Hydroalcoholic extract of Withania somnifera leaf and α-tocopherol acetate in diets containing oxidised oil: effects on growth performance, immune response, and oxidative status in broiler chickens

Figures & data

Table 1. Composition of the basal diets (g/kg as fed).

Ingredient	Starter (1–10 days)	Grower (11–23 days)	Finisher (24–42 days)
Corn	522.50	523.7	570.8
Soybean meal	340	378.6	324.50
Corn gluten	48.2	–	–
Non oxidised soybean oil^a	12.2	35	43
Oxidised soybean oil^b	20	20	20
Dicalcium phosphate	17.8	15.6	15.6
Limestone	12.5	11.8	11.5
Salt	4	4.5	4.3
DL-Methionin	3.4	3.2	2.9
L-Lysine	4.6	1.8	1.8
Vitamin premix^c	2.5	2.5	2.5
Mineral premix^d	2.5	2.5	2.5
L-Threonine	1.4	0.8	0.6
Sand	8.4	–	–
Calculated nutrients and energy
AME, (kcal/kg)	3000	3100	3210
Crude protein (g/kg )	230.2	215.2	195.1
L-Lysine (g/kg)	14.4	12.9	11.6
DL-Methionin (g/kg)	7	6.4	5.9
TSAA (g/kg)	10.7	9.8	9.1
Calcium (g/kg)	9.6	9	8.8
Nonphytate P (g/kg)	4.8	4.5	4.4
Total P (g/kg)	7.3	6.9	6.6

^a Peroxide value <1 mEqO₂/kg.

^b Peroxide value 75 mEqO₂/kg.

^c Vitamin premix provided per kilogram of diet: retinyl acetate, 11,000 U; cholecalciferol, 1800 U; DL-α-tocopheryl acetate, 11 mg; menadione sodium bisulphate, 2 mg; riboflavin, 5.7 mg; pyridoxine hydrochloride, 2 mg; cyanocobalamin, 0.024 mg; nicotinic acid, 28 mg; folic acid, 0.5 mg; pantothenic acid, 12 mg; choline chloride, 250 mg.

^d Mineral premix provided per kilogram of diet: Mn, 100 mg; Zn, 65 mg; Cu, 5 mg; Se, 0.22 mg; I, 0.5 mg; and Co, 0.5 mg.

AME: apparent metabolizable energy.

Table 2. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on body weight gain (BWG), feed intake (FI), and feed conversion efficiency (FCE) at 42 days of age.

Treatment			FI (g)	BWG (g)	FCE (g/g)
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)
0	0	0	3933.0	2380.75^abc	1.65
0	0	200	4105.0	2449.65^a	1.67
0	100	0	4048.5	2411.70^a	1.67
0	100	200	4055.3	2366.01^abc	1.71
0	200	0	3882.1	2284.33^abc	1.70
0	200	200	3864.9	2211.72^bc	1.75
2	0	0	3831.2	2193.06^c	1.74
2	0	200	3818.2	2197.00^c	1.73
2	100	0	3822.2	2272.81^abc	1.68
2	100	200	3864.9	2323.62^abc	1.66
2	200	0	3825.2	2261.12^abc	1.69
2	200	200	3938.8	2398.98^ab	1.64
SEM			100.406	57.124	0.036
Main effect
Oxidised oil		0	3981.33^a	2350.69^a	1.69
		2	3850.09^b	2274.43^b	1.69
WS		0	3921.86	2305.11	1.70
		100	3947.70	2343.54	1.68
		200	3877.57	2289.04	1.69
α-Toc		0	3890.37	2300.63	1.69
		200	3941.05	2324.50	1.69
				P
Oxidised oil			0.028	0.025	0.937
WS			0.610	0.389	0.780
α-Toc			0.386	0.472	0.740
Oxidised oil × WS			0.241	0.002	0.250
Oxidised oil × α-Toc			0.960	0.227	0.138
WS × α-Toc			0.928	0.900	0.961
Oxidised oil × WS × α-Toc			0.525	0.240	0.859

^a–cMeans within each column with no common superscript differ significantly (p < .05).

Table 3. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on primary and secondary antibody responses.

