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Comparison of two commercial yeast cultures (Saccharomyces cerevisiae) on ruminal fermentation and digestion in sheep fed on corn-stover diet
Small Ruminant Research 31 (1998) 45±50
Comparison of two commercial yeast cultures (Saccharomyces cerevisiae) on ruminal fermentation and digestion in sheep fed on corn-stover diet S.C. Angeles C.a,*, G.D. Mendoza M.b, M.A. Cobos P.b, M.M. Crosby G.b, F.A. CastrejoÂn P.a,1 a
Universidad Nacional AutoÂnoma de MeÂxico, Facultad de Medicina Veterinaria y Zootecnia, Departamento de NutricioÂn Animal, Cd. Universitaria, MeÂxico, D.F. 04510, Mexico b Colegio de Postgraduados, Programa de GanaderõÂa, Montecillo, Km 35.5 Carr, MeÂxico-Texcoco Estado de Mexico, 56230, Mexico Accepted 4 March 1998
Abstract A metabolism trial was conducted to evaluate the effect of two direct-fed microbial cultures containing Saccharomyces cerevisiae on ruminal fermentation, digestibility and duodenal amino acid ¯ow. Nine Suffolk ewes with ruminal and duodenal cannula (29 kg BW) were assigned to a completely randomized design, where treatments were: (1) control group (CG); (2) 3 g/day of Yea±Sacc1026 (YS, 1108 CFU/g); and (3) 1 g/day of Levucell (LC, 20109 CFU/g). The diet consisted of corn stover (66.5%), sorghum grain (23.5%), molasses (8.6%) and urea (1.4%). Ruminal pH was lower (p<.01) with yeast culture (CG 6.81a, YS 6.51b, LC 6.61b). No effects were detected in VFA concentration, molar proportion, ammonia-N, and protozoa population. Total tract digestibility of DM, OM, NDF, and ADF were not affected by treatments. Duodenal amino acid ¯ow was not affected by the inclusion of yeast cultures. None of the direct-fed microbial cultures with Saccharomyces cerevisiae improved either fermentation, digestion or amino acid ¯ow to the duodenum in sheep fed on corn-stover diet. # 1998 Elsevier Science B.V. All rights reserved. Keywords: Saccharomyces cerevisiae; Fermentation; Digestion; Corn-stover diet
1. Introduction Direct-fed microbial products with Saccharomyces cerevisiae have been used to manipulate ruminal fermentation and animal production (Mutsvangwa et al., 1992; Walli, 1994). Bene®cial effects of these microbial compounds are associated with an increase *Corresponding author. Responsible for project PAPIIT No IN505994.
1
in cellulolytic bacteria (Dawson and Newman, 1987), with a concomitant improvement in ®ber digestibility in low quality forages (Ayala et al., 1992; Gonzalez, 1993). Fiber source is another factor which affects the response to yeast culture additions (Roa et al., 1997). Some studies indicated that amino acid pro®le and duodenal ¯ow are improved by the inclusion of yeast culture (Erasmus et al., 1992). However, results in ruminant production with yeast cultures have been inconsistent (Miranda et al., 1994; Mir and Mir, 1994),
0921-4488/98/$ ± see front matter # 1998 Elsevier Science B.V. All rights reserved. PII: S0921-4488(98)00115-1
46
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
and factors associated with the differences in commercial yeast cultures have received little attention. Several commercial feed additives with yeast cultures have been developed; however, comparisons of different products are scarce (Mendoza et al., 1995). Since feed additives are increasing production costs, it is important to compare the response with different yeast cultures. Also, strains of Saccharomyces cerevisiae differ in their effects on ruminal population (Newbold et al., 1995). In a previous trial (Corona et al., 1995), two yeasts cultures with Saccharomyces cerevisiae, dosed at similar colony-forming units (CFU) showed no effect on either fermentation or digestion in sheep fed on corn-stover diet. Therefore, this trial was conducted to evaluate the effect of two products containing Saccharomyces cerevisiae administered at commercial dosage rates on ruminal fermentation and digestibility. 