Full length articleWithania somnifera dietary supplementation improves lipid profile, intestinal histomorphology in healthy Nile tilapia (Oreochromis niloticus), and modulates cytokines response to Streptococcus infection
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.
References (89)
- et al.
Effect of Sophora flavescens on non-specific immune response of tilapia (GIFT Oreochromis niloticus) and disease resistance against Streptococcus agalactiae
Fish Shellfish Immunol.
(2013) - et al.
The use of caraway seed meal as a feed additive in fish diets: growth performance, feed utilization, and whole-body composition of Nile tilapia, Oreochromis niloticus (L.) fingerlings
Aquaculture
(2011) - et al.
Effects of dietary Astragalus polysaccharides (APS) on growth performance, immunological parameters, digestive enzymes, and intestinal morphology of Nile tilapia (Oreochromis niloticus)
J Fish Shellfish Immunol.
(2014) - et al.
A standardized root extract of Withania somnifera and its major constituent withanolide-A elicit humoral and cell-mediated immune responses by up regulation of Th1-dominant polarization in BALB/c mice
Life Sci.
(2007) - et al.
Immune modulation and apoptosis induction: two sides of antitumoural activity of a standardised herbal formulation of Withania somnifera
Eur. J. Cancer
(2009) Effect of ashwagandha on lipid peroxidation in stress-induced animals
J. Ethnopharmacol.
(1998)- et al.
Hypocholesteremic and antioxidant effects of Withania somnifera (Dunal) in hypercholesteremic rats
Phytomedicine
(2007) - et al.
Molecular insight into the immune up-regulatory properties of the leaf extract of Ashwagandha and identification of Th1 immunostimulatory chemical entity
Vaccine
(2009) - et al.
Dietary Withania sominefera root confers protective and immunotherapeutic effects against Aeromonas hydrophila infection in Nile tilapia (Oreochromis niloticus)
Fish Shellfish Immunol.
(2018) - et al.
Immunomodulatory role of Withania somnifera root powder on experimental induced inflammation: an in vivo and in vitro study
Vascul. Pharmacol.
(2006)
Anti-inflammatory activity of Withania somnifera leaf extract in stainless steel implant induced inflammation in adult zebrafish
J. Genetic Eng. Biotechnol.
Withaferin A strongly elicits IκB kinase β hyperphosphorylation concomitant with potent inhibition of its kinase activity
J. Biol. Chem.
Withaferin A inhibits NF-kappaB activation by targeting cysteine 179 in IKKβ
Biochem. Pharmacol.
Tilapia piscidin 4 (TP4) enhances immune response, antioxidant activity, intestinal health and protection against Streptococcus iniae infection in Nile tilapia
Aquaculture
Dietary effects of garlic (Allium sativum) on haemato-immunological parameters, survival, growth, and disease resistance against Vibrio harveyi infection in Asian sea bass, Lates calcarifer (Bloch)
Aquaculture
Effects of soybean meal and salinity on intestinal transport of nutrients in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss)
J. Comp. Biochem. Physiol. Part B Biochem. Mol. Biol. Evol.
Competition between selenomethionine and methionine absorption in the intestinal tract of green sturgeon (Acipenser medirostris)
J. Aquatic Toxicol.
Enhanced intestinal epithelial barrier health status on European sea bass (Dicentrarchus labrax) fed mannan oligosaccharides
Fish Shellfish Immunol.
Development of immunity in rainbow trout (Oncorhynchus mykiss, Walbaum) to Aeromonas hydrophila after the dietary application of garlic
J. Fish Shellfish Immunol.
Immunostimulants, adjuvants, and vaccine carriers in fish: applications to aquaculture
Annu. Rev. Fish Dis.
Early antiviral response and virus-induced genes in fish
Dev. Comp. Immunol.
Oral administration of recombinant epinecidin-1 protected grouper (Epinephelus coioides) and zebrafish (Danio rerio) from Vibrio vulnificus infection and enhanced immune-related gene expressions
J. Fish Shellfish Immunol.
Aeromonas salmonicida induced immune gene expression in Aloe vera fed steelhead trout, Oncorhynchus mykiss (Walbaum)
Aquaculture
Genomic organization, gene duplication, and expression analysis of interleukin-1β in channel catfish (Ictalurus punctatus)
Mol. Immunol.
Cloning, characterization and mRNA expression of interleukin-6 in blunt snout bream (Megalobrama amblycephala)
Fish Shellfish Immunol.
Gene expression profiling in naïve and vaccinated rainbow trout after Yersinia ruckeri infection: insights into the mechanisms of protection seen in vaccinated fish
Vaccine
The bioactivity of teleost IL-6: IL-6 protein in orange-spotted grouper (Epinephelus coioides) induces Th2 cell differentiation pathway and antibody production
Dev. Comp. Immunol.
Oxidative stress, inflammation, and cancer: how are they linked?
J. Free Rad. Biol. Med. Glob. Surv.
Oxidative stress and pro-inflammatory responses induced by silica nanoparticles in vivo and in vitro
J. Toxicol. Lett.
Differential gene expression and immune response of Nile tilapia (Oreochromis niloticus) challenged intraperitoneally with Photobacterium damselae and Aeromonas hydrophila demonstrating immunosuppression
J. Aquacult.
Aquaculture in Egypt: status, constraints and potentials
Aquacult. Int.
Review on the progress in the role of herbal extracts in tilapia culture
Cogent Food Agric.
Protection against heterologous Streptococcus iniae isolates using a modified bacterin vaccine in Nile tilapia, Oreochromis niloticus (L.)
J. Fish. Dis.
Current research on the use of plant‐derived products in farmed fish
Aquacult. Res.
Herbal biomedicines: a new opportunity for aquaculture industry
Aquacult. Int.
Plant review Withania somnifera (Ashwagandha): a review
Pharmacogn. Rev.
Steroidal lactones from Withania somnifera, an ancient plant for novel medicine
Molecules
Chemistry and pharmacology of Withania somnifera: an update
TANG
Antimicrobial properties of a non‐toxic glycoprotein (WSG) from Withania somnifera (Ashwagandha)
J. Basic Microbiol.
Scientific basis for the use of Indian ayurvedic medicinal plants in the treatment of neurodegenerative disorders: 1. Ashwagandha
Central Nervous Sys. Agents Med. Chem. (Form. Curr. Med. Chem. Central Nervous Sys. Agents)
Prophylactic effect of Withania somnifera on inflammation in a non-autoimmune prone murine model of lupus
J. Drug Discoveries Therap. Clin. Risk Manage.
An overview on ashwagandha: A Rasayana (Rejuvenator) of Ayurveda
Afr. J. Tradit., Complementary Altern. Med.
Effect of supplementation of Withania somnifera (Linn.) dunal roots on growth performance, serum biochemistry, blood hematology, and immunity of broiler chicks
J. Herbs Spices Med. Plants
Dietary medicinal plant extracts improve growth, immune activity and survival of tilapia Oreochromis mossambicus
J. Fish. Biol.
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