Table 1
Composition of experimental diets (as fed basis)
Ingredients, %
|
Pre-care period
|
Late-care period
|
A
|
B
|
C
|
D
|
E
|
A
|
B
|
C
|
D
|
E
|
Waxy corn starch
|
54.80
|
45.21
|
38.36
|
33.43
|
29.32
|
53.54
|
44.17
|
37.48
|
32.66
|
28.64
|
High-Maize 1043
|
-
|
9.59
|
16.44
|
21.37
|
25.48
|
-
|
9.37
|
16.06
|
20.88
|
24.90
|
Soybean meal
|
9.00
|
9.00
|
9.00
|
9.00
|
9.00
|
12.00
|
12.00
|
12.00
|
12.00
|
12.00
|
Full fat expanded soybean meal
|
9.00
|
9.00
|
9.00
|
9.00
|
9.00
|
10.00
|
10.00
|
10.00
|
10.00
|
10.00
|
Fermented soybean meal
|
8.00
|
8.00
|
8.00
|
8.00
|
8.00
|
10.00
|
10.00
|
10.00
|
10.00
|
10.00
|
Fish meal
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.80
|
5.80
|
5.80
|
5.80
|
5.80
|
Whey, dried
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
5.00
|
Plasma protein powder
|
4.84
|
4.84
|
4.84
|
4.84
|
4.84
|
-
|
-
|
-
|
-
|
-
|
Monocalcium Phosphate
|
1.17
|
1.17
|
1.17
|
1.17
|
1.17
|
1.33
|
1.33
|
1.33
|
1.33
|
1.33
|
Soybean oil
|
1.00
|
1.00
|
1.00
|
1.00
|
1.00
|
-
|
-
|
-
|
-
|
-
|
Premix1
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
Choline chloride
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
0.08
|
Limestone
|
0.74
|
0.74
|
0.74
|
0.74
|
0.74
|
0.72
|
0.72
|
0.72
|
0.72
|
0.72
|
DL-lysine
|
0.23
|
0.23
|
0.23
|
0.23
|
0.23
|
0.21
|
0.21
|
0.21
|
0.21
|
0.21
|
DL Methionine
|
0.22
|
0.22
|
0.22
|
0.22
|
0.22
|
0.21
|
0.21
|
0.21
|
0.21
|
0.21
|
Salt
|
-
|
-
|
-
|
-
|
-
|
0.19
|
0.19
|
0.19
|
0.19
|
0.19
|
TOTAL
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
Calculated nutrient content2
|
|
|
|
|
|
|
|
|
|
|
Amylose/amylopectin ratio
|
0.00
|
0.20
|
0.40
|
0.60
|
0.80
|
0.00
|
0.20
|
0.40
|
0.60
|
0.80
|
Digestive energy, kcal/kg
|
3500
|
3500
|
3500
|
3500
|
3500
|
3408
|
3408
|
3408
|
3408
|
3408
|
Crude protein,%
|
18.50
|
18.50
|
18.50
|
18.50
|
18.50
|
18.00
|
18.00
|
18.00
|
18.00
|
18.00
|
Ca,%
|
0.85
|
0.85
|
0.85
|
0.85
|
0.85
|
0.92
|
0.92
|
0.92
|
0.92
|
0.92
|
Av.P,%
|
0.42
|
0.42
|
0.42
|
0.42
|
0.42
|
0.48
|
0.48
|
0.48
|
0.48
|
0.48
|
Salt,%
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
0.50
|
Amino acids, %SID3
|
|
|
|
|
|
|
|
|
|
|
Lys
|
1.45
|
1.45
|
1.45
|
1.45
|
1.45
|
1.31
|
1.31
|
1.31
|
1.31
|
1.31
|
TSAA
|
0.79
|
0.79
|
0.79
|
0.79
|
0.79
|
0.71
|
0.71
|
0.71
|
0.71
|
0.71
|
Thr
|
0.83
|
0.83
|
0.83
|
0.83
|
0.83
|
0.74
|
0.74
|
0.74
|
0.74
|
0.74
|
Trp
|
0.25
|
0.25
|
0.25
|
0.25
|
0.25
|
0.23
|
0.23
|
0.23
|
0.23
|
0.23
|
1Vitamin-mineral premix supplied per kilogram of feed: 10,000 IU of Vitamin A, 1,000 IU of Vitamin D3, 80 IU of Vitamin E, 2.0 mg of Vitamin K3, 0.03 mg of Vitamin B12, 12 mg of riboflavin, 40 mg of niacin, 25 mg of d-pantothenic acid, 0.25 mg of biotin, 1.6 mg of folic acid, 3.0 mg of thiamine, 2.25 mg of pyridoxine, 300 mg of choline chloride, 150 mg of Fe (FeSO4), 100 mg of Zn (ZnSO4), 30 mg of Mn (MnSO4), 25 mg of Cu (CuSO4), 0.5 mg of I (KIO3), 0.3 mg of Co (CoSO4), 0.3 mg of Se (Na2SeO3), and 4.0 mg of ethoxyquin
2Nutrient content of diets based on estimted nutrient contents of ingredients according to NRC(2012).
