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86 (2000) 205219
Abstract
Seven adult mixed breed female dogs (17:4 2:9 kg) surgically tted with ileal T-cannulas were
used in a 4 7 incomplete Latin square design experiment to evaluate oligosaccharide
supplementation on dry matter (DM), nitrogen (N), ammonia, volatile fatty acid (VFA), bacteria,
blood glucose concentrations, ileal pH, and fecal consistency. Fructooligosaccharide (FOS),
mannanoligosaccharide (MOS), and xylooligosaccharide (XOS) were added at 5 g/kg of diet DM.
There were no differences in DM digestibility, diet or fecal N, N digestibility, ileal or fecal
ammonia, fecal consistency, ileal bacteria colony forming units, blood glucose, or ileal pH. Ileal
butyrate proportion tended to be greater (P 0:07) in the control diet (0.076 of total VFA)
compared with the oligosaccharide supplemented diets and lower (P 0:07) for the MOS diet
compared with the FOS and XOS diets. Ileal propionate tended to be higher (P 0:09) in MOS
(0.198 of total VFA) than FOS and XOS. Fecal bidobacteria numbers were unaffected by dietary
treatment. Fecal Clostridium perfringens tended to be lower (P 0:09) in MOS when compared to
FOS and XOS. Oligosaccharides had relatively minor effects on bacterial growth in the large
intestine and VFA proportions in the small intestine of the canine. For oligosaccharide feeding to
cause microbial changes in the canine greater amounts of oligosaccharide may be required, or it
may require application in select dietary situations. # 2000 Elsevier Science B.V. All rights
reserved.
$
Approved by the director of the Kentucky Agricultural Experimental Station as publication 99-07-75.
*
Corresponding author. Tel.: 1-859-257-7516; fax: 1-859-257-3412.
E-mail address: dharmon@ca.uky.edu (D.L. Harmon).
0377-8401/00/$ see front matter # 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 3 7 7 - 8 4 0 1 ( 0 0 ) 0 0 1 7 5 - 9
206 J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219
1. Introduction
2.1. Dogs
Seven adult, mixed breed female dogs with body weights of 17:4 2:9 kg were used in
this experiment to evaluate the effect of oligosaccharide addition to diets on nutrient
digestion and microbial populations. All seven dogs had been surgically tted with a
polyvinyl chloride T-cannula 610 cm from the ilealcecal junction (Walker et al., 1994).
J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219 207
All procedures described herein were approved by the Institutional Animal Care and Use
Committee.
The animals were located in the Division of Laboratory Animal Resources at the
University of Kentucky, Lexington, in an environmentally controlled room at 228C with a
14 h:10 h light:dark schedule. The dogs were exercised daily for a minimum of 25 min
except on blood sampling and ileal collection days. Each dog was housed in an individual
cage. Five dogs were in stainless steel cages (1:2 m 1:8 m) with a raised step
(1:01 m 0:46 m). The elevated oor and the step were diagonally grated, coated, and
removable. The sixth dog was housed in a cage (0:78 m 1:8 m) with chain link sides
and ceiling. The berglass oor and raised step (0:78 m 0:51 m) were solid and not
grated with a small circular drain opening in the center. The seventh dog was in a
(1:3 m 0:90 m) crate without a step. The oor was plastic coated and diagonally grated.
The sidewalls were made of stainless steel.
All diets (Table 1) were prepared by the Hil's Science and Technology Center (Topeka,
KS). Diets were formulated using current guidelines for dogs (American Association of
Table 1
Composition of the experimental diets
Maize 400
Poultry meal 200
Soybean meal 150
Maize starch 91.05
Choice white grease 73
Corn gluten 50
Soy oil 10
Flavora 10
Salt 6
Chromic oxide 2
Vitamins, mineralsa 2.75
Ethoxyquin 0.2
Cornstarch or oligosaccharideb 5.0
Nutrient (dry matter basis)
Protein 300
Fat 150
Nitrogen-free extract 480
Crude ber 15
Calcium 8
Phosphorus 7
Magnesium 1
Sodium 4
Potassium 7
a
Composition of avor and vitamin-mineral premixes are condential information.
b
Amounts added prior to extrusion. No direct measures of oligosaccharide concentrations are available.
