ß-1,3-glucan improved the health and immunity of juvenile African catfish (Clarias gariepinus) and neutralized the histological changes caused by lead and fipronil pollutants.

BACKGROUND: Water pollutants cause adverse effects in aquatic ecosystems. The immunomodulatory and mitigating effects of dietary 1,3-glucan on fipronil and lead-induced intoxication in African catfish (Clarias gariepinus) were investigated. Two hundred forty catfish were randomly divided into four equal groups: those in the first group were fed basic diet and served as controls; those in the second group were supplemented with ß-1,3-glucan (0.1%); those in the third group were exposed to combination of lead nitrate at 0.041 mg/L (1/10 96 h LC50) and fipronil at 2.8 mg/l (1/10 96 h LC50); and those in the fourth group were exposed to combination of fipronil, lead, and ß-1,3-glucan. The health status, haematological, immunological, and histological changes were all evaluated. RESULT: Swelling on the dorsolateral side, spinal column deviation, sluggish movement, skin bleaching, excessive mucus secretion, significant variations in blood indices-related measures, and a 45% death rate were observed in the third group. There was a significant reduction in interleukin-1 (IL-1) and interleukin-6 (IL-6) and immunoglobulin M (IgM) concentrations, as well as decrease in their corresponding gene expression, indicating that fipronil and lead had immunosuppressive activity. Severe catarrhal enteritis and mucinous degeneration of the lining epithelium, and notable depletion of white pulp, congested red pulp and hemosiderosis were common pathological findings in the spleen. ß-1,3-glucan alone or in combination with fipronil and lead provoked physical activity, blood indices, with elevations in IL-1ß, IL-2, IL-6, and IgM concentrations, as well as up-regulation in their genes' expression in splenic tissues, when compared to the third group. The spleen and intestine had normal histological architecture with 5% mortalities. There were no fish deaths in the ß-1,3-glucan-alone or control groups. CONCLUSION: The use of ß-1,3-glucan (0.1%) as dietary supplement could be implemented to protect against the toxic effects of fipronil and lead toxicity by improving the health and immunological parameters of intoxicated catfish.

Melaleuca alternifolia essential oil prevents alterations to purinergic enzymes and ameliorates the innate immune response in silver catfish infected with Aeromonas hydrophila.

Aeromonas hydrophila infection represents a major impediment to the development of aquaculture, leading to important economic losses. Over the last few years, different methods have been used to counteract and minimize the negative effects of this infection, such as the use of Melaleuca alternifolia essential oil, popularly known as tea tree oil (TTO), that possess a bactericide action against A. hydrophila. The purinergic system develops an important role in the inflammatory response, principally due to involvement of adenosine triphosphate (ATP) in the inflammatory process, as well as by the anti-inflammatory properties of adenosine (Ado), a molecule that is controlled by NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) enzymes. Thus, the aim of this study was to investigate the involvement of purinergic enzymes in the pathogenesis of A. hydrophila infection, and whether the purinergic pathway and innate immune response are involved in the protective effects of TTO in silver catfish (Rhamdia quelen) experimentally infected with A. hydrophila. Our results revealed that A. hydrophila infection increased seric NTPDase and 5'-nucleotidase activity, while ADA activity decreased. Also, the seric levels of pro-inflammatory cytokines such as interleukin-1 (IL-1), IL-6, tumor necrosis factor alpha (TNF-α) and interferon gamma (INF-γ) increased in the infected fish, while the seric level of anti-inflammatory interleukin-10 (IL-10) decreased. Treatment with TTO was able to prevent the impairment of purinergic enzymes and improve the innate immune response through the modulation of cytokine response during A. hydrophila infection. In summary, prophylactic therapy with TTO can be considered an important approach to improve the immune response and consequently avoid the inflammatory process in fish infected with A. hydrophila.

Effects of recombinant flagellin B and its ND1 domain from Vibrio anguillarum on macrophages from gilthead seabream (Sparus aurata L.) and rainbow trout (Oncorhynchus mykiss, W.).

Flagellin is the principal component of flagellum in Gram negative and positive bacteria, and it is also the ligand that activates the Toll-like receptor 5 (TLR5) in mammals and fish. In higher vertebrates, flagellin induces the activation of the membrane-bound TLR5 (TLR5M), which promotes the expression of proinflammatory cytokines and chemokines and the co-stimulatory molecules present in antigen-presenting cells needed for the activation of T cells. In the present study, we report the production of two recombinant proteins of Vibrio anguillarum: i) a full length flagellin B (FlaB) (rFla) and ii) the amino-terminus of the D1 domain (rND1) of the same protein, the region mainly responsible for binding to TLR5 and for the immunostimulatory activity of flagellin. The effects of these recombinant proteins were assessed in vitro using head kidney macrophages of gilthead seabream (Sparus aurata L., Perciformes, Sparidae) and rainbow trout (Oncorhynchus mykiss W., Salmoniformes, Salmonidae). In both species, 3 h of stimulation with rFla and rND1 induced expression of the proinflammatory cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α), and of the chemokine IL-8. In gilthead seabream macrophages stimulated with rFla and rND1, a 900- and 6-fold increase were observed for IL-1ß transcription, while a 900- and 3-fold increase were recorded for IL-8 transcription, respectively, as compared to non-stimulated macrophages. In rainbow trout, rFla increased expression of IL-8 40-fold in macrophages, whereas rND1 increased expression of the chemokine 3-fold, as compared to non-stimulated cells. The results obtained for rFla and rND1 demonstrate their modulatory capabilities in vitro, suggesting that rFla and rND1 could be evaluated as immunostimulatory candidates for use in farmed fish. However, further in vivo studies are needed to confirm and expand on the present results.