Different combinations of peppermint, chamomile and a yeast prebiotic have different impacts on production and severity of intestinal and bursal abnormalities of broilers challenged with coccidiosis

Figures & data

Table 1. Composition of the basal starter, grower and finisher diets as g/100 g (as fed basis).

Ingredients	Starter	Grower	Finisher
Corn	53.06	56.9	61.8
Soybean meal	31.48	32.89	28.23
Fish meal	4.0	–	–
Vegetable oil	3.0	4.48	4.6
Limestone	2.0	1.39	1.4
Dicalciun phosphate	2.72	0.95	0.82
Salt	0.11	0.19	0.10
DL-methionine	0.38	0.32	0.24
L-lysine HCl	0.5	0.26	0.21
L-threonine	0.25	0.13	0.09
Broiler premix^a	2.5	2.5	2.5
Nutrient composition
Metabolisable energy (kcal/kg)	3000	3150	3200
Crude protein (%)	23	21	19.16
Crude fibre (%)	2.25	2.38	2.33
Digestible arginine (%)	1.29	1.14	0.99
Digestible lysine (%)	1.29	1.14	0.99
Digestible meth + cyst (%)	0.87	0.84	0.73
Digestible tryptophan (%)	0.226	0.24	0.21
Digestible isoleucine (%)	0.87	0.81	0.73
Digestible threonine (%)	0.82	0.73	0.63
Digestible valine (%)	0.99	0.92	0.83
Calcium (%)	1.60	0.9	0.85
Available phosphorus (%)	0.844	0.45	0.42
Sodium (%)	0.16	0.16	0.16
Chloride (%)	0.35	0.312	0.23
Linoleic acid (%)	2.18	2.64	2.73

^a The broiler premix contained per kg: Vitamin A, 400,000 U; Vitamin D_3, 160,000 U; Vitamin E, 1200 mg; Vitamin B₁, 120 mg; Vitamin B₂, 280 mg; Vitamin B₆, 160 mg; Vitamin B₁₂, 1400 mcg; Biotin, 4 mg; Niacin, 1600 mg; Folic acid, 40 mg; Vitamin K₃, 100 mg; Calcium-D-pantothenate, 600 mg; Choline chloride, 12,000 mg; Choline, 10411.2 mg; Cu, 0.4 g; Mn, 3.2 g; Zn, 2.4 g; Fe, 2.0 g; I, 40 mg; Se, 10 mg; Lysine, 113.2 g; Methionine, 113.5 g; Meth + Cyst, 113.9 g; Tryptophan, 0.4 g; Threonine, 58.9 g; Valine, 1.4 g; Arginine, 2.2 g; Calcium, 62.0 g; Available Phosphorus, 121.3 g; Sodium, 50.0 g; Chloride, 64.0 g; Endo-1,3(4)-Beta-glucanase, 2800 U activity; Endo-1,4-Beta-xylanase, 10,800 U activity; BHT (E321) 1.34 g; Propyl gallate (E310) 0.112 g; Citric acid (E330) 0.2 g; Mycotoxin binder 40 g.

Table 2. Effect on weight gain (WG), feed intake (FI) and feed conversion ratio (FCR) at 10, 24 and 35-d-of-age in Eimeria challenged broilers given different dietary supplements (mean, pooled SEM, n = 4 pens).

Period	0–10 d			0–24 d			0–35 d
Treatments*	WG (g)	FI (g)	FCR	WG (g)	FI (g)	FCR	WG (g)	FI (g)	FCR
Negative control	248	296^a	1.20	982^b	1369	1.39^b	1895^a	2794	1.48^b
Positive control	234	293^a	1.25	882^c	1467	1.67^a	1698^b	2859	1.69^a
C P + CH	246	297^a	1.21	1050^a	1430	1.36^bc	1853^a	2767	1.49^b
C P + Y	249	303^a	1.22	1079^a	1394	1.29^c	1859^a	2802	1.51^b
C CH + Y	244	296^a	1.22	1069^a	1456	1.36^bc	1865^a	2814	1.51^b
C P + CH + Y	243	270^b	1.11	1068^a	1405	1.32^bc	1872^a	2799	1.50^b
C Sal	249	297^a	1.19	1054^a	1418	1.35^bc	1903^a	2784	1.46^b
SEM	2.45	2.71	0.02	14.36	11.16	0.02	15.37	14.88	0.02
p Value	.72	.02	.35	<.0001	.23	<.0001	.001	.82	<.0001