208 J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219
Feed Control Ofcials, 1994). Chromic oxide was added directly to the diet at 2 g/kg (dry
basis) to serve as an indigestible marker of digesta ow. The daily maintenance ration
was based on weight at the beginning of each period as follows:
The rations were calculated based on 16.7 MJ/g feed, with one exception, and that diet
was at 18.8 MJ/g feed to accommodate the eating habit of that particular dog. Daily
rations were divided into two equal meals and fed at 07.00 and 17.00 h. Any food not
consumed within 45 min was removed from the cage and recorded as orts. Water was
provided on a free choice basis. Diets were weighed into stainless steel bowls for each
feeding. From day 2 through day 21, grab samples of each diet were collected and pooled
for diet analysis.
The dogs were allotted randomly to treatments using a 4 7 incomplete Latin square
design structure. The four treatments were based on 5 g/kg of each oligosaccharide
product at the expense of cornstarch as follows: control, fructooligosaccharide (FOS),
mannanoligosaccharide (MOS), and xylooligosaccharide (XOS).
The fructooligosaccharide product used was Raftifeed1 P75 (Encore Technologies,
Eden Prairie, MN), a powder prepared from the chicory root that contains 0.75
oligofructose. In addition, this product also contains 0.15 fructose, glucose, and
sucrose.
The mannanoligosaccharide product was DP607 (Alltech, Inc., Nicholasville, KY).
These spray-dried mannanoligosaccharides are derived from the yeast cell wall of
Saccharomyces cerevisiae. They are harvested by centrifugation from a lysed yeast
culture.
The xylooligosaccharide product Xylo-oligo 95P (Suntory Limited, Consumer Health
Products Department, Pharmaceutical Division, Tokyo, Japan), used in this experiment
was composed of at least 0.95 xylooligosaccharide in the solid form. The main
components of this xylooligosaccharide product are xylobiose and xylotriose, which are
dimers and trimers of xylose, respectively.
2.3. Sampling
Each experimental period was 21 days. Adaptation diets, half of the diet to be fed for
the new period plus half of the diet fed the previous period, were fed the rst day of each
period. The remaining 20 days were at 100% of the experimental diet.
Jugular blood samples were collected on day 7 to test the glucose tolerance of the dogs
to their respective diets. The neck area was shaved the evening prior to the sampling day.
Beginning at 06.00 h catheters (18.5 gauge 5 cm, needle 17 gauge 5 cm; Char-
terMed, Inc., Lakewood, NJ), a 50 cm extension (Baxter Healthcare Corp., Deereld, IL),
and a three way stopcock were inserted into the jugular vein of each dog. After all
catheters were inserted, a blood sample was taken (10 min) to establish the amount of
glucose in the bloodstream prior to diet consumption. In order to ensure that the entire
meal would be consumed within 5 min, half of the normal morning ration was fed. As
J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219 209
soon as the meal was complete, another blood sample was taken (0 min) and the time
recorded. Additional samples were taken at 30, 45, 60, 90, 120, 180, 240, 300, and
360 min post 0 min. The rst 3 ml of heparinized saline and blood removed was
discarded as waste, after which 5 ml blood samples were taken. Catheters were then
ushed with sterile heparinized saline (0.9% NaCl; 20 U/ml heparin, 0.1% benzyl
alcohol). All syringes were heparinized by rinsing with heparin solution (1000 U/ml,
Elkins-Sinn, Inc., Cherry Hill, NJ) prior to sample collection. All blood samples were
immediately stored on ice (maximum 2 h) until they could be brought to the lab,
centrifuged (15 min at 6913 g), plasma removed, and frozen (208C) for later analysis.
For any dog that removed its catheter during the sampling period, blood was collected
with an 18 gauge needle and syringe for the rest of the day. At the end of the sampling
day catheters were removed.
Fecal collections began at 07.00 h on day 9, ending at 07.00 h on day 13 (5 days). All
feces were removed from the cages and placed in plastic bags, keeping each dog separate.
