various pharmacological effects, including antiviral, anti-oxidative, and immunomodulatory activities. Herein Bombyx mori silkworm was used as a model animal to evaluate the immunomodulatory effects of PS via detecting the changes of innate immune parameters and explore the underlying molecular mechanism of the immunoregulatory effect of PS using Illumina HiSeq Xten platform. The results presented here demonstrated that a hemocoel injection of PS significantly enhanced the cellular immunity of silkworm, including hemocyte phagocytosis, microaggregation, and spreading ability. A total of 335 differentially expressed genes (DEGs) were screened, including 214 upregulated genes and 121 downregulated genes by differential expression analysis. Gene annotation and enrichment analyses showed that many DEGs related to immune signal recognition, detoxification, proPO activation, carbohydrate metabolism, and lipid metabolism were significantly upregulated in the treatment group. The Kyoto Encyclopedia of Genes and Genomes-based Gene Set Enrichment Analysis also revealed that the more highly expressed gene sets in the PS treatment silkworm were mainly related to immune signal transduction pathways and energy metabolism. In addition, the activity of four enzymes related to immunity and energy metabolism-including phenoloxidase, glucose-6-phosphate dehydrogenase, hexokinase, and fatty acid synthetase-were all significantly increased in the larvae injected with PS. We performed qRT-PCR to examine the expression profile of immune and metabolic-related genes, which further verified the reliability of our transcriptome data and suggested that PS can regulate the immunity of silkworm by enhancing the cellular immunity and modulating the expression levels of genes related to immune responses and physiological metabolism. These findings will lay a scientific foundation for the use of PS as an immunomodulator in disease prevention in human beings or animals.

Integrated analysis of immune parameters, miRNA-mRNA interaction, and immune genes expression in the liver of rainbow trout following infectious hematopoietic necrosis virus infection.

Rainbow trout (Oncorhynchus mykiss) is an important economical cold-water fish worldwide. However, infection with infectious hematopoietic necrosis virus (IHNV) has severely restricted the development of aquaculture and caused huge economic losses. Currently, little is known about the immune defense mechanisms of rainbow trout against IHNV. In this study, we detected the changes of immune parameters over different post-infection periods (6-, 12-, 24-, 48-, 72-, 96-, 120-, and 144 hours post-infection (hpi)), mRNA and miRNA expression profiles under 48 hpi (T48L) compared to control (C48L), and key immune-related genes expression patterns in rainbow trout liver following IHNV challenge through biochemical methods, RNA-seq, and qRT-PCR, and the function of miR-330-y was verified by overexpression and silencing in vitro and in vivo. The results revealed that alkaline phosphatase (AKP), alanine aminotransferase (ALT), catalase (CAT), and total superoxide dismutase (T-SOD) activities, and lysozyme (LZM) content showed significant peaks at 48 hpi, whereas malondialdehyde (MDA) content and aspartate aminotransferase (AST) activity decreased continuously during infection, and acid phosphatase (ACP) activity varied slightly. From RNA-seq, a total of 6844 genes and 86 miRNAs were differentially expressed, and numerous immune-related differentially expressed genes (DEGs) involved in RIG-I-like receptor signaling pathway, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, cytokine-cytokine receptor interaction, and antigen processing and presentation were significantly upregulated in T48Lm group, including IFIH1, DHX58, MAVS, TRAF3, IRF3, IRF7, MX1, TLR3, TLR8, MYD88, NOD1, NOD2, IL-8, CXCR1, CD209, CD83, and TAP1. Integrated analysis identified seven miRNAs (miR-425-x, miR-185-x, miR-338-x, miR-330-y, miR-361-x, miR-505-y, and miR-191-x) that target at least three key immune-related DEGs. Expression analysis showed that IFIH1, DHX58, IRF3, IRF7, MX1, TLR3, TLR8, and MYD88 showed a marked increase after 24 hpi during infection. Further research confirmed TAP1 as one of the targets of miR-330-y, overexpression of miR-330-y with mimics or agomir significantly reduced the expression levels of TAP1, IRF3, and IFN, and the opposite effects were obtained by inhibitor. These results facilitate in-depth understanding of the immune mechanisms in rainbow trout against IHNV.