Dietary Bacillus velezensis R-71003 and sodium gluconate improve antioxidant capacity, immune response and resistance against Aeromonas hydrophila in common carp.

This study aimed to evaluate the effects of dietary supplementation with Bacillus velezensis R-71003 combined with sodium gluconate on antioxidant capacity, immune response and resistance against Aeromonas hydrophila in common carp. In addition, the biocontrol potential of the secondary metabolites of B. velezensis R-71003 was also evaluated to analyze the possible mechanism of B. velezensis R-71003 against A. hydrophila. The results indicated that the antibacterial crude extract of B. velezensis R-71003 can destroy the cell wall of A. hydrophila. Moreover, the results showed that dietary B. velezensis R-71003 could promote antioxidant capacity, which significantly increased the activities of CAT and SOD and decreased the content of MDA. Additionally, B. velezensis R-71003 supplementation significantly enhanced the immunity of common carp, as measured by the mRNA expression levels of cytokine-related genes (TNF-α, TGF-ß, IL-1ß and IL-10). In addition, dietary B. velezensis R-71003 exhibited an upregulation of IL-10 and a downregulation of IL-1ß, coupled with higher survival rates when challenged with A. hydrophila compared to the positive group. Furthermore, compared to prechallenge, the mRNA expression levels of TLR-4, MyD88, IRAK1, TRAF6, TRIF and NF-κB in the head kidney of common carp were significantly increased after challenge. The fish fed the B. velezensis R-71003 diet showed lower expression of TLR-4, MyD88, IRAK1, TRAF6, TRIF and NF-κB after the challenge than those fed the control diet. Thus, this study revealed that B. velezensis R-71003 can improve the resistance of common carp to pathogenic bacteria by destroying bacterial cell walls and improving fish immunity by activating the TLR4 signaling pathway. Importantly, this study indicated that sodium gluconate has a positive effect on B. velezensis R-71003 in enhancing the anti-infection ability of common carp. The results of this study will lay the foundation for the application of B. velezensis R-71003 in combination with sodium gluconate as an alternative to antibiotics in aquaculture.

Effects of an exopolysaccharide from Lactococcus lactis Z-2 on innate immune response, antioxidant activity, and disease resistance against Aeromonas hydrophila in Cyprinus carpio L.

Microbial exopolysaccharides (EPS) from Lactococcus have been found to have an important role in the probiotic activity of this bacterium; however, the immunomodulatory and antioxidant activities have not been fully explored in aquaculture. In the present study, we investigated EPS-2 from Lactococcus lactis Z-2, isolated from healthy common carp, for its immunomodulatory and antioxidant effects and disease resistance against Aeromonas hydrophila in Cyprinus carpio L. We found that the molecular weight of EPS-2 was 18.65 KDa. The monosaccharide composition of this polymer was rhamnose, xylose, mannose, glucose, and galactose at a molar percentage of 13.3%, 14.1%, 18.5%, 27.4%, and 26.7%, respectively. EPS-2 treatment could modulate the immune responses in vitro and in vivo. In vitro tests showed that EPS-2 could significantly enhance the proliferation and phagocytosis activities (P < 0.05) as well as induce the production of nitic oxide (NO), pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6), and anti-inflammatory cytokines (IL-10, TGF-ß) (P < 0.05) in head kidney cells. When the fish were gavaged with three different concentrations of EPS-2 (250, 500, 1000 µg/mL) for 7 days and infected with A. hydrophila, different expression patterns of the NO, cytokines, lysozyme (LZM), and alkaline phosphatase (AKP) in the serum and of antioxidants (T-AOC, SOD, CAT, GSH, GSH-Px and MDA) in hepatopancreas were observed. Before infection with A. hydrophila, EPS-2 supplementation significantly up-regulated the NO production, protein levels of pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6), LZM and AKP activities, and levels of antioxidant molecules compared to those in the negative (G1) group (P < 0.05), whereas levels of NO and pro-inflammatory cytokines and LZM and AKP activities were significantly lower than those in the positive (G2) group after infection (P < 0.05). However, whether infected or not, the expression levels of anti-inflammatory cytokines (IL-10, TGF-ß) were significantly increased in the EPS-2 treatment groups (P < 0.05). These results indicate that EPS-2 has immunomodulatory and antioxidant effects on common carp, both in vitro and/or in vivo, and can be applied as a common carp feed supplement to enhance fish immunity and disease resistance against A. hydrophila.