In addition, dogs were watched carefully during exercise to ensure that feces were
correctly allocated to the proper animal. All feces collected were individually weighed
for each dog and immediately frozen.
Ileal collections began on day 15 and ended on day 17. Dogs were fed at 07.00 h,
cages were cleaned, but the dogs were not exercised. Prior to collection each dog was
tted with an Elizabethan collar to prevent them from removing the collection bags.
Sterile, 1 oz Whirl-pak1 bags (Nasco, Fort Atkinson, WI) were placed on each dog
at 08.00, 10.00, 12.00, 14.00, 16.00 h, on day 15 and 16, and at 09.00, 11.00, 13.00,
and 15.00 h on day 17. The bags were removed 1 h later. The dogs were watched
while wearing the collection bags. When a bag became full or started to leak it was
removed and another bag attached. The pH of each collection bag was taken using
an Accumet Basic pH meter (Fisher Scientic, Pittsburgh, PA) and recorded for each
dog. Individual ileal samples were weighed, then pooled by dog, and immediately
frozen.
The last 4 days of each period were used to analyze bacteria found in the ileal digesta
and feces. Beginning at 06.00 h on collection days, polyethylene 100 15 mm BioPro1
petri dishes (International Bioproducts, Inc., Redmond, WA) were labeled in triplicate.
Previously prepared media was melted in an autoclave and placed in a waterbath to
maintain the temperature at 478F. When lab preparation was complete, ileal and fecal
samples were collected from the dogs. The dog's chosen to sample on a given day were
those that voided fresh feces following the morning feeding. Feces in cages upon arrival
each morning were not used. Samples were collected into either ziplock bags (fecal) or
sterile 1 oz Whirl-pak1 bags (ileal) and immediately brought back to the lab and diluted.
Samples were prepared within 2 h of collection. One dog was not sampled because she
had been on antibiotics at the onset of this experiment for treatment of an infection.
Antibiotics were discontinued prior to the start of any sample collection; however, to
avoid any lingering effects on microorganisms these measurements were not made. It was
felt that the previous antibiotic treatment would have no inuence on the digestibility
measures and these were included.
Fecal consistency scores were recorded every day throughout the experiment as
follows.
210 J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219
Grade 1 more than two-thirds of the feces in a defecation is liquid. The feces have
lost all form, appearing as a puddle or squirt.
Grade 2 soft-liquid feces; an intermediate between soft and liquid feces.
Approximately equal amounts of feces in a defecation are soft and liquid.
Grade 3 more than two-thirds of the feces in a defecation is soft. The feces retain
enough form to pile but have lost their cylindrical appearance.
Grade 4 firm-soft feces; an intermediate between the grades of firm and soft.
Approximately equal amounts of feces in a defecation are firm and soft.
Grade 5 more than two-thirds of the feces in a defecation are firm. They have a
cylindrical shape with little flattening.
The rating was subjective and performed by the three different people who fed the dogs
during the experiment.
Diet palatability was rated and recorded on day 2 through day 21 of each period.
Grading was divided into four categories as follows.
1. Ate entire meal without hesitation.
2. Ate a portion of the meal but was more concerned with the activity outside of their
cage than the meal itself, but, total meal completed within 1 h.
3. Totally unconcerned about the meal, ate a portion, but, did not consume the entire
ration.
4. Would not or refused to eat.
The rating was subjective and performed by the three different people who fed the dogs
during the experiment.
2.4. Analyses
Both ileal and fecal samples were later thawed and thoroughly mixed so that each
sample was representative of the period. Fresh portions of ileal digesta which were
required for VFA (1 g) and ammonia (1 g) analysis, and fresh portions of fecal samples
(1 g) used for ammonia analysis, were removed. The remainder of the samples were then
weighed into a pan and lyophilized. Weights post-lyophilization were recorded and the
DM coefcient was calculated. Lyophilized ileal and fecal samples were then ground
with a mortar and pestle until a ne, uniform consistency was obtained. Samples were
then dried, in duplicate, in a 708C vacuum oven overnight to a constant weight for DM
determination. Samples were then stored in sealed plastic bags at room temperature.
The diet samples were rst thoroughly mixed in a large bowl. Representative samples
of each diet for the four periods were dried in a 558C oven for 46 h for DM determination.
