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Modulatory Effects of Probiotics or Bio-Selenium Nanoparticles on Performance, Immunity, Redox Balance, and Microbial Load in Acrylamide-Challenged Oreochromis niloticus Cover

Modulatory Effects of Probiotics or Bio-Selenium Nanoparticles on Performance, Immunity, Redox Balance, and Microbial Load in Acrylamide-Challenged Oreochromis niloticus

Annals of Animal Science
Volume 26 (2026): Issue 2 (April 2026)
By: ,  ,  ,  ,   and    
Open Access
|Apr 2026
Figures & tables

Figures & Tables

Figure 1.

A transmission electron microscope (TEM) image that depicts selenium nanoparticles (SeNPs). The image showcases the nanoparticles' nearly spherical shape, smooth surface, and uniform size distribution (from 32.1 to 66.7 nm)

Table 1.

Ingredients and chemical analysis of basal diets (on air-dry weight basis)

Ingredients%
Fish meal18
Soybean meal 44%17
Wheat bran26.2
Maize24.2
Corn gluten meal 60%10
Wheat meal2.1
Vegetable oil1
Fish oil1
Vitamin premix10.25
Mineral premix20.25
Total100
Proximate chemical composition
Crude protein (N×6.25)31.23
Crude lipids8.49
Ash4.89
Crude fiber4.92
Nitrogen free extract (NFE)350.47
Gross energy (GE, MJ/Kg diet)418.39

1 Composition of mineral premix kg−1: manganese, 53 g; zinc, 40 g; iron, 20 g; copper, 2.7 g; iodine, 0.34 g; selenium, 70 mg; cobalt, 70 mg and calcium carbonate as carrier up to 1 kg.

2 Composition of vitamin premix kg−1: vitamin A, 8000000 IU; vitamin D3, 2000000 IU; vitamin E, 7000 mg; vitamin K3, 1500 mg; vitamin B1, 700 mg; vitamin B2, 3500 mg; vitamin B6, 1000 mg; vitamin B12, 7 mg; biotin, 50 mg; folic acid, 700 mg; nicotinic, 20000 mg; pantothenic acid, 7000 mg.

3 NFE = 100 − (crude protein % + crude lipids% + ash % + crude fiber%).

4 Gross energy (GE) was calculated from NRC (2011) as 23.6 kJ/g, 39.4 kJ/g, and 17.2 kJ/g for protein, lipid, and carbohydrates, respectively.

Table 2.

Growth performance and feed efficiency of Nile tilapia fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) (1, 2 and 3 g kg−1) or bacterial synthesized selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( graphic/j_aoas-2025-0076_ingr_001.png 96 h LC50= 34.6 mg L−1) for 30 days

IBW (g)FBW (g)WG (g)DWG (g)DWG (%)TFI (g)FCR (g/g)RGR (g/g)SGR (% d−1)
CTR24.25±0.03 a71.8±0.25 ab47.55±0.37 ab0.84±0.04 ab84.91±0.13 ab33.65±0.31 b1.41±0.1 ab66.15±0.111.93±0.16 ab
PM123.85±0.03 b64.2±0.11 b40.35±0.12 a0.72±0.73 a72.05±0.42 a35.10±0.0 1 ab1.14±0.12 b62.54±0.401.76±0.11 b
PM223.75±0.09 b71.73±0.96 ab47.98±0.83 ab0.85±0.32 ab85.68±0.33 ab35.10±0.01 ab1.46±0.1 ab66.84±0.811.97±0.04 ab
PM323.6±0.06 b71.33±0.86 ab47.73±0.93 ab0.85±0.10 ab85.24±0.60 ab35.15±0.03 ab1.45±0.2 ab66.44±2.91.96±0.2 ab
SeNPs123.83±0.14 b88.73±0.84 ab64.90±0.91 a1.15±0.03 a115.89±0.42 a36.65±0.83 a1.87±0.04 a73.11±0.642.34±0.04 a
SeNPs223.85±0.03 b87.2±0.83 a63.35±0.81 a1.13±0.06 a113.13±0.82 a35.10±0.01 ab1.80±0.10 a72.54±0.212.31±0.18 a
SeNPs323.8±0.06 b70.0±0.91 ab46.20±0.98 ab0.82±0.14 ab82.50±0.14 ab35.10±0.01 ab1.31±0.22 ab65.10±0.011.90±0.20 ab
P value
  linear0.7000.0070.0060.0060.0060.0040.0130.0200.010
  quadratic0.1000.5040.5550.5550.5550.0010.3010.7020.684
  cubic0.3000.5130.5370.5370.5370.9060.5550.6570.620