Characterization of a polysaccharide HP-02 from Honeysuckle flowers and its immunoregulatory and anti-Aeromonas hydrophila effects in Cyprinus carpio L.

Aquaculture systems are known to be major reservoirs of Aeromonas hydrophila. The use of Chinese medicinal polysaccharides has become a successful strategy to prevent bacterial diseases in aquaculture. A water-soluble polysaccharide, HP-02, was purified from honeysuckle flowers, and the present study was aimed to examine its immunomodulatory and anti-A. hydrophila effects in Cyprinus carpio L. The results showed that the molecular weight of HP-02 was estimated to be 38 kDa. Monosaccharide composition was determined to be arabinose, rhamnose, mannose, glucose, and galactose in a molar ratio of 2.5: 1.8: 3.6: 3.7: 1.9. HP-02 not only distinctly improved the proliferation and phagocytosis of head kidney cells, but also exerted significant immunoregulation activity by increasing the secretion of the pro-inflammatory cytokines TNF-α, IL-1ß, IL-6, and IL-12 and the anti-inflammatory cytokines IL-10 and TGF-ß at the protein level in head kidney cells and serum. Moreover, during pathogen infection in vivo, HP-02 significantly reduced the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines. These results indicated that HP-02 had immunomodulatory effects on common carp both in vitro and in vivo, and HP-02 can be applied as a common carp feed supplement to enhance fish immunity and disease resistance against A. hydrophila.

Effects of dietary Radix Rehmanniae Preparata polysaccharides on the growth performance, immune response and disease resistance of Luciobarbus capito.

This work explores the effects of dietary Radix Rehmanniae Preparata polysaccharide (RRPP) supplementation on the growth performance, nonspecific immune responses, immune- and growth-related gene expression and disease resistance to Aeromonas hydrophila in Luciobarbus capito. Diets containing five concentrations of 0%, 0.05%, 0.1%, 0.2% and 0.4% RRPP were fed to fish for 60 d. The results indicated that the growth performance significantly increased in the 0.1%, 0.2% and 0.4% RRPP groups compared with that in the control (P < 0.05). The activities of serum lysozyme (LAZ), acid phosphatase (ACP), superoxide dismutase (SOD), alkaline phosphatase (AKP) and total protein (TP) were significantly increased in the appropriate RRPP supplemented groups (P < 0.05). With respect to immune- and growth-related genes, such as interleukin (IL)-1ß, IL-8, tumor-necrosis factor (TNF)-α, interferon (IFN)-γ, growth hormone (GH), insulin-like growth factor (IGF)-I and IGF-II, up-regulation were observed in the three organs (kidney, spleen, gut) of the fish fed with RRPP, compared with the control. In contrast, the mRNA expression of IL-10 and transforming-growth factor (TGF)-ß were downregulated. After challenge with A. hydrophila, the final survival rate was significantly higher in fish fed the RRPP supplement than that in the control group (P < 0.05). In conclusion, RRPP enhanced the growth performance, immune response and disease resistance of Luciobarbus capito, with the greatest effects at 0.2% RRPP.

Dietary supplementation with Rehmannia glutinosa affects the composition of intestinal microorganisms in common carp.

A feeding experiment was conducted to investigate the effects of Rehmannia glutinosa (RG) on the bacterial, archaeal, and fungal community composition in the gut of common carp (Cyprinus carpio L.). Common carp were given a control diet and an RG-supplemented diet (basal diet plus 4% prepared Rehmannia root powder) over a period of 80 days. Our results indicated that the bacteria Fusobacteria, Proteobacteria, and Bacteroidetes; the archaea Crenarchaeota and Euryarchaeota; and the fungi Basidiomycota are the most abundant microbial taxa in the gut of common carp. Compared with the common carp fed a control diet, the common carp fed an RG-supplemented diet contained a higher content of Akkermansia sp., and a lower proportion of Aeromonas sp. These results indicate that the consumption of a diet containing RG can lead to the accumulation of more beneficial microorganisms while inhibiting the growth of potential pathogens. Moreover, Crenarchaeota, the methanogenic, and Basidiomycota were detected in this study, these populations may be of high physiological relevance in carp because they have been implicated in human health and disease. Our results suggest that an RG-supplemented diet changes the intestinal microbial composition of common carp, which could have positive effects on the immune response of carp.