Beruflich Dokumente
Kultur Dokumente
www.elsevier.com/locate/nutres
Abstract
Because increasing viscosity of gastrointestinal tract contents has been shown to alter physiologic
responses in many species, and no single standardized method of intestinal viscosity measurement
exists, effects of diet and measurement techniques were studied in a canine model. Three
experiments were conducted to evaluate the use of mixer viscometry, effects of freezing,
centrifugation, time of sampling, and dilution of digesta, and effects of diet on viscosity of canine
ileal digesta and simulated small intestinal digesta viscosity. Digesta viscosity values measured at
218C were within 5% of those at 218C after a 24-hour freeze. Viscosity constants of whole digesta
sampled between 8:00 am and 2:00 pm daily ranged from 6575 to 32 692 mPa d s and 7.47 to 9.03
mPa d s after centrifugation. Digesta sampled between 2:00 pm and 8:00 pm daily had viscosity
constants of whole digesta ranging from 7137 to 17 345 mPa d s and 6.42 and 9.46 mPa d s after
centrifugation. Digesta samples diluted with Millipore filtered water, to determine whether a dilution
factor could be used to estimate viscosity, had drastically underestimated viscosity constants. Ileal
digesta viscosity constants were similar for dogs fed test diets varying in carbohydrate source and
ranged from 6901 to 12 590 mPa d s. During simulated small intestinal simulation, viscosity peaked
between 6 and 9 hours. Data indicate that alteration of digesta by centrifugation or dilution
underestimates viscosity data. Viscosity of whole digesta can be measured using mixer viscometry.
Variations in diet ingredients appear to alter intestinal digesta viscosity, indicating further need for
investigation of the role of diet in viscosity.
D 2007 Elsevier Inc. All rights reserved.
Keywords:
1. Introduction
Increasing the viscosity of digesta in the gastrointestinal
tract through diet manipulation with dietary fiber has been
shown to significantly reduce postprandial glucose response
in humans, pigs, canines, and rodents, indicating a potential
mechanism for management of diabetes and abnormal
carbohydrate metabolism [1-4]. Although it has been
demonstrated that ingestion of viscous fibers added to the
diets of animals (humans, canines, pigs, rodents) will lower
4 Corresponding author. Tel.: +1 217 333 2361; fax: +1 217 333 7861.
E-mail address: gcfahey@uiuc.edu (G.C. Fahey).
0271-5317/$ see front matter D 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.nutres.2006.12.005
57
58
2.2.1. Animals
Purpose-bred female dogs (n = 6; Butler Farms USA)
with hound bloodlines, an average initial body weight of
26.1 kg (range, 19.4-28.2 kg), and an average age of
4.2 years (range, 3.0-7.8 years) were used. Dogs had
previously been surgically prepared with an ileal cannula
and were housed as previously described. Dogs were fed the
same commercial diet as was used in Experiment 1. The diet
met or exceeded the Association of American Feed Control
Officials [21] recommendations for dogs at weight maintenance and had an ME concentration of 20628 kJ/kg. Dogs
were fed a total of 400 g of each diet divided equally
between 2 feedings occurring at 8:00 am and 8:00 pm. This
study was conducted over a 14-day period. Initially, a 6-day
diet adaptation phase preceded an 8-day collection of ileal
digesta. Ileal digesta was collected once daily during the
8-day collection period, and each sample collection lasted
1.5 hours in length. Sample times rotated by 1.5 hours each
day with the first-day collection beginning at 8:00 am.
Collected ileal digesta was immediately frozen at 208C.
Before the viscosity measurement, ileal digesta was
composited for days 1 to 4 (8:00 am-2:00 pm) and days
5 to 8 (2:00 pm-8:00 pm) and thawed in a 558C water bath
until samples reached room temperature (238C). Water was
offered ad libitum by providing 1500 mL at 8:00 am and
8:00 pm daily into 2000-mL stainless steel bowls attached
to the cages. Before offering new water at 8:00 am and
8:00 pm, remaining water from the previous offering was
measured with a graduated cylinder and amounts recorded.
