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Effect of Lactobacillus acidophilus Supplementation of Milk Replacer on Preweaning Performance of Calves
Effect of Lactobacillus acidophilus Supplementation of Milk Replacer on Preweaning Performance of Calves C. W. CRUYWAGEN,' INA JORDAAN, and L. VENTER Animal Nutrition and Animal Products Institute, Irene, Private Bag X2, 1675 Irene, South Africa
ABSTRACT
Forty Holstein-Friesian calves were used to evaluate the effect on young calves of daily dietary supplementation with Lactobacillus acidophilus. Calves were randomly assigned at 2 d of age to one of two treatments; 1) milk replacer with no additives or 2 ) milk replacer supplemented with 1 ml (5 x lo7) of viable L. acidophilus bacteria at each of two feedings per day. Milk replacer was reconstituted to 12% DM and fed at 10% of BWld for the duration of the 6-wk trial. A commercial starter pellet was offered for ad libitum intake from 7 d of age. Treatment had no effect on actual BW at any stage or on total BW gain; however, average daily gain during wk 2 was affected by L. acidophilus supplementation. Calves receiving L. acidophilus maintained initial BW, and the control calves lost BW until 2 wk of age, at an average rate of 112 g/d. Starter intake, total DMI, feed efficiency, and occurrence of diarrhea were unaffected by treatment. Therefore, L. acidophilus supplementation for calves fed milk replacer may be beneficial during the first 2 wk of life. ( Key words: calves, milk replacers, Lactobacillus acidophilus, probiotics)
Abbreviation key: L A = Lactobacillus acidophilus added to the MR, MR = milk replacer alone. INTRODUCTION
The widespread and prolonged use of antibiotics as therapeutic agents and growth stimulants since the 1950s has resulted in growing concern regarding the development of resistant bacteria populations that complicate subsequent antibiotic therapy ( 9 ) . Increased pressure against the use of antibiotics as growth stimulants by antiadditive lobbies, as well as reaction against the use of antibiotics as therapeutic agents because of intestinal disorders that often fol-
Received March 6, 1995. Accepted November 14, 1995. 1Present address: Department of Animal Science, University of Stellenbosch, 7600 Stellenbosch, South Africa. 1996 J Dairy Sci 79:483486
low oral administration, have prompted both consumer and manufacturer t o look for alternatives. Already, for a number of years, probiotics have been used t o replace antibiotics in certain applications. Probiotics have been defined in many ways. One clear definition is that a probiotic is a live microbial feed supplement that improves the intestinal microbial balance of the host animal ( 9 1. Lactic acid bacteria form part of the natural microbial population of the digestive tract of animals and are regarded as probiotics. The microbial population of lactic acid bacteria in the intestine depends on the type of animal and type of feeding regimen but generally consists of various genera and species. The following genera are classified under the family Lactobacillaceae ( 2 5 1: Lactobacillus (62 species), Leuconostoc (6 species), Pediococcus (8 species), Streptococcus (29 species), and Lactococcus ( 3 species). The salutary effect of naturally fermented milk (yogurt) has long been recognized. In 1908, Metchnikoff [cited by Atherton and Robbins ( 111 postulated that the effect of detrimental microbes in the intestinal tract could be alleviated by the ingestion of beneficial microbes. Even today, the probiotic concept is based on claims that ingestion of those organisms results i n colonization of the digestive tract, prevention of pathogen proliferation, neutralization of enterotoxins produced in situ, modulation of certain bacterial enzyme activity, enhancement of the small intestine digestive capacity, and the exertion of adjuvant effects on the immune system (21). Reports on Lactobacillus supplementation for calves are limited and contradictory. Calf performance was enhanced in some studies (2, 10, 20, 241, but others (8,11, 15, 17) showed no effect. Probiotics normally consist of cultures containing either one or a variety of different microbial species, especially Lactobacillus, usually produced by fermentation processes (12). Lactose (glucose plus galactose) is the only carbohydrate that can be digested by young calves, and because Lactobacillus acidophilus is a homofermenter that ferments lactose efficiently to lactic acid as an end product, we decided to investigate the effect of daily oral supplementation of L. acidophilus on the preweaning performance of calves.
483
484
CRUYWAGEN ET AL.
