In-Ovo injection of melittin into Alexandria chicken eggs: a way for early immune acceleration

Figures & data

Table 1. Effect of Melittin in-ovo injection on hatchability traits of Alexandria chick’s strain at hatch.

Item	Treatments					SME	P-value
	NC	PC	MLT (µg/egg)
			5	10	15
Egg weight, g	45.40	44.85	45.23	45.45	44.65	0.14	0.273
Chick weight, g	35.77^ab	35.33^ab	34.40^b	36.75^a	34.25^b	0.32	0.038
Chick yield, %	78.78^ab	78.76^ab	76.05^b	80.85^a	76.70^b	0.56	0.021
Hatchability, %	91.25^a	91.50^a	92.50^a	92.50^a	84.74^b	0.80	0.0001

NC = negative control (non-injected). PC = positive control (injected with normal saline). MLT = Melittin. SEM = standard error of mean.

Different superscripts in each row indicate significant differences (P ≤ 0.05).

Table 2. Effect of Melittin in-ovo injection on relative organs weight (%) of Alexandria chick’s strain at hatch.

	Treatments					SME	P-value
Item	NC	PC	MLT (µg/egg)
			5	10	15
Heart	0.67	0.73	0.58	0.67	0.62	0.02	0.343
Liver	2.31	2.56	1.82	2.80	2.20	0.16	0.375
Gizzard	4.25	4.71	4.30	5.46	4.31	0.25	0.564
Intestine	3.54^b	3.96^ab	3.15^b	4.59^a	3.42^b	0.15	0.033
Yolk sac	19.69^a	17.73^a	9.20^c	9.01^c	12.37^b	1.21	0.0001

NC = negative control (non-injected). PC = positive control (injected with normal saline). MLT = Melittin. SEM = standard error of mean.

Different superscripts in each row indicate significant differences (P ≤ 0.05).

Table 3. Effect of Melittin in-ovo injection on hematological traits and pH of Alexandria chick’s strain at hatch.

Item	Treatments					SME	P-value
	NC	PC	MLT (µg/egg)
			5	10	15
WBCs, 10^3/mL	16.53	14.62	15.57	14.96	15.89	3.08	0.333
Lymphocyte, %	84.67	84.00	80.00	82.67	81.67	0.83	0.422
Monocyte, %	4.67	5.00	5.67	5.00	5.00	0.51	0.988
Heterophils, %	7.67^c	8.33^bc	9.67^abc	11.50^a	10.33^ab	0.38	0.008
Eosinophils, %	3.00	2.67	3.00	3.00	3.00	0.30	0.907
H/L ratio	9.20	10.03	12.20	11.67	12.70	0.96	0.805
RBCs, 10^6/uL	2.78	2.15	2.46	2.30	2.43	0.06	0.067
Hb, g/dL	11.93^b	10.53^b	12.43^b	14.90^a	12.57^b	0.44	0.008
PCV, %	35.79 ^b	31.59^b	37.29^b	44.7^a	37.71^b	1.36	0.022
MCV, mm^3/RBC	169.33^a	160.40^bc	157.27^c	162.90^b	160.50^bc	1.02	0.000
MCH, pg	50.00	51.20	50.87	51.90	51.67	0.34	0.445
MCHC, %	36.33^a	32.00^b	32.37^b	31.97^b	32.23^b	0.39	0.000
pH	7.67^a	7.00^b	7.11^b	7.11^b	7.29^b	0.06	0.002

NC = negative control (non-injected). PC = positive control (injected with normal saline). MLT = Melittin. SEM = standard error of mean. WBC’s = white blood cells count. H/L ratio = heterophils to lymphocyte ratio. RBC’s = red blood cells. Hb = hemoglobin. PCV = picked cell volume; MCV = mean corpuscular volume. MCH = mean corpuscular hemoglobin. MCHC = mean corpuscular hemoglobin concentration. Different superscripts in each row indicate significant differences (P ≤ 0.05).

Table 4. Effect of Melittin in-ovo injection on T₃, T₄, T₃/T₄ ratio and glucose levels of Alexandria chick’s strain at hatch.

	Treatments					SME	P-value
Item	NC	PC	MLT (µg/egg)
			5	10	15
T3, nmol/L	1.78^c	1.99^bc	2.02^bc	2.59^a	2.16^b	0.08	0.004
T4, ug/dL	10.86^c	8.87^d	11.60^bc	14.80^a	12.50^b	0.55	0.0001
T3/ T4 ratio	16.37^b	22.67^a	17.43^b	17.70^b	17.33^b	0.77	0.039
Glucose, mg/dl	101.67^b	133.67^a	108.00^b	133.33^a	129.00^a	3.51	0.000

NC = negative control (non-injected). PC = positive control (injected with normal saline). MLT = Melittin. SEM = standard error of mean. T₃ = triiodothyronine; T₄ = thyroxine.

Different superscripts in each row indicate significant differences (P $\le$0.05).

Table 5. Effect of Melittin in-ovo injection on relative weight of lymphoid organs (spleen and bursa), protein profile, and immune indices of Alexandria chick’s strain at hatch.

Item	Treatments					SME	P-value
	NC	PC	MLT (µg/egg)
			5	10	15
Spleen, %	0.02^c	0.05^ab	0.03^c	0.06^a	0.04^ab	0.00	0.001
Bursa, %	0.13^b	0.18^b	0.14^b	0.31^a	0.12^b	0.02	0.027
Total Protein, g/dL	2.89	2.59	2.84	3.20	2.71	0.08	0.192
Albumin, g/dL	1.67^b	1.58^b	1.60^b	2.07^a	1.74^b	0.05	0.005
Globulin, g/dL	1.22	1.00	1.24	1.13	0.97	0.05	0.462
IgG, g/dL	18.83^c	22.57^bc	30.40^b	44.00^a	29.27^b	2.50	0.0001
IgM, g/dL	9.23^b	6.57^b	7.83^b	19.23^a	8.40^b	1.32	0.0001
IgA, g/dL	5.97^bc	4.14^d	7.47^a	4.87^cd	6.66^ab	0.28	0.000
T cells, %	6.00^c	6.00^c	11.67^b	10.33^b	16.67^a	0.81	0.000
B cells, %	11.33^c	12.67^bc	12.33^bc	14.00^ab	15.50^a	0.41	0.013

NC = negative control (non-injected). PC = positive control (injected with normal saline). MLT = Melittin. SEM = standard error of mean.

Different superscripts in each row indicate significant differences (P ≤ 0.05).

Table 6. The changing values (%) in Melittin in-ovo injection treatments compared to control (NC) treatment.

Traits	MLT (µg/egg)
	5	10	15
Hatchability	1.25	1.25	- 6.51
Spleen	0.01	0.04	0.02
Bursa	0.01	0.18	−0.01
IgG	61.45	133.67	55.44
IgM	−15.17	108.34	−8.99
IgA	25.13	−18.43	11.56
T cells	94.50	72.17	177.83
B cells	8.33	23.57	36.80

Italic numbers indicate the significance with the control (NC) group.