2.3.2. Procedures
Dogs were fed 1 of 5 dietary treatments that consisted
of commercially available canine extruded diets. Data
have shown that various carbohydrates impact the
viscosity of intestinal fluid of many species including dogs
[2,4,6,8-10,13]. Therefore, the diets chosen differed in
carbohydrate ingredients and included either ground wheat
(GW) (Kibbles n Bits Original, DLM Foods L.L.C, San
Francisco, CA), brewers rice (BR) (Purina One, Nestle
Purina, St Louis, Mo), corn meal (CM) (Eukanuba Adult
Premium Performance Formula, The Iams Company), corn
meal + ground sorghum + ground wheat (Mixed) (Science
Diet Adult Original, Hills Pet Nutrition, Inc, Topeka, Ks),
or oatmeal (OatM) (Cycle Custom Fitness, DLM Foods,
LLC, San Francisco, Calif). Dogs were fed a total of 500 g
of each diet daily divided equally between 2 feedings
occurring at 8:00 am and 8:00 pm.
The experimental design was a 5 5 Latin square
consisting of 10-day periods. A 6-day adaptation phase
preceded a 4-day collection of ileal digesta. Ileal digesta was
collected 3 times a day, with an interval of 4 hours between
collections. Ileal collections were 1 hour in length.
Sampling times on the remaining 3 days rotated 1 hour
from the previous days collection time. Ileal samples were
collected as described above. Ileal digesta collected at each
time was immediately frozen at 208C. At the end of each
period, ileal digesta was composited for each individual
dog. Composited ileal samples were allowed to thaw in a
558C water bath until they reached room temperature
(238C). Samples were mixed by hand with a metal spatula
for 60 seconds. Approximately 30-mL duplicates of ileal
digesta were placed in glass beakers (5 cm in diameter).
Viscosity of ileal digesta was analyzed as described
above. If samples were too viscous to measure directly, they
were diluted 1:1 (w/w basis) with Millipore filtered water
(viscosity, 1 mPa d s).
2.2.2. Viscosity
Viscosity was measured in duplicate as described in
Experiment 1. After the initial viscosity measurement, ileal
2.2. Experiment 2
59
Table 1
Chemical analyses of dietary treatmentsa
Diet
DM
OM
Diet A
BR
CM
GW
Mixed
OatM
91.0
93.3
93.3
91.1
92.5
92.2
93.4
92.8
91.7
91.6
95.0
93.9
CP
AHF
TDF
IDF
SDF
11.7
19.4
22.6
11.9
15.4
13.1
10.3
5.4
8.5
10.9
6.7
9.8
8.6
3.3
5.9
9.3
5.1
8.4
1.7
2.1
2.6
1.6
1.6
1.4
20628
22190
22948
20105
21436
20704
23.4
28.2
33.5
24.9
26.8
24.7
Diet A used in Experiments 1 and 2; BR, CM, GW, Mixed, and OatM diets used in Experiments 3 and for in vitro digestion simulation.
3. Results
3.1. Chemical analyses
Chemical analyses of diets tested in all 3 experiments are
presented in Table 1. Dry matter and OM concentrations
60
Table 2
Nonlinear regression viscosity parameters for ileal digesta fresh and after a 24-hour freeze from 3 dogs consuming a standard dieta
Dog
Day
Fresh digesta
Viscosity constant, mPa d s
1
2
3
1
2
3
1
2
3
13 931
15 563
15 663
23 726
16 206
11 416
12 512
20 813
16 518
Frozen digesta
Exponent
1.26
1.30
1.77
1.44
1.91
1.52
1.22
0.83
1.71
0.99
0.99
0.99
0.99
0.99
0.99
0.99
0.99
0.99
Exponent
13 241
16 084
15 171
24 878
17 503
10 846
12 410
22 270
16 301
1.18
1.31
1.62
1.41
1.87
1.50
1.23
1.00
1.69
R2
0.99
0.99
0.99
0.99
0.99
0.99
0.99
0.99
0.99
Nonlinear viscosity parameters are based on the power law equation ( y = a * x b ), where y is shear stress, a is the viscosity constant, x is shear rate, and
b is a dimensionless exponent indicating deviation from Newtonian flow (exponent b 1 for pseudoplastic fluids); R 2 indicates the proportion of variation
explained by the nonlinear regression model.
