Antioxidant and immune system status, plasma lipid, abdominal fat, and growth performance of broilers exposed to heat stress and fed diets supplemented with pomegranate pulp (Punica granatum L.)

Figures & data

Figure 1. The experimental pattern of heat stress used for broilers (25–42 d). A constant of 21°C was applied for thermoneutral (TN) chamber and, whereas daily heat stress (21–37°C) was applied for heat-stressed chambers.

Table 1. The chemical composition, phenolic compounds of pomegranate seed pulp.

Chemical composition	% of dry matter
Metabolisable energy (kcal/kg)	2320.51
Dry matter (%)	91.42
Crude protein (%)	11.87
Ether extract (%)	9.69
Crude fibre (%)	29.97
Ash (%)	4.63
Calcium (%)	0.53
Phosphorous (%)	0.17
Total phenols	3.68
Total tannins	2.89
Condensed tannins	0.13
Hydrolyzable tannins	2.76
Gallic acid	0.42
Tannic acid	0.21
Ellagic acid	1.53
Punicalin	0.11
Punicalagin A	0.10
Punicalagin B	0.39

Table 2. Composition of starter (1–10 d), grower (11–24 d) and finisher (25–42 d) diets.

	Starter diets (1–10 d)				Grower diets (11–24d)				Finisher diets (25–42 d)
Ingredients (%)	Percentage of Pomegranate Pulp (%)				Percentage of Pomegranate Pulp (%)				Percentage of Pomegranate Pulp (%)
	0	4	7	10	0	4	7	10	0	4	7	10
Corn	53.31	50.45	48.58	46.41	60.31	57.41	55.38	52.82	62.10	59.31	57.87	55.70
Soybean meal	36.70	35.24	34.03	33.12	31.69	30.63	30.12	29.34	30.31	29.40	28.01	27.05
Fishmeal	5.00	5.00	4.91	4.77	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
Pomegranate pulp	0.00	4.00	7.00	10.00	0.00	4.00	7.00	10.00	0.00	4.00	7.00	10.00
Oil	1.97	1.96	2.49	2.71	4.64	4.64	4.64	4.64	4.56	4.34	4.15	4.30
Oyster shell	1.16	1.18	1.15	1.15	1.28	1.27	1.25	1.22	1.19	1.08	1.09	1.09
Dicalcium phosphate	1.01	1.02	1.00	1.00	1.18	1.17	1.15	1.13	1.01	1.01	1.01	1.00
Salt	0.30	0.30	0.29	0.29	0.30	0.30	0.29	0.29	0.22	0.22	0.22	0.22
Vitamin Premix	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25
Mineral Premix	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25	0.25
DL-Methionine	0.050	0.050	0.050	0.050	0.090	0.089	0.088	0.086	0.117	0.117	0.117	0.117
Nutrient composition
Metabolizable energy (kcal/kg)	2,930	2,930	2,930	2,930	3,050	3,052	3,051	3,049	3,150	3,152	3,154	3,149
Crude protein (%)	22.51	22.51	22.51	22.51	20.14	20.13	20.12	20.12	18.98	18.99	19.00	19.00
Lysine (%)	1.26	1.26	1.26	1.26	1.16	1.15	1.16	1.16	1.02	1.02	1.02	1.02
Met + Cys (%)	0.94	0.93	0.93	0.93	0.88	0.88	0.88	0.88	0.81	0.80	0.80	0.80
Crude fibre (%)	3.92	5.12	6.04	6.94	3.81	5.01	5.95	6.83	3.78	4.97	5.88	6.73
Calcium (%)	1.01	1.02	1.01	1.01	0.96	0.96	0.96	0.96	0.89	0.90	0.90	0.90
Available phosphorous (%)	0.50	0.51	0.50	0.50	0.48	0.48	0.48	0.48	0.45	0.45	0.45	0.45

Supplied the following per kilogram of diet (Manufactured by Javan Daru Co, Iran): Vitamin A, 25000 IU; Vitamin D, 5000 IU; Vitamin E, 12.5 IU; Vitamin K, 2.5 IU; Vitamin B1, 1 mg; Vitamin B2, 8 mg; Vitamin B6, 3 mg; Vitamin B12, 0.015 mg; Folic acid, 0.025 mg; nicotinic acid, 17.5 mg; calcium pantothenate, 12.5 mg; Fe, 80 mg; Cu, 10 mg; Mn, 80 mg; Se, 0.15 mg; I, 0.35 mg.

Table 3. Effects of pomegranate pulp on growth production parameters of broilers reared in heat-stressed conditions.