[i] CTR, fish group fed un-supplemented diets;

[ii] PM1, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 1 mg kg−1;

[iii] PM2, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 2 mg kg−1;

[iv] PM3, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 3 mg kg−1;

[v] SeNPs1, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.001 mg kg−1;

[vi] SeNPs2, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.002 mg kg−1;

[vii] SeNPs3, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.003 mg kg−1.

[viii] Values within the same row having different letters are significantly different (P<0.05). Data were presented as the mean ± standard error.

[ix] IBW: initial body weight (g); FBW: final body weight (g); WG: weight gain (g); TFI: total feed intake (g); FCR: feed conversion ratio (g/g); RGR, relative growth rate (g/g); SGR: specific growth rate (% d−1).

Table 3.

Blood biochemical profile of Nile tilapia fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 g kg−1) or selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( graphic/j_aoas-2025-0076_ingr_001.png 96 h LC50 = 34.6 mg L−1) for 30 days

TreatmentsParameters
TP (g dL−1)ALB (g dL−1)GLOB (g dL−1)A/G (%)CR (mg dL−1)UA (mg dL−1)AST (UL)ALT (UL)T.G (mg dL−1)
CTR6.08±0.19 ab3.68±0.58 ab2.40±0.291.64±0.450.53±0.03 a1.69±0.13 a211.80±0.61 a24.5±0.81 a182.28±0.91 a
PM17.09±0.39 a4.04±0.29 a3.05±0.011.32±0.440.33±0.03 b1.36±0.33 b55.64±0.31 e18.04±0.21 b98.56±0.29 e
PM26.51±0.09 ab3.91±0.11 ab2.60±0.191.54±0.140.24±0.01 c1.05±0.61 d102.30±0.58 b25.05±0.73 a162.2±0.99 b
PM35.61±0.29 b2.91±0.21 b2.70±0.291.10±0.120.19±0.01 cd0.95±0.11 e43.34±0.21 f13.25±0.58 c163.7±0.01 b
SeNPs16.75±0.28 ab3.64±0.31 ab3.11±0.391.19±0.110.12±0.01 e0.52±0.21 g28.29±0.58 g10.25±0.01 d148.5±0.47 c
SeNPs26.32±0.25 ab3.71±0.01 ab2.61±0.591.45±0.260.16±0.01 de0.68±0.01 f77.27±0.51 d18.11±0.22 b146.7±0.33 c
SeNPs35.41±0.39 b3.03±0.14 b2.40±0.091.29±0.160.32±0.01 b1.24±0.31 c82.24±0.41 c13.54±0.25 c140.11±0.28 d
P value
  linear0.0480.0390.7970.334<0.001<0.001<0.001<0.0010.526
  quadratic0.0930.7130.1310.313<0.001<0.001<0.001<0.001<0.001
  cubic0.8460.9080.9160.8070.053<0.001<0.0010.230<0.001

[i] PM1, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 1 mg kg−1;

[ii] PM2, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 2 mg kg−1;

[iii] PM3, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 3 mg kg−1;

[iv] SeNPs1, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.001 mg kg−1;

[v] SeNPs2, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.002 mg kg−1;

[vi] SeNPs3, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.003 mg kg−1.