2. Material and methods Nine Suffolk ewes (29 kg BW) with rumen and duodenal cannulae were randomly assigned to the following treatments: (1) control group; (2) 3 g/day of Yea±Sacc1026 (Alltech, Nicholasville; 1108 CFU/ g), and (3) 0.015 g/day of Levucell (Agrimerica, Northbrook, IL; 20109 CFU/g). The dosages of treatments 2 and 3 were based on the commercial information for each product, aiming at obtaining 3108 CFU for both the Yea±Sacc treatment and the Levucell treatment. Yeast cultures were dosed intraruminally. The diet (dry basis) was formulated with: corn stover (66.5%), sorghum grain (23.5%), molasses (8.6%) and urea (1.4%). The diet had the following composition: DM (82.57%), OM (94.37%), CP (10.15%), NDF (70.97%), and ADF (26.13%). Mineral premix was offered ad libitum (Ca 10%, P 12%, S 1.5%, Mg 0.2%, 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 to 90% of the ad libitum intake during the collection period (7 days). The animals were fed twice daily (7 and 16 h). Ruminal ¯uid samples were collected on day 7 of the collection period at 2, 6, 10, 14, 18 and 22 h after feeding. Ruminal ¯uid pH was measured immediately
after sampling and then a 50 ml aliquot of ruminal ¯uid was acidi®ed with 1 ml of HCl 6N, and stored in a freezer (ÿ208C) for further analysis. VFA were determined by gas chromatography in 4 ml of the acidi®ed samples prepared with 1 ml metaphosphoric acid (25%) (Erwin et al., 1961). Ammonia-N was also measured in the acidi®ed samples using the indophenol method (McCullough, 1967). A 5 ml ruminal ¯uid sample mixed with 5 ml of an iodine solution was used to count protozoa (count/ml) in a hemocytometer (Benjamin, 1984). Chromic oxide, used to calculate rate of passage, was dosed intraruminally (1 g/day) during the last 10 days of the adaptation period and during the collection period; chromium was measured by atomic absorption spectrophotometry (Williams et al., 1962). During the ®rst 4 days of the collection period, duodenal and faecal grab samples were collected as recommended by Stock et al. (1987). Feed and faecal samples were oven-dried (558C, 24 h) and ground to pass a 1 mm screen and composited by site and animal. Dry matter and organic matter were analyzed by standard methods (AOAC, 1985). NDF and ADF were determined by procedures outlined by Goering and Van Soest (1970). Duodenal samples were freeze-dried and analyzed for amino acid content using a Beckman 7300 analyzer. The samples were prepared as described by LadroÂn de Guevara et al. (1995). Data were analyzed as a completely randomized design (Steel and Torrie, 1980). If the response variable was measured at different times, data were analyzed with repeated measurements (Wilcox et al., 1990). Treatment means were compared by orthogonal contrasts using the GLM procedure of SAS (1985). 3. Results and discussion Ruminal pH was lower (p<.01) with yeast cultures than in the control group (Table 1), as observed by Harrison et al. (1988). No differences were observed between cultures, as reported by Corona et al. (1995) who used the same commercial cultures but at different dosages. Although most studies have shown that ruminal pH is not affected by Saccharomyces cerevisiae (Chademana and Offer, 1990; Carro et al., 1992a), some researchers (Jones and Dawson, 1993; Newbold
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
47
Table 1 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on pH, ammonia-N and fermentation pattern and ruminal protozoa in Suffolk ewes fed on corn-stover diet Item
Treatments CG
Ruminal pH N-NH3, mg/dl Total VFA, mM Molar proportion, % Acetate Propionate Butyrate Protozoa, organisms104/ml Entodinidae Holotrichidae Total
SEM YS
LC
Contrasta I
II
6.81 11.01 82.8
6.51 10.16 87.9
6.61 10.71 94.5
0.04 1.72 4.79
0.002 0.59 0.10
0.22 0.65 0.38
66.6 21.3 11.8
67.4 21.4 11.2
67.5 21.1 11.4
0.92 0.98 0.54
0.76 0.70 0.22
0.82 0.59 0.18
1.48 0.79 1.53
0.55 0.23 0.34
0.30 0.90 0.93
843 9 852
1120 25 1145
838 23 859
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell.
