Effect of mannan-rich fractions and a mixture of sodium butyrate and zinc proteinate on performance, intestinal morphometry, and gene expression in broilers

Figures & data

Table 1. Negative control diets for experimental ages, as-fed basis.

Ingredients %	Age, days
	1 to 7	8 to 21	22 to 35	36 to 42
Corn	57.28	59.53	67.21	69.98
Soybean meal 45.5%	37.83	31.11	25.11	25.84
Extruded full-fat soybean	---	5.77	4.71	---
Meat and bone meal	1.55	0.91	0.28	---
Poultry fat	0.41	---	---	1.69
common salt	0.57	0.54	0.52	0.47
Limestone	1.18	1.17	1.15	1.14
Sodium bicarbonate	0.24	0.18	0.18	0.18
Biolys 55	0.19	0.14	0.25	0.15
DL-methionine	0.32	0.27	0.24	0.20
Threonine	0.04	0.01	0.01	0.01
Allzyme SSF	0.05	0.05	0.05	0.05
Anticoccidial^†	0.05	0.05	0.03	0.03
Vitamin premix^‡	0.05	0.05	0.05	0.05
Mineral premix^§	0.10	0.08	0.07	0.06
Starch^¶	0.14	0.14	0.14	0.14
Chemical composition
Metabolizable energy (kcal/kg)	3,000	3,080	3,150	3,220
Crude protein^††	23.67	23.17	19.88	19.19
Crude Fibre^††	2.40	2.47	2.50	2.62
Calcium	0.970	0.880	0.770	0.719
Total phosphorus^††	0.541	0.488	0.405	0.369
Phosphorus, available	0.435	0.393	0.344	0.321
Potassium	0.757	0.622	0.502	0.517
Sodium	0.230	0.220	0.210	0.190
Digestible arginine	1.53	1.39	1.77	1.08
Digestible isoleucine	0.89	0.86	0.72	0.69
Digestible lysine	1.26	1.17	1.05	0.93
Digestible methionine	0.64	0.58	0.51	0.47
Digestible Met + Cys	0.94	0.87	0.78	0.72
Digestible threonine	0.78	0.82	0.71	0.68
Digestible tryptophan	0.25	0.24	0.20	0.19
Digestible leucine	1.69	1.65	1.46	1.40
Digestible valine	0.98	1.37	0.81	0.77

^†Anticoccidial: Semduramycin + Nicarbazine at ages 1 to 21 days and Diclazuril 0.5% from 22 to 42 days. ^‡Vitamin Premix: Folic Acid 800 mg, Pantothenic Acid 12000 mg, Sodium Selenite 250 mg, Vitamin A 9000 IU, Vitamin B1 1500 mg, Vitamin B12mg, Vitamin B2 6000 mg, Vitamin B6 3000 mg, Vitamin D3 2500 IU, Vitamin E 20000 IU, nicotinic acid 25,000mg, vitamin K3 2500 mg, biotin 60 mg, inert material q.s.p. 1000 g. ^§Mineral premix: Iron sulphate 100 mg, copper sulphate 20 mg, zinc oxide 100 mg, manganese oxide 160 mg, cobalt 2 mg, calcium iodate 2 mg, inert material q.s.p. 1000 g. ^¶In the medicated treatments, 55 ppm of Methylene Disalicylate Bacitracin was used at the age of 1 to 21 days and 15 ppm of Virginiamycin from 22 to 42 days. In the other treatments, the prebiotic (Actigen) and the Blend (Viligen). Both additives were used in partial substitution of starch. ^††Analyzed values were obtained by the infra-red-reflectance-spectroscopy method (NIRS-005), by the EvonikBrazil laboratory.

Table 2. Sequence of primers used in the qRT-PCR reaction.

Genes^†	No. Acess	Gene sequence (5'→ 3')	Reference
TLR2	NM_204278.1	F: ACTGCCTGCAACGGTCAT R: CATCAGCTTCATTGTTGGTTTCTGT	Quinteiro-Filho et al. (2017)
IL1β	NM_204524.1	F: GTCAACATCGCCACCTACAA R: CGGTACATACGAGATGGAAACC	Del Vesco et al. (2020)
IL-2	GU119890.1	F: CCTCAAGAGTCTTACGGGTCTA R: AGTTGGTCAGTTCATGGAGAAA	El-Deep et al. (2019)
GLUT2	BI392505	F: CACACTATGGGCGCATGCT R: ATTGTCCCTGGAGGTGTTGGTG	Gilbert et al. (2007)
SGLT1	NM_207178	F: GCCATGGCCAGGGCTTA R: CAATAACCTGATCTGTGCACCAGTA	Gilbert et al. (2007)
y + LAT-2	XM_418326.6	F: CAGAAAACCTCAGAGCTCCCTTT R: TGAGTACAGAGCCAGCGCAAT	Gilbert et al. (2007)
β-Actina	L08165.1	F: GCCAACAGAGAGAAGATGAC R: CACCAGAGTCCATCACAATAC	Del Vesco et al. (2020)

^†TLR2 = Toll-like receiver 2; IL-1β = Interleukin 1 beta; IL-2 = Interleukin 2; GLUT2 = Glucose transporter 2; SGLT1 = Glucose transporter 1 Na+/dependent; y + LAT-2 = Neutral and cationic amino acid transporter.

Table 3. Performance of broilers fed experimental diets.

