Evaluation of dietary supplementation of frankincense oil on broiler chicken growth performance, hepatic histomorphology, antioxidant activity, blood biochemical parameters, and inflammatory responses

Figures & data

Table 1. The proximate chemical constituents of the consumed basal diet (fed basis, g Kg⁻¹).

Ingredients	Starter period (4–10 d)	Grower period (11–23 d)	Finisher period (24–35 d)
Yellow corn 7.5% CP	559	593	620
Corn gluten, 60% CP	39.35	52.75	60.7
Soybean meal, 48% CP	334.2	281	238
Soybean oil	22	30	40
Calcium dibasic phosphate	15	14	13
Calcium carbonate	12	12	11
Common salt	1.5	1.5	1.5
DL-methionine, 98%	4	3	3.3
Premix*	3	3	3
Lysine HCl, 78%	4.7	4.5	4
Phytase	0.05	0.05	0.05
Threonine	1	1	1
Choline	0.7	0.7	0.7
Antimycotoxin	1	1	1
Na2Co3	2.5	2.5	2.5
Chemical composition (g kg^-1)
Crude protein	230.5	215.2	201.5
ME (Kcal/kg)	3004	3101	3201
Calcium	9.41	9.04	8.32
Available phosphorus	4.81	4.48	4.17
Lysine	14.6	13.1	11.6
Methionine	7.21	6.10	6.26
Threonine	8.24	7.65	7.13

* Premix per kg of diet: (Vit. A); 1 500 IU- (Vit. E); 10 mg- (Vit. D3); 200 IU- (Vit. K3); 0.5 mg (thiamine); 1.8 mg- (pantothenic acid); 10 mg- (riboflavin); 3.6 mg- (pyridoxine); 3.5 mg- (niacin); 35 mg- (folic acid); 0.55 mg- (biotin); 0.15 mg- (cobalamin); 0.01 mg- (Cu); 8 mg- (Fe); 80 mg- (Mn); 60 mg- (Zn); 40 mg- (Se); 0.15 mg- (I); 0.35 mg. CP: crude protein.

ME: metabolizable energy.

Table 2. The retention time (RT) and peak area (%) of the chemical constituents contained in frankincense oil (FKO), as evidenced by GC-MS.

Compound	RT (min)	Peak area %	Molecular formula	Molecular weight
1,6,10-dodecatrien-3-OL, 3,7,11-Trimethyl-, [S-(Z)]-	43.53	12.42	C15H26O	222
à-Farnesene	43.53	12.42	C15H24	204
Phenol,2,4-bis(1,1-dimethylethyl)-	17.78	7.15	C14H22O	206
2,4-Di-tert-butylphenol	17.78	7.15	C14H22O	206
10-Undecyn-1-ol	30.56	3.50	C11H20O	168
2-Furanmethanol,5-ethenyltetrahydro-à,à,5-trimethyl-,cis-	29.97	2.97	C10H18O2	170
Dodecanoic acid,2,3-bis(acetyloxy)propyl ester	27.91	2.44	C19H34O6	358
1,3-Butanediol, 2TMS derivative	33.07	2.36	C10H26O2Si2	234
(4-Isopropenyl-1-Cyclohexen-1-Yl)methanol	27.48	2.08	C10H16O	152
d-Mannose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, o-methyloxyme, (1E)-	26.13	2.06	C22H55NO6Si5	569
Silane, (butoxymethyl)trimethyl-	26.13	2.06	C8H20OSi	160
2,3-Butanediol	33.55	1.81	C4H10O2	90
4′-Hydroxy-4-methoxy-2′-methylchalcone	36.37	1.59	C17H16O3	268
Carbitol, TBDMS derivative	25.95	1.57	C12H28O3Si	248
Benzeneethanamine, N,à-dimethyl-	34.11	1.54	C10H15N	149
Thiocyanic acid	34.11	1.54	CHNS	59
Octadecanoic acid,trimethylsilyl ester	37.43	1.51	C21H44O2Si	356
Guanosine 3′,5′-cyclicmonophosphate	37.43	1.51	C10H12N5O7P	345
Furazolidone	35.73	1.51	C8H7N3O5	225
Propanoic acid, ethyl ester	36.56	1.51	C5H10O2	102
4-Methylcinnamic acid	37.14	1.50	C10H10O2	162
GLYCINE	37.71	1.41	C2H5NO2	75
3-Nonyn-1-ol	30.35	1.30	C9H16O	140
Silane, trimethyl(1-phenylethyl)-	48.50	1.18	C11H18Si	178
L-THREONINE	38.44	1.17	C4H9NO3	119
Idebenone	18.63	1.16	C19H30O5	338
Z-THUJENOL	18.63	1.16	C10H16O	152
2′,4′-Dihydroxy-4-methoxychalcone	38.04	1.14	C16H14O4	270
6,9,12-Octadecatrienoic acid,phenylmethyl ester, (Z,Z,Z)-	30.65	1.08	C25H36O2	368
L-(-)-Fucose	38.22	1.06	C6H12O5	164
1,3-Butanediol, 2TMS derivative	48.40	1.04	C10H26O2Si2	234
S-Methyl-L-cysteine	34.38	1.04	C4H9NO2S	135
Arachidonyl trifluoromethyl ketone	35.47	0.94	C21H31F3O	356
Sedoheptulosan	34.17	0.84	C7H12O6	192
Heptadecanoic acid, 16-methyl-,methyl ester	34.53	0.63	C19H38O2	298