a
et al., 1995) suggest that yeast cultures may stabilize ruminal pH. However, quantitative evidence of this has not yet been presented. There was no effect of microbial culture on ruminal ammonia concentration (Table 1). In most of the studies, microbial cultures based on Saccharomyces cerevisiae have no effect on ammonia-N (Carro et al., 1992a, b; Plata et al., 1994) and in some studies increments (Martin et al., 1989; Ayala et al., 1992) or reductions (Newbold et al., 1990; Moloney and Drennan, 1994) have been reported. Yeast cultures did not affect total VFA concentrations or fermentation patterns. Changes in molar proportions of VFA by additions of Saccharomyces cerevisiae were inconsistent (Offer, 1990; Chademana and Offer, 1990; Carro et al., 1992a, b; Mutsvangwa et al., 1992). Ruminal protozoa population was not affected by Saccharomyces cerevisiae (Table 1) in contrast to other studies with low quality forages (Ayala et al., 1992; Plata et al., 1994). According to Crosby (1995), there is a quadratic (p<0.001) effect of yeast culture dosage on protozoa population showing a greater population using 3 g/day of yeast culture in sheep fed on corn-stover diet. Dry matter intake was restricted in this study, and there were no differences among treatments on ruminal ¯uid volume or fractional rate of passage (Table 2). Chademana and Offer (1990) did not detect
changes in ¯uid kinetics, and ruminal volume was not affected by yeast culture addition (Weidmeier et al., 1987; Olson et al., 1994). Total tract digestibility of DM, OM, NDF and ADF was not improved by either yeast culture (Table 2) which agrees with previous reports (Arambel and Kent, 1990; Carro et al., 1992b; Mutsvangwa et al., 1992; Smith et al., 1993; Moloney and Drennan, 1994). On the Contrary, digestibility was improved in other cases (Chademana and Offer, 1990; Ayala et al., 1992; Mendoza et al., 1995). Results from Roa et al. (1997) suggest that yeast culture does not improve NDF digestibility in low quality roughages, but it does in alfalfa hay. Identi®cation of dietary factors which in¯uence the animal response to direct-fed microbial products with Saccharomyces cerevisiae is needed.Values of duodenal ¯ow of amino acids are given in Table 3. The addition of yeast culture did not affect amino acid ¯ow. Although some researchers have found a tendency to increase the ¯ow of non-ammonia-N at the duodenum with yeast culture (Williams et al., 1990; Carro et al., 1992b), only Erasmus et al. (1992) have reported an increase in methionine ¯ow. Our results did not support the hypothesis that Saccharomyces cerevisiae improves amino acid ¯ow. According to the results of this trial, the addition of microbial cultures with Saccharomyces cerevisiae at commercial dosages in diets based on corn stover did not have a signi®cant impact on digestibility, pattern
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S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
Table 2 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on intake, ruminal fluid kinetics and digestibility of nutrients in Suffolk ewes fed on corn-stover diet Item
Treatments
n dry matter intake, g/day Ruminal fluid Molar proportion, % Volume, L kp, %hb Digestibility, % Protozoa, organisms104/ml Dry matter Organic matter Neutral detergent fiber Acid detergent fiber
CG
YS
LC
3 811
3 963
3 800
0.44 8.03
0.35 10.93
46.6 57.0 60.6 28.7
Contrasta
SEM
II
0.59
0.32
0.05 1.12
0.46 0.38
0.51 0.33
2.91 1.11 1.46 1.97
0.94 0.83 0.71 0.73
0.51 0.77 0.57 0.52
60
0.36 9.61
55.9 56.2 57.8 25.3
I
55.6 58.9 59.8 29.0
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell. b Fractional rate of fluid passage. a
Table 3 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on amino acid duodenal flow (g/day) in Suffolk ewes fed on corn-stover diet Amino acid
Asp Thr Ser Glu Gly Ala Val Ile Leu Tyr Phe His Lys Arg Met
Treatments
SEM
CG
YS
LC
10.34 5.51 6.51 14.37 4.84 9.82 6.87 6.53 8.76 4.66 4.99 1.68 5.33 3.61 9.01
15.20 8.11 9.61 20.59 7.24 13.63 10.34 9.57 13.49 7.33 7.68 2.54 9.05 5.87 14.32
8.72 5.02 7.45 10.19 4.37 7.96 6.03 5.62 7.59 4.10 4.37 1.51 5.04 3.30 7.44
2.27 1.30 1.57 3.47 1.10 2.20 1.54 1.48 2.16 1.07 1.17 0.41 1.37 0.88 3.85
Contrasta I
II
0.75 0.72 0.58 0.90 0.70 0.84 0.71 0.75 0.72 0.67 0.70 0.72 0.63 0.63 0.45
0.43 0.46 0.46 0.50 0.42 0.52 0.41 0.45 0.42 0.36 0.40 0.45 0.36 0.36 0.18