a, b, c – Means within the same column with different superscripts differ significantly (p < .05).

* Negative control = unchallenged, un-supplemented diet; Positive control = Eimeria challenged, un-supplemented treatment diet; C P + CH = Eimeria challenged, peppermint and chamomile supplemented diet; C P + Y = Eimeria challenged, peppermint and yeast supplemented diet; C CH + Y = Eimeria challenged, chamomile and yeast supplemented diet; C P + CH + Y = Eimeria challenged, peppermint, chamomile and yeast supplemented diet; C Sal = Eimeria challenged, salinomycin supplemented diet.

Table 3. Effect on relative internal organs weights (g/100g body weight) in 24 d-old Eimeria challenged broilers given different dietary supplements (mean, pooled SEM, n = 4 pens).

Treatments*	Liver	Heart	Spleen	Gizzard	Small intestine	Bursa of fabricius
Negative control	3.11	0.78^a	0.10	3.73	7.08^bc	0.15
Positive control	2.82	0.61^b	0.10	3.96	6.54^bcd	0.21
C P + CH	2.65	0.71^a^b	0.11	3.93	6.44 ^cd	0.15
C P + Y	2.87	0.60^b	0.09	3.78	8.28^a	0.16
C CH + Y	3.07	0.60^b	0.09	3.65	7.32^b	0.17
C P + CH + Y	2.99	0.61^b	0.10	4.24	8.25^a	0.16
C Sal	2.77	0.57^b	0.10	3.73	6.06^d	0.21
SEM	0.076	0.020	0.003	0.083	0.180	0.007
p Value	.70	.03	.85	.58	<.0001	.08

a, b, c, d – Means within the same column with different superscripts differ significantly (p < .05).

* See Table 2 for details of treatments.

Table 4. Effect on villus height (VH), crypt depth (CD), VH:CD, apparent villus widths and surface area and jejunum muscle thickness in 24 d-old Eimeria challenged broilers given different dietary supplements (mean, pooled SEM, n = 4 pens).

Treatments*	VH (µm)	CD (µm)	VH:CD ratio	Width villus tip (µm)	Width villus base (µm)	Villus surface area (mm^2)	Jejunal muscle thickness (µm)
Negative control	945^a^b	183^b	5.55^a^b	241	247	0.230	204^bc
Positive control	762^c	402^a	1.91^c	178	207	0.146	330^a
C P + CH	912^b	203^b	4.57^a^b	143	149	0.136	177^c
C P + Y	907^b	188^b	4.83^a^b	145	162	0.139	258^b
C CH + Y	946^a^b	160^b	5.90^a	136	141	0.130	156^c
CP + CH + Y	877^b	171^b	5.29^a^b	169	174	0.150	172^c
C Sal	1008^a	311^a	3.82^b	182	242	0.211	262^b
SEM	12.70	14.83	0.22	11.57	14.49	0.011	9.79
p Value	<.0001	<.0001	.0002	.30	.20	.05	<.0001

a, b, c – Means within the same column with different superscripts differ significantly (p < .05).

* See Table 2 for details of treatments.

Table 5. Effect on histomorphology of bursa of Fabricius follicles in 24 d-old Eimeria challenged broilers given different dietary supplements (mean, pooled SEM, n = 4 pens).