Table 3. The impact of dietary frankincense oil (FKO) supplementation on the growth of broiler chickens.

Variables	FKO 0 mg/kg	FKO 200 mg/kg	FKO 400 mg/kg	FKO 600 mg/kg	SEM	P-values
						ANOVA	Linear	Quadratic
IBW (g)	100.56	100.63	100.00	100.42	0.14	0.43	0.42	0.55
Starter period
BW(g)	328.33	338.54	317.08	331.04	3.12	0.08	0.56	0.71
BWG(g)	227.78	237.92	217.08	230.63	3.10	0.10	0.60	0.74
FI (g)	269.63	270.63	256.46	264.58	2.54	0.18	0.18	0.45
FCR	1.18	1.14	1.18	1.15	0.01	0.60	0.66	0.85
Grower period
BW(g)	1169.70^b	1269.67^a	1224.44^ab	1260.22^a	13.45	0.01	0.01	0.07
BWG(g)	841.37^b	931.13^a	907.36^a	929.18^a	12.39	0.01	< 0.01	0.04
FI (g)	1201.85	1212.29	1121.46	1172.08	17.77	0.29	0.26	0.56
FCR	1.43	1.30	1.24	1.26	0.03	0.08	0.03	0.15
Finisher period
BW(g)	1888.89^b	2121.73^ab	2009.56^ab	2146.67^a	38.81	0.04	0.03	0.41
BWG(g)	719.19	852.06	785.12	886.44	27.55	0.13	0.07	0.74
FI(g)	1483.33	1602.08	1447.92	1450.00	31.31	0.28	0.36	0.35
FCR	2.062^a	1.878^ab	1.849^ab	1.656^b	0.05	0.03	0.01	0.95
Overall performance
BW(g)	1888.89^b	2121.73^ab	2009.56^ab	2146.67^a	38.81	0.04	0.03	0.41
BWG(g)	1788.33^b	2021.10^ab	1909.56^ab	2046.25^a	38.83	0.04	0.03	0.41
FI (g)	2954.81	3085.00	2825.83	2886.67	47.23	0.26	0.27	0.70
FCR	1.65^a	1.53^ab	1.48^ab	1.42^b	0.03	0.02	< 0.01	0.48
PER	2.90^b	3.15^ab	3.23^ab	3.39^a	0.07	0.03	0.01	0.60
RGR	994.72^b	1111.18^ab	1054.78^ab	1123.54^a	19.39	0.04	0.03	0.41

SEM = total standard error of means (n = 10).

^a,b,c Means with different superscripts are significantly different (p < 0.05). initial body weight- (IBW); body weight- (BW); body weight gain (BWG); feed intake- (FI); feed conversion ratio- (FCR); protein efficiency ratio- (PER); relative growth rate-(RGR).

Table 4. The impact of dietary frankincense oil (FKO) supplementation on broiler chickens’ hepatic and renal function variables.

Variables	FKO0 mg/kg	FKO 200 mg/kg	FKO400 mg/kg	FKO 600 mg/kg	SEM	P-Values
						ANOVA	Linear	Quadratic
ALT (U/L)	5.00^b	6.67^ab	7.00^ab	7.67^a	0.38	0.05	0.01	0.39
AST (U/L)	47.67	53.00	52.33	54.67	1.53	0.47	0.18	0.64
Creatinine (mg/dL)	1.11^b	1.16^ab	1.15^ab	1.21^a	0.01	< 0.01	< 0.01	1.00
Uric acid (mg/dL)	2.50	2.57	2.77	2.65	0.05	0.24	0.14	0.32

SEM total standard error of means (n = 10).