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. a Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell
of ruminal fermentation or amino acid ¯ow to the duodenum. Acknowledgements This study was supported by PAPIIT (Support Program for Research Projects and Technological
Innovation) from DGAPA-UNAM (Direction for Academic Affairs, University of Mexico). Support from the laboratory of the Department of Biotechnology from the Institute of Biomedical Research, UNAM is recognized. The technical assistance from Ma. Antonieta Aguirre, AndreÂs Lee-HernaÂndez, Antonio YaÂnÄez RamõÂrez and AndreÂs Toledo Bartolo is greatly
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
appreciated. The help of Jose Luis Cordero in surgeries is recognized. Appreciation is also extended to Edsel Bixtler, General Manager of ANGLO-CORP, Mexico. References Arambel, M.J., Kent, B.A., 1990. Effect of yeast culture on nutrient digestibility and milk yield response in early to mid lactation dairy cows. J. Dairy Sci. 73, 1560±1563. Association of Official Analytical Chemists, 1985. Official Methods of Analysis. AOAC, 14th edn. Washington, DC, pp. 152±157. Ayala, O.J., Gonzalez, S.S., Herrera, R., Mendoza, G.D., 1992. Effect of a probiotic and a molasses±urea supplement on fiber digestibility of sesame straw. J. Anim. Sci. 70 (Suppl. 1), 307. Benjamin, M.N., 1984. Manual of Veterinary clinical pathology. In: Limusa, S.A. (Ed.), ClõÂnica Veterinaria, de C.V. MeÂxico, pp. 93. Carro, M.D., Lebzien, P., Rohr, K., 1992a. Influence of yeast culture on the in vitro fermentation (Rusitec) of diets containing variable portions of concentrates. Anim. Feed Sci. Technol. 37, 209±220. Carro, M.D., Lebzien, P., Rohr, K., 1992b. Effects of yeast culture on rumen fermentation, digestibility and duodenal flow in dairy cows fed a silage-based diet. Livest. Prod. Sci. 32, 219±229. Chademana, Y., Offer, N.W., 1990. The effect of dietary inclusion of yeast culture on digestion in the sheep. Anim. Prod. 50, 483± 489. Corona, G.L., F. CastrejoÂn, P., G. Mendoza, M., y M. Cobos, P., 1995. Ruminal degradability of NDF in corn stover using two cultures of Saccharomyces cerevisiae. Vet. MeÂx. 26 (Suppl. 1), 276. Crosby, M.M.G., 1995. Efecto de la doÂsis de un cultivo de levadura (Saccharomyces cerevisiae) en la fermentacioÂn y en la digestibilidad ruminal de la fibra en borregas. Tesis MaestrõÂa. Colegio de Postgraduados. Programa de ganaderõÂa. Montecillo, Estado de MeÂxico. Dawson, K.E., Newman, K.E., 1987. Fermentation in rumen stimulating continuous cultures receiving probiotic supplements. J. Anim. Sci. 66 (Suppl. 1), 500. Erasmus, L.J., Botha, P.M., Kistner, A., 1992. Effect of yeast culture supplement on production, rumen fermentation, and duodenal nitrogen flow in dairy cows. J. Dairy Sci. 75, 3056± 3065. Erwin, E.S., Marco, G.J., Emery, E., 1961. Volatile fatty acid analysis of blood and rumen fluid by gas chromatography. J. Dairy Sci. 44, 1768±1776. Goering, H.K., Van Soest, P.J., 1970. Forage fiber analysis (apparatus, reagents, procedures and some applications). USDA Agr. Handbook No. 379, Washington, DC, 90 pp. Gonzalez, M.S.S., 1993. Improving utilization of poor quality forages with yeast culture. Biotechnology in the Feed Industry: Proceedings of Alltech's 9th Annual Symposium. Alltechs Technical Publications, Nicholasville, KY, USA.
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Harrison, G.A., Hemken, R.W., Dawson, K.A., Harmon, R.J., Barker, K.B., 1988. Influence of addition of yeast culture supplement to diets of lactating cows on ruminal fermentation and microbial populations. J. Dairy Sci. 71, 2967±2975. Jones, C.R., Dawson, K.A., 1993. Differentiating yeast strains in feed supplements using plasmid polymorphisms and arbitraryprimed polymerase chain reaction techniques. J. Anim. Sci. 71 (Suppl. 