Treatment			Primary antibody response				Secondary antibody response
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)	IgG	IgM	Total		IgG	IgM	Total
0	0	0	2.00	5.60	7.60		2.40	5.40	7.80^e
0	0	200	2.20	5.60	7.80		2.20	6.40	8.60^bcde
0	100	0	2.20	6.00	8.40		3.00	6.80	9.80^ab
0	100	200	2.00	6.20	8.20		2.80	6.80	9.60^abc
0	200	0	2.80	6.00	8.80		3.00	7.00	10.00^a
0	200	200	2.60	5.60	8.20		3.00	6.80	9.80^ab
2	0	0	2.00	5.00	7.00		2.20	6.00	8.20^de
2	0	200	2.00	5.20	7.20		2.20	6.20	8.40^cde
2	100	0	2.00	6.20	8.20		260	6.00	8.60^bcde
2	100	200	2.00	5.20	7.20		2.40	5.60	8.00^de
2	200	0	2.00	5.40	7.40		2.20	5.80	8.00^de
2	200	200	2.00	5.00	7.00		2.40	6.80	9.20^abcd
SEM			0.200	0.316	0.336		0.230	0.378	0.380
Main effect
Oxidised oil		0	2.30^a	5.83^a	8.16^a		2.73^a	6.53^a	9.26^a
		2	2.00^b	5.33^b	7.33^b		2.33^b	6.06^b	8.40^b
WS		0	2.05	5.35^b	7.40^b		2.25^b	6.00^b	8.25^b
		100	2.05	5.90^a	8.00^a		2.70^a	6.30^ab	9.00^a
		200	2.35	5.50^ab	7.85^ab		2.65^a	6.60^a	9.25^a
α-Toc		0	2.16	5.70	7.90		2.56^a	6.16	8.73
		200	2.13	5.46	7.60		2.50^a	6.43	8.93
						P
Oxidised oil			0.012	0.008	0.0001		0.004	0.037	<0.001
WS			0.059	0.048	0.040		0.015	0.091	0.001
α-Toc			0.774	0.207	0.129		0.619	0.228	0.367
Oxidised oil × WS			0.059	0.905	0.246		0.195	0.084	0.012
Oxidised oil × α-Toc			0.774	0.365	0.609		0.619	1.00	0.763
WS × α-Toc			0.718	0.440	0.197		0.648	0.307	0.166
Oxidised oil × WS × α-Toc			0.718	0.248	0.543		0.939	0.152	0.134

^a-eMeans within each column with no common superscript differ significantly (p < .05).

Table 4. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on swelling response to phytohematoglutinin.

Treatment			Swelling response (mm)
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)	12 h	24 h
0	0	0	0.296	0.302
0	0	200	0.306	0.306
0	100	0	0.316	0.312
0	100	200	0.310	0.314
0	200	0	0.312	0.330
0	200	200	0.314	0.304
2	0	0	0.302	0.317
2	0	200	0.306	0.314
2	100	0	0.310	0.316
2	100	200	0.308	0.318
2	200	0	0.314	0.318
2	200	200	0.328	0.316
SEM			0.005	0.005
Main effect
Oxidised oil		0	0.309	0.311
		2	0.311	0.316
WS		0	0.302^b	0.309
		100	0.311^a	0.315
		200	0.317^a	0.317
α-Toc		0	0.308	0.315
		200	0.312	0.312
			P
Oxidised oil			0.459	0.122
WS			0.003	0.215
α-Toc			0.273	0.247
Oxidised oil × WS			0.314	0.358
Oxidised oil × α-Toc			0.627	0.414
WS × α-Toc			0.255	0.108
Oxidised oil × WS × α-Toc			0.513	0.143

^a–bMeans within each column with no common superscript differ significantly (p < .05).

Table 5. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on relative weight of the thymus, spleen, and bursa of Fabricius of broilers at 42 days of age.