The diet samples were then ground through a 1 mm screen in a Wiley Mill. Dry matters
of ground samples were obtained at 708C as described above, using a 1 g sample of each
diet.
Samples were ashed at 5008C for 16 h and then prepared for Cr analysis as described
previously (Williams et al., 1962) and stored in amber bottles. Chromium analysis was
performed by atomic absorption spectroscopy (Unicam 929 Spectrometer, Thermo Jarell
Ash, Franklin, MA).
J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219 211
Previously pooled, fresh, ileal samples were prepared for VFA analysis by mixing 1 g of
each sample with 1 ml of 25% (w/v) metaphosphoric acid and centrifuging (16; 000 g) for
10 min. The supernatant of each sample was removed and analyzed on a Hewlett Packard
5890 Gas Chromatograph (Avondale, PA) with a 1.8 m 4 mm glass column packed
with 10% sp-1000/1% H3PO4 on 100/120 Chromsorb W AW (Supelco, Bellefonte, PA).
For ammonia analysis, the ileal samples previously mixed with metaphosphoric acid for
the VFA analysis, were further diluted with nanopure water to attain a 1:75 dilution. Fresh
pooled fecal samples (0.0125 g) were extracted with 1 ml 0.12N HCl, centrifuged
(13; 000 g) for 4 min, and the supernatant removed. Ammonia was analyzed using
glutamate dehydrogenase (Ammonia Analysis Enzymatic Kit (#171-B), Sigma, St. Louis,
MO) on a Cobas Fara II (Roche Diagnostic Systems, Branchburg, NJ).
Fecal and ileal samples (11 g) were added to 99 ml 0.1% peptone (Difco Labs, Detroit,
MI) diluent (1 g Bacto1 peptone/l distilled water) and mixed in a blender (101).
Dilution sequences representing from 102 through 107 g/ml were made using 1 ml of
the previous dilution mixed with 9 ml of peptone diluent. Samples (0.1 ml) were
measured into plates, media was added, plates were stirred for several minutes to assure
that the sample was blended thoroughly into the media, then set aside to gel. The plates
were placed in either the aerobic incubator or the anaerobic chamber, depending on which
bacteria were to be grown. The specic bacteria cultured were; C. perfringens (Oxoid
Agar, including supplement A, SR76 and supplement B, SR77, Unipath Ltd.,
Basingstoke, Hampshire, England), Bidobacteria (Bacto1 Liver Veal Agar, Difco Labs,
Detroit, MI) as described previously (McCann et al., 1996), E. coli and total coliforms
(Bacto1 Violet Red Bile Agar with Mug, Difco Labs, Detroit, MI), Lactobacilli (Bacto1
Rogosa SL Agar, Difco Labs, Detroit, MI), and anaerobes (Bacto1 Reinforced Clostridial
Agar, Difco Labs, Detroit, MI). Plates for E. coli and total coliforms were read after a
24 h incubation period using an ultraviolet transilluminator (Fotodyne, Inc., New Berlin,
WI), while the remaining plates were read after a 48 h incubation period. All plates were
digitally counted by the same person to avoid variation in the counting procedure. Dry
matter analysis was performed on both the ileal and fecal samples from the rst dilution
(101).
Jugular plasma samples were thawed, transferred to sample cups and analyzed for
glucose using the Cobas Fara II (Roche Diagnostic Systems, Branchburg, NJ). Plasma
glucose concentrations were determined by enzymatic kit (Glucose (HK) 20, Sigma, St.
Louis, MO).
plasma collected 11 times post-feeding was also analyzed using the Mixed procedure
with a heterogeneous Compound Symmetry covariance structure for the repeated variable
time, and Satterthwaite's approximation used for the degrees of freedom. Differences
were considered signicant with P < 0:05.
