The effects of hydroalcoholic extracts of Withania somnifera root, Withania coagulans fruit and 1,25-dihydroxycholecalciferol on immune response and small intestinal morphology of broiler chickens

Figures & data

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

Ingredient	Starter (1–10 days)		Grower (11–23 days)		Finisher (24–42 days)
	−	+	−	+	−	+
Maize	520.0	520.0	532.0	532.0	530.0	530.0
Soybean meal	350.0	350.0	370.0	370.0	369.0	369.0
Gluten	50.0	50.0	–	–	–	–
Vegetable oil	32.7	32.7	58.0	58.0	65.6	65.6
Limestone	5.3	13.1	4.0	10.7	3.7	10.3
Dicalcium phosphate	17.5	17.5	15.5	15.5	14.0	14.0
Salt	3.5	3.5	4.7	4.7	4.1	4.1
Methionin	3.2	3.2	2.8	2.8	2.0	2.0
Lysine	4.0	4.0	1.3	1.3	–	–
Threonine	1.0	1.0	–	–	–	–
Vitamin premix^a	2.5	2.5	2.5	2.5	2.5	2.5
Mineral premix^b	2.5	2.5	2.5	2.5	2.5	2.5
Sand	7.8	–	6.7	–	6.6	–
Calculated nutrients and energy
AME, (MJ/kg)	12.6	12.6	13.1	13.1	13.3	13.3
Crude protein (g/kg)	235.2	235.2	211.5	211.5	209.1	209.1
Lysine (g/kg)	14.4	14.4	12.4	12.4	11.3	11.3
Methionine (g/kg)	7.0	7.0	6.1	6.1	5.2	5.2
TSAA (g/kg)	10.7	10.7	9.5	9.5	8.6	8.6
Calcium (g/kg)	7.3	10.4	6.3	9.0	5.9	8.5
Nonphytate P (g/kg)	5.0	5.0	4.5	4.5	4.2	4.2
Total P (g/kg)	7.0	7.0	6.8	6.8	6.5	6.5
Analysed Ca and total P concentration
Calcium (g/kg)	7.5	10.7	6.5	9.3	6.1	8.8
Total P (g/kg)	7.4	7.4	7.1	7.1	6.7	6.7

^aVitamin premix provided per kilogram of diet: retinyl acetate, 11,000 IU; cholecalciferol, 1,800 IU; 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.

^bMineral 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.

Table 2. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on broiler BWG, FI, FCE (kg feed per kg gain) at 42 d of age.

Treatment			BWG (g)	FI (g)	FCE (g/g)
Control	Plant extract (mg/kg)	1,25(OH)2D3 (μg/kg)
+	0	0	2339	3927	1.679
−	0	0	2235	3883	1.741
−	0	0.5	2219	3868	1.742
−	75 WS	0	2368	3880	1.639
−	75 WS	0.5	2396	3939	1.648
−	150 WS	0	2299	3888	1.693
−	150 WS	0.5	2441	4006	1.642
−	75 WC	0	2357	4019	1.706
−	75 WC	0.5	2291	3812	1.666
−	150 WC	0	2284	3828	1.676
−	150 WC	0.5	2428	3923	1.614
SEM			56.273	90.822	0.037
p-Value			0.108	0.869	0.298

Table 3. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on primary and secondary antibody responses.

Treatment			Primary antibody response			Secondary antibody response
Control	Plant extract (mg/kg)	1,25(OH)2D3 (μg/kg)	IgG	IgM	Total	IgG	IgM	Total
+	0	0	1.600^abc	6.400	8.000	1.600^c	6.200	7.800
−	0	0	2.200^ab	5.600	7.800	2.000^bc	5.600	7.600
−	0	0.5	1.400^bc	5.600	7.000	2.400^a	6.000	8.400
−	75 WS	0	1.000^c	6.200	7.200	2.000^bc	6.000	8.000
−	75 WS	0.5	1.600^abc	5.000	6.600	2.200^ab	5.600	7.800
−	150 WS	0	3.200^a	4.800	8.000	2.250^ab	6.000	8.250
−	150 WS	0.5	1.600^abc	5.600	7.200	2.200^ab	5.400	7.600
−	75 WC	0	2.200^ab	6.000	8.200	2.000^bc	5.200	7.200
−	75 WC	0.5	1.600^abc	6.000	7.600	2.000^bc	6.200	8.200
−	150 WC	0	1.600^abc	5.800	7.400	2.400^a	5.400	7.800
−	150 WC	0.5	1.600^abc	6.400	8.000	2.200^ab	6.200	8.400
SEM			o.5169	0.7273	0.6494	0.3106	0.5282	0.4990
p-Value			0.0092	0.7098	0.5794	0.0155	0.4452	0.2539

Note: Means within a column with no common superscript are significantly different (p < .05).

Table 4. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on swelling response to phytohematoglutinin.