MATERIALS AND METHODS
Forty Holstein-Friesian bull calves were used in the trial. Calves were obtained either from the Irene herd a t the Animal Nutrition and Animal Products Institute, Irene, South Africa or purchased from a nearby farm at 2 d of age. Only healthy calves that weighed 730 kg at birth and that had at least 2 L of colostrum during the first 6 h after birth were used. Treatment for the first calf was randomly chosen; subsequent calves were alternately allocated to one of two treatments. Because all calves could not be obtained at the same time, mean initial BW was determined after 20 calves had been entered into the trial and, subsequently, after every second calf had been entered. It was sometimes necessary t o reverse the allocating order to minimize the difference in mean initial BW between groups. The first two feedings after arrival of calves consisted solely of electrolyte solutions. The composition of the electrolyte solution is shown in Table 1. Calves were housed in a semi-open barn in individual wooden crates (0.8 x 1.5 m ) with slatted wooden floors. All calves received a commercial milk replacer diet (Denkavit, Johannesburg, South Africa) containing, as a percentage of DM, 22% CP, 12% fat, <0.5% crude fiber, 1.6% Ca (maximum), and 0.8% P (minimum). The milk replacer was reconstituted to 12% DM and fed at a rate of 10% of initial BW/d. The daily ration was presented in two feedings (0730 and 1600 h ) and was fed at approximately 35°C. One group received the milk replacer alone ( MR), and the other group received L. acidophilus bacteria added to the MR a t each feeding ( LA). The L. acidophilus culture was cultivated at the Animal Nutrition and Animal Products Institute by culturing the bacteria in Bios 2000 (lactose milk) medium (Laboratorium Wiesby, Niebiill, Germany). Samples of the culture were stored in 1-ml ampules at -18°C until required. Each
ampule contained at least 5 x lo7 viable L. acidophilus cells, and one ampule per feeding was used for each calf in the treatment. All calves received a commercial calf starter pellet (Meadow Feeds, Delmas, South Africa) containing 18% CP and 11.0 M J of metabolizable energykg of DM. Starter pellets were offered for ad libitum intake from 7 d of age. Fecal consistency was scored and recorded daily according to the following definitions: 1 = firm, 2 = normal, 3 = soft but not running, 4 = soft and running, and 5 = watery. Scores of 4 and 5 were considered t o be diarrhea. If diarrhea occurred, milk and starter pellets were removed for 24 h while calves received 1 L of the commercial electrolyte solution three times per day. Milk was then reintroduced at 5% of BW/d for another 24 h before returning to normal feeding. If diarrhea persisted beyond 36 h, calves received two daily oral administrations (1 mV 10 kg of BW per administration) of Diazo1 Doser, a commercial preparation containing 100 mg/ml of furazolidone and 180 mg/ml of sulfamonomethoxine (Elvet, Johannesburg, South Africa) for 3 consecutive d. Calves were weighed weekly, and orts were weighed back at the same time. The experimental period was 6 wk. A completely randomized experimental design was used. A one-way ANOVA was performed on the data (16), except for treatment x time interactions, for which trends over time were tested using polynomial contrasts (27). Data were tested for normality and homogeneity of variances between treatments ( 27 ). Significance was declared at P < 0.05, unless otherwise noted. RESULTS AND DISCUSSION
The BW gain of calves is presented in Figure 1. No significant differences occurred between treatment means in actual BW at any stage during the trial. Trends over the first 3 wk differed ( P < 0.054) between treatments. Total BW gain 113.9kg ( It 0.8 S E ) and 12.9 kg ( k 0.8 SE) for calves on the MR and LA TABLE 1. Composition of commercial electrolyte solution1 used for treatments, respectively] did not differ between treatfirst t w o feedings and for the treatment of diarrhea. ments. However, average daily gain differed and was Sachet B highly significant ( P < 0.01) during the 2nd wk of life Item Sachet A2 . ,-, (Table 21,resulting in a difference in BW gain for the 31.8 Glycerine first 2-wk period. Calves on the MR treatment lost 4% Electrolytes and of initial BW during the first 2 wk, but calves in the citric acid 68.2 ... LA group almost completely maintained BW and lost Dextrose ... 100 only 0.8% of initial BW during the first 2 wk of age 1SmithKline Beecham Animal Health (Johannesburg, South 1). Loss of BW during the 1st or 2nd wk of (Figure Africa). is a phenomenon often observed for calves fed life *Each treatment consisted of two sachets ( A = 19.4 g; B = 44.6 (3,5, 7, 14). Nutrient digestibility was milk replacer g) mixed in 2 L of warm water. Journal of Dairy Science Vol. 79, No. 3, 1996
485
LACTOBAClLLUS SUPPLEMENTATION FOR CALVES
--
,
0
1
2
3
5
4
6
Age IwU
Figure 1. Mean (*SEMI change in BW of calves fed milk replacer (MR; ). or MR with Lactobacillus acidophilus supplementation ( A). No differences ( P > 0.05)occurred between treatment means at any stage.