Table 2
Evaluation of fructooligosaccharide (FOS), xylooligosaccharide (XOS) and mannanoligosaccharide (MOS)
feeding on dry matter (DM) digestibilities in dogs
Control FOS MOS XOS S.E.M.b Control vs. MOS vs. FOS vs.
others FOS, XOS XOS
Body weight (kg) 17.3 17.3 17.6 17.7 0.2 0.30 0.70 022
Dry matter intake (g/d) 222 224 222 223 3.0 0.93 0.70 0.88
Fecal moisture (g/d) 350 340 330 350 6 0.09 0.13 0.25
Feces (g DM/d) 33.4 34.9 35.5 36.7 1.3 0.14 0.86 0.33
Ileal ow (g DM/d) 54.2 52.7 52.2 53.2 2.1 0.54 0.78 0.88
DM digestibility coefcients
Ileal 0.759 0.767 0.772 0.769 0.008 0.29 0.73 0.92
Large intestinec 0.377 0.330 0.310 0.302 0.034 0.13 0.89 0.57
Total tract 0.852 0.847 0.843 0.839 0.005 0.17 0.95 0.33
a
Probability of greater F-value.
b
Standard error of the mean, n 7.
c
Coefcients calculated as fraction of ileal ow.
J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219 213
considered to be nondigestible in the small intestine, therefore, it was expected that the
ileal digestibilities in the supplemented diets would be similar to each other. The control
was similar since oligosaccharides were included at only 5 g/kg of the diet.
There were no differences in large intestinal and total tract DM digestibility. Zuo et al.
(1996) fed a low oligosaccharide soybean meal diet to dogs and found it to be more
digestible than a conventional (12 g/kg oligosaccharides; 11 g/kg stachyose and 1 g/kg
rafnose) soybean meal diet. Although the Zuo et al. (1996) oligosaccharide sources are
different from those used in this experiment, their results indicate that oligosaccharides
can affect digestibility. The 5 g/kg addition of oligosaccharides may not have been
sufcient to measure effects on DM digestion.
3.2. Nitrogen
There were no differences in N intake or excretion (Table 3). It was expected that
oligosaccharide fermentation in the large intestine would increase fecal N excretion.
Levrat et al. (1993) found that oligosaccharides with relatively high degrees of
polymerization, promoted fecal N excretion in the rat when the level of protein is
moderate. The diet in this experiment was high in protein for the canine and the level of
inclusion was fairly low.
Ileal, large intestinal and total tract N digestibility were unaffected by treatment.
Fructooligosaccharides and XOS supplemented to rats decreased blood urea and renal N
excretion, while there was a corresponding increase in fecal N excretion (Younes et al.,
1995). These effects are dependent on adequate fermentable substrate being presented to
the large intestine for fermentation. The lack of a signicant increase in fecal N excretion
in the present study suggests that greater dietary concentrations of oligosaccharides may
be necessary to see such effects in the dog.
Table 3
Evaluation of fructooligosaccharide (FOS), xylooligosaccharide (XOS) and mannanoligosaccharide (MOS)
feeding on nitrogen (N) concentrations in dogs
Treatments Contrastsa
Control FOS MOS XOS S.E.M.b Control vs. MOS vs. FOS vs.
others FOS, XOS XOS
N intake (g/d) 10.8 10.8 10.9 11.0 0.1 0.35 0.79 0.30
Fecal N (g/kg) 52 53 52 52 1 0.49 0.50 0.17
Fecal N excreted (g/d) 1.8 1.9 1.9 1.9 0.1 0.16 0.82 0.69
Ileal N (g/kg) 48 48 47 48 1 0.93 0.09 0.92
Ileal N ow (g/d) 2.6 2.6 2.5 2.6 0.1 0.66 0.55 0.91
N digestibility coefcients
Ileal 0.752 0.758 0.769 0.762 0.010 0.37 0.50 0.83
Large intestinec 0.319 0.259 0.243 0.254 0.041 0.18 0.79 0.94
Total tract 0.834 0.824 0.825 0.822 0.007 0.20 0.84 0.84
a
Probability of greater F-value.
b
Standard error of the mean, n 7.
c
Coefcients calculated as fraction of ileal ow.