1
2
3
4
5
6
DMI, g/d
313
296
365
232
361
198
1066
1022
1095
568
902
628
Centrifuged
15 711
8808
22 588
7631
12 360
10 104
8.04
7.70
7.86
7.64
9.23
6.96
a
All nonlinear regression viscosity constants for whole digesta were
higher ( P b .05) compared with centrifuged digesta (pooled SEM,
1397.25).
b
Correlations between DMI, water intake, and nonlinear viscosity
constants measured on whole and centrifuged digesta were not significant
( P N .05).
61
Table 4
Nonlinear regression viscosity parameters for digesta measured whole and after centrifugation in morning and afternoon hours from 6 dogs consuming a
standard dieta,b,c,d
Dog
1
Constant, mPa
Exponent
R2
2
Constant, mPa
Exponent
R2
3
Constant, mPa
Exponent
R2
4
Constant, mPa
Exponent
R2
5
Constant, mPa
Exponent
R2
6
Constant, mPa
Exponent
R2
Morning hours
Afternoon hours
Whole digesta
Centrifuged digesta
Whole digesta
Centrifuged digesta
d s
14 077
1.58
0.99
9.03
1
NS
17 345
1.20
0.99
7.05
0.61
0.84
d s
6575
1.38
0.99
7.47
1
NS
11 041
1.46
0.99
7.94
1
NS
d s
32 692
1.27
0.99
8.67
0.96
0.98
12 483
1.30
0.99
7.06
1
NS
d s
8124
1.29
0.99
8.23
1
NS
7137
1.36
0.99
7.05
1
NS
d s
12 236
1.46
0.99
8.99
1
NS
12 483
1.70
0.99
9.46
1
NS
d s
10 283
1.38
0.99
7.50
1
NS
9925
1.30
0.99
6.42
1
NS
Nonlinear viscosity parameters are based on the power law equation ( y = a * x b ), where y is shear stress, a is the viscosity constant, x is shear rate, and
b is a dimensionless exponent indicating deviation from Newtonian flow (exponent b 1 for pseudoplastic fluids); exponents were given as 1 if nonlinear
regression was not significant (NS; P N .05); R 2 indicates the proportion of variation explained by the nonlinear regression model.
b
Constant mean estimates were used in the case of nonsignificant nonlinear regression analysis.
c
Ileal digesta sampled in the morning hours between 8:00 am and 2:00 pm and ileal digesta sampled in the afternoon hours between 2:00 pm and 8:00 pm.
d
All nonlinear regression viscosity constants for whole digesta were higher ( P b .05) compared with centrifuged digesta (pooled SEM, 1397.25).
Table 5
Nonlinear regression viscosity parameters, apparent total tract dry matter and organic matter digestibilities, and ileal dry matter concentrations for dogs
consuming diets containing various carbohydrate ingredients1,2
Item
BR
CM
GW
Mixed
OatM
SEM
P value
12 590
1.32
358
13.0a,b
11 291
1.19
353
14.6a
9431
1.19
303
12.4b
7934
1.21
372
12.7a,b
10 228
1.17
379
11.7b
3134.7
0.09
38.17
0.59
NS
NS
NS
b .05
a,b
Least squares means in the same row that do not have common superscript letters differ ( P b .05).
Nonlinear viscosity parameters are based on the power law equation ( y = a * x 2 ), where y is shear stress, a is the viscosity constant, x is shear rate, and
b is a dimensionless exponent indicating deviation from Newtonian flow (exponent b 1 for pseudoplastic fluids).