Treatments*	Follicle length (µm)	Follicle width (µm)	Follicle area (µm)
Negative control	5018^bc	2985	12,909^b
Positive control	6576^a	3995	21,532^a
C P + CH	4485^c	3023	10,803^b
C P + Y	4622^bc	2724	10,314^b
C CH + Y	4924^bc	2925	11,498^b
C P + CH + Y	5572^a^b	2904	12,712^b
C Sal	5610^a^b	3604	16,538^a^b
SEM	169.9	132.4	967.0
p Value	.004	.1	.01

a, b, c – Means within the same column with different superscripts differ significantly (p < .05).

* See Table 2 for details of treatments.

Table 6. Effect on serum biochemistry in 24 d-old Eimeria challenged broilers given different dietary supplements (mean, pooled SEM, n = 4 pens).

Treatments*	Cholesterol (mg/dL)	ALT (µL/dL)	AST (µL/dL)	Total Protein (g/dL)	Albumen (g/dL)	Globulin (g/dL)
Negative control	91^bc	2.43^c	158	2.53	1.06	1.48
Positive control	141^a	4.70^a	244	2.35	1.02	1.34
C P + CH	128^a^b	4.43^a^b	188	2.34	1.08	1.26
C P + Y	83^c	3.25^bc	169	2.08	0.86	1.22
C CH + Y	114^a^bc	3.88^a^b	180	2.45	1.05	1.40
C P + CH + Y	106^a^bc	3.48^a^bc	182	2.53	1.05	1.48
C Sal	120^a^bc	3.80^a^b	213	2.57	1.07	1.51
SEM	5.412	0.187	8.270	0.080	0.035	0.054
p Value	.03	.01	.07	.73	.73	.74

a, b, c – Means within the same column with different superscripts differ significantly (p < .05).

ALT: alanine aminotransferase; AST: aspartate aminotransferase.

* See Table 2 for details of treatments.

Table A2. Antioxidant inhibition activities in water and methanol extracts of peppermint (P) and chamomile (CH).

	DPPH analysis (inhibition %)				CUPRAC analysis (inhibition %)
	Water		Methanol		Water		Methanol
Concentration (µg/mL)	P	CH	P	CH	P	CH	P	CH
1000	81.03	84.09	85.19	88.17	3.74	1.11	3.63	1.11
500	64.78	70.33	74.78	73.11	2.32	0.74	2.31	0.76
250	47.75	65.22	56.75	67.20	1.26	0.42	1.25	0.42
125	34.57	40.96	42.57	48.03	0.81	0.27	0.80	0.27
62.5	27.85	31.63	31.85	32.65	0.54	0.19	0.54	0.19

DPPH: diphenyl-2-picryl-hydrazil inhibition (antioxidant capacity); CUPRAC: CUPric Reducing Antioxidant Capacity.

Table A1. Phenolic and flavonoid contents and neuroprotective enzyme activities in water and methanol extracts of peppermint (P) and chamomile (CH) (mean ± SD).

		Water extract	Methanol extract
Total phenolic (mg gallic acid equivalents/g dried plant material)	P	102.67 ± 0.98	90.88 ± 1.02
	CH	42.12 ± 0.62	26.38 ± 1.05
Total flavonoid (mg quercetin equivalents/g of dried plant material)	P	21.46 ± 0.36	26.02 ± 0.61
	CH	4.19 ± 0.02	6.19 ± 1.08
AChE (Inhibition % ± SD)	P	90.28 ± 0.36	94.16 ± 0.25
	CH	49.35 ± 1.21	32.16 ± 1.05
BChE (Inhibition % ± SD)	P	86.06 ± 1.12	88.55 ± 0.19
	CH	60.86 ± 0.14	56.50 ± 0.92

AChE: Acetylcholinesterase inhibition; BChE: Butyrylcholinesterase inhibition.

Data availability statement

The data that support the findings of this study are available from the corresponding author, [SMH], upon reasonable request.