1), 288. LadroÂn de Guevara, O., Padilla, P., GarcõÂa, L., Pino, J.M., RamosElorduy, J., 1995. Amino acid determination in some edible Mexican insects. Amino acids 9, 161±173. Martin, S.A., Nisbet, D.J., Dean, R.G., 1989. Influence of a commercial yeast supplement on the in vitro ruminal fermentation. Nutr. Rep. Int. 40, 395±403. McCullough, H., 1967. The determination of ammonia in whole blood by a direct colorimetric method. Clin. Chem. Acta 17, 297±304. Mendoza, M.G.D., R. Ricalde, V., H. Esparza, B. y L., Velazquez, T., 1995. Technical note: Effect of two cultures of Saccharomyces cerevisiae on neutral detergent fiber degradation of wheat straw. Invest. Agr.: Prod. Sanid. Anim. 10, 33±38. Mir, Z., Mir, P.S., 1994. Effect of the addition of live yeast (Saccharomyces cerevisiae) on growth and carcass quality of steers fed high-forage or high-grain diets on feed digestibility and in situ degradation. J. Anim. Sci. 72, 537±545. Miranda, R.L.A., G. Mendoza, M., GonzaÂlez, S.S., R. BaÂrcena, G., 1994. Effect of Saccharomyces cerevisiae and Aspergillus oryzae on NDF level on ruminal fermentation. J. Anim. Sci. 72 (Suppl. 1), 12. Moloney, A.P., Drennan, M.J., 1994. The influence of the basal diet on the effects of yeast culture on ruminal fermentation and digestibility in steers. Anim. Feed Sci. Technol. 50, 55±73. Mutsvangwa, T., Edwards, I.E., Topps, J.H., Peterson, G.F.M., 1992. The effect of dietary inclusion of yeast culture (Yea± Sacc) on patterns of rumen fermentation, food intake and growth of intensively fed bulls. Anim. Prod. 55, 35±40. Newbold, C.J., Williams, P.E.V., McKain, N., Walker, A., Wallace, R.J., 1990. The effect of yeast culture on yeast numbers and fermentation in the rumen of sheep. Proc. Nutr. Soc. 49, 47. Newbold, C.J., Wallace, R.J., Chen, X.B., McIntosh, F.M., 1995. Different strains of Saccharomyces cerevisiae differ in their effects on ruminal bacterial numbers in vitro and in sheep. J. Anim. Sci. 73, 1811±1818. Offer, N.W., 1990. Maximizing fibre digestion in the rumen: The role of yeast culture. In: Lyons, T.P. (Ed.), Biotechnology in the Feed Industry. Alltech's Technical Publications, Nicholasville, KY, USA, pp. 79±93. Olson, K.C., Caton, J.S., Kirby, D.R., Norton, P.L., 1994. Influence of yeast culture supplementation and advancing season on steers grazing mixed-grass prairie in the northern great plains: II. Ruminal fermentation, site of digestion and microbial efficiency. J. Anim. Sci. 72, 2158±2170. Plata, P.F., Mendoza, M.G.D., BaÂrcena-Gama, J.R., GonzaÂlez, M.S., 1994. Effect of a yeast culture (Saccharomyces cerevisiae) on neutral detergent fiber digestion in steers fed oat straw based diets. Anim. Feed. Sci. Technol. 49, 203± 210.
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Roa, M.L., BaÂrcena-Gama, R., GonzaÂlez, M.S., Mendoza, M.G., Ortega, M.E., GarcõÂa, B.C., 1997. Effect of fiber source and a yeast culture (Saccharomyces cerevisiae1026) on digestion and the environment in the rumen of cattle. Anim. Feed Sci. Technol. 64, 327±336. Smith, W.A., Harris Jr., B., HornVan, H., Wilcox, C.J., 1993. Effects of forage type on production of dairy cows supplemented with whole cottonseed, tallow and yeast. J. Dairy Sci. 76, 205±215. Statistical Analysis System, 1985. S.A.S. User's Guide: Statistics. version 5, SAS Institute Inc., Cary, NC. Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics: A Biometrical Approach. 2nd edn. McGraw-Hill, New York. Stock, R.A., Brink, D.R., Britton, R.A., Goedeken, F.K., Sindt, M.H., Kreikemeier, K.K., Brauer, M.L., Smith, K.K., 1987. Feeding combinations of high moisture corn and dry-rolled grain sorghum to finishing steers. J. Anim. Sci. 65, 290±302. Walli, T.K., 1994. Role of yeast culture in rumen ecosystem and animal performance. Int. J. Anim. Sci. 9, 117±121.
Weidmeier, R.R.D., Arambel, M.J., Walters, J.L., 1987. Effect of yeast culture and Aspergillus oryzae fermentation extract on ruminal characteristics and nutrient digestibility. J. Dairy Sci. 70, 2063±2068. Wilcox, C.J., Thatcher, W.W., Martin, F.G., 1990. Statistical analysis of repeated measurements in physiology experiments. In: International Atomic Energy Agency, Proc. Final Research Co-ordination meeting of the FAO/IAEA/ARCAL III. Regional Network for Improving the Reproductive Management of Meat-and-Milk-Producing Livestock in Latin America. BogotaÂ, Colombia, pp. 141±155. Journal Series No. 9552 of the Florida Agricultural Experiment Station. 141. Williams, C.H., David, D.J., Iismaa, O., 1962. The determination of chromic oxide in faeces samples by atomic spectrophotometry. J. Agric. Sci. 59, 381±385. Williams, P.E.V., Walker, A., MacRae, J.C., 1990. Rumen probiosis: The effects of addition of yeast culture (viable yeast, Saccharomyces cerevisiae, plus growth medium) on duodenal protein flow in wheter sheep. Proc. Nutr. Soc. 49, 128A (Abstr.).