3SID, Standardized ileal digestible.
Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.
Table 2
RT-PCR primers related to tight connection and inflammation
Name1
|
F/R
|
Primer
|
Claudin
|
F
|
TTTCCTCAATACAGGAGGGAAGC
|
|
R
|
CCCTCTCCCCACATTCGAG
|
Occludin
|
F
|
CAGGTGCACCCTCCAGATTG
|
|
R
|
GGACTTTCAAGAGGCCTGGAT
|
ZO-1
|
F
|
CTGAGGGAATTGGGCAGGAA
|
|
R
|
TCACCAAAGGACTCAGCAGG
|
IL-1β
|
F
|
ACCTGGACCTTGGTTCTC
|
|
R
|
GGATTCTTCATCGGCTTC
|
TNF-α
|
F
|
ACGCTCTTCTGCCTACTGC
|
|
R
|
TCCCTCGGCTTTGACATT
|
18S
|
F
|
GAGCGAAAGCATTTGCCAAG
|
|
R
|
GGCATCGTTTATGGTCGGAAC
|
ZO-1: tight junction protein zonula occluden-1, IL-1 β: interleukin 1 β, TNFα: tumor necrosis factor, alpha.
Table 3
Effect of dietary amylose/amylopectin ratio on diarrhea occurrence of weaned piglets challenged with E.coli LPS
Items
|
A
|
B
|
C
|
D
|
E
|
SEM
|
P value
|
Diarrhea rate
|
|
|
|
|
|
|
|
First week
|
19.64
|
17.86
|
9.52
|
11.90
|
13.69
|
2.08
|
0.517
|
Second week
|
13.89
|
24.31
|
29.86
|
21.53
|
20.14
|
2.76
|
0.464
|
Third week
|
23.21AB
|
11.31BC
|
29.17A
|
18.45ABC
|
7.74C
|
2.25
|
0.024
|
Fourth week1
|
19.64
|
8.33
|
24.40
|
19.05
|
9.52
|
2.79
|
0.293
|
Total four weeks1
|
19.29
|
15.12
|
22.99
|
17.59
|
12.50
|
1.51
|
0.238
|
Diarrhea degree
|
|
|
|
|
|
|
|
First week
|
7.42
|
6.75
|
4.00
|
5.67
|
5.75
|
0.90
|
0.790
|
Second week
|
5.83
|
10.42
|
13.25
|
10.50
|
8.33
|
1.31
|
0.466
|
Third week
|
10.42AB
|
4.67B
|
14.25A
|
9.17AB
|
3.08B
|
1.19
|
0.029
|
Fourth week1
|
8.92
|
3.67
|
11.92
|
10.17
|
4.42
|
1.45
|
0.295
|
Total four weeks1
|
32.58
|
25.50
|
43.42
|
35.50
|
21.58
|
3.19
|
0.224
|
1Statistical analysis ended in day 27. Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.
