The effect of the in ovo injection of some carbohydrates and antioxidants on incubating parameters, blood and immunological parameters, intestinal morphometry and post-hatching production performance in broiler chickens

Figures & data

Table 1. The ingredients of the basal diet.

Ingredients (% on dry matter)	Starter (1–21 days old)	Grower (22–42 days old)
Corn	61.5	63.00
Soybean meal (CP 44 %)	31.00	30.20
Soybean oil	3.00	3.00
Calcium bicarbonate	1.40	1.50
Dicalcium phosphate	1.5	1.00
Salt	0.35	0.20
Mineral premix^a	0.50	0.50
Vitamin premix^b	0.50	0.50
DL-methionine	0.25	0.10
Chemical analyses
Metabolizable energy (kcal/kg)	3027	3036
Crud protein (DM)	21.07	19.02
Lysine (%)	1.10	1.00
Methionine + Cysteine (%)	0.90	0.72
Arginine	1.19	0.31
Methionine (%)	0.48	0.34
Threonine (%)	0.80	0.74
Ca (%)	0.94	0.86
Available phosphorus (%)	0.41	0.38
Cl (%)	0.18	0.17

^aMineral mixture per kg of diet: Mn: 64 mg; Zn: 75 mg; Fe: 40 mg; Cu: 10 mg; Se: 0.3 mg, and I: 1.85 mg.

^bVitamin premix per kg of diet: vitamin A: 11013 U; vitamin D₃: 3525 U; vitamin E: 33 U; vitamin K₃: 2.75 mg; vitamin B₆: 5 mg; vitamin B₁₂: 0.028 mg; Thiamine: 2.2 mg; Riboflavin: 7.7 mg; Pantothenate: 17.6 mg; Nicotinic acid: 55.1 mg; Folic acid: 1.1 mg; vitamin H2: 0.22 mg and Choline chloride: 378 mg.

Ca: calcium; Cl: chlorine

Table 2. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on hatching parameters.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Hatchability (%)	76.72^b	76.41^b	78.89^ab	74.46^ab	89.66^a	4.068	.0340
BW of hatched chicks (g)	46.18^c	46.30^bc	47.21^a	47.11^ab	47.75^a	0.299	.0008
BW of hatched chicks / Egg Weight	69.30^b	69.11^b	70.42^a	67.69^b	72.69^a	2.076	<.0010
Losses (%)	5.64^a	5.65^a	4.98^ab	4.52^ab	1.82^b	1.255	.0212

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 3. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the physical quality of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Chicken activity	4.71^b	4.56^b	5.35^a	5.62^a	5.72^a	0.154	<.001
Appearance	9.33^b	9.32^b	9.73^a	9.78^a	9.85^a	0.137	<.001
Status of eyes	13.93^b	13.45^b	15.30^a	15.57^a	15.90^a	0.337	<.001
Status of legs	12.56^b	12.82^b	15.27^a	15.17^a	15.20^a	0.421	<.001
Status of navel	8.85^b	8.08^b	11.08^a	10.93^a	11.12^a	0.299	<.001
Status of residual membrane	9.29^b	9.26^b	11.22^a	10.93^a	11.25^a	0.278	<.001
Amount of residual yolk	12.44^b	12.78^b	14.82^a	14.40^a	15.02^a	0.368	<.001
Status of yolk stretching	8.74^c	8.65^c	11.18^ab	10.83^b	11.58^a	0.252	<.001
Total Tona score	79.67^b	79.69^b	94.20^a	91.32^a	95.60^a	1.663	<.001

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 4. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the red blood cell count of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Red blood cells count (10^6/mm^3)	3.17^b	3.10^b	3.36^ab	3.55^ab	3.76^a	0.135	.0138
Haemoglobin (g/dL)	10.11^b	10.03^b	11.18^a	12.21^a	12.55^a	0.059	.0200
Haematocrit (%)	34.11	34.24	34.78	34.36	34.63	0.5611	.9006
Mean corpuscle volume (µm^3)	130.78	130.93	130.50	129.98	130.87	10.53	.509

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 5. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the blood biochemical parameters of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Glucose (mg/dL)	192.00^b	192.70^b	218.00^a	193.88^b	204.80^ab	5.9618	.0345
Total Protein (mg/dL)	1.93	1.90	1.98	2.04	2.04	0.0991	.8123
Triasilgliserol (mg/dL)	72.85	69.90	66.30	69.15	69.50	3.315	.7398

NC:negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 6. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the immune system of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
White blood cells count (10^3/mm^3)	15.99	15.53	15.46	16.37	17.35	0.889	.5392
Neutrophil (%)	42.60	43.88	45.13	45.69	45.94	1.629	.6354
Lymphocyte (%)	45.25^b	45.27^b	47.20^ab	48.88^ab	50.10^a	1.458	.0418
Monocyte (%)	2.94	3.00	3.00	3.12	3.13	0.316	.9893
Eosinophil (%)	2.70	3.71	3.94	3.93	4.13	0.467	.3491
Basophil (%)	0.53	0.53	0.60	0.60	0.75	0.191	.9104

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 7. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the liver glycogen store and muscle tissue of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Liver glycogen (mg/g)	44.45^c	47.19^c	62.95^a	52.14^b	59.22^ab	5.100	.0459
Glycogen of breast muscle (mg/g)	8.19^b	8.21^b	10.48^a	9.27^ab	9.90^a	0.444	<.0010
Glycogen of thigh muscle (mg/g)	5.48^b	5.39^b	9.35^a	5.67^b	9.23^a	0.400	<.0010

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > 0.05).