[vii] Values within the same row having different letters are significantly different (P<0.05). Data were presented as the mean ± standard error.

[viii] TP, total protein; ALB, albumin; GLOB, globulin; A/G ratio, albumin//globulin ratio; CR, creatinine; UA, uric acid; ALT, alanine transaminase; AST, aspartate transaminase; TG, triglycerides.

Table 4.

Immune parameters of Nile tilapia fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 g kg−1) or selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( graphic/j_aoas-2025-0076_ingr_001.png 96 h LC50 = 34.6 mg L−1) for 30 days

TreatmentsParameters
IGA (ng ml−1)IGM (ng ml−1)AChE (pg ml−1)C3 (mg dL−1)LZM (ng ml−1)IL-1β (pg ml−1)
CTR1045.90±0.34 a775.35±0.89 a687.52±0.99 b38.34±0.99 b0.81 ±0.01 f827.54 ±0.28 b
PM1522.25±0.73 c223.14±0.44 g583.18±0.02 c33.50±0.15 c1.58±0.03 d630.02 ±0.97 d
PM2411.14±0.21 d720.11±0.91 b291.19±0.95 e18.58±0.11 e0.66±0.33 g1005.37±0.51 a
PM3355.25±0.11 e343.34±0.84 e895.25±0.61 a52.25±0.01 a4.24±0.41 a286.68±0.22 g
SeNPs1168.86±0.10 g281.11±0.31 f650.50±0.89 b39.39 ±0.81 b3.55±0.28 b757.64±0.27 c
SeNPs2282.28±0.41 f530.25±0.51 c525.25±0.91 c31.13 ±0.71 c2.71±0.46 c535.35±0.41 e
SeNPs3653.35±0.51 b445.25±0.64 d470.11±0.79 d25.25 ±0.01 d0.99±0.75 e464.28±0.61 f
P value
  linear<0.001<0.001<0.001<0.001<0.001<0.001
  quadratic<0.001<0.0010.8920.0120.1750.780
  cubic0.014<0.0010.0070.470<0.0010.358

[i] PM1, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 1 mg kg−1;

[ii] PM2, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 2 mg kg−1;

[iii] PM3, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum ) 3 mg kg−1;

[iv] SeNPs1, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.001 mg kg−1;

[v] SeNPs2, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.002 mg kg−1;

[vi] SeNPs3, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.003 mg kg−1.

[vii] Values within the same row having different letters are significantly different (P<0.05). Data were presented as the mean ± standard error.

[viii] IGA, immunoglobulin A; IGM, immunoglobulin M; ACHE, acetylcholinesterase; C3, complement C3; LZM, lysozyme; IL-1β, interleukin-1β.

Table 5.

Antioxidant activities of Nile tilapia fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 g kg−1) or selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( graphic/j_aoas-2025-0076_ingr_001.png 96 h LC50 = 34.6 mg L−1) for 30 days

SOD (U mL−1)GSH (ng mL−1)TAC (ng mL−1)MDA (nmol mL−1)
CTR44.68±1.99 f61.25±0.56 f0.79±0.16 f3.85±0.39 b
PM199.75±0.39 d116.24±0.39 d1.29±0.03 d2.20±0.03 d
PM228.53±0.51 g38.52±0.36 g0.58 ±0.96 g4.52±0.91 a
PM3180.25±0.63 a220.21±0.64 a3.54 ±0.52 a0.85±0.35 e
SeNPs182.28±0.78 e95.59±0.24 e0.91±0.54 e2.82±0.43 c
SeNPs2121.15±0.77 c134.34±0.63 c2.16 ±0.21 c1.82±0.11 d
SeNPs3149.67±0.41 b152.36±0.75 b2.67±0.23 b1.16±0.01 e
P value
  linear˂0.001˂0.001˂0.001˂0.001
  quadratic˂0.001˂0.001˂0.0010.695
  cubic˂0.0010.002˂0.0010.039

[i] PM1, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 1 mg kg−1;

[ii] PM2, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 2 mg kg−1;

[iii] PM3, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 3 mg kg−1;

[iv] SeNPs1, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.001 mg kg−1;

[v] SeNPs2, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.002 mg kg−1;

[vi] SeNPs3, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.003 mg kg−1.

