Arginine: lysine ratio influences on performance, egg quality, haematology, biochemistry, antioxidant status and immunity of dual-purpose breeding hens exposed to cyclic heat stress

Figures & data

Table 1. Ingredient and chemical composition of the experimental diets.

Treatments	Tr1 + 2	Tr3	Tr4
Diets	Diet 1	Diet 2	Diet 3
Ingredients, (kg/ton)
Yellow corn 8.30% (CP)	630	630	630
Soybean meal 44% (CP)	260	260	260
Vit-Min Premix^a	3	3	3
NaHCO3	0	0	0
KCO3	0.23	0.23	0.23
KCl	0	0	0
NaCl	3.9	3.80	3.8
Dicalcium phosphate	19.4	19.4	19.4
Limestone	82.47	81.59	80.47
DL-Methionine	1.0	1.0	1.0
Arginine	0	0.98	2.1
Total	1000	1000	1000
Calculated composition,
CP, (g/kg)	165.4	165.4	165.4
ME, (kcal/Kg)	2706	2706	2706
Calcium, (g/kg)	32.7	32.4	3.20
Av. Phos, (g/kg)	4.90	4.90	4.90
Argnine (g/kg)	10.5	11.5	12.6
Lysine, (g/kg)	8.4	8.4	8.4
Arg/Ly	1.25	1.37	1.50
Methionine, (g/kg)	3.5	3.5	3.5
TSAA, (g/kg)	6.3	6.3	6.3
Na+, (g/kg)	1.70	1.65	1.65
K+, (g/kg)	7.20	7.20	7.20
Cl-, (g/kg)	2.80	2.70	2.70
DEB, (mEq)^b	180	180	180
Ether extract, (g/kg)	26.4	26.9	26.4
Crude fibre, (g/kg)	28.2	28.2	28.2
Chemical analysis,
CP, (g/kg)	161.3	163.3	163.1
Lysine, (g/kg)	8.2	8.2	8.2
Arginine, (g/kg)	10.3	11.4	12.2
Ether extract, (g/kg)	25.0	25.8	26.9
Crude fibre, (g/kg)	29.0	27.4	28.2
Ash, (g/kg)	57.8	62.6	63.1

Tr1 + 2: diets with treatment Arg/Ly 1.25; Tr3: diet with treatment Arg/Ly 1.37; Tr4: diet with treatment Arg/Ly 1.50.

^a Vit-Min premix provides per kilogram of diet: vitamin A, 12000 IU, vitamin E, 10 IU, menadione, 3 mg, Vit. D3, 2200 ICU, riboflavin, 10 mg, Ca pantothenate, 10 mg, nicotinic acid, 20 mg, choline chloride, 500 mg, vitamin B12, 10 µg, vitamin B6, 1.5 mg, vitamin B1, 2.2 mg, folic acid, 1 mg, biotin, 50 µg. Trace mineral (milligrams per kilogram of diet): Mn, 55, Zn, 50, Fe, 30, Cu, 10, Se, 0.10, Anti oxidant, 3 mg.

^b Dietary electrolyte balance (DEP): (Na++K+–Cl-).

CP: Crude Protein.

Table 2. The temperature degrees (Tdb^oC) and relative humidity (RH, %) during the experimental periods.

	Before CHS			During CHS
	Tdb	RH	THI	Tdb	RH	THI
January	19.67	48.83	61.37	37.17	35.67	89.73
February	22.00	50.25	65.77	37.42	41.58	91.92
March	23.00	48.75	67.56	38.08	40.83	92.89
mean ± SE	21.6 ± 1.71	49.3 ± 0.84	64.9 ± 3.18	37.6 ± 0.47	39.4 ± 3.22	91.5 ± 1.62

THI: temperature-humidity index Mild (72 to 79 THI) Moderate (80 to 89 THI) Severe (90 THI or greater); Tdb: dry bulb temperature; RH: relative humidity calculated according to Berman (2016); CHS: Cyclic Heat Stress. 

Table 3. Effect of dietary arginine/lysine ratio (Arg/lys) on final and change body weight (BW) and egg production, egg weight and egg mass of Mandara breeding hens under cyclic heat stress.

Arg /lys ratio		Body weight (BW)			Egg production (%)	Egg weight (g)	Egg mass (g/day)
		Initial BW (g)	Final BW (g)	BW Change (g)
1.25 (Control+)		1519.4	1630.8	111.4	50.90^c	55.81^a	26.32^b
Cyclic heat stress	1.25	1566.1	1693.3	127.2	43.86^d	51.70^b	24.5^b
	1.37	1507.3	1632.6	125.3	65.18^a	53.46^ab	34.82^a
	1.50	1499.6	1614.7	115.1	61.40^b	53.65^ab	32.94^a
SEM		110.35	120.15	33.25	1.18	1.17	0.91
p Value		0.1618	0.0945	0.5561	0.0001	0.0381	0.0001

^a,b,c,dmeans having different superscripts in the same columns are significantly different (p < 0.05).