Comparison of two commercial yeast cultures (Saccharomyces cerevisiae) on ruminal fermentation and digestion in sheep fed on corn-stover diet S.C. Angeles C.a,*, G.D. Mendoza M.b, M.A. Cobos P.b, M.M. Crosby G.b, F.A. CastrejoÂn P.a,1 a
Universidad Nacional AutoÂnoma de MeÂxico, Facultad de Medicina Veterinaria y Zootecnia, Departamento de NutricioÂn Animal, Cd. Universitaria, MeÂxico, D.F. 04510, Mexico b Colegio de Postgraduados, Programa de GanaderõÂa, Montecillo, Km 35.5 Carr, MeÂxico-Texcoco Estado de Mexico, 56230, Mexico Accepted 4 March 1998
Abstract A metabolism trial was conducted to evaluate the effect of two direct-fed microbial cultures containing Saccharomyces cerevisiae on ruminal fermentation, digestibility and duodenal amino acid ¯ow. Nine Suffolk ewes with ruminal and duodenal cannula (29 kg BW) were assigned to a completely randomized design, where treatments were: (1) control group (CG); (2) 3 g/day of Yea±Sacc1026 (YS, 1108 CFU/g); and (3) 1 g/day of Levucell (LC, 20109 CFU/g). The diet consisted of corn stover (66.5%), sorghum grain (23.5%), molasses (8.6%) and urea (1.4%). Ruminal pH was lower (p<.01) with yeast culture (CG 6.81a, YS 6.51b, LC 6.61b). No effects were detected in VFA concentration, molar proportion, ammonia-N, and protozoa population. Total tract digestibility of DM, OM, NDF, and ADF were not affected by treatments. Duodenal amino acid ¯ow was not affected by the inclusion of yeast cultures. None of the direct-fed microbial cultures with Saccharomyces cerevisiae improved either fermentation, digestion or amino acid ¯ow to the duodenum in sheep fed on corn-stover diet. # 1998 Elsevier Science B.V. All rights reserved. Keywords: Saccharomyces cerevisiae; Fermentation; Digestion; Corn-stover diet
1. Introduction Direct-fed microbial products with Saccharomyces cerevisiae have been used to manipulate ruminal fermentation and animal production (Mutsvangwa et al., 1992; Walli, 1994). Bene®cial effects of these microbial compounds are associated with an increase *Corresponding author. Responsible for project PAPIIT No IN505994.
1
in cellulolytic bacteria (Dawson and Newman, 1987), with a concomitant improvement in ®ber digestibility in low quality forages (Ayala et al., 1992; Gonzalez, 1993). Fiber source is another factor which affects the response to yeast culture additions (Roa et al., 1997). Some studies indicated that amino acid pro®le and duodenal ¯ow are improved by the inclusion of yeast culture (Erasmus et al., 1992). However, results in ruminant production with yeast cultures have been inconsistent (Miranda et al., 1994; Mir and Mir, 1994),
0921-4488/98/$ ± see front matter # 1998 Elsevier Science B.V. All rights reserved. PII: S0921-4488(98)00115-1
46
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
and factors associated with the differences in commercial yeast cultures have received little attention. Several commercial feed additives with yeast cultures have been developed; however, comparisons of different products are scarce (Mendoza et al., 1995). Since feed additives are increasing production costs, it is important to compare the response with different yeast cultures. Also, strains of Saccharomyces cerevisiae differ in their effects on ruminal population (Newbold et al., 1995). In a previous trial (Corona et al., 1995), two yeasts cultures with Saccharomyces cerevisiae, dosed at similar colony-forming units (CFU) showed no effect on either fermentation or digestion in sheep fed on corn-stover diet. Therefore, this trial was conducted to evaluate the effect of two products containing Saccharomyces cerevisiae administered at commercial dosage rates on ruminal fermentation and digestibility. 