Treatment			Thymus (g)	Spleen (g)	Bursa (g)
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)
0	0	0	0.658	0.152	0.113
0	0	200	0.607	0.140	0.195
0	100	0	0.514	0.152	0.120
0	100	200	0.642	0.160	0.100
0	200	0	0.663	0.151	0.138
0	200	200	0.665	0.153	0.098
2	0	0	0.490	0.129	0.119
2	0	200	0.639	0.122	0.119
2	100	0	0.692	0.154	0.111
2	100	200	0.584	0.151	0.146
2	200	0	0.529	0.150	0.104
2	200	200	0.566	0.158	0.083
SEM			0.071	0.010	0.034
Main effect
Oxidised oil		0	0.625	0.151	0.128
		2	0.583	0.144	0.113
WS		0	0.599	0.135^b	0.136
		100	0.608	0.154^a	0.119
		200	0.606	0.153^a	0.106
α-Toc		0	0.591	0.148	0.117
		200	0.617	0.147	0.124
				P
Oxidised oil			0.321	0.266	0.496
WS			0.981	0.049	0.464
α-Toc			0.533	0.915	0.772
Oxidised oil × WS			0.211	0.354	0.517
Oxidised oil × α-Toc			0.993	0.962	0.947
WS × α-Toc			0.920	0.638	0.355
Oxidised oil × WS × α-Toc			0.105	0.835	0.354

^a–bMeans within each column with no common superscript differ significantly (p < .05).

Table 6. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on antioxidant enzymes activity, and thigh meat malondialdehyde (MDA) content at 42 days of age.

Treatment			MDA content (mg/kg)	GPx activity (U/mL)	SOD activity (U/mL)
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)
0	0	0	0.235^c	106.80	126.80
0	0	200	0.232^c	133.20	122.80
0	100	0	0.220^c	141.20	145.80
0	100	200	0.222^c	160.60	146.00
0	200	0	0.235^c	136.80	148.80
0	200	200	0.205^c	152.40	147.80
2	0	0	0.387^a	108.00	130.50
2	0	200	0.335^ab	119.00	126.00
2	100	0	0.222^c	121.80	132.40
2	100	200	0.167^c	130.80	133.75
2	200	0	0.262^bc	122.20	130.00
2	200	200	0.192^c	133.00	131.75
SEM			0.030	12..841	6.343
Main effect
Oxidised oil		0	0.225^b	138.50^a	139.66^a
		2	0.261^a	122.10^b	130.76^b
WS		0	0.297^a	116.75^b	126.33^b
		100	0.208^b	138.60^a	139.78^a
		200	0.223^b	136.26^a	140.00^a
α-Toc		0	0.260	122.80^b	135.89
		200	0.225	138.34^a	135.14
				P
Oxidised oil			0.047	0.037	0.023
WS			<0.001	0.040	0.010
α-Toc			0.057	0.045	0.787
Oxidised oil × WS			0.002	0.609	0.077
Oxidised oil × α-Toc			0.172	0.499	0.882
WS × α-Toc			0.826	0.950	0.839
Oxidised oil × WS × α-Toc			0.979	0.959	0.985

^a–cMeans within each column with no common superscript differ significantly (p < .05).

GPx: glutathione Peroxidase; SOD: superoxide dismutase.

Table 7. Effect of Oxidised oil, Withania somnifera (WS), and α-tocopherol (α-Toc) on relative length of small intestine.

Treatment			Relative length (cm/100g of BW)
Oxidised oil (%)	WS (mg/kg)	α-Toc (mg/kg)	Duodenum	Jejunum	Ileum
0	0	0	0.726	1.63	1.57
0	0	200	0.826	1.60	1.71
0	100	0	0.732	1.66	1.64
0	100	200	0.788	1.64	1.60
0	200	0	0.733	1.64	1.51
0	200	200	0.708	1.67	1.62
2	0	0	0.731	1.68	1.61
2	0	200	0.733	1.72	1.59
2	100	0	0.719	1.60	1.66
2	100	200	0.729	1.53	1.47
2	200	0	0.672	1.51	1.51
2	200	200	0.707	1.66	1.62
SEM			0.044	0.059	0.073
Main effect
Oxidised oil		0	0.752	1.64	1.61
		2	0.715	1.62	1.58
WS		0	0.754	1.66	1.62
		100	0.742	1.61	1.59
		200	0.705	1.62	1.56
α-Toc		0	0.719	1.62	1.58
		200	0.748	1.64	1.60
				P
Oxidised oil			0.161	0.469	0.493
WS			0.282	0.451	0.541
α-Toc			0.254	0.619	0.719
Oxidised oil × WS			0.979	0.091	0.865
Oxidised oil × α-Toc			0.589	0.516	0.209
WS × α-Toc			0.766	0.282	0.085
Oxidised oil × WS × α-Toc			0.447	0.551	0.689