Table 4
Effect of dietary amylose/amylopectin ratio on pH value of digest of weaned piglets challenged with E.coli LPS
Items1
|
A
|
B
|
C
|
D
|
E
|
SEM
|
P value
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
DAR
|
STRESS
|
D*S
|
Stomach
|
2.77
|
3.15
|
3.40
|
3.46
|
3.54
|
3.22
|
3.79
|
2.62
|
3.39
|
2.79
|
0.11
|
0.91
|
0.35
|
0.69
|
Jejunum
|
5.85
|
5.57
|
5.76
|
6.35
|
6.14
|
5.79
|
5.54
|
6.16
|
5.28
|
6.13
|
0.08
|
0.92
|
0.33
|
0.57
|
Ileum
|
6.84
|
6.50
|
6.21
|
7.17
|
6.49
|
7.20
|
6.92
|
6.66
|
6.84
|
7.10
|
0.05
|
0.83
|
0.15
|
0.11
|
Cecum
|
6.14B
|
6.13
|
6.71A
|
6.34
|
6.43AB
|
6.15
|
6.55AB
|
6.06
|
6.03B
|
6.10
|
0.04
|
0.02
|
0.02
|
0.22
|
Colon
|
6.55
|
6.76
|
6.80
|
6.56
|
6.92
|
6.32
|
6.93
|
6.37
|
6.91
|
6.40
|
0.05
|
1.00
|
0.00
|
0.14
|
1LPS:lipopolysaccharide, SAL:saline, DAR:dietary amylose/amylopectin ratio.Means within each row with different superscripts differ significantly (P<0.05).
Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.
Table 5
Effect of dietary amylose/amylopectin ratio on concentrations of volatile fatty acids in digest of weaned piglets challenged with E.coli LPS
Items1
|
A
|
B
|
C
|
D
|
E
|
SEM
|
P value
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
DAR
|
STRESS
|
D*S
|
Jejunum
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Acetic acid
|
41.84
|
30.39
|
300.98
|
56.52
|
58.57
|
17.84
|
69.97
|
99.37
|
49.07
|
27.86
|
10.63
|
0.24
|
0.28
|
0.33
|
Propionic acid
|
11.14
|
7.23
|
77.46
|
28.46
|
22.61
|
7.96
|
13.32
|
20.08
|
24.19
|
10.67
|
2.95
|
0.28
|
0.33
|
0.70
|
Iso-Butyrate acid
|
0.00
|
0.00
|
12.42
|
0.00
|
0.00
|
0.00
|
0.00
|
3.88
|
0.00
|
0.00
|
0.45
|
0.55
|
0.59
|
0.83
|
Butyrate acid
|
10.35B
|
0.00
|
62.50AB
|
0.00
|
0.00B
|
0.00
|
0.00B
|
6.57
|
37.87A
|
86.79
|
3.37
|
0.00
|
0.77
|
0.04
|
Iso-Valeric acid
|
6.68
|
7.69
|
20.16
|
0.00
|
0.00
|
4.64
|
9.11
|
25.37
|
15.59
|
13.49
|
1.84
|
0.38
|
0.81
|
0.28
|
Valeric acid
|
27.52
|
9.05
|
46.52
|
0.00
|
27.48
|
33.69
|
7.90
|
18.08
|
37.25
|
23.59
|
1.88
|
0.23
|
0.04
|
0.00
|
Total amount of SCFA
|
97.53
|
54.37
|
520.04
|
84.98
|
108.66
|
64.13
|
100.30
|
173.35
|
163.97
|
162.40
|
13.92
|
0.25
|
0.26
|
0.13
|
Ileum
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Acetic acid
|
594.22
|
927.69
|
786.89
|
631.66
|
541.72
|
928.44
|
634.98
|
544.22
|
942.19
|
848.87
|
37.54
|
0.18
|
0.33
|
0.08
|
Propionic acid
|
44.18
|
58.87
|
91.71
|
41.11
|
81.28
|
73.66
|
48.28
|
60.25
|
77.71
|
104.72
|
11.91
|
0.83
|
0.97
|
0.88
|
Iso-Butyrate acid
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
4.25
|
0.00
|
0.00
|
0.00
|
0.00
|
0.47
|
0.55
|
0.39
|
0.55
|
Butyrate acid
|
48.89
|
52.47
|
51.66
|
28.96
|
44.53
|
62.24
|
27.83
|