not measured in this trial; therefore, the apparently beneficial effect of LA cannot be fully explained. Improvement of absorption of nutrients in the lower intestine has been suggested by Sissons (261, and evidence of beneficial effects during the early stages of life in calves supplemented with probiotics has also been reported by others (2, 10, 13, 19, 22, 24). Calves on the LA treatment did not retain the initial advantage from LA, and calves in the MR group appeared to compensate for initial BW loss by tending to grow faster than calves in the LA group ( P < 0.10) during
TABLE 2.Average daily gain of calves fed milk replacer ( M R ) or MR with Lactobacillus acidophilus supplementation (LA).
wk 3 and 4, but weaning BW was not higher for calves on the MR treatment at 6 wk. Calves are vulnerable and quite susceptible to diseases during the first few weeks of life; therefore, the apparent beneficial effect of LA during wk 1 and 2 of this trial would support recommendation of the prophylactic use of probiotics in calves 52 wk of age. Feed intake and occurrence of diarrhea data are presented in Table 3. The MR DMI was determined by initial BW and, therefore, could not be affected by treatment. Treatment had no effect on DMI from starter pellets or on total DMI. This result is in contrast with those reported by Higginbotham and Bath ( 13) and by Ruppert et al. ( 23 ) . According to Ruppert et al. (231, when the diet was supplemented with a probiotic and when calves were kept under stressful conditions, feed intake of calves ( 2 to 28 d ) was higher than intake for calves in the negative control group. The probiotic in that experiment contained a combination of L. acidophilus, Streptococcus cervisiae, and Streptococcus faecium. The higher feed intake observed by those researchers also resulted in faster BW gains than those of the control group. Morrill et al. (171, however, reported lower starter intakes for calves receiving cultured LA than for calves receiving pasteurized milk. The lack of response to LA in our study was probably because the calves were not stressed. Because total BW gain and total DMI did not differ between treatment means, feed efficiency was the same for both treatments. Feed efficiency in the current study corresponded with reported data on MR fed at the same level (4, 5 , 15) but was lower than that reported for MR fed at higher levels ( 6 ) or for whole milk (4, 5). Rose11 ( 2 2 ) reported improved feed efficiency in calves fed LA, S. faecium, and a yeast culture. Treatment had no effect on the occurrence of diarrhea. Also, there was no relationship between occur-
Treatment Item Calves, no. Average gain, g/d wk 1 wk 2 wk 1 and 2 wk 3 wk 4 wk 3 and 4 wk 5 wk 6 wk 5 and 6 Total period, g/d
LA 20 -43.9 -1.3 -22.6 265.7 421.5 343.7 555.0 650.7 602.9 306.9
MR
SEM
20 -57.9 -165.1** -111.5* 381.6 517.8 449.7t 659.3 645.8 652.5 330.2
*Means within a row tended to differ ( P < 0.10). *Means within a row differ ( P < 0.05). **Means within a row differ ( P < 0.01).
34.9 30.8 21.6 39.8 38.6 29.8 34.4 40.4 27.0 13.4
TABLE 3. Feed intake, feed efficiency, and occurrence of diarrhea in calves fed milk replacer ( M R ) or MR with Lactobacillus acidophilus supplementation (LA).1 Treatment
LA
Item
-~
Calves, no. Milk replacer intake, kg of DM Starter intake, kg of DM Total intake, kg of DM Feed efficiency, kg of DMYkg of gain Diarrhea, mean days ~
MR
SEM ~
~
20
20
21.0 20.2 41.2
20.7 17.4 38.0
0.4 1.0 1.3
3.1 2.2
3.1 2.3
0.1 0.3
~
'Differences were not significant ( P > 0.05) Journal of Dairy Science Vol. 79, No. 3, 1996
486
CRUYWAGEN ET AL.
