Vitamin B5 supplementation enhances intestinal development and alters microbes in weaned piglets

Figures & data

Table 1. Ingredient and chemical composition of piglet diets, as-fed basis.

Phase I		Phase II
Ingredient	Content, %	Ingredient	Content, %
Corn	40.92	Corn	40.10
Extruded corn	20.00	Extruded corn	22.00
Soy protein concentrate	8.00	Soybean meal (43%)	20.50
Soybean meal (43%)	8.20	Fishmeal (63%)	4.00
Fishmeal (63%)	5.00	Whey powder	5.00
Whey powder	10.00	Glucose	3.00
Glucose	3.00	Soybean oil	1.50
Soybean oil	0.50	Limestone	0.58
Limestone	0.88	Dicalcium phosphate	0.88
Dicalcium phosphate	0.50	Choline chloride	0.10
Choline chloride	0.10	Antioxidants	0.05
Antioxidants	0.05	Acidifiers	0.50
Acidifiers	0.80	Salt	0.10
Salt	0.10	Mineral premix^a	0.15
Mineral premix^a	0.15	Vitamin premix^b	0.50
Vitamin premix^b	0.50	Lys (98%)	0.53
Lys (98%)	0.64	dl-Met	0.27
dl-Met	0.36	l-Thr	0.19
l-Thr	0.24	l-Trp	0.05
l-Trp	0.06
Total	100	Total	100
Calculated nutrition level
Net energy, Mcal/kg	2.49	Net energy, Mcal/kg	2.45
CP, %	18.61	CP, %	18.02
Ca, %	0.80	Ca, %	0.70
Available P, %	0.40	Available P, %	0.40
Lys^c, %	1.35	Lys^c, %	1.24
Met + Cys^c, %	0.77	Met + Cys^c, %	0.68
Thr^c, %	0.79	Thr^c, %	0.74
Trp^c, %	0.22	Trp^c, %	0.22

^aMineral premix: 100 mg/kg Zn (ZnSO₄), 30 mg/kg Mn (MnSO₄), 0.3 mg/kg Co (CoSO₄), 150 mg/kg Fe (FeSO₄), 25 mg/kg Cu (CuSO₄), 0.3 mg/kg Se (Na₂SeO₃) and 0.5 mg/kg I (KIO₃).

^bVitamin premix: 2200 IU/kg vitamin A, 17.5 μg/kg vitamin B12, 220 IU/kg vitamin D3, 16 IU/kg vitamin E, 0.5 mg/kg vitamin K3, 30 mg/kg niacin, 3.5 mg/kg riboflavin, 0.05 mg/kg biotin, 0.3 mg/kg folate acid, 1.0 mg/kg thiamin, 7 mg/kg pyridoxine, 4.0 mg/kg ethoxyquin.

^cStadardized ideal digestible.

Table 2. The growth performance in piglet.^a

Item	L-VB5	Control	H-VB5	SEM	p Value
BW, kg
Initial BW	7.21	7.10	7.20	0.25	.984
14 days BW	8.01	8.21	8.18	0.33	.970
Final BW	12.81	12.90	13.06	0.54	.982
ADG, g
Day 0–14	57.65	79.17	69.90	11.79	.780
Day 14–28	342.35	335.12	348.98	20.30	.967
ADFI, g/d
Day 0–14	194.39	208.33	211.08	10.63	.801
Day 14–28	398.36	380.46	405.74	16.26	.832
FCR, g/g
Day 0–14	3.06	2.42	2.73	0.21	.513
Day 14–28	1.83	1.72	1.74	0.06	.488
Diarrhea score
Day 0–14	0.88	0.84	0.82	0.09	.966
Day 14–28	0.61	0.61	0.72	0.13	.936

^an = 7.

BW, body weight; ADG, average daily gain; ADFI, average daily feed intake; FCR, feed conversion rate.