Treatments		Body weight gain(g)			Feed intake (g)			Feed: gain			PBI
		0–10 days	11–24 days	25–42 days	0–10 days	11–24 days	25–42 days	0–10 days	11–24 days	25–42 days	25–42days
Temperature
	TN**	162.02	667.72	1530.8^a	225.15	959.44	2957.4^a	1.429	1.445	1.937^a	4.42^a
	HS	157.71	641.60	1294.2^b	222.30	960.31	2682.2^b	1.373	1.506	2.100^b	3.50^b
SEM^#		3.689	13.605	17.605	2.619	8.997	32.48	0.046	0.033	0.032	0.096
Pomegranate
	0	158.68	636.67^b	1258.1^c	224.68	968.66	2761.8	1.416	1.532	2.222^c	3.20^c
	4	160.64	648.49^b	1402.6^b	223.80	955.75	2856.3	1.395	1.478	2.054^b	3.86^b
	7	160.82	682.56^a	1476.7^a	226.29	972.10	2896.8	1.410	1.436	1.963^a,b	4.19^b
	10	159.33	690.92^a	1512.6^a	220.13	942.99	2764.3	1.384	1.458	1.835^a	4.61^a
	SEM	2.389	13.240	24.897	3.704	12.73	45.936	0.0236	0.0468	0.0452	0.135
Temperature*Pomegranate
	Control	159.41	638.82	1437.0^b	227.74	971.86	2968.6	1.428	1.534	2.067^a,b	3.87^b
	4% PP	162.68	653.23	1542.5^a	222.60	960.00	2940.1	1.369	1.472	1.905^b	4.51^a,b
	7%PP	161.83	722.53	1531.9^a,b	228.32	980.23	3032.1	1.415	1.362	1.987^a,b	4.32^a,b
	10%PP	164.18	756.33	1612.0^a	210.52	925.68	2888.7	1.281	1.412	1.793^a	5.01^a
	Control + HS	157.94	634.52	1079.1^c	221.62	965.45	2555.0	1.403	1.529	2.377^b	2.54^c
	4% PP+ HS	158.60	643.75	1262.7^c	225.00	963.50	2772.5	1.420	1.483	2.203^b	3.21^b,c
	7% PP+ HS	159.81	712.59	1421.6^b	224.25	963.98	2761.6	1.406	1.509	1.942^a,b	4.07^b
	10% PP+ HS	154.48	745.52	1437.0^b	229.73	960.30	2639.9	1.487	1.504	1.877^a	4.22^a,b
	SEM	3.378	27.209	35.210	5.238	17.993	64.963	0.033	0.066	0.0638	0.292
P-value
Temperature		0.9038	0.1871	<0.0001	0.4486	0.9464	<0.0001	0.0569	0.2054	0.0014	<0.0001
Pomegranate		0.0832	0.0330	<0.0001	0.6879	0.3728	0.1184	0.7734	0.5238	<0.0001	<0.0001
Temperature*Pomegranate		0.6093	0.4681	0.0115	0.0932	0.5011	0.3164	0.0800	0.6323	0.0288	0.031

Values are means, n = 5.

^# SEM = standard error of the mean.

** TN = Thermoneutral (21°C); HS = Heat stress temperature (37 ± 1°C).

^a,b,cMeans within a column with different letters are significantly different (P < 0.05).

Table 4. Effects of pomegranate pulp on some blood parameters of broilers reared in heat-stressed conditions.

