Livestock Production Science 63 (2000) 153–157

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Effect of two commercial yeast cultures with Saccharomyces

cerevisiae on ruminal fermentation and digestion in sheep fed
sugar cane tops
´ b , F.A. Castrejon
J.L. Arcos-Garcıa ´ a , *, G.D. Mendoza b , E.P. Perez-Gavilan
´ ´ a
a
´
Universidad Nacional Autonoma ´
de Mexico ´ Animal,
, Facultad de Medicina Veterinaria y Zootecnia, Departamento de Nutricion
´
Cd. Universitaria, Mexico , D.F. 04510, Mexico
b
Colegio de Postgraduados, Programa de Ganaderıa´ , Montecillo, km 35.5 Carr. Mexico-Texcoco
´ ´
Estado de Mexico ´
, 56230 Mexico,
Mexico

Received 14 September 1998; received in revised form 19 April 1999; accepted 6 May 1999

Abstract

A feeding trial was conducted to evaluate the effect of two direct-fed microbial cultures containing Saccharomyces
cerevisiae, on ruminal fermentation and digestibility of diets based on sugar cane tops. Three Suffolk ewes (30 kg BW) with
ruminal cannula were used in a latin square design, where treatments were control group (CG); 3 g / day of Yea-Sacc 1026
(YS, 1 3 10 8 CFU / g) and 1 g / day of Levucell (LC, 20 3 10 9 CFU / g). The cultures were added to the rumen. Diet was
based on sugar cane tops (50%), sorghum grain (21%), wheat bran (15%), molasses (12%) and urea (2%). Ruminal pH was
highest (P , 0.05) in CG (6.05), and lower (P , 0.05) with YS than with LC (5.85 vs. 5.96). Total VFA concentration was
greater (P , 0.05) with yeast cultures (LC, 107.6 mM; YS 105.5 mM) than in CG (97.3 mM). However, no effects were
detected in VFA molar proportion, protozoa population, or total tract digestibility. In situ DM and NDF degradation was not
affected by treatments. Neither direct-fed microbial culture with Saccharomyces cerevisiae improved either fermentation or
digestion in sheep fed sugar cane tops.  2000 Elsevier Science B.V. All rights reserved.

Keywords: Yeast culture; Saccharomyces cerevisiae; Probiotics; Rumen fermentation; Digestion

1. Introduction inconsistent (Chiquette, 1995). Some of the possible

Direct-fed microbial products containing Sac-
charomyces cerevisae have been used to improve
daily gain and milk production in ruminants (Wal-
lace, 1994); however, results have been variable and
*Corresponding author. Fax: 1 52-5-500-057.
´
E-mail address: fcp@servidor.unam.mx (F.A. Castrejon)
causes for the inconsistency could be associated with
characteristics of the strain (Newbold et al., 1995),
differences between commercial additives (Mendoza
et al., 1995), and diet composition (Wallace, 1994).
Improvement of ruminal NDF digestion using
Saccharomyces cerevisae with low quality forages
has been reported previously (Plata et al., 1994;
Sommart et al., 1993) and benefits are explained by a