rence or severity of diarrhea and BW changes. The occurrence of diarrhea was low and at an acceptable level; LA, therefore, did not have any additional effect. Contrasting reports (2, 8, 15, 17, 18, 22, 24) on the effect of probiotics on the occurrence of diarrhea have been documented. A significant reduction in mortality rate has been reported by Rosell ( 22 ). The effects of feeding regimen and stress appeared to have complicated the observation of absolute effects of probiotics on calf performance, and, according to Fuller (9 1, probiotics are only effective when animals are stressed by the presence of a microbial population that depresses growth. Management and feeding conditions in our study were good, and LA might have had different effects under suboptimal conditions. CONCLUSIONS
In our study, LA had no effect on weaning BW, feed efficiency, or general health of calves. There was no difference between groups in general appearance or in response to treatment for diarrhea. However, LA calves almost maintained initial BW during the first 2 wk of life, but the MR group lost 4% of BW. Calves are vulnerable and susceptible to diseases during the first few weeks of life, and maintenance of BW during that stage could improve resistance against diseases; therefore, the prophylactic use of probiotics during the first 2 wk of life is recommended, especially for calves fed MR or for calves reared under suboptimal management conditions. ACKNOWLEDGMENTS
The authors gratefully acknowledge the assistance of Amos Shabangu and Jerry Shongwe in the care of the animals. Denkavit SA ( P t y . ) Ltd. is also thanked for supplying the milk replacer. The assistance of Cobus Brayshaw with the statistical analyses is also appreciated. REFERENCES 1Atherton, D., and S. Robbins. 1987. Probiotics-European perspective. Page 167 in Biotechnology in the Feed Industry. T. P. Lyons, ed. Alltech Tech. Publ., Nicholasville, KY. 2 Bechman, T. J., J. V. Chambers, and M. D. Cunningham. 1977. Influence of Lactobacillus acidophilus on the performance of young calves. J. Dairy Sci. Gl(Supp1. 1):74.(Abstr.) 3 Cruywagen, C. W. 1982. Optimizing soybean flour, whey powder and colostrum ratios for rearing dairy calves. S. Afr. J. h i m . Sci. 12:103. 4Cruywageq C. W., G. J. Brisson, G. F. Tremblay, and H. H. Meissner. 1990. Effect of curd suppression in a milk replacer on physiological parameters in calves. I. Digestibility of nutrients and body mass gain. S. Afr. J. Anim. Sci. 20~234. 5 Cruywagen, C. W., and J. G. Horn. 1985. Pre-weaning growth and feed intake of dairy calves receiving different combinations Journal of Dairy Science Vol. 79, No. 3, 19%
of soybean flour, whey powder and colostrum. S. Afr. J. Anim. Sci. 15:ll. 6 Cruywagen, C. W., and J. G. Horn-Quass. 1991. Effect of curd suppression in a calf milk replacer fed a t increasing levels on nutrient digestibility and body mass gain. S. Afr. J. Anim. Sci. 21:153. 7Downes, T.E.H., C. W. Cruywagen, G. A. Smith, and J. C. Pelster. 1982. The use of whey, whey protein concentrate and spray dried blood powders in the manufacture of milk replacers for calves. S. A&. J . Dairy Technol. 14:81. 8 Ellinger, D. K, L. D. Muller, and P. J. Glantz. 