Treatments		Cholesterol (mg/dl)	Triglyceride (mg/dl)	HDL (mg/dl)	LDL (mg/dl)	LDH (U/L)	AST (U/L)	Total protein (mg/dl)	Uric acid (mg/dl)
Temperature
	TN**	140.68	44.35	58.15	73.66	827	176.1^b	3.22^a	54.05^b
	HS	138.55	41.43	56.77	73.50	1299	225.7^a	2.97^b	80.19^a
	SEM^#	2.916	2.185	2.374	2.628	176.58	13.582	0.0829	0.7849
Pomegranate
	0	164.1^a	41.44	59.54	96.26^a	872	189.2	3.17	73.28^a
	4	136.8^b	43.24	57.43	70.77^b	1148	201.4	3.20	69.81^a
	7	133.1^b	42.89	59.40	65.07^b	1042	188.7	3.03	63.76^b
	10	124.5^b	44.00	53.47	62.21^b	1188	224.2	2.99	61.61^b
	SEM	4.123	3.090	3.569	3.796	249.72	19.209	0.177	1.110
Temperature*PP
	Control	165.88	41.08	62.25	95.41	750	161.91	3.275	53.83^c
	4% PP	144.28	42.83	61.18	74.54	731	175.28	3.515	54.26^c
	7%PP	127.55	45.60	59.80	58.63	784	159.22	3.190	53.70^c
	10%PP	125.00	47.90	49.38	66.05	1042	206.90	2.918	54.40^c
	Control + HS	162.3	41.80	56.83	97.12	994	215.45	3.058	92.72^a
	4% PP+ HS	129.40	43.65	53.68	66.99	1565	227.50	2.892	85.37^a
	7% PP+ HS	138.55	40.18	59.00	71.52	1301	218.23	2.868	73.82^b
	10% PP+ HS	123.97	40.10	57.57	58.38	1334	241.5	3.060	68.82^b
	SEM	5.831	4.370	4.747	5.365	353.16	27.17	0.166	1.5699
P-value-
Temperature		0.6117	0.3543	0.6839	0.9680	0.0708	0.0164	0.0398	<0.0001
PP		<0.0001	0.9469	0.5557	<0.0001	0.8113	0.3339	0.5120	<0.0001
Temperature*PP		0.2027	0.6853	0.3772	0.2049	0.1321	0.4732	0.1721	<0.0001

Values are means, n = 10.

^# SEM = standard error of the mean.

** TN = Thermoneutral (21°C); HS = Heat stress temperature (37 ± 1°C).

^a,bMeans within a column with different letters are significantly different (P < 0.05).

Table 5. Antioxidant status of broilers fed pomegranate pomace reared under thermoneutral or heat-stressed conditions.

Treatments		Superoxide dismutase (U/L)	Glutathione peroxidase (U/mgHb)	Malondialdehyde (ng/L)	Total antioxidant capacity
Temperature
	TN**	436.44^a	92.36^a	0.597^b	0.776^a
	HS	255.4^b	30.59^b	1.841^a	0.545^b
	SEM^#	0.0569	0.699	0.0320	0.0135
Pomegranate
	0	237.6^b	58.67^b	1.708^a	0.541^c
	4	328.2^b	61.69^a,b	1.181^b	0.636^b
	7	346.6^b	62.53^a,b	1.025^b,c	0.718^a
	10	371.1^a	63.02^a	0.961^c	0.746^a
	SEM	5.737	0.990	0.0454	0.0191
Temperature*PP
	Control	410.0	81.82^c	0.755^d	0.605^b,c
	4% PP	424.5	91.77^b	0.597^d	0.740^b
	7%PP	439.0	96.73^a,b	0.532^d	0.853^a
	10%PP	472.3	100.14^a	0.503^d	0.905^a
	Control + HS	205.7	36.53^d	2.66^a	0.478^c
	4% PP+ HS	246.5	61.30^d	1.765^b	0.533^c
	7% PP+ HS	254.3	28.34^d	1.518^b,c	0.583^c
	10% PP+ HS	270.0	28.91^d	1.420^c	0.588^c
	SEM	8.114	1.399	0.0641	0.0270
P-value
Temperature		<0.0001	<0.0001	<0.0001	<0.0001
PP		0.0002	0.0215	<0.0001	<0.0001
Temperature*PP		0.1285	<0.0001	<0.0001	0.0109

Values are means, n = 10.

^# SEM = standard error of the mean.

** TN = Thermoneutral (21°C); HS = Heat stress temperature (37 ± 1°C).

^a,b,cMeans within a column with different letters are significantly different (P < 0.05).

Table 6. Effects of pomegranate pomace on growth production parameters of broilers reared in heat-stressed conditions.