0301-6226 / 00 / $ – see front matter  2000 Elsevier Science B.V. All rights reserved.
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154 ´ et al. / Livestock Production Science 63 (2000) 153 – 157
J.L. Arcos-Garcıa
more active microbial population associated with the
ability of the yeast to remove O 2 in ruminal fluid
(Newbold et al., 1993), improving anaerobiosis in
the rumen (Wallace, 1994).
This trial was designed to compare the effect of
two commercial yeast cultures with Saccharomyces
cerevisae on ruminal fermentation and digestibility,
in sheep fed a diet based on sugar cane tops, which
is an important lignocellulosic by-product in de-
veloping countries.
2. Material and methods
Three ruminally fistulated Suffolk ewes (30 kg
BW) were assigned randomly to the following
treatments: control group, 3 g / day of Yea-Sacc 1026
(Alltech, Nicholasville; YS, 1 3 10 8 CFU / g) and 1
g / day of Levucell [Agrimerica (Northbrook, IL,
USA) LC, 20 3 10 9 CFU / g). Yeast cultures were
dosed intraruminally once daily at 8:00 h, immedi-
ately after feed was offered. Viability of Yea-Sacc 1026
and Levucell was measured by counts of CFU during
incubation of yeasts in dextrose agar culture with a
penicillin–estreptomicin (200 ppm) at 308C for 48 h
(Cruisckshank et al., 1975); CFU values were
3.4260.58 3 10 8 for Yea-Sacc and 20.563.75 3 10 9
Levucell.
The diet (dry basis) consisted of sugar cane tops
(50%), sorghum grain (21%), wheat bran (15%),
molasses (12%) and urea (2%) with the following
composition: DM, 90.23%; OM, 92.37%; CP,
11.47%; NDF, 62.57% and ADF, 28.45%. A mineral
premix was offered ad libitum (Ca, 10%; P, 12%; S,
1.5%; Mg, 2%; K, 2%; Co, 0.0015%; Cu, 0.07%; Fe,
0.15%; I, 0.005%; Mn, 0.25%; Se, 0.0008%; Zn,
0.25%).
Sheep were fed ad libitum during the adaptation
period (10 days) and then restricted at 90% of intake
during the collection period (5 days). Feed was
offered in two meals (8:00 and 16:00).
Ruminal fluid samples were collected at 0, 4, 8
and 12 h on days 1 and 2. Ruminal fluid pH was
measured immediately after sampling and then 50 ml
of ruminal fluid were acidified with 1 ml 6 M HCl,
and stored in a freezer (2208C) for further analysis.
Volatile fatty acids (VFA) were determined by gas
chromatography in samples prepared with metaphos-
phoric acid (Erwin et al., 1961). Ammonia-N was
measured by the indophenol method (McCullough,
1967). A ruminal fluid sample was also used to count
protozoa, mixing 5 ml of ruminal fluid with a 5 ml
iodine solution (Coleman, 1978) stored at 108C and
counted per milliliter of ruminal fluid with a
hemocytometer.
In situ disappearance of neutral detergent fiber
(NDF) was measured incubating 3 g of sugar cane
tops ground through a 1-mm screen, in polyester
bags (7 3 15 cm; 40-mm pore size). Duplicate bags
were incubated at 12, 24, 48, 72 and 96 h starting on
day 2. NDF was determined by procedures outlined
by Goering and Van Soest (1970).
Chromic oxide was dosed intraruminally (1 g /
day). Fecal grab samples were collected for 4 days,
as recommended by Stock et al. (1987). Feed and
fecal samples were oven-dried (558C, 24 h) and
ground to pass a 1-mm screen and composited by
site and animal. Dry matter, organic matter and
nitrogen were analyzed by standard methods
(AOAC, 1980) and chromium was measured by
atomic absorption spectrophotometry (Williams et
al., 1962).
Data were analyzed as a 3 3 3 latin square design
(Steel and Torrie, 1980). Orthogonal contrasts were
used to compare control group vs. microbial cultures,
and Yea-Sacc 1026 vs. Levucell, using the GLM
procedure of SAS (1985).
3. Results and discussion
Addition of Saccharomyces cerevisae reduced
(P , 0.01) ruminal pH compared to the control group
(Table 1), as reported by other authors using low
quality forages (Angeles et al., 1995). In some
experiments addition of yeast culture did not affect
ruminal pH (Newbold et al., 1995; Plata et al.,
1994). In this experiment, ruminal pH was higher
(P , 0.01) with Levucell than with Yea-Sacc, proba-
bly due to an effect of yeast strain, as has been also
noted by Newbold et al. (1995). Angeles et al.
(1995) did not report differences comparing the
same direct-fed microbial cultures.
Ruminal ammonia concentration was higher (P ,
0.02) with Yea-Sacc 1026 than Levucell (Table 1) as
observed by Angeles et al. (1995). In most of the
 ´ et al. / Livestock Production Science 63 (2000) 153 – 157
J.L. Arcos-Garcıa 155

Table 1
Effect of two yeast cultures (Saccharomyces cerevisae) on variables related to rumen fermentation in sheep fed sugar cane tops
Item Treatments S.E.M.b Contrast a
CG YS LC I II
Ruminal pH 6.05 5.85 5.96 0.05 0.003 0.01
NH 3 -N (mg / dl) 9.24 10.5 9.38 0.65 0.10 0.02
Total VFA (mM) 97.3 105.6 107.6 3.11 0.02 0.05
Molar proportion (%)
Acetate 66.6 66.6 67.3 0.72 0.81 0.99
Propionate 22.2 22.2 20.4 0.72 0.53 0.96
Butyrate 11.3 11.2 12.3 0.33 0.27 0.92
Acetate:propionate 3.07 3.20 3.41 0.11 0.33 0.64
Protozoa organisms ( 3 10 4 / ml)
Entodinidae 53 57 77 5.86 0.08 0.65
Holotrichidae 2 2 2 0.33 0.19 0.17
Total 55 59 79 5.98 0.08 0.62
a
Probability of Type I error. Mean comparisons: I control group (CG) vs. yeast cultures mean [(YS 1 LC) / 2]; II Yea-Sacc 1026 vs.
Levucell (YS vs. LC).
b
Standard error of mean.