[vii] Values within the same row having different letters are significantly different (P<0.05). Data were presented as the mean ± standard error.

[viii] SOD, superoxide dismutase; GSH, glutathione peroxidase; TAC, total antioxidant capacity; MDA, malondialdehyde

Table 6.

Body chemical analysis of Nile tilapia fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 g kg−1) or selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( graphic/j_aoas-2025-0076_ingr_001.png 96 h LC50 = 34.6 mg L−1) for 30 days

MOTDMAshOMEECP
CTR4.10±0.11 c95.89±0.13 a16.48±0.1783.51±1.2726.51±0.23 a60.67±1.30
PM15.12±0.81 a94.87±0.81 a20.49±0.8179.50±1.8119.45±1.89 c64.01±2.81
PM25.16±0.24 c94.83±0.24 a18.77±0.6981.22±1.6922.89±0.89 abc61.97±1.36
PM34.53±0.22 c95.46±0.22 a16.86±0.9683.13±1.9625.19±1.50 ab62.40±1.67
SeNPs17.47±0.83 b92.53±1.03 b19.93±0.3980.06±1.2920.89±0.72 c62.50±0.58
SeNPs25.43±0.21 c94.56±0.21 a17.28±0.5382.71±1.5423.05±0.64 abc57.68±5.69
SeNPs33.76±0.29 c96.23±0.29 a18.64±1.1681.35±1.1625.99±1.50 a61.09±0.59
P value
  linear0.0020.0020.8360.8360.5250.327
  quadratic0.0030.0030.4410.4410.0070.456
  cubic0.0290.0290.1400.1400.4260.232

[i] PM1, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 1 mg kg−1;

[ii] PM2, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 2 mg kg−1;

[iii] PM3, fish group fed diets supplemented with probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) 3 mg kg−1;

[iv] SeNPs1, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.001 mg kg−1;

[v] SeNPs2, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.002 mg kg−1;

[vi] SeNPs3, fish group fed diets supplemented with bacterial synthesized selenium nanoparticles 0.003 mg kg−1.

[vii] Values within the same row having different letters are significantly different (P<0.05). Data were presented as the mean ± standard error.

[viii] MOT, moisture; DM, dry matter; OM, organic matter; EE, ether extract; CP, crude protein.

Figure 2.

Effects of enriched tilapia diets with several levels of probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 mg kg−1) or bacterial synthesized selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( 96 h LC50 = 34.6 mg L−1) for 30 days on total and lactic acid bacterial count and coliform counts of experimental feed. The oneway ANOVA analysis was applied to identify the significant differences (P<0.05) between treated and non-treated groups. NS indicated that there were no significant differences identified between tested groups

Figure 3.

Effects of enriched tilapia diets with several levels of probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 mg kg−1) or bacterial synthesized selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( 96 h LC50 = 34.6 mg L−1) for 30 days on total and lactic acid bacterial count and coliform counts (ND; not detected) of intestinal samples. The one-way ANOVA analysis was applied to identify the significant differences (P<0.05) between treated and non-treated groups

Figure 4.

Effects of enriched tilapia diets with several levels of probiotic mixture (Bacillus coagulans and Lactobacillus paraplantarum) (1, 2 and 3 mg kg−1) or bacterial synthesized selenium nanoparticles (SeNPs) (1, 2 and 3 mg kg−1) for 26 days and then exposed to sublethal dose of acrylamide ( 96 h LC50 = 34.6 mg L−1) for 30 days on total and lactic acid bacterial count and coliform counts (ND; not detected) of fish rearing water samples. The one-way ANOVA analysis was applied to identify the significant differences (P<0.05) between treated and non-treated groups

Figure 5.