		Primary response to SRBCs			Secondary response to SRBCs			NDV titre		H/L ratio
		Total	IgM	IgG	Total	IgM	IgG	Day 24	Day 42	Day 24	Day 42
Temperature
	TN**	7.77	5.75	2.02	7.37^a	2.72^a	4.65	7.91	7.30^a	0.41	0.37^b
	HS	7.63	5.71	1.92	6.44^b	2.25^b	4.21	7.93	5.67^b	0.38	0.64^a
	SEM^#	0.121	0.091	0.083	0.328	0.126	0.252	0.728	0.738	0.011	0.092
Levels of PP
	0	7.0^b	5.05	1.95	6.29^b	2.15^b	4.14	7.19^b	5.66^b	0.43	0.59^a
	4	7.6^a,b	5.67	1.93	6.82^a,b	2.37^a,b	4.45	7.71^a,b	6.25^a,b	0.40	0.53^a,b
	7	7.9^a,b	5.90	2.00	7.05^a,b	2.55^a,b	4.50	8.10^a,b	6.72^a,b	0.38	0.46^b
	10	8.3^a	6.22	2.08	7.47^a	2.88^a	4.59	8.64^a	7.31^a	0.37	0.44^b
	SEM	0.382	0.493	0.125	0.409	0.206	0.308	0.532	0.490	0.009	0.045
Temperature*PP
	Control	7.02	4.96	2.06	6.62	2.33	4.29	7.15	6.58	0.45	0.42
	4% PP	7.68	5.67	2.01	7.18	2.55	4.63	7.68	7.01	0.43	0.39
	7%PP	8.0	5.96	2.04	7.59	2.77	4.82	8.12	7.52	0.37	0.34
	10%PP	8.39	6.27	2.12	8.08	3.23	4.85	8.62	8.08	0.39	0.33
	Control + HS	6.98	5.14	1.84	5.95	1.97	3.98	7.22	4.73	0.41	0.76
	4% PP+ HS	7.52	5.67	1.85	6.45	2.18	4.27	7.74	5.49	0.37	0.67
	7% PP+ HS	7.8	5.85	1.95	6.51	2.33	4.18	8.08	5.92	0.39	0.58
	10% PP+ HS	8.21	6.17	2.04	6.85	2.52	4.33	8.66	6.54	0.35	0.55
	SEM	0.625	0.583	0.215	1.542	0.957	0.592	0.783	1.548	0.091	0.209
P-value
Temperature		0.1456	0.087	0.3240	0.021	0.0124	0.1245	0.683	0.0123	0.7564	0.0020
PP		0.0325	0.124	0.2652	0.0423	0.0325	0.2630	0.042	0.0234	0.8524	0.0003
Temperature*PP		0.2543	0.325	0.4521	0.2403	0.4258	0.4290	0.543	0.0654	0.7562	0.0809

Values are means, n = 10.

^# SEM = standard error of the mean.

** TN = Thermoneutral (21°C); HS = Heat stress temperature (37 ± 1°C).

^a,bMeans within a column with different letters are significantly different (P < 0.05).

Table 7. Effects of pomegranate pulp (PP) on carcass relative weights (percentages of live weight) of broilers reared in heat-stressed conditions (42 d).

Treatments		Carcass	Breast	Thigh	Abdominal Fat	Liver	Bursa	Spleen
Temperature
	TN**	61.88	21.90	19.61	1.789	2.187	0.179	0.131
	HS	62.95	22.37	20.31	1.771	2.102	0.177	0.122
	SEM^#	0.479	0.380	0.215	0.0677	0.0450	0.0066	0.0111
Levels of PP
	0	62.56	22.30	20.02	2.312^a	2.163	0.136^b	0.099^b
	4	62.87	21.60	20.17	1.768^b	2.103	0.188^a	0.135^a,b
	7	61.57	22.38	19.53	1.588^b,c	2.169	0.180^a	0.122^a,b
	10	62.66	22.26	20.11	1.445^c	2.143	0.207^a	0.150^a
	SEM	0.6778	0.5383	0.3043	0.0942	0.0636	0.0093	0.0158
Temperature*PP
	Control	60.88	21.64	19.62	2.623^a	2.454^a	0.137	0.096
	4% PP	63.03	21.45	19.69	1.824^b	2.022^b,c	0.168	0.157
	7%PP	61.54	22.58	19.29	1.453^c	2.053^b,c	0.191	0.115
	10%PP	62.07	21.93	19.83	1.256^c	2.216^b	0.220	0.154
	Control + HS	64.24	22.96	20.42	2.001^b	1.871^c	0.135	0.102
	4% PP+ HS	62.72	21.74	20.64	1.711^b	2.183^b,c	0.208	0.113
	7% PP+ HS	61.60	22.18	19.77	1.722^b	2.284^a,b	0.169	0.128
	10% PP+ HS	63.26	22.60	20.39	1.652^b,c	2.070^b,c	0.194	0.145
	SEM	0.9585	0.7613	0.4304	0.1332	0.0899	0.0132	0.0223
P-value
Temperature		0.2291	0.3857	0.0552	0.8505	0.1913	0.8111	0.5888
PP		0.1167	0.7144	0.4404	<0.0001	0.8828	<0.0001	0.0149
Temperature*PP	0.5376	0.7179	0.9466	0.0014	<0.0001	0.0573	0.5838

Note: Values are means, n = 10.

^# SEM = standard error of the mean.

** TN = Thermoneutral (21°C); HS = Heat stress temperature (37 ± 1°C).

^a,b,cMeans within a column with different letters are significantly different (P < 0.05).