Parameters	Treatments^†	Age
		1–7	1–21	22–42	1–42
Weight gain (g)	NC	143.5^b	888.3^b	1642.8^b	2535.2^b
	PC	160.6^a	942.1ª	1673.4^ab	2617.2^ab
	Prebio	142.2^b	892.3^b	1773.0^a	2669.3^a
	Blend	146.5^b	896.1^b	1768.9^a	2666.7^a
	Prebio + blend	142.2^b	894.9^b	1741.1^ab	2636.1^ab
SEM		1.756	7.699	27.323	27.462
P-value		<0.001	<0.001	0.007	0.017
Feed intake (g)	NC	173.7	1335.0^ab	3192.9	4536.3
	PC	178.8	1373.1^a	3212.5	4592.2
	Prebio	174.3	1285.2^b	3258.0	4542.1
	Blend	178.2	1326.9^ab	3252.7	4581.0
	Prebio + blend	177.2	1316.0^ab	3245.1	4561.1
SEM		1.445	15.266	39.333	48.339
P-value		0.063	0.006	0.748	0.914
Feed conversion (g/g)	NC	1.21ª	1.50	1.95	1.79^a
	PC	1.12^b	1.46	1.92	1.76^ab
	Prebio	1.23ª	1.44	1.84	1.70^b
	Blend	1.22ª	1.48	1.84	1.72^b
	Prebio + blend	1.25ª	1.47	1.87	1.73^ab
SEM		0.016	7.699	0.028	0.017
P-value		<0.001	0.204	0.060	0.013

^†NC: diets without the use of antibiotics; PC: antibiotic diets; Prebio: diets enriched with Actigen Prebiotic; Blend = diets enriched with Viligen; Prebio + Blend: diets combined with additives. a,b Means in the same column with different superscript letters are significantly different by Tukey’s Test (p < 0.05). SEM: Standard error of the mean.

Table 4. C. perfringens count (CFU/g) present in the aviary litter.

Treatments^†	7 days	21 days	42 days
NC	820	55	160,000
PC	180	<10	250
Prebio	82	<10	<10
Blend	200,000	<10	<10
Prebio + blend	55,000	<10	<10

^†NC: diets without the use of antibiotics; PC: antibiotic diets; Prebio: diets enriched with Actigen Prebiotic; Blend: diets enriched with Viligen; Prebio + Blend: diets combined with additives.

Figure 1. Effects of experimental treatments on the intestinal lesion (duodenum to ileum) score of broilers at 42 days (n = 40). NC: Negative control diets without antibiotics; CP: Positive control diets with antibiotics; Prebio - diets with actigen prebiotic; blend – diets with Viligen; Prebio + blend: diets combined with additives.

Table 5. Intestinal morphometry of broilers fed experimental diets at 42 days of age.

Treatments^†	Villus height (µm)	Crypt depth (µm)	Villus: crypt
	Duodenum
NC	1523.6^ab	228.8ª	7.0^c
PC	1453.0^b	165.8^d	9.2^a
Prebio	1627.7ª	190.0^bc	8.6^ab
Blend	1632.9ª	186.3 ^cd	9.1^a
Prebio + blend	1598.7ª	210.7^ab	7.8^bc
SEM	30.721	5.350	0.275
P-value	<0.0001	<0.0001	<0.0001
	Jejunum
NC	1269.5	216.1ª	6.1^c
PC	1302.6	187.3^b	7.9^b
Prebio	1246.5	144.2^d	8.9^a
Blend	1279.8	165.2^c	7.8^b
Prebio + blend	1267.1	161.1^c	8.0^b
SEM	21.259	4.288	0.212
P-value	0.496	<0.0001	<0.0001
	Ileum
NC	869.8^b	196.9ª	4.7^d
PC	887.6^b	171.8^b	5.5^c
Prebio	906.1^b	118.3^c	8.0^a
Blend	781.6^c	158.1^b	5.3 ^cd
Prebio + blend	1079.5ª	164.1^b	6.6^b
SEM	22.420	5.232	0.192
P-value	<0.0001	<0.0001	<0.0001

^†NC: diets without the use of antibiotics; PC: antibiotic diets; Prebio: diets enriched with Actigen Prebiotic; Blend: diets enriched with Viligen; Prebio + Blend: diets combined with additives. a,b,c,d Means in the same column with different superscripted letters are significantly different by Tukey’s Test (p < 0.05). SEM: Standard error of the mean.

Figure 2. Relative expression of the immune system genes, Toll-like receptor 2 (TLR2), Interleukin 1β (IL-1β) and Interleukin 2 (IL-2) from the jejunum of broilers at 42 days of age (n = 25). NC: diets without the use of antibiotics; PC: antibiotic diets; Prebio: diets enriched with actigen prebiotic; blend: diets enriched with Viligen; Prebio + blend: diets combined with additives.

Figure 3. Relative expression of nutrient uptake genes, glucose transporter 2 (GLUT2), sodium-dependent glucose transporter (SGLT1), and neutral and cationic amino acid transporter (y-LAT2) from the jejunum of broilers aged 42 days (n = 25). Means with different letters a, b differ significantly (p < 0.05). NC: diets without the use of antibiotics; PC: antibiotic diets; Prebio: diets enriched with actigen prebiotic; blend = diets enriched with Viligen; Prebio + blend: diets combined with additives.

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Data availability statement

The data that support this study cannot be publicly shared due to ethical or privacy reasons and may be shared upon reasonable request to the corresponding author if appropriate.