214 J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219
Table 4
Evaluation of fructooligosaccharide (FOS), xylooligosaccharide (XOS) and mannanoligosaccharide (MOS)
feeding on ammonia and ileal volatile fatty acid (VFA) concentrations in dogs
Treatments Contrastsa
Control FOS MOS XOS S.E.M.b Control vs. MOS vs. FOS vs.
others FOS, XOS XOS
Ileal ammonia (mmol/gDM) 5.1 5.0 4.6 4.4 0.4 0.39 0.87 0.35
Fecal ammonia (mmol/gDM) 25.9 20.9 20.2 25.3 2.0 0.12 0.25 0.14
Ileal VFA (mol/100 mol)
Acetate 68.8 69.2 69.5 69.5 0.5 0.25 0.73 0.68
Propionate 19.1 18.9 19.8 19.1 0.3 0.70 0.09 0.61
Isobutyrate 2.3 2.4 2.1 2.2 0.1 0.77 0.47 0.39
Butyrate 7.6 7.3 6.5 7.0 0.3 0.07 0.07 0.50
Isovalerate 1.5 1.4 1.3 1.3 0.1 0.25 0.26 0.44
Valerate 0.8 0.8 0.8 0.8 0.0 0.67 0.44 0.54
Total ileal VFA (mmol/g DM) 597.6 600.8 554.7 556.3 22.3 0.31 0.40 0.18
Total ileal VFA (mmol/g wet) 79.2 79.5 71.8 73.5 3.1 0.24 0.24 0.19
a
Probability of greater F-value.
b
Standard error of the mean, n 7.
J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219 215
also found no difference in cecal VFA concentrations when neonatal pigs were fed 3 g/l
(liquid diet) fructooligosaccharide.
One might expect to see a higher acetate concentration (Mitsuoka and Kaneuchi, 1977;
Wang and Gibson, 1993), an end-product from the break down of oligosaccharides by
bidobacteria. However, the diets had no effect on acetate proportion. Mannanoligo-
saccharides also did not affect cecal concentrations of VFA in chicks from 3 to 10 days of
age (Spring et al., 2000).
There were no effects from dietary oligosaccharide addition on any of the bacterial
colony forming units (CFU) plated from the ileal digesta (Table 5). It was hypothesized
that differences in bacterial counts from oligosaccharide feeding found in the ileum
would be minimal since this is not the major site of fermentation. However, counts of
bacteria between the ileum and feces were similar.
In fecal samples (Table 5) the growth of C. perfringens tended to be lower (P 0:09)
for MOS versus FOS and XOS. Feeding of MOS has been shown to reduce the
colonization of Salmonella in chicks (Spring et al., 2000), but comparable reports for
Clostridial species are lacking.
Literature suggests that an increase in bidobacterial counts should be expected with
the addition of fructooligosaccharide or xylooligosaccharide (Wang and Gibson, 1993;
Okazaki et al., 1990). However, others (Mitsuoka and Kaneuchi, 1977) have isolated
Table 5
Evaluation of fructooligosaccharide (FOS), xylooligosaccharide (XOS) and mannanoligosaccharide (MOS)
feeding on concentration of specic bacteria (log CFU/g DM)a
Control FOS MOS XOS S.E.M.c Control vs. MOS vs. FOS vs.
others FOS, XOS XOS
Ileal
C. perfringens 4.80 5.21 5.28 5.25 0.4 0.36 0.91 0.94
Bidobacteria 10.44 10.60 10.56 10.73 0.1 0.15 0.42 0.44
Lactobacilli 9.37 9.46 10.11 9.60 0.4 0.50 0.31 0.84
Aerotolerant anaerobes 10.50 10.43 10.49 10.36 0.2 0.75 0.68 0.80
E. coli 6.27 6.22 6.42 6.19 0.5 1.00 0.71 0.96
Coliforms 6.70 6.61 7.15 6.86 0.5 0.80 0.55 0.76
Fecal
C. perfringens 4.73 4.74 4.48 5.16 0.2 0.80 0.07 0.16
Bidobacteria 10.87 11.05 10.79 10.87 0.1 0.76 0.19 0.23
Lactobacilli 9.32 9.80 10.34 10.15 0.5 0.17 0.54 0.62
Aerotolerant anaerobes 10.59 10.71 10.63 10.61 0.1 0.71 0.88 0.61
E. coli 5.96 5.94 5.80 5.97 0.3 0.83 0.64 0.93
Coliforms 6.28 6.46 6.49 6.61 0.4 0.60 0.92 0.79
a
CFU: colony forming unit.
b
Probability of greater F-value.
c
Standard error of the mean, n 6.