Table 3. Effect of vitamin B5 on intestinal organ indices.^a

Item	L-VB5	Control	H-VB5	SEM	p Value
Small intestine length, m	11.2	10.41	11.5	0.24	.195
Small intestine weight, g	552.77	534.81	589.91	24.05	.662
Large intestine length, m	2.13	2.02	2.31	0.05	.083
Large intestine weight, g	241.29	225	269.29	10.2	.212
Small intestine length/ BW, m/kg	0.9	0.82	0.91	0.04	.653
Small intestine weight/BW, g/kg	43.9	41.1	45.48	1.24	.379
Large intestine length/ BW, m/kg	0.17	0.16	0.18	0.01	.332
Large intestine weight/BW, g/kg	18.84a^b	17.57^b	20.85a	0.49	.016
Small intestine weight/small intestine length, g/m	49.32	50.96	51.21	1.68	.322
Large intestine weight/large intestine length, g/m	112.44	110.6	116.22	2.98	.758

^an = 7.

a, b Values in the same row with different superscript letters are significantly different (p < .05).

Table 4. Effects of vitamin B5 on intestinal morphology.^a

Item	L-VB5	Control	H-VB5	SEM	p Value
Villus height, μm
Duodenum, μm	284.21	308.69	281.95	8.32	.420
Jejunum, μm	293.14	294.11	265.53	8.03	.252
Ileum, μm	242.23^b	304.86a	306.00a	11.34	.016
Crypt depth, μm
Duodenum, μm	365.63	341.83	378.85	9.81	.353
Jejunum, μm	289.28	275.99	309.21	8.82	.325
Ileum, μm	305.77	281.13	281.62	9.13	.348
Cecum, μm	418.16a^b	383.46^b	469.21a	12.98	.017
Colon, μm	458.12a^b	416.57^b	513.05a	14.59	.020
Villus height: Crypt depth
Duodenum	0.8	0.91	0.75	0.04	.240
Jejunum	0.99	1.07	0.88	0.05	.289
Ileum	0.90^b	1.30a	1.21a	0.06	.011

^an = 7.

a, b Values in the same row with different superscript letters are significantly different (p < .05).

Figure 1. Intestinal morphological structure and immunohistochemical analysis of weaned piglets fed with different dietary concentrations of VB5 (magnification 100×). (A) ileum morphology. (B) Colon morphology. (C) Cecum morphology. (D) Immunohistochemistry staining with a Ki67 antibody in the colon. (E) Immunohistochemistry staining with a Ki67 antibody in the cecum. (F) Alcian blue-periodic acid-shiff staining in the colon. (G)Immunohistochemistry of the ileum. (H) Immunohistochemistry of the cecum. (I) Immunohistochemistry of the colon. n = 7. *p < .05, **p < .01, ***p < .001.

Figure 2. Effect of vitamin B5 on the composition of colonic bacterial. (A) NMDS plot of functional profiles among groups. (B) PCoA plot of functional profiles among groups. (C) Colon bacterial communities at the genus level. (D) Colonic differential bacteria at the genus level. n = 7. *p < .05, **p < .01, ***p < .001.

Table 5. Alpha diversity index.^a

Items	L-VB5	Control	H-VB5	SEM	p Value
Sobs	446.71	456.00	381	22.04	.330
ACE	509.84	531.40	452.49	23.63	.392
Shannon	3.67	3.99	3.42	0.11	.291
Simpson	0.11	0.06	0.11	0.14	.259
Chao1	520.33	533.98	451.85	23.42	.321

^an = 7.

Figure 3. Abundances of KEGG pathways in the functional prediction by Tax4Fun. (A) Prediction of colonic microbial function. (B) Metabolic function annotation map. (C) and (D) Amino acid metabolism. n = 7. *p < .05, **p < .01, ***p < .001.

Figure 4. Spearman correlation analysis of gut morphology and microbes (genus level). *p < .05, **p < .01, ***p < .001.

Table 6. Effect of VB5 on colonic SCFAs.^a

Item, μg/g	L-VB5	Control	H-VB5	SEM	p Value
Acetic acid	3451.79	3018.71	3176.17	135.50	.470
Propionic acid	1257.29	1327.61	1118.37	79.31	.577
Isobutyric acid	142.34^a	161.97^a	82.70^b	12.29	.007
Butyric acid	688.77	611.99	404.49	55.10	.073
Isovaleric acid	201.80^a	92.62^b	87.62^b	21.64	.034
Valeric acid	208.95	97.43	118.58	29.21	.281

^an = 7.

a, b Values in the same row with different superscript letters are significantly different (p < .05).