Alleviation of glyphosate-induced toxicity by Horseradish tree (Moringa oleifera) Leaf extract and phytase in Nile Tilapia (Oreochromis niloticus) highlighting the antioxidant, anti-inflammatory, and anti-apoptotic activities

Abstract

The danger posed by waterborne toxicity from herbicides endangers the aquatic ecosystem. Using dietary medicinal herbs is a useful approach to mitigate the effects of herbicide toxicity on aquatic animals. This study attempts to examine the consequences and potential mechanisms behind the dietary addition of horseradish tree (Moringa oleifera) leaf extract (MOLE) with the help of phytase addition to check the overall growth performance, biochemical changes, histological alteration, and gene expression in normal and after glyphosate challenge in Nile tilapia. A total number of 135 Nile tilapia fish (7.93 ± 0.03 g) were randomly assigned into three groups each in triplicate. The first group is the control group and fed basal diet; the second group supplied with MOLE (200 mg of extract/kg), and the third group was supplied with MOLE (200 mg /kg), and phytase (0.2g/ kg) for 8 weeks. After the feeding trial, each experimental group was divided into two subgroups to be unchallenged and challenged with glyphosate (30 mg/L of water). The results declared significant enhancements (P < 0.05) in Weight Gain Percent (WG%), Specific growth rate (SGR), and Protein efficiency ratio (PER) and reducing feed conversion ratio (FCR) with up-regulating hepatic gh, igf1,myogenine, intestinal ghrelin and NPY in fish groups fed MOLE and phytase compared with the control group. Moreover, improving the hepatic antioxidant capacity while down-regulating hepatic igf1bp, myostatin. Interstingly, MOLE and phytase lightened glyphosate-induced biochemical alterations, antioxidants, apoptosis, and inflammation-associated genes compared to the glyphosate-challenged group. Interestingly, UPLC-ESI-MS/MS analysis recognized 16 compounds encompasing two glucosinolates, three flavonoids, one phenolic and three alkaloids in addition to four fatty acids, a terpenoid, one phytate and an aromatic glycoside. These components might be accountable for the potential effects exerted by MOLE. Therefore, the current study suggests that dietary supplementation to MOLE and phytase can be used as substitute feed supplements in sustainable farming of Nile tilapia to defend against glyphosate challenges and enhance growth, antioxidant capacity, exerting anti-inflammatory and antiapoptotic effects under normal health conditions or post glyphosate challenge.

Keywords: Glyphosate; Horseradish; Nile tilapia; Phytase.

Fig. 1

UPLC-ESI-MS base peak ion chromatograms of MOLE (negative ion mode: top, positive ion mode: bottom)

Fig. 2

Representative photomicrograph of gills from different treatment groups. A) Control gills showing normal histological appearance of primary and secondary lamellae. B) Glyphosate Challenged group showing diffuse lamellar fusion (thin arrow) with lamellar congestion and hemorrhage (thick arrow). C) MOLE showing normal histological appearance. D) MOLE challenged with glyphosate group showing focal lamellar thickening (thin arrow) with interlamellar congestion (thick arrow). E) MOLE and phytase showing lamellar congestion with focal lamellar fusion (thin arrow). F) MOLE and phytase challenged with glyphosate showing moderate to severe lamellar thickening (thin arrow) with lamellar fusion besides lamellar lifting. Image magnification = 100x

Fig. 3

Representative photomicrograph of liver from different treatment groups. A) Control liver showing normal histological appearance of hepatic parenchyma and hepatopancrease. B) Glyphosate Challenged group showing diffuse swollen hepatocytes (thick arrow) with multifocal coalescing hepatocyte necrosis (arrowhead) and inflammatory aggregates (thin arrow). C) higher magnification showing necrotic hepatocytes (arrowhead) surrounded with inflammatory aggregates (thin arrow) beside diffuse hepatic vacuolation (thick arrow) and sinusoidal congestion. D) Glyphosate Challenged group showing diffuse extensive hepatic vacuolation (thick arrow) and minimal sinusoidal congestion (thin arrow). E) MOLE showing moderate hepatic vacuolation (thick arrow) with sinusidal congestion (thin arrow). F) MOLE challenged with glyphosate showing diffuse hepatic vacuolation (thick arrow). G) MOLE and phytase showing moderate hepatic vacuolation (thick arrow). H) MOLE and phytase challenged with glyphosate showing diffuse severe hepatic vacuolation (thick arrow). Image magnification = 100x

Fig. 4

Representative photomicrograph of intestinal section from different treatment groups. A) Control group showing normal histological appearance of intestinal villi. B, C) Glyphosate Challenged group showing extensive apical desquamation (thin arrows) with mild lamina proprial inflammatory aggregates (arrowhead). D) Glyphosate-Challenged group showing diffuse shortened, stunted fused villi (thin arrow) with marked lamina proprial cellular infilterates (arrowhead) and submucosal edema. E) MOLE showing normal histological appearance. F) MOLE challenged with glyphosate showing mild intestinal vacuolation (thick arrow). G) MOLE and phytase showing few intestianl vacuolation (thick arrow) with few invaded inflammatory cells (arrowhead). H) MOLE and phytase challenged with glyphosate showing few vacuolation and apoptotic bodies (thick arrow) beside minimal submucosal edema (arrowhead). Image magnification = 100x

Fig. 5

Relative gene expression of hepatic growth-related genes. GH: growth hormone gene, igf: insulin-like growth factor, igfbp: insulin-like growth factor binding protein, NPY: Neuropeptide Y. Columns with different superscript letters in the same figure are significantly different (p ≤ 0.05)

Fig. 6

Relative gene expression of antioxidant-related (sod), and inflammatory-related genes (cox2) and apoptotic gene (caspase-3). Columns with different superscript letters in the same figure are significantly different (p ≤ 0.05)