Representative photomicrograph of H&E stained sections from gills (scale bar 100 μm) showing: A (positive control group): destruction of most secondary gill lamellae (curved arrow), dilated capillaries within primary gill lamellae (arrowhead) and some secondary lamellae fused with inflammatory cells (arrow). B (CTR), C (PM1), E (PM3),G (SeNP2): normal morphological striations of primary filaments (arrowheads) and secondary filament (arrow) in CTR (control–ve), PM1, PM3, SeNP2, respectively. D (PM2): focal thickened area of both primary gill lamellae (arrowhead) and secondary gill lamellae (arrow) by inflammatory cells infiltrates in PM2 treated group. F (SeNp1): some capillaries engorged at secondary filaments (arrowhead) and aggregates of lymphocytes in a few secondary filaments (arrow) in SeNp1 treated group. H (SeNP3): invasionof some secondary gill lamellae by inflammatory cells (arrow) and apparent normal primary gill filaments (arrowhead) in SeNP3 treated group

Figure 6.

Representative photomicrograph of H&E-stained sections from hepatopancreas (scale bar 100 μm) showing: A: focal necrotic area of the pancreatic acini (arrowhead) with fatty cysts (arrow) and congestion of vasculatures (curved arrow) in control+ve group. B, C, E, G: normal architectures of hepatic cells (star), sinusoids, and hepatoportal pancreas (arrow) in CTR (control–ve), PM1, PM3, SeNP2, respectively. D: fatty change of pancreatic acini (arrow), congested hepatic blood vessels (arrowhead) and moderate areas of vacuolated hepatocytes (star) in PM2 treated group. F: peripancreatic fatty change (arrow), congested portal vein (arrowhead) and apparently normal most hepatic parenchyma (star) in SeNp1 treated group. H: Preserved structures of hepatic acini (star), pancreatic acini (arrow) and vascular tissue in in SeNP3 treated group

Figure 7.

Representative photomicrograph of H&E stained sections from spleen (scale bar 20 μm) showing: A: marked depleted areas of white pulp (star) with wide areas of melanomacrophages centers (arrowhead) in control+ve group. B, C, E, G: normal histological architectures of white pulps (arrow) with ellipsoids arterioles (curved arrows), few melanomacrophages centers (arrowhead) and red pulps (stars) in CTR (control–ve), PM1, PM3, SeNP2, respectively. D: high density of melanomacropahges centers (arrowhead), dilated splenic sinusoids (arrow) in PM2 treated group. F: widespread areas of melanomacropahges centers (arrowhead) and alternated white and red pulps (star) in SeNp1 treated group. H: moderate area of melanomacropahges centers within white pulp areas (arrowhead), dilated splenic vasculatures (arrow) and aggregations of different hemopiotic series within red pulp (star) in SeNP3 treated group

DOI: https://doi.org/10.2478/aoas-2025-0076 | Journal eISSN: 2300-8733 (formerly 1642-3402) | Journal ISSN: 1642-3402
Journal RSS Feed
Language: English
Page range: 555 - 573
Submitted on: Mar 2, 2025
Accepted on: Jun 27, 2025
Published on: Apr 15, 2026
Published by: National Research Institute of Animal Production
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
sustainability,
detoxification,
organic fish,
biological nano-synthesis,
decontamination
Related subjects:
Medicine,
Veterinary medicine

© 2026 Samar S. Mohamed, Samir A. Mahgoub, Maha Nader, Nadin G. El-Din, Xiaolin Meng, Mohammed A.E. Naiel, published by National Research Institute of Animal Production
This work is licensed under the Creative Commons Attribution 4.0 License.

Volume 26 (2026): Issue 2 (April 2026)
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