1978. Influence of feeding fermented colostrum and Latobacillus acidophilus on fecal flora and selected blood parameters of young dairy calves. J. Dairy Sci. 61(Suppl. 1):126.(Abstr.) 9 Fuller, R. 1989. Probiotics in man and animals. A review. J. Appl. Bacteriol. 66365. 10 Gilliland, S. E., B. B. Bruce, L. J. Bush, and T. E. Staley. 1980. Comparison of two strains of Lactobacillus acidophilus as dietary adjuncts for young calves. J. Dairy Sci. 63:964. 11Hatch, R. C., R. 0. Thomas, and W. V. Thayne. 1973. Effect of adding Bacillus acidophilus to milk fed to baby calves. J. Dairy Sci. 56682. 12 Hertrampf, J . 1979. Wachstumsforderer der Zukunft. Mdhle und Mischfuttertechnik 116:611.(Abstr.) 13Higginbotham, G. E., and D. L. Bath. 1993. Evaluation of Lactobacillus fermentation cultures in calf feeding systems. J. Dairy Sci. 76:615. 14Jaster, E. H., G. C. McCoy, and R. L. Fernando. 1990. Dietary fat in milk or milk replacers for dairy calves raised during the winter. J. Dairy Sci. 73:1843. 15Jenny, B. F., H. J. Vandijk, and J. A. Collins. 1991. Performance and fecal flora of calves fed a Bacillus subtilis concentrate. J. Dairy Sci. 74:1968. 16Little, T. M., and F. J. Hills. 1975. Statistical Methods in Agricultural Research. Univ. California, Davis. 17 Morrill, J. L., A. D. Dayton, and R. Mickelsen. 1977. Cultured milk and antibiotics for young calves. J. Dairy Sci. 60:1105. laOwen, F. G., and L. L. Larsen. 1984. Effect of Probiocin and starter preparations on calf performance. J. Dairy Sci. 61(Suppl. l):139.(Abstr.) 19 Quintero-Gonzalez, C. I., J. W. Comerford, G. A. Varga, and T. W. Cassidy. 1994. Effects of direct-fed microbials on productivity and blood parameters in young - calves. J . Dairy Sci. 77(Suppl. 1):89.(Abstr.) 20Radulovic. D.. P. Krdzalic. M. Marinac. D. Bresianac. and S. Stojicevic.’1976. The effect’of Lactobacilius on thd development of intestinal flora of the healthy and affected calves. Vet. Glas. 30:911.(Abstr.) 21 Raibaud, P., and J. P. Raynaud. 1991. Experimental data on the modes of action of probiotics. Page 269 in New Trends in Veal Calf Production. Proc. Int. Symp. Veal Calf Prod., Wageningen, The Netherlands. Eur. Assoc. h i m . Prod. Publ. No. 52. J.H.M. Metz and C. M. Groenestein, ed. Pudoc, Wageningen, The Netherlands. 22 Rosell, V. 1987. Acidification and probiotics in Spanish pig and calf rearing. Page 177 in Biotechnology in the Feed Industry. T. P. Lyons, ed. Alltech Tech. Publ., Nicholasville, KY. 23Ruppert, L. D., G. C. McCoy, and M. F. Hutjens. 1994. Feeding of probiotics to calves. J. Dairy Sci. 77(Suppl. 1):296.(Abstr.) 24Schwab, C. G., J. J. Moore, 111, P. M. Hoyt, and J. L. Prentice. 1980. Performance and fecal flora of calves fed a nonviable Lactobacillus bulgaricus fermentation product. J. Dairy Sci. 63:1412. 25 Sharpe, M. E. 1986. Identification of lactic acid bacteria. Page 1218 in Bergey’s Manual of Systematic Bacteriology. Vol. 11. Williams and Wilkins, London, England. 26 Sissons, J. W. 1989. Potential of probiotic organisms tu prevent diarrhea and promote digestion in farm animals-a review. J . Sci. Food Agric. 49:l. 27 Snedecor, G. W., and W. G. Cochran. 1980. Statistical Methods. 7th ed. Iowa State Univ. Press, Ames.
