Elsevier

Fish & Shellfish Immunology

Volume 106, November 2020, Pages 133-141
Fish & Shellfish Immunology

Full length article
Withania somnifera dietary supplementation improves lipid profile, intestinal histomorphology in healthy Nile tilapia (Oreochromis niloticus), and modulates cytokines response to Streptococcus infection

https://doi.org/10.1016/j.fsi.2020.07.056Get rights and content

Highlights

  • Withania somnifera (WS) can be incorporated in fish diets as safe functional feeds.

  • WS did not have any negative impacts on lipid profile and tissue histology of Nile tilapia.

  • WS enhanced high density lipoprotein in Nile tilapia, particularly at dose of 5%.

  • WS supplementation improves gut health in Nile tilapia.

  • WS protect against the inflammation induced during streptococcal infection in tilapia.

Abstract

Despite Withania somnifera (WS), stimulating effects have been investigated on many animal species, its role on lipid profile and intestinal histomorphology in healthy animals, and its modulating role on pro-inflammatory cytokines following infection in fish are yet scarce. In this context, lipid profile, liver, and intestinal histomorphology were measured in Nile tilapia fed with a basal diet or diets containing 2.5 and 5% of supplementary WS for 60 days. Besides, cytokines response was measured at 1, 3,7, and 14 days following Streptococcus iniae (S. iniae) infection after the feeding trial. All lipid profile parameters were nominally lowered, excluding high-density lipoprotein (HDL) that exhibited a significant increase in WS 5% group compared to other groups. Improved gut health integrity was observed, especially in WS 5% group in terms of increased goblet cell numbers, villous height, the width of lamina propria in all parts of the intestine, and a decrease in the diameter of the intestinal lumen of the distal intestine only. A significant down-regulation in the mRNA transcript level of cytokine genes (interleukin 1β/IL-1β, tumor necrosis factor α/TNFα, and interleukin 6/IL-6) was demonstrated in the kidney and spleen of WS-supplemented groups following S. iniae infection compared with the control infected (positive control/PC) group. Our findings give new insights for the potential roles of WS dietary inclusion not only on lipid profile and intestinal health integrity improvement in healthy fish under normal rearing but also as a prophylactic against the infection. Thus, WS can be incorporated as a promising nutraceutical in aquaculture.

Introduction

Nile tilapia, Oreochromis niloticus is one of the most paramount species within tilapias for the sake of its impressive global production performance that is attributed to its rapid growth, adequate survival in different grow-out strategies, and diseases tolerance [1,2]. However, high stocking density in the intensive grow-out system accompanied by poor water quality is considered major stress to fish, which results in increased susceptibility to bacterial pathogens with a severe decrease in the economic gains [3]. In particular, S. iniae is one of the most serious bacterial pathogens in finfish aquaculture, causing 30–75% mortality in Nile tilapia [4].

The adoption of various antibiotics to control bacterial diseases in aquaculture gives rise to the emergence of drug-resistant bacteria, immunosuppression, and bioaccumulation of hazardous residues raising a public health concern [5]. In this context, alternative and preventive herbal-based therapeutics are becoming increasingly vital to halt the progression of aquatic diseases. In tilapia aquaculture, many natural medicinal herbal extracts are used in fish as feed supplements to enhance feed utilization, growth performance, antioxidation, and immune response activation [2,[6], [7], [8]].

Withania somnifera (WS) is a herbal plant that belongs to the family Solanaceae, and most pharmacological activities of WS have been attributed to their roots [9]. The main bioactive constituents of Ashwagandha extracts are alkaloids (isopellertierine and anferine), steroidal lactones (withanolides), which withanone, withaferin A, withanolide A, and withanolide D comprise the major fractions, and saponins containing an additional acyl 76 groups (sitoindoside VII and VIII) [10,11]. Therefore, WS is used extensively as a medicinal herbal-drug [[12], [13], [14], [15]], and particularly, the roots are believed to be most potent for therapeutic purposes [16]. Besides, antioxidant properties of WS in stress-exposed animals, concerning its hypolipidemic effect, have been reported in many previous studies [[17], [18], [19], [20], [21], [22]], and also its anti-stress effects in Oreochromis mossambicus [23].