216 J.A. Strickling et al. / Animal Feed Science and Technology 86 (2000) 205219
various species of bidobacteria and found only three species in the canine as follows: (1)
Bidobacteria longum (4.9% of strains isolated), (2) B. adolescentis (70.7% of strains
isolated), and (3) B. pseudolongum (24.4% of strains isolated). When Wang and Gibson
(1993) compared bidobacteria growth using oligofructose as a substrate, they found that
all species grew well with the highest specic growth rates recorded with B.
pseudolongum, B. infantis, and B. catenulatum, and the lowest growth rate with B.
adolescentis. Two of the highest growth rate species are not found in the canine, and B.
pseudolongum strains, fast growers and high fermenters (Desjardins and Roy, 1990),
comprise only 24.4% of the total bidobacteria strains isolated in the canine. In addition,
the lowest growing species, B. adolescentis, comprises 70.7% of the total strains of
bidobacteria found in the dog. Therefore, one would expect an increase in the amount of
bidobacteria when oligofructose is added to the diet, but not as dramatic as that found in
other experimental animals. Also, Roberfroid and Delzenne (1998) summarized available
literature for humans and reported that although Bidobacteria increase their numbers in
response to oligosaccharide feeding, their numbers rarely exceed 109.5, values
comparable to ours when expressed on an `as is' basis (data not shown).
Diet composition is probably the single most important control factor for microbial
activity in the gastrointestinal tract of non-ruminant animals. Digesta reaching the large
intestine determines the fate of the microbial population via the amount that arrives and
the type of substrate that it provides. Diets fed in the present study consisted of 150 g
soyabean meal/kg. The soyabean itself contains a-galactooligosaccharides which pass
along to the large intestine in an intact form where they are fermented by bacteria. These
oligosaccharides may also be used as a selective substrate for the growth of bacteria in the
large intestine. It is possible that the supplemented oligosaccharides were, therefore,
under an additive or masking effect, and that true results of the specic oligosaccharides
alone can only be seen if soya products are not used.
Yazawa et al. (1978) reported that B. infantis readily utilized rafnose and stachyose as
a substrate. However, this bidobacteria species is not specically found in the canine and
information was not available on other species. If other bidobacteria acted in a similar
fashion to B. infantis, there was approximately 10 g/kg (conventional soyabean meal diet
used by Zuo et al. (1996) contained 185 g soyabean meal, 11 g stachyose, and 1 g
rafnose/kg) of naturally occurring oligosaccharides included in the base diet which
could have affected the control populations. The theory of a dilution, additive, or masking
effect is even more profound if the log10 CFU of bidobacteria and lactobacilli results
from this experiment are compared to the counts normally found in the canine. According
to Mitsuoka and Kaneuchi (1977) lactobacilli CFU (9:3 1:3) are greater than
bidobacteria CFU (6:6 2:7). In both the ileal and the fecal samples from this
experiment, the bidobacteria CFU were greater than the lactobacilli CFU.
3.6. Ileal pH
There was no treatment effect on ileal pH. The overall mean ileal pH was 7.1. Similar
results have also been found in the supplementation of oligosaccharides (2 g/kg), for 6
days to weanling pigs that did not alter ileal pH over a 3-day collection (Gabert et al.,
1995). Similarly, neonatal pigs supplemented with 3 g/l of FOS for 15 days resulted in no
change in cecal pH (Howard et al., 1993).
There was no effect of treatment on fecal consistency or palatability scores. The fecal
consistency scores ranged from 4.9 in the FOS and MOS diets to 5.0 in the CON and
XOS diets (data not shown). Oligosaccharides are readily fermentable in the colon.
Supplementation in large quantities could cause acidic fermentation leading to extensive
gas, cramping, and diarrhea. There were no side effects observed indicating that in the
canine, supplementation of oligosaccharides at 5 g/kg is not excessive.
4. Conclusion
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