SEM: standard error of the means.

Table 8. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the intestinal morphology of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
Duodenum length (cm)	6.80	6.39	7.04	6.76	7.03	0.318	.6060
Jejunum length (cm)	15.34^b	15.76^b	16.64^a	16.15^a	16.82^a	0.768	.0493
Ileum length (cm)	9.25	9.35	9.84	9.67	9.98	0.646	.9200
Duodenum length/entire length of the small intestine (%)	21.89^ab	22.10^ab	21.23^ab	20.13^b	23.38^a	1.074	.0470
Jejunum length/entire length of the small intestine (%)	45.56^b	44.59^b	49.06^a	48.93^a	50.40^a	1.470	.0487
Ileum length/entire length of the small intestine (%)	28.96	29.47	30.55	29.71	32.03	1.407	.5880
Villus height (µm)	209.92^b	207.44^b	280.95^a	280.45^a	286.45^a	10.700	<.0010
Villus width (µm)	66.47	61.62	64.11	63.45	69.77	3.547	.1010
Crypt depth (µm)	46.38^b	49.67^b	44.28^b	45.05^b	29.58^c	2.321	<.0410
Villus height/villus width	4.17	3.98	4.16	4.12	4.10	0.700	.1270
Villus height/crypt depth	4.62^b	4.36^b	6.36^ab	6.22^ab	9.16^a	0.259	.0451

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 9. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the antioxidant activity of the liver tissue of the newly hatched chickens.

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
CAT (U/mg protein liver)	21.61^bc	21.56^bc	18.53^c	27.87^a	24.80^b	2.056	.0480
MDA (nmol/mL)	0.11^a	0.11^a	0.09^ab	0.06^b	0.08^ab	0.008	.0080
SOD (U/mg protein liver)	7.31^b	7.11^b	8.65^ab	10.03^a	8.90^ab	0.775	.0445
GPx (U/mg protein liver)	1.23	1.22	1.29	1.48	1.28	0.091	.3230
Free radical scavenging power DPPH (mmol/L)	97.08^b	97.00^b	137.11^ab	212.94^a	138.94^ab	21.626	.0450
TAC (mmol/L)	0.76^b	0.75^b	0.77^b	1.27^a	1.10^ab	0.134	.0270

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

CAT: catalase; MDA: malondialdehyde; SOD: superoxide dismutase; GPx: glutathione peroxidase; TAC: total antioxidant capacity.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Table 10. The effect of the in ovo injection of carbohydrates and antioxidants into the stored incubating eggs on the body weight gain (BWG), feed intake (FI), conversion ratio (FCR).

Items			Treatments			SEM	p-Value
	NC	PC	CS	AS	CS + AS
1–21 Days of age
BWG (g/chicken/period)	27.89^b	28.21^b	35.47^a	34.28^a	36.07^a	1.124	.049
FI (g/chicken/period)	48.13^b	48.60^b	54.14^a	53.42^a	56.34^a	1.305	.007
FCR	1.71^a	1.72^a	1.51^b	1.56^b	1.58^b	0.038	.001
22–42 days of age
BWG (g/chicken/period)	76.52^b	75.36^b	78.89^a	78.72^a	79.28^a	1.28	.030
FI (g/chicken/period)	148.99^b	149.11^b	154.29^ab	157.13^ab	158.09^a	1.44	.027
FCR	1.98	1.99	1.96	1.99	1.98	1.44	.127
1-42 days of age
BWG (g/chicken/period)	53.51^b	53.13^b	61.01^a	59.11^a	62.18^a	1.19	.045
FI (g/chicken/period)	105.85^b	107.27^b	114.21^a	114.08^a	114.96^a	1.6	.002
FCR	1.98^a	2.00^a	1.86^b	1.91^b	1.85^b	0.15	.035

NC: negative control; PC: positive control; CS: carbohydrates solution; AS: antioxidants solution and CS + AS: carbohydrates solution + antioxidants solution.

The means within the same row with at least one common letter do not have significant difference (p > .05).

SEM: standard error of the means.

Data availability statement

I certify that all data in the article are true and reliable. All public data generated or analysed during this study are included in this article and the results can be shared with others.