The liver and intestines are the most important organs involved in the digestion and absorption process of nutrients from feed. Therefore, their histological analysis, combined with other health parameters, is considered as biomarkers for evaluating the dietary incorporation of natural herbal plants, including WS, on fish health [24].

Cytokines- and chemokines-related genes have been used to monitor the immediate and early stages of fish immune response during infection [25]. Recently, many studies evaluated the anti-inflammatory effects of WS concerning its immunomodulatory role to be developed as a therapeutic for inflammatory diseases [[26], [27], [28], [29]]. Withaferin A and withanolide D have been well studied as a natural agent for suppressing the inflammatory responses via inhibiting IkappaB phosphorylation and the nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) signaling cascade [[30], [31], [32]], with subsequent inhibition of many pro-inflammatory mediators such as TNF-α, IL-1β, and proteases [29,33].

As pointed earlier, several research works have been conducted to evaluate WS effects in many animal species exposed to stressors. However, studies concerned the role of dietary WS on the non-stressed animal species with respect mainly to its hypolipidemic effect and its influence on the liver and intestinal morphology; besides, its immunomodulatory role on the expression of the pro-inflammatory cytokines in fish are yet very few. Therefore, the current study was conducted to evaluate the impact of WS root dietary incorporation on lipid profile, liver, and intestines histological indices, besides its immunomodulatory role of pro-inflammatory cytokines to protect against S. iniae induced infection in Nile tilapia.

Section snippets

Materials and methods

Two successive experiments were carried out in this study. The first experiment investigated the effects of WS supplementation at a level of 2.5 and 5% on lipid profile, intestinal, and liver histopathology of Nile tilapia. The second experiment was conducted to explore the immunomodulatory effects of WS on pro-inflammatory cytokines response following the S. iniae infection.

Lipid profile analysis

WS supplementation at 5% resulted in a significant increase in HDL level compared to other groups. Meanwhile, no statistical changes were noticed on other lipid profile parameters in WS-supplemented groups compared to the control one (Fig. 1).

Histomorphometric analysis

Microscopical examination of intestine and liver from WS-supplemented diets did not reveal any histopathological alterations and appeared similar to that of the control group Fig. 3. Histomorphometric measurements were presented in Table 3. Nile tilapia

Discussion

Lipid metabolism regulation by WS root extract, including hypolipidemic effects over blood biochemistry, was documented in rats and broiler chicks [18,[37], [38], [39]]. As observed, WS had no side effects on the lipid profile parameters. However, it showed a favorable effect, particularly at the level of 5%, where a significant increase in HDL was noticed compared to other groups. These observations might be justified in the view of the hypolipidemic activity of WS owing to its high flavonoid

Conclusion

In summary, the findings documented herein indicated that WS dietary supplementation has significantly improving effects on lipid profile and histomorphometric indices of liver and intestine. Therefore, it can be incorporated as a safe, functional feed in the fish diet. Furthermore, WS exhibited a prophylactic effect by modulating the induced expression of pro-inflammatory cytokines during the infections. Thus, WS can be considered as a promising nutraceutical in aquaculture.

CRediT authorship contribution statement

Eman Zahran: Conceptualization, Methodology, Investigation, Validation, Supervision, Writing - review & editing. Mahmoud G. El Sebaei: Methodology, Investigation, Visualization, Writing - original draft. Walaa Awadin: Methodology, Investigation, Visualization, Writing - original draft. Samia Elbahnaswy: Writing - original draft. Engy Risha: Methodology, Investigation, Visualization. Youssef Elseady: Methodology, Investigation, Visualization.

Acknowledgments

This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

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