30.76
|
78.89
|
68.53
|
5.35
|
0.14
|
0.87
|
0.82
|
Iso-Valeric acid
|
0.00
|
0.00
|
0.00
|
10.03
|
0.00
|
17.68
|
0.00
|
0.00
|
0.00
|
3.04
|
1.25
|
0.15
|
0.02
|
0.15
|
Valeric acid
|
1.32
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
0.00
|
0.15
|
0.60
|
0.39
|
0.60
|
Total amount of SCFA
|
688.62
|
1039.03
|
930.26
|
711.75
|
667.53
|
1086.28
|
711.09
|
635.23
|
1098.79
|
1025.16
|
39.02
|
0.06
|
0.33
|
0.06
|
Cecal
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Acetic acid
|
2746.25B
|
3610.85
|
3433.77AB
|
4426.02
|
4198.72A
|
4643.30
|
3093.49B
|
3187.17
|
2901.98B
|
3329.84
|
148.66
|
0.03
|
0.08
|
0.90
|
Propionic acid
|
1242.81B
|
1476.83
|
1631.64AB
|
1805.10
|
2204.71A
|
1844.70
|
1391.95B
|
1414.73
|
1473.23B
|
1279.86
|
69.77
|
0.02
|
0.87
|
0.67
|
Iso-Butyrate acid
|
86.89BC
|
89.75
|
211.57A
|
135.97
|
124.75B
|
117.36
|
132.88BC
|
63.96
|
82.11C
|
70.14
|
5.42
|
<0.0001
|
0.01
|
0.10
|
Butyrate acid
|
585.71
|
965.03
|
637.10
|
712.82
|
820.75
|
966.94
|
802.65
|
778.01
|
668.31
|
758.83
|
45.61
|
0.65
|
0.17
|
0.75
|
Iso-Valeric acid
|
113.68B
|
110.53
|
309.97A
|
184.45
|
201.58 B
|
122.02
|
255.10 B
|
80.98
|
117.09 B
|
83.84
|
10.09
|
0.00
|
0.00
|
0.10
|
Valeric acid
|
192.85B
|
200.68
|
379.11A
|
292.34
|
295.05AB
|
232.47
|
245.74B
|
142.09
|
211.27B
|
159.77
|
14.57
|
0.02
|
0.06
|
0.82
|
Total amount of SCFA
|
4968.18B
|
6453.67
|
6603.15AB
|
7556.70
|
7845.56A
|
7926.79
|
5921.81B
|
5666.94
|
5453.97B
|
5682.28
|
260.26
|
0.03
|
0.37
|
0.86
|
Colon
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Acetic acid
|
2862.32
|
3032.80
|
2292.78
|
3213.04
|
2378.01
|
3367.35
|
2104.43
|
3360.75
|
1701.21
|
3347.96
|
65.92
|
0.40
|
<0.0001
|
0.04
|
Propionic acid
|
1225.44
|
1169.05
|
1080.91
|
1280.66
|
1030.95
|
1375.93
|
940.75
|
1397.74
|
772.60
|
1348.47
|
33.87
|
0.71
|
0.00
|
0.09
|
Iso-Butyrate acid
|
126.31
|
132.60
|
185.56
|
144.87
|
161.69
|
110.50
|
134.54
|
128.63
|
161.95
|
125.40
|
5.36
|
0.26
|
0.03
|
0.45
|
Butyrate acid
|
607.55
|
750.58
|
561.84
|
758.93
|
685.08
|
923.08
|
475.83
|
861.94
|
488.16
|
918.83
|
31.77
|
0.60
|
0.00
|
0.62
|
Iso-Valeric acid
|
201.67
|
200.77
|
343.89
|
216.31
|
273.89
|
148.26
|
220.49
|
186.95
|
298.49
|
173.75
|
11.99
|
0.24
|
0.00
|
0.36
|
Valeric acid
|
229.56
|
225.36
|
355.00
|
240.91
|
259.62
|
241.07
|
190.01
|
236.07
|
235.32
|
241.87
|
9.30
|
0.08
|
0.39
|
0.12
|
Total amount of SCFA
|
5252.84
|
5511.16
|
4819.99
|
5854.72
|
4789.24
|
6166.18
|
4066.05
|
6172.07
|
3657.73
|
6156.28
|
124.10
|
0.64
|
<0.0001
|
0.09
|
1LPS:lipopolysaccharide, SAL:saline, DAR:dietary amylose/amylopectin ratio.Means within each row with different superscripts differ significantly (P<0.05).
Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.
Tabel 6
Effect of dietary amylose/amylopectin ratio on crypt depth of intestinal of weaned piglets challenged with E.coli LPS
Items1
|
A
|
B
|
C
|
D
|
E
|
SEM
|
P value
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
DAR
|
STRESS
|
D*S
|
Cecal
|
429.98A
|
424.59
|
401.36A
|
429.59
|
396.38AB
|
385.18
|
396.34A
|
414.19
|
372.74B
|
362.18
|
5.26
|
0.01
|
0.73
|
0.69
|
Colon
|
436.03
|
475.67
|
470.36
|
461.69
|
450.36
|
474.08
|
446.81
|
477.60
|
464.20
|
434.81
|
7.12
|
0.96
|
0.45
|
0.55
|
1LPS:lipopolysaccharide, SAL:saline, DAR:dietary amylose/amylopectin ratio.Means within each row with different superscripts differ significantly (P<0.05).
Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.
Table7
Effect of dietary amylose/amylopectin ratio on gene expression of intestinal mucosa of weaned piglets challenged with E.coli LPS
Items1
|
A
|
B
|
C
|
D
|
E
|
SEM
|
P value
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
LPS
|
SAL
|
DAR
|
STRESS
|
D*S
|
Jejunal mucosa
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
claudin
|
0.75B
|
1.04
|
0.98B
|
1.03
|
1.01B
|
1.02
|
1.11A
|
1.51
|
0.79B
|
1.16
|
0.04
|
0.045
|
0.01
|
0.49
|
zo-1
|
0.92
|
1.03
|
0.96
|
1.04
|
0.96
|
1.00
|
0.82
|
0.71
|
0.61
|
0.96
|
0.04
|
0.15
|
0.22
|
0.46
|
IL-1β
|
0.84
|
1.11
|
0.60
|
0.91
|
0.39
|
0.73
|
0.65
|
0.77
|
0.37
|
0.87
|
0.08
|
0.53
|
0.06
|
0.97
|
TNFa
|
1.25
|
1.06
|
0.75
|
1.63
|
0.61
|
0.83
|
0.65
|
0.73
|
0.47
|
0.79
|
0.13
|
0.50
|
0.32
|
0.78
|
Ileal mucosa
|
|
|
|
|
|
|
|
|
|
|
0.00
|
|
|
|
Occludin
|
1.49
|
1.06
|
1.27
|
1.93
|
1.08
|
1.44
|
0.98
|
1.05
|
1.08
|
1.62
|
0.10
|
0.55
|
0.26
|
0.51
|
claudin
|
1.15B
|
1.04
|
2.37A
|
2.20
|
1.25B
|
1.53
|
1.13B
|
1.24
|
1.22B
|
1.38
|
0.05
|
<0.0001
|
0.60
|
0.60
|
ZO-1
|
1.07
|
1.06
|
1.07
|
1.25
|
1.21
|
1.02
|
0.82
|
0.92
|
0.90
|
1.11
|
0.06
|
0.63
|
0.65
|
0.84
|
IL1β
|
0.80
|
1.14
|
1.12
|
1.60
|
0.79
|
1.16
|
0.62
|
2.45
|
1.00
|
1.16
|
0.18
|
0.82
|
0.09
|
0.60
|
TNFa
|
0.76C
|
1.02
|
0.97BC
|
1.02
|
1.96A
|
1.25
|
1.30A
|
1.84
|
1.46AB
|
1.42
|
0.07
|
0.01
|
0.87
|
0.10
|
1LPS:lipopolysaccharide, SAL:saline, DAR:dietary amylose/amylopectin ratio.Means within each row with different superscripts differ significantly (P<0.05).
ZO-1: tight junction protein zonula occluden-1, IL-1 β: interleukin 1 β, TNFα: tumor necrosis factor, alpha. Dietary amylose/amylopectin ratio of A, B, C, D, E was 0.00, 0.20, 0.40,0.60 and 0.80 respectively.