ABSTRACT
Forty Holstein-Friesian calves were used to evaluate the effect on young calves of daily dietary supplementation with Lactobacillus acidophilus. Calves were randomly assigned at 2 d of age to one of two treatments; 1) milk replacer with no additives or 2 ) milk replacer supplemented with 1 ml (5 x lo7) of viable L. acidophilus bacteria at each of two feedings per day. Milk replacer was reconstituted to 12% DM and fed at 10% of BWld for the duration of the 6-wk trial. A commercial starter pellet was offered for ad libitum intake from 7 d of age. Treatment had no effect on actual BW at any stage or on total BW gain; however, average daily gain during wk 2 was affected by L. acidophilus supplementation. Calves receiving L. acidophilus maintained initial BW, and the control calves lost BW until 2 wk of age, at an average rate of 112 g/d. Starter intake, total DMI, feed efficiency, and occurrence of diarrhea were unaffected by treatment. Therefore, L. acidophilus supplementation for calves fed milk replacer may be beneficial during the first 2 wk of life. ( Key words: calves, milk replacers, Lactobacillus acidophilus, probiotics)
Abbreviation key: L A = Lactobacillus acidophilus added to the MR, MR = milk replacer alone. INTRODUCTION
The widespread and prolonged use of antibiotics as therapeutic agents and growth stimulants since the 1950s has resulted in growing concern regarding the development of resistant bacteria populations that complicate subsequent antibiotic therapy ( 9 ) . Increased pressure against the use of antibiotics as growth stimulants by antiadditive lobbies, as well as reaction against the use of antibiotics as therapeutic agents because of intestinal disorders that often fol-
Received March 6, 1995. Accepted November 14, 1995. 1Present address: Department of Animal Science, University of Stellenbosch, 7600 Stellenbosch, South Africa. 1996 J Dairy Sci 79:483486
low oral administration, have prompted both consumer and manufacturer t o look for alternatives. Already, for a number of years, probiotics have been used t o replace antibiotics in certain applications. Probiotics have been defined in many ways. One clear definition is that a probiotic is a live microbial feed supplement that improves the intestinal microbial balance of the host animal ( 9 1. Lactic acid bacteria form part of the natural microbial population of the digestive tract of animals and are regarded as probiotics. The microbial population of lactic acid bacteria in the intestine depends on the type of animal and type of feeding regimen but generally consists of various genera and species. The following genera are classified under the family Lactobacillaceae ( 2 5 1: Lactobacillus (62 species), Leuconostoc (6 species), Pediococcus (8 species), Streptococcus (29 species), and Lactococcus ( 3 species). The salutary effect of naturally fermented milk (yogurt) has long been recognized. In 1908, Metchnikoff [cited by Atherton and Robbins ( 111 postulated that the effect of detrimental microbes in the intestinal tract could be alleviated by the ingestion of beneficial microbes. Even today, the probiotic concept is based on claims that ingestion of those organisms results i n colonization of the digestive tract, prevention of pathogen proliferation, neutralization of enterotoxins produced in situ, modulation of certain bacterial enzyme activity, enhancement of the small intestine digestive capacity, and the exertion of adjuvant effects on the immune system (21). Reports on Lactobacillus supplementation for calves are limited and contradictory. Calf performance was enhanced in some studies (2, 10, 20, 241, but others (8,11, 15, 17) showed no effect. Probiotics normally consist of cultures containing either one or a variety of different microbial species, especially Lactobacillus, usually produced by fermentation processes (12). Lactose (glucose plus galactose) is the only carbohydrate that can be digested by young calves, and because Lactobacillus acidophilus is a homofermenter that ferments lactose efficiently to lactic acid as an end product, we decided to investigate the effect of daily oral supplementation of L. acidophilus on the preweaning performance of calves.
483
484
CRUYWAGEN ET AL.
MATERIALS AND METHODS
Forty Holstein-Friesian bull calves were used in the trial. Calves were obtained either from the Irene herd a t the Animal Nutrition and Animal Products Institute, Irene, South Africa or purchased from a nearby farm at 2 d of age. Only healthy calves that weighed 730 kg at birth and that had at least 2 L of colostrum during the first 6 h after birth were used. Treatment for the first calf was randomly chosen; subsequent calves were alternately allocated to one of two treatments. Because all calves could not be obtained at the same time, mean initial BW was determined after 20 calves had been entered into the trial and, subsequently, after every second calf had been entered. It was sometimes necessary t o reverse the allocating order to minimize the difference in mean initial BW between groups. The first two feedings after arrival of calves consisted solely of electrolyte solutions. The composition of the electrolyte solution is shown in Table 1. Calves were housed in a semi-open barn in individual wooden crates (0.8 x 1.5 m ) with slatted wooden floors. All calves received a commercial milk replacer diet (Denkavit, Johannesburg, South Africa) containing, as a percentage of DM, 22% CP, 12% fat, <0.5% crude fiber, 1.6% Ca (maximum), and 0.8% P (minimum). The milk replacer was reconstituted to 12% DM and fed at a rate of 10% of initial BW/d. The daily ration was presented in two feedings (0730 and 1600 h ) and was fed at approximately 35°C. One group received the milk replacer alone