SEM: standard error of means; P value: probability level; Arg/Lys = arginine/lysine ratio.

Table 4. Effect of dietary arginine/lysine ratio (Arg/lys) on feed intake, feed conversion ratio, fertility and hatchability of total and fertile eggs and survival rate of Mandara breeding hens under cyclic heat stress condition.

Arg /lys ratio		FI (g/day)	FCR (g feed/g egg)	Fertility%	Hatchability of total eggs, %	Hatchability of fertile eggs, %	Survival rate
							No	%
1.25 (Control+)		120^a	4.34^b	91.3^ab	89.2^a	97.6^a	40/0	0
Cyclic heat stress	1.25	120^a	5.12^a	89.4^b	76.1^b	85.0^c	40/2	2.5
	1.37	113^c	3.36^c	95.4^a	90.0^a	94.4^a	40/0	0
	1.50	116^b	3.42^c	86.94^c	79.17^b	90.99^b	40/0	0
SEM		0.80	0.17	5.90	4.60	3.56	ND	ND
p Value		0.0001	0.0001	0.0241	0.0364	0.0421	ND	ND

^a,b,cmeans having different superscripts in the same columns are significantly different (p < 0.05). SEM:standard error of means; p value: probability level; FI: feed intake; FCR: feed conversion ratio; Arg/Lys = arginine/lysine ratio; ND: not done.

Table 5. Effect of dietary arginine/lysine ratio on nutrient digestibility traits of Mandara breeding hens under cyclic heat stress condition.

Arg /lys ratio		Organic Matter %	Dry matter%	protein%	Ether extract%	Fiber%
1.25 (Control+)		58.11	82.56^b	81.84^a	77.28	27.67
Cyclic heat stress	1.25	57.87	80.62^c	79.11^b	77.97	26.52
	1.37	59.38	83.62^a	81.35^a	78.00	26.21
	1.50	54.68	84.42^a	81.69^a	78.34	26.25
SEM		0.363	0.077	0.235	0.695	0.753
p Value		0.5525	0.0001	0.0223	0.6757	0.9875

^a,b,cmeans having different superscripts in the same column are significantly different (p < 0.05).

SEM: standard error of means; p value: probability level; Arg/Lys: arginine/lysine ratio.

Table 6. Effect of dietary arginine/lysine ratio (Arg/lys) on some external and internal egg quality traits of Mandara breeding hens under cyclic heat stress condition.

(Arg /lys ratio)		Egg shape index, %	Shell weight, %	Shell^1 Thic, mm	SWUSA, mg/cm^2	Yolk index	Yolk color	Haugh unit score
1.25 (Control+)		75.23	10.20^b	34.09	103.45	47.3^a	6.64	84.92
Cyclic heat stress	1.25	74.51	9.30^c	32.62	110.72	44.0^b	6.23	83.98
	1.37	75.61	10.23^a	33.21	117.97	43.0^b	6.76	84.84
	1.50	73.71	10.22^a	33.98	112.13	43.2^b	6.65	84.45
SEM		1.17	0.693	1.13	6.68	1.62	0.30	2.82
P Value		0.8467	0.0001	0.7447	0.6435	0.0001	0.5513	0.5064

^a,b,cmeans having different superscripts in the same columns are significantly different (p < 0.05).

SEM: standard error of means; P value: probability level; Arg/Lys: arginine/lysine ratio; 1-ShelThic: Shell Thickness; 2-SWUSA: Shell weight per unit of surface area.

Table 7. Effect of dietary arginine/lysine ratio (Arg/lys) on some hematological parameters of Mandara breeding hens under cyclic eat stress condition.

Arg /lys ratio		RBCs	Hgb mg	WBCs x10^3/mm^3	Lym %	Het %	H/L ratio	pH
1.25 (Control+)		2.93^a	13.85^a	63.95^a	78.80^b	14.8^b	19.0^b	7.71^b
Cyclic heat stress	1.25	2.40^b	11.23^b	53.27^c	64.33^c	28.8^a	45.1^a	8.26^a
	1.37	2.65^ab	12.70^a	61.33^b	85.67^a	9.00^c	10.7^b	7.74^b
	1.50	2.79^a	12.75^a	62.85^ab	71.50^ab	10.6^c	14.8^b	7.63^b
SEM		0.09	1.34	1.27	3.40	1.22	1.564	0.46
P Value		0.0690	0.0026	0.0072	0.0092	0.0356	0.0001	0.0155

^a,b,cmeans having different superscripts in the same row are significantly different (p < 0.05).