Treatment			Swelling response (mm)
Control	Plant extract(mg/kg)	1,25(OH)2D3(μg/kg)	12 h	24 h
+	0	0	0.312	0.420
−	0	0	0.350	0.324
−	0	0.5	0.164	0.140
−	75 WS	0	0.250	0.172
−	75 WS	0.5	0.238	0.204
−	150 WS	0	0.384	0.516
−	150 WS	0.5	0.316	0.350
−	75 WC	0	0.204	0.150
−	75 WC	0.5	0.424	0.294
−	150 WC	0	0.230	0.230
−	150 WC	0.5	0.378	0.304
SEM			0.0766	0.1023
p-Value			0.3227	0.4192

Table 5. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on relative weight of the thymus, spleen, and bursa of fabricius of broilers at 21 and 42 days of age.

Treatment			21 day			42 day
Control	Plant extract (mg/kg)	1,25(OH)2D3 (μg/kg)	Thymus	Spleen	Bursa	Thymus	Spleen	Bursa
+	0	0	0.932	0.095	0.254	0.632	0.142^ab	0.117
−	0	0	0.971	0.111	0.252	0.520	0.125^abc	0.095
−	0	0.5	0.765	0.104	0.235	0.632	0.105^c	0.098
−	75 WS	0	0.811	0.125	0.243	0.691	0.145^ab	0.121
−	75 WS	0.5	0.797	0.094	0.210	0.584	0.127^abc	0.153
−	150 WS	0	0.756	0.111	0.238	0.530	0.129^abc	0.105
−	150 WS	0.5	0.865	0.096	0.232	0.662	0.156^a	0.083
−	75 WC	0	0.802	0.106	0.291	0.693	0.112^bc	0.116
−	75 WC	0.5	0.730	0.095	0.214	0.619	0.114^bc	0.113
−	150 WC	0	0.859	0.108	0.247	0.906	0.116^bc	0.098
−	150 WC	0.5	0.926	0.117	0.246	0.857	0.132^abc	0.084
SEM			0.0700	0.0104	0.0347	0.0998	0.0108	0.0185
p-Value			0.2699	0.5095	0.9468	0.4196	0.0424	0.4965

Note: Means within a column with no common superscript are significantly different (p < .05).

Table 6. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on the small intestinal morphology of broilers at 21 days of age.

Treatment			Villus height (μm)	Villus width (μm)	Crypt depth (μm)	Muscle depth (μm)	Villus height:crypt depth ratio (μm.μm^−1)
Control	Plant extract (mg/kg)	1,25(OH)2D3 (μg/kg)
+	0	0	1153.9	96.18	234.94	272.20	4.93
−	0	0	1136.0	110.08	250.58	394.05	4.56
−	0	0.5	990.7	98.27	276.56	255.95	3.60
−	75 WS	0	1072.4	100.56	226.88	264.15	4.84
−	75 WS	0.5	900.9	100.48	188.53	230.00	4.86
−	150 WS	0	1094.4	105.90	208.56	276.15	5.38
−	150 WS	0.5	1093.7	104.00	240.42	241.45	5.14
−	75 WC	0	1008.8	99.50	232.14	210.40	4.52
−	75 WC	0.5	1013.1	89.74	235.62	247.90	4.36
−	150 WC	0	1060.3	85.88	244.76	282.75	4.48
−	150 WC	0.5	1175.5	99.21	234.78	297.65	5.27
SEM			82.227	8.903	20.286	46.272	0.592
p-Value			0.627	0.889	0.450	0.672	0.708

Table 7. Effect of WS, WC, and 1,25(OH)₂D₃ in negative control diets on the small intestinal morphology of broilers at 42 days of age.

Treatment			Villus height (μm)	Villus width (μm)	Crypt depth (μm)	Muscle depth (μm)	Villus height:crypt depth ratio (μm.μm^−1)
Control	Plant extract (mg/kg)	1,25(OH)2D3 (μg/kg)
+	0	0	1369.32	115.22^bc	321.34	432.76	4.45
−	0	0	1649.22	111.75^bc	360.72	383.88	4.65
−	0	0.5	1344.04	120.58^abc	360.78	354.16	3.74
−	75 WS	0	1380.14	115.65^bc	300.26	405.88	4.64
−	75 WS	0.5	1426.74	100.76^c	278.46	431.84	5.12
−	150 WS	0	1432.20	160.07^a	326.88	338.28	4.38
−	150 WS	0.5	1488.62	111.31^bc	357.30	466.20	4.17
−	75 WC	0	1241.90	111.16^bc	347.78	449.48	3.65
−	75 WC	0.5	1231.08	102.03^c	277.00	379.20	4.51
−	150 WC	0	1279.22	116.42^bc	318.34	337.76	4.16
−	150 WC	0.5	1458.86	134.61^ab	347.10	395.88	4.26
SEM			105.845	9.018	26.940	33.710	0.3570
p-Value			0.264	0.008	0.252	0.132	0.228

Note: Means within a column with no common superscript are significantly different (p < .05).