Mitigating multiple stresses in Pangasianodon hypophthalmus with a novel dietary mixture of selenium nanoparticles and Omega-3-fatty acid

Abstract

Effects of a novel dietary mixture of selenium nanoparticles (Se-NPs) and omega-3-fatty acids i.e., Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on mitigating arsenic pollution, high-temperature stress and bacterial infection were investigated in Pangasianodon hypophthalmus. To aim this, four isocaloric and iso-nitrogenous diets were prepared: control feed (no supplementation), Se-NPs at 0.2 mg kg^-1 diet with EPA + DHA at 0.2, 0.4 and 0.6% as supplemented diets. Fish were reared under normal condition or concurrent exposure to arsenic (2.65 mg L^-1), and temperature (34 °C) (As + T) stress for 105 days. The experiment was conducted with eight treatments in triplicates. Response to various stresses i.e., primary (cortisol), secondary (oxidative stress, immunity, and stress biomarkers) and tertiary stress response (growth performance, bioaccumulation and mortality due to bacterial infection) were determined. Supplementation of dietary Se-NPs at 0.2 mg kg^-1 diet and EPA + DHA at 0.2 and 0.4% reduced the primary stress level. Exposure to arsenic and temperature (As + T) and fed with control diet and EPA + DHA at 0.6% aggravated the cortisol level. Anti-oxidative enzymes (Catalase, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase) and immunity (Nitroblue tetrazolium, total protein, albumin, globulin, A:G ratio, total immunoglobulin and myeloperoxidase) of the fish were augmented by supplementation of Se-NPs and EPA + DHA at 0.2 and 0.4%. Neurotransmitter enzyme, HSP 70, Vitamin C were significantly enhanced (p < 0.01) with supplementation of Se-NPs at 0.2 mg kg^-1 and EPA + DHA at 0.2 and 0.4%. Whereas total lipid, cholesterol, phospholipid, triglyceride and very low-density lipoprotein (VLDL) were reduced (p < 0.01) with the supplementation of Se-NPs at 0.2 mg kg^-1 diet and EPA + DHA at 0.2 and 0.4%. Tertiary stress response viz. growth performance was also significantly enhanced with supplementation of Se-NPs at 0.2 mg kg^-1 and EPA + DHA at 0.2 and 0.4% reared under As + T. Whereas arsenic bioaccumulation in fish tissues was significantly reduced with dietary supplementation of Se-NPs and EPA + DHA. Cumulative mortality and relative percentage survival were reduced with Se-NPs at 0.2 mg kg^-1 and EPA + DHA at 0.2 and 0.4%. The investigation revealed that a novel combination of Se-NPs at 0.2 mg kg^-1 and EPA + DHA at 0.4% followed by 0.2% has the potential to alleviate temperature stress, bacterial infection and arsenic pollution. Whereas diet containing Se-NPs at 0.2 mg kg^-1 diet and EPA + DHA at 0.6% was noticeably enhanced the stress in P. hypophthalmus.

Figure 1

(A) Size of selenium-nanoparticles (203.5 nm) and (B) zeta potential − 41.8 mV.

Figure 2

(A, B) Mitigation of primary and secodary stress response (Cortisol and HSP 70) through dietary supplementation of selenium nanoparticles, eicosapentanoic acid and dicosahexanoic acid fed to P. hypophthalmus reared in control or under arsenic and high temperature (As + T) for 105 days. Within endpoints and groups, bars with different superscripts differ significantly (a–d) cortisol (p = 0.032), HSP-L (p = 0.026), HSP-G (p = 0.014). Data expressed as Mean ± SE (n = 3).

Figure 3

(A, B) Mitigation of secondary stress response (acetylcholine esterase and vitamin C) through dietary selenium nanoparticles, eicosapentanoic acid and dicosahexanoic acid fed to P. hypophthalmus reared in control or under arsenic and high temperature (As + T) for 105 days. Within endpoints and groups, bars with different superscripts differ significantly (a–f) AChE-B (p = 0.0028), AChE-M (p = 0.029), VitC-B (p = 0.0056), VitC-M-G (p = 0.038). Data expressed as Mean ± SE (n = 6).

Figure 4

(A,B) Mitigation of secondary stress response (total immunoglobulin and myeloperoxiadase) through dietary supplementation of selenium nanoparticles, eicosapentanoic acid and dicosahexanoic acid fed to P. hypophthalmus reared in control or under arsenic and high temperature (As + T) for 105 days. Within endpoints and groups, bars with different superscripts differ significantly (a–f) Total immunoglobulin (p = 0.0042), Myeloperoxidase (p = 0.017). Data expressed as Mean ± SE (n = 6).

Figure 5

(A,B) Mitigation of tertiary stress response (relative survival (%) and cumulative mortality) through dietary supplementation of selenium nanoparticles, eicosapentanoic acid and dicosahexanoic acid fed to P. hypophthalmus reared in control or under arsenic and high temperature (As + T) for 105 days. Data expressed as Mean ± SE (n = 30).