2
Diluted samples were not included in the statistical analysis.
1
62
Table 6
Nonlinear regression viscosity constants calculated for digesta samples of
dogs fed diets containing various carbohydrate ingredientsa
Dog
BR
CM
GW
Mixed
OatM
1
2
3
4
5
21 926
9216
10 331
4715
16 760
1585a
9349
3018a
6712
17 813
5362
5505
12 369
7713
16 211
9412
6835
5894a
7557
4811a
9094
8431
3026a
2943
20 446
Table 7
Nonlinear regression viscosity parameters for solutions containing diets with various carbohydrate ingredients during small intestinal digestion simulation1
Sample
Time, h
0
BR
Constant, mPa
Exponent
R2
CM
Constant, mPa
Exponent
R2
GW
Constant, mPa
Exponent
R2
Mixed
Constant, mPa
Exponent
R2
OatM
Constant, mPa
Exponent
R2
c, d, e
12
15
18
d s
27d
1.04
0.95
47c,d
1.53
0.99
130c
0.87
0.97
22d
1.19
0.97
36c,d
1.07
0.99
23d
0.57
0.90
75c,d
0.84
0.98
d s
254c,d
0.97
0.99
148c,d
1.04
0.99
262c,d
0.85
0.99
84c,d
0.98
0.98
95c,d
0.94
0.99
34e
1.15
0.99
43e
1.10
0.98
d s
14d
1.50
0.99
63c,d
0.92
0.99
77c
0.78
0.99
29c,d
0.97
0.98
34c,d
1.31
0.97
31c,d
1.52
0.98
48c,d
1.00
0.93
d s
33c,d
1.30
0.98
14d,e
1.62
0.96
4e
2.18
0.97
89c
1.09
0.99
31c,d
1.06
0.99
4e
1.24
0.93
17d,e
1.66
0.98
d s
32
1.05
0.99
29
0.95
0.99
17
0.69
0.95
18
1.18
0.99
24
1.06
0.97
25
0.94
0.96
26
0.87
0.97
Least squares means (5 diets, 7 time points; n = 35) in the same row that do not have common superscript letters differ ( P b .05) (pooled SEM, 21.20).
Nonlinear viscosity parameters are based on the power law equation ( y = a * x b ), where y is shear stress, a is the viscosity constant, x is shear rate, and
b is a dimensionless exponent indicating deviation from Newtonian flow (exponent b 1 for pseudoplastic fluids); R 2 indicates the proportion of variation
explained by the nonlinear regression model.
1
63
64
65
[39] Carr TP, Gallaher DD, Yang C-H, Hassel CA. Increased intestinal
contents viscosity reduces cholesterol absorption efficiency in
hamsters fed hydroxypropylmethylcellulose. J Nutr 1996;126:1463 - 9.
[40] Gallaher DD, Hassel CG, Lee K-J, Gallaher CM. Viscosity and
fermentability as attributes of dietary fiber responsible for the
hypocholesterolemic effect in hamsters. J Nutr 1993;123:244 - 52.
[41] Gallaher DD, Hassel CA, Lee K-J. Relationships between viscosity
of hydroxypropylmethylcellulose and plasma cholesterol in hamsters.
J Nutr 1993;123:1732 - 8.
[42] Wang L, Newman RK, Newman CW, Hofer PJ. Barley b-glucans alter
intestinal viscosity and reduce plasma cholesterol concentrations in
chicks. J Nutr 1992;122:2292 - 7.
[43] Ikegami S, Tsuchihashi F, Harada H, Tsuchihashi N, Nishide E,
Innami S. Effect of viscous indigestible polysaccharides on pancreatic-biliary secretion and digestive organs in rats. J Nutr 1990;120:
353 - 60.
[44] Piel C, Montagne L, Se`ve B, Lalle`s J-P. Increasing digesta viscosity
using carboxymethylcellulose in weaned piglets stimulates ileal goblet
cell numbers and maturation. J Nutr 2005;135:86 - 91.