SEM: standard error of means; p value: probability level; RBC: Red blood cells; Hgb: haemoglobin; WBCs: White blood cell; Lym: Lymphocyte; Het: Heterophi; H/L ratio: Heterophil/lympho; Arg/Lys: arginine/lysine ratio.

Table 8. Effect of dietary arginine/lysine ratio (Arg/lys) on some biochemical parameters of Mandara breeding hens under cyclic heat stress condition.

(Arg /lys ratio)		Glucose (mg/dL)	Tpr (g/dL)	Alb (g/dL)	AST, U/L	ALT, U/L	ALP, U/L	Urea (mg/dL)	Creat (mg/dL)
1.25 (Control+)		176^a	6.26	3.32	51.8^b	60.4^b	102^b	2.57	1.22
Cyclic heat stress	1.25	171^b	6.46	3.40	55.4^a	64.2^a	119^a	2.50	1.16
	1.37	175^a	6.22	3.2	57.0^a	65.4^a	112^a	2.33	1.16
	1.50	177^a	6.22	3.2	57.1^a	65.2^a	114^a	2.40	1.19
SEM		0.325	0.284	0.34	0.3421	0.354	1.658	0.165	0.028
P Value		0.0001	0.0752	0.421	0.0005	0.0006	0.0024	0.7251	0.2172

^a,b,cmeans having different superscripts in the same row are significantly different (p < 0.05).

SEM: standard error of means; p value: probability level; Arg/Lys: arginine/lysine ratio; Tpr: Total protein; Alb: Albumin; ALT: Alanine amino transferase; AST: Aspartate amino transferase; Creat: creatinine; ALP: alkaline phosphatase.

Table 9. Effect of dietary arginine/lysine ratio (Arg/lys) on lipid profile of Mandara breeding hens under cyclic heat stress condition.

(Arg /lys ratio)		T lipid g/dL	Cholest mg/dL	HDL mg/dl	LDL mg/dL	Trig mg/dL	CAT mg/dL	MDA mg/dL
1.25 (Control+)		4.66^b	210^b	44.2	94.8^ab	183^b	39.5^a	0.168^b
Cyclic heat stress	1.25	5.18^a	217^a	42.6	97.3^a	187^a	32.3^c	0.224^a
	1.37	4.38^b	212^b	46.6	91.3^b	181^b	37.8^b	0.167^b
	1.50	4.30^b	211^b	45.3	93.3^b	183^b	38.2^b	0.181^b
SEM		0.055	0.512	1.321	0.325	0.884	0.124	0.213
P Value		0.0001	0.0001	0.0954	0.0001	0.0001	0.0001	0.0001

^a,b,cmeans having different superscripts in the same columns are significantly different (p < 0.05).

SEM: standard error of means; P value: probability level; Arg/Lys: arginine/lysine ratio; Cholest: Cholesterol; HDL: high density lipoprotein; LDL: low density lipoprotein; Trig: Triglyceride; CAT: Catalase; MDA: Malondialdehyde.

Table 10. Effect of dietary arginine/lysine ratio (Arg/lys) on some hormone’s activity and immune parameters of Mandara breeding hens under cyclic heat stress condition.

(Arg /lys ratio) + EDB		T3 µg/dL	T4 µg/dL	TAC mg/dL	IgG, mg/L	IgM, mg/mL	IgA, mg/L	Ca mg/dL	P mg/dL
1.25 (Control+)		0.229	11.7^c	411	9.78	2.33	0.833^a	30.75^a	4.97^ab
Cyclic heat stress	1.25	0.224	17.0^a	405	9.80	2.36	0.663^c	24.5.0^c	4.24^c
	1.37	0.225	15.0^b	412	9.71	2.38	0.837^a	28.15^b	4.67^b
	1.50	0.224	15.1^bc	410	9.79	2.39	0.830^a	28.75^b	4.78^b
SEM		0.002	0.086	13.62	0.854	0.532	0.031	0.245	0.042
P Value		0.3215	0.0001	0.542	0.075	0.2541	0.0217	0.0001	0.0001

^a,b,cmeans having different superscripts in the same row are significantly different (p < 0.05).

SEM: standard error of means; P value: probability level; TAC: Total antioxidant capacity; IgG: Gamma immunoglobulin; IgM: Mu immunoglobulin; IgA: Alpha immunoglobulin; T3: Triiodothyronine; T₄: Thyroxine; Arg/Lys: arginine/lysine ratio; Ca: Calcium; P: Phosphores.

Berman A, Horovitz T, Kaim M, Gacitua H. 2016. A comparison of THI indices leads to a sensible heat-based heat stress index for shaded cattle that aligns temperature and humidity stress. Int J Biometeorol. 60(10):1453–1462. doi: 10.1007/s00484-016-1136-9.

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Data availability statement

The data are available upon official request from the principal investigator and with the permission of the funding agent.