Effect of alfalfa supplementary change dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio on rumen fermentation and microbial function in Gansu alpine fine wool sheep (Ovis aries)

Figures & data

Table 1. Dietary formula and proximate nutrition content.

Items	Groups
	H	M	L
Ingredients (%)
alfalfa hay	15	25	35
Alfalfa silage	15	25	35
Corn	40.15	30.46	16.28
Rapeseed meal	14.23	7.25	2.34
Cottonseed meal	6.29	3.25	2.12
Soybean meal	2.33	2.04	2.26
NaCl	0.5	0.5	0.5
Limestone	0.5	0.5	0.5
NaHCO3	0.5	0.5	0.5
CaHPO4	0.5	0.5	0.5
Premix^a	5	5	5
Total	100	100	100
Nutrient levels^b
DE, MCal/kg	3.022	3.013	3.005
Crude protein, %	13.88	13.82	13.79
Ash, %	4.66	4.64	4.65
Ether extract, %	4.01	4.91	4.97
Neutral detergent fiber (NDF), %	26.53	36.29	45.54
Non-fiber carbohydrates (NFC), %	50.92	40.34	31.05
NFC/NDF^c	1.92	1.11	0.68
Ca, %	1.12	1.12	1.12
P, %	0.23	0.24	0.24

^a Provided per kilogram of diets: Cu 15 mg, Fe 55 mg, Zn 25 mg, Mn40 mg, Se 0.30 mg, I 0.5 mg, Co 0.20 mg, VA 20,000 IU, VD 4000 IU, VE 40 IU.

^b ME and NFC were calculated values according to the formula in Nutritional Requirements of Sheep for Meat in China, while other nutrient levels were measured values.

^c NFC was calculated according to the following equation: NFC = 100 − (% CP + %NDF + %EE + %mineral matter) (Mertens, 1997).

Table 2. Effect of dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio with body weight.

Items	H (1.92)	M (1.11)	L (0.68)
Initial Weight (Kg)	27.97 ± 0.95	28.31 ± 0.45	28.03 ± 0.72
Final Weight (Kg)	35.08 ± 0.84b	34.67 ± 0.64b	33.57 ± 0.77a
Average daily gain (g/d)	78.89 ± 4.03a	70.37 ± 3.25b	61.56 ± 4.43c

Table 3. Effect of dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio with Blood biochemical indices.

Items	H (1.92)	M (1.11)	L (0.68)	Normal range
TP g/L	65.67 ± 1.55	62.25 ± 0.75	63.28 ± 1.18	47.30–78.03
Alb g/L	32.75 ± 0.48	32.33 ± 0.33	32.42 ± 0.51	15.48–38.11
AST U/L	133.48 ± 3.08	130.25 ± 4.31	129.81 ± 4.11	106.5–142.5
ALT U/L	30.85 ± 2.14	30.25 ± 1.85	30.18 ± 1.59	19.5–34.8
BUN mmol/L ↑	7.45 ± 0.06a	7.21 ± 0.15b	7.25 ± 0.17b	2.9–7.3
TG mmol/L	0.26 ± 0.01	0.23 ± 0.02	0.02 ± 0.01	0.14–1.09
CHO mmol/L	2.93 ± 0.04a	2.62 ± 0.02b	2.54 ± 0.06b	2.0–3.4
LDH mmol/L	303.00 ± 9.51a	286.00 ± 5.72b	289.37 ± 7.98b	233.3–341.5
GLU mmol/L	2.31 ± 0.22	2.22 ± 0.27	2.31 ± 0.17	1.5–2.5

Table 4. Effect of dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio with rumen fermentation parameters.

Items	pH	Ammonia-N	Urea-N	Total-N	Soluble protein-N
H (1.92)	6.44 ± 0.11	24.24 ± 0.70a	1.15 ± 0.04	134.29 ± 0.56a	112.65 ± 0.92a
M (1.11)	6.41 ± 0.07	20.79 ± 0.15b	1.09 ± 0.05	125.31 ± 0.12b	102.40 ± 0.05b
L (0.68)	6.43 ± 0.58	21.23 ± 0.71b	1.11 ± 0.03	129.54 ± 1.23a	106.37 ± 0.73a

Table 5. Effect of dietary NFC/NDF with ruminal VFAs.

Items	TVFA(mmol/L)	Acetate(%)	Propionate(%)	Butyrate(%)	Acetate/ Propionate
H (1.92)	59.49 ± 3.28a	27.23 ± 4.03	21.39 ± 0.91a	7.34 ± 0.78a	1.28 ± 0.12c
M (1.11)	55.97 ± 1.96a	28.46 ± 3.15	17.29 ± 0.54b	7.64 ± 0.38a	1.64 ± 0.13b
L (0.68)	48.71 ± 3.16b	29.90 ± 2.49	12.93 ± 0.62c	4.80 ± 0.26b	2.32 ± 0.08a

Figure 1. Ruminal fluid samples microbiota venn diagram.

Figure 2. Percent of community abundance of microorganism.

Figure 3. Ruminal fluid samples microbiota beta-diversity analysis.

Figure 4. Ruminal fluid samples microbiota PCoA analysis.

Table 6. Effect of dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio with ruminal fluid samples alpha diversity.

Items	A (1.92)	C (1.11)	D (0.68)
Shannon	4.73 ± 0.21a	4.13 ± 0.28b	4.71 ± 0.29a
Simpson	0.027 ± 0.08b	0.064 ± 0.019a	0.032 ± 0.007b
ACE	758.33 ± 9.99b	878.87 ± 3.47a	747.25 ± 5.35b
Chao1	702.43 ± 17.47b	873.69 ± 15.07a	747.99 ± 16.33b
Coverage/%	98.33	98.67	98.36

Figure 5. The Linear discriminant analysis effect size (LEfSe) method identifies the significantly different abundant taxa of bacteria.

Figure 6. Ruminal fluid samples microbiota KEGG analysis.