reviewed studies, microbial cultures based on Sac-
charomyces cerevisae had no effect on ruminal
ammonia nitrogen (Plata et al., 1994; Newbold et al.,
1995), although in some others increases were
reported (Martin and Nisbet, 1992) and in others
reductions were found (Newbold et al., 1995). Even
when Yea-Sacc 1026 presented a greater ammonia-N,
which should increase concentration gradient and
more absorption, the lower pH in ruminal fluid, and
the pK may result in similar absorption through the
ruminal wall. Differences observed in ammonia
nitrogen could be associated with a stimulation of
proteolytic bacteria. More information is needed on
this subject because most of the studies involve
changes on cellulolytic bacteria population (Newbold
et al., 1995) and protozoa (Plata et al., 1994;
Miranda et al., 1996).
Total VFA concentration was increased (P , 0.02)
with yeast cultures, and was more elevated (P ,
0.05) with Levucell than with Yea-Sacc (Table 1).
The molar proportion of acetate, propionate and
butyrate was similar (P . 0.05). Differences between
commercial yeasts have not been detected with low
quality forages (Angeles et al., 1995; Plata et al.,
1994). As observed by Angeles et al. (1995), rumi-
nal protozoa population tended (P 5 0.08) to be
reduced with yeast cultures, particularly the en-
todiniomorphids (Table 1). In some studies with low
quality diets, protozoa counts were elevated with
Saccharomyces cerevisae (Plata et al., 1994), but not
in others (Miranda et al., 1996).
Although intake was restricted, it tended (P ,
0.10) to be higher in sheep fed yeast cultures (Table
2). Total tract digestibility of DM, OM, NDF and
ADF were not affected by Saccharomyces cerevisae
(Table 2). Improvement in NDF digestibility using
yeast cultures in low quality forages has been
reported previously (Plata et al., 1994). In contrast,
other studies show no effect (Sommart et al., 1993).
In situ disappearance of DM and NDF at 12 and
24 h were not changed by addition of yeast culture or
by type of culture (Table 2). In situ DM disappear-
ance at 48 h was higher (P , 0.01) in yeast cultures
than in CG and was higher for LC than for YS. In
situ NDF disappearance was higher (P , 0.002) for
YS than LC at 48 h. This effects disappearance at 72
h. Similar results were observed by others authors
comparing the same commercial yeast cultures
(Mendoza et al., 1995; Angeles et al., 1995).
Inconsistency in results with yeast cultures could
be associated to forage differences. Results from Roa
et al. (1997) showed that forage quality may affect
the response to yeast culture in NDF digestion, and
that more benefits can be obtained with good quality
roughages.
In conclusion, yeast cultures with Saccharomyces
156 ´ et al. / Livestock Production Science 63 (2000) 153 – 157
J.L. Arcos-Garcıa

Table 2
Effect of two yeast cultures (Saccharomyces cerevisae) on intake and total tract digestibility of nutrients in sheep fed sugar cane tops
Item Treatments S.E.M.b Contrast a
CG YS LC I II
DMI (g / day) 1183 1379 1401 34.4 0.10 0.15
Digestibility (%)
DM 77.3 77.7 79.3 0.79 0.54 0.86
OM 87.1 87.2 89.1 0.99 0.66 0.97
NDF 39.0 36.0 32.1 2.22 0.40 0.62
ADF 16.7 16.3 18.6 2.32 0.90 0.95
In situ DM disappearance (%)
12 c 27.8 27.3 27.4 0.22 0.37 0.37
24 35.6 35.3 35.1 0.39 0.63 0.63
48 39.6 42.7 46.4 0.28 0.01 0.0009
72 46.3 46.2 49.6 0.47 0.98 0.89
96 48.4 49.1 49.6 0.46 0.37 0.55
In situ NDF disappearance (%)
12 c 10.0 8.0 9.7 0.64 0.41 0.23
24 20.1 17.0 17.6 0.84 0.15 0.16
48 24.6 30.0 23.7 0.54 0.08 0.002
72 28.8 31.5 32.8 0.86 0.10 0.24
96 37.2 35.1 38.3 0.75 0.74 0.27
a
Probability of Type I error. Mean comparisons: I control group (CG) vs. yeast cultures mean [(YS 1 LC) / 2]; II Yea-Sacc 1026 vs.
Levucell (YS vs. LC).
b
Standard error of mean.
c
Hours of incubation.
DMI 5 Dry matter intake.
OM 5 Organic matter.
NDF 5 Neutral detergent fiber.
ADF 5 Acid detergent fiber.

cerevisae, Yea-Sacc 1026 and Levucell, did not im- Anglo-Corp, Mexico, who provided one yeast culture
prove digestibility or fermentation in diets based on for this study.
sugar cane tops.

Acknowledgements
This study was financed by PAPIIT (Programa de
Apoyo a Proyectos de Investigacion ´ e Innovacion´
´
Tecnologica) project IN505994 from DGAPA-
UNAM, with technical support from the Animal
Nutrition Department of the Veterinary Faculty and
Biotechnology Department of the Biomedical Re-
search Institute of the National University of Mex-
ico. The help of Jose´ Luis Cordero with surgeries is
recognized. The technical assistance of Ma. An-
tonieta Aguirre, Magda Crosby, Antonio Yanez´ ˜ and
Andres Toledo is greatly appreciated. Thanks are
also extended to Edsel Bixtler, General Manager of
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