^a,bMeans with different superscripts significantly differ (p < 0.05). Aspartate aminotransferase (AST); Alanine aminotransferase (ALT).

Table 5. The impact of dietary frankincense oil (FKO) supplementation on broiler chickens’ serum antioxidant activity and inflammatory responses.

Variables	FKO0 mg/kg	FKO 200 mg/kg	FKO 400 mg/kg	FKO 600 mg/kg	SEM	P-Values
						ANOVA	Linear	Quadratic
TAC (U/mL)	10.22^c	11.51^b	13.05^a	13.72^a	0.42	0.000	< 0.01	0.22
CAT (U/mL)	3.19^c	4.16^bc	5.55^ab	6.30^a	0.40	0.002	< 0.01	0.77
SOD (U/mL)	135^c	145^bc	154^ab	161^a	3.14	0.001	< 0.01	0.58
MDA (nmol/mL)	6.28^a	3.71^b	2.89^b	3.10^b	0.43	0.000	< 0.01	< 0.01
IL1β ug/mL	144^c	157^b	161.67^b	170^a	2.90	0.000	< 0.01	0.14
IFN- γ (pg/mL)	7.13^c	11.10^b	13.33^ab	14.73^a	0.90	0.000	< 0.01	0.05

SEM: total standard error of means (n = 10).

^a,b,c Means with different superscripts are significantly different (p < 0.05). total antioxidant capacity-(TAC); catalase-(CAT); superoxide dismutase-(SOD); malondialdehyde-(MDA); interleukin 1 beta-(IL1β); interferon-γ- (IFN-γ).

Table 6. The impact of dietary frankincense oil (FKO) supplementation on serum lipid profile.

Variables	FKO0 mg/kg	FKO 200 mg/kg	FKO 400 mg/kg	FKO 600 mg/kg	SEM	P-values
						ANOVA	Linear	Quadratic
TC (mmol/L)	3.56^a	3.39^b	3.42^b	3.41^b	0.02	0.01	0.01	0.02
TG (mmol/L)	1.31^a	1.19^b	1.18^b	1.16^b	0.02	<0.01	<0.01	0.01
LDL-C (mmol/L)	1.35^a	1.06^b	1.03^b	0.96^b	0.05	<0.01	<0.01	0.03
VLDL-C (mmol/L)	0.22	0.23	0.23	0.23	0.00	0.66	0.29	0.57
HDL-C (mmol/L)	1.99^b	2.10^ab	2.15^a	2.21^a	0.03	<0.01	<0.01	0.43

SEM: total standard error of means (n = 10).

^a,bMeans with different superscripts significantly differ (p < 0.05). Total cholesterol-(TC); Triglycerides-(TG); Low-density lipoprotein cholesterol-(LDL-C); High-density lipoprotein cholesterol-(HDL-C); Very low-density lipoprotein cholesterol-(VLDL-C).

Figure 1. Photomicrographs of liver sections from chickens administered frankincense oil (FKO) at 0 mg kg⁻¹ (A), 200 mg kg⁻¹ (B), 400 mg kg⁻¹ (C), and 600 mg kg⁻¹ (D) exhibited typical histomorphological structures, like portal area (PA, yellow arrow), hepatocytes (HC, black arrow) which are perceived as a small masses around the Central veins (CV), a few round cells are seen as a natural immune response around the portal area (PA, arrow). mild to moderate portal lymphoplasmacytic aggregations (red arrows) and biliary proliferation (BD, yellow arrow) were seen in the FKO600 group. Each group with two magnification powers; H&E × 100, × 400.

Figure 2. The liver sections were immunostained for TGF-β positive cells (shown as red arrows) in the four experimental groups: (A) frankincense oil (FKO)0, (B) FKO200, (C) FKO400, and (D) FKO600. × 400 magnification power. (E) Quantitative analysis of the morphological features is as follows: 0.70%, 0.54%, 1.17%, and 1.46% for FKO0, FKO200, FKO400, and FKO600 groups, respectively.

Figure 3. The spleen sections were immunostained for TGF-β positive cells (showed as red arrows) in the four experimental groups: (A) frankincense oil (FKO)0, (B) FKO200, (C) FKO400, and (D) FKO600. × 400 magnification power. (E) Quantitative analysis of the morphological features is as follows: 0.50%, 1.72%, 1.73%, and 2.77% for FKO0, FKO200, FKO400, and FKO600 groups, respectively.

Data availability statement

The datasets generated or analyzed during the current study are not publicly available but are available from the corresponding author at reasonable request.