2. Material and methods Nine Suffolk ewes (29 kg BW) with rumen and duodenal cannulae were randomly assigned to the following treatments: (1) control group; (2) 3 g/day of Yea±Sacc1026 (Alltech, Nicholasville; 1108 CFU/ g), and (3) 0.015 g/day of Levucell (Agrimerica, Northbrook, IL; 20109 CFU/g). The dosages of treatments 2 and 3 were based on the commercial information for each product, aiming at obtaining 3108 CFU for both the Yea±Sacc treatment and the Levucell treatment. Yeast cultures were dosed intraruminally. The diet (dry basis) was formulated with: corn stover (66.5%), sorghum grain (23.5%), molasses (8.6%) and urea (1.4%). The diet had the following composition: DM (82.57%), OM (94.37%), CP (10.15%), NDF (70.97%), and ADF (26.13%). Mineral premix was offered ad libitum (Ca 10%, P 12%, S 1.5%, Mg 0.2%, 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 to 90% of the ad libitum intake during the collection period (7 days). The animals were fed twice daily (7 and 16 h). Ruminal ¯uid samples were collected on day 7 of the collection period at 2, 6, 10, 14, 18 and 22 h after feeding. Ruminal ¯uid pH was measured immediately
after sampling and then a 50 ml aliquot of ruminal ¯uid was acidi®ed with 1 ml of HCl 6N, and stored in a freezer (ÿ208C) for further analysis. VFA were determined by gas chromatography in 4 ml of the acidi®ed samples prepared with 1 ml metaphosphoric acid (25%) (Erwin et al., 1961). Ammonia-N was also measured in the acidi®ed samples using the indophenol method (McCullough, 1967). A 5 ml ruminal ¯uid sample mixed with 5 ml of an iodine solution was used to count protozoa (count/ml) in a hemocytometer (Benjamin, 1984). Chromic oxide, used to calculate rate of passage, was dosed intraruminally (1 g/day) during the last 10 days of the adaptation period and during the collection period; chromium was measured by atomic absorption spectrophotometry (Williams et al., 1962). During the ®rst 4 days of the collection period, duodenal and faecal grab samples were collected as recommended by Stock et al. (1987). Feed and faecal samples were oven-dried (558C, 24 h) and ground to pass a 1 mm screen and composited by site and animal. Dry matter and organic matter were analyzed by standard methods (AOAC, 1985). NDF and ADF were determined by procedures outlined by Goering and Van Soest (1970). Duodenal samples were freeze-dried and analyzed for amino acid content using a Beckman 7300 analyzer. The samples were prepared as described by LadroÂn de Guevara et al. (1995). Data were analyzed as a completely randomized design (Steel and Torrie, 1980). If the response variable was measured at different times, data were analyzed with repeated measurements (Wilcox et al., 1990). Treatment means were compared by orthogonal contrasts using the GLM procedure of SAS (1985). 3. Results and discussion Ruminal pH was lower (p<.01) with yeast cultures than in the control group (Table 1), as observed by Harrison et al. (1988). No differences were observed between cultures, as reported by Corona et al. (1995) who used the same commercial cultures but at different dosages. Although most studies have shown that ruminal pH is not affected by Saccharomyces cerevisiae (Chademana and Offer, 1990; Carro et al., 1992a), some researchers (Jones and Dawson, 1993; Newbold
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47
Table 1 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on pH, ammonia-N and fermentation pattern and ruminal protozoa in Suffolk ewes fed on corn-stover diet Item
Treatments CG
Ruminal pH N-NH3, mg/dl Total VFA, mM Molar proportion, % Acetate Propionate Butyrate Protozoa, organisms104/ml Entodinidae Holotrichidae Total
SEM YS
LC
Contrasta I
II
6.81 11.01 82.8
6.51 10.16 87.9
6.61 10.71 94.5
0.04 1.72 4.79
0.002 0.59 0.10
0.22 0.65 0.38
66.6 21.3 11.8
67.4 21.4 11.2
67.5 21.1 11.4
0.92 0.98 0.54
0.76 0.70 0.22
0.82 0.59 0.18
1.48 0.79 1.53
0.55 0.23 0.34
0.30 0.90 0.93
843 9 852
1120 25 1145
838 23 859
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell.
a
et al., 1995) suggest that yeast cultures may stabilize ruminal pH. However, quantitative evidence of this has not yet been presented. There was no effect of microbial culture on ruminal ammonia concentration (Table 1). In most of the studies, microbial cultures based on Saccharomyces cerevisiae have no effect on ammonia-N (Carro et al., 1992a, b; Plata et al., 1994) and in some studies increments (Martin et al., 1989; Ayala et al., 1992) or reductions (Newbold et al., 1990; Moloney and Drennan, 1994) have been reported. Yeast cultures did not affect total VFA concentrations or fermentation patterns. Changes in molar proportions of VFA by additions of Saccharomyces cerevisiae were inconsistent (Offer, 1990; Chademana and Offer, 1990; Carro et al., 1992a, b; Mutsvangwa et al., 1992). Ruminal protozoa population was not affected by Saccharomyces cerevisiae (Table 1) in contrast to other studies with low quality forages (Ayala et al., 1992; Plata et al., 1994). According to Crosby (1995), there is a quadratic (p<0.001) effect of yeast culture dosage on protozoa population showing a greater population using 3 g/day of yeast culture in sheep fed on corn-stover diet. Dry matter intake was restricted in this study, and there were no differences among treatments on ruminal ¯uid volume or fractional rate of passage (Table 2). Chademana and Offer (1990) did not detect
changes in ¯uid kinetics, and ruminal volume was not affected by yeast culture addition (Weidmeier et al., 1987; Olson et al., 1994). Total tract digestibility of DM, OM, NDF and ADF was not improved by either yeast culture (Table 2) which agrees with previous reports (Arambel and Kent, 1990; Carro et al., 1992b; Mutsvangwa et al., 1992; Smith et al., 1993; Moloney and Drennan, 1994). On the Contrary, digestibility was improved in other cases (Chademana and Offer, 1990; Ayala et al., 1992; Mendoza et al., 1995). Results from Roa et al. (1997) suggest that yeast culture does not improve NDF digestibility in low quality roughages, but it does in alfalfa hay. Identi®cation of dietary factors which in¯uence the animal response to direct-fed microbial products with Saccharomyces cerevisiae is needed.Values of duodenal ¯ow of amino acids are given in Table 3. The addition of yeast culture did not affect amino acid ¯ow. Although some researchers have found a tendency to increase the ¯ow of non-ammonia-N at the duodenum with yeast culture (Williams et al., 1990; Carro et al., 1992b), only Erasmus et al. (1992) have reported an increase in methionine ¯ow. Our results did not support the hypothesis that Saccharomyces cerevisiae improves amino acid ¯ow. According to the results of this trial, the addition of microbial cultures with Saccharomyces cerevisiae at commercial dosages in diets based on corn stover did not have a signi®cant impact on digestibility, pattern
48
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
Table 2 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on intake, ruminal fluid kinetics and digestibility of nutrients in Suffolk ewes fed on corn-stover diet Item
Treatments
n dry matter intake, g/day Ruminal fluid Molar proportion, % Volume, L kp, %hb Digestibility, % Protozoa, organisms104/ml Dry matter Organic matter Neutral detergent fiber Acid detergent fiber
CG
YS
LC
3 811
3 963
3 800
0.44 8.03
0.35 10.93
46.6 57.0 60.6 28.7
Contrasta
SEM
II
0.59
0.32
0.05 1.12
0.46 0.38
0.51 0.33
2.91 1.11 1.46 1.97
0.94 0.83 0.71 0.73
0.51 0.77 0.57 0.52
60
0.36 9.61
55.9 56.2 57.8 25.3
I
55.6 58.9 59.8 29.0
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell. b Fractional rate of fluid passage. a
Table 3 Effect of two commercial yeast cultures (Saccharomyces cerevisiae) on amino acid duodenal flow (g/day) in Suffolk ewes fed on corn-stover diet Amino acid
Asp Thr Ser Glu Gly Ala Val Ile Leu Tyr Phe His Lys Arg Met
Treatments
SEM
CG
YS
LC
10.34 5.51 6.51 14.37 4.84 9.82 6.87 6.53 8.76 4.66 4.99 1.68 5.33 3.61 9.01
15.20 8.11 9.61 20.59 7.24 13.63 10.34 9.57 13.49 7.33 7.68 2.54 9.05 5.87 14.32
8.72 5.02 7.45 10.19 4.37 7.96 6.03 5.62 7.59 4.10 4.37 1.51 5.04 3.30 7.44
2.27 1.30 1.57 3.47 1.10 2.20 1.54 1.48 2.16 1.07 1.17 0.41 1.37 0.88 3.85
Contrasta I
II
0.75 0.72 0.58 0.90 0.70 0.84 0.71 0.75 0.72 0.67 0.70 0.72 0.63 0.63 0.45
0.43 0.46 0.46 0.50 0.42 0.52 0.41 0.45 0.42 0.36 0.40 0.45 0.36 0.36 0.18
CG: control group; YS: 3 g/day of Yea±Sacc1026; LC: 0.015 g/day of Levucell. a Probability of Type I error. Mean comparisons: I control group vs. yeast cultures; II Yea±Sacc1026 vs. Levucell
of ruminal fermentation or amino acid ¯ow to the duodenum. Acknowledgements This study was supported by PAPIIT (Support Program for Research Projects and Technological
Innovation) from DGAPA-UNAM (Direction for Academic Affairs, University of Mexico). Support from the laboratory of the Department of Biotechnology from the Institute of Biomedical Research, UNAM is recognized. The technical assistance from Ma. Antonieta Aguirre, AndreÂs Lee-HernaÂndez, Antonio YaÂnÄez RamõÂrez and AndreÂs Toledo Bartolo is greatly
S.C. Angeles C. et al. / Small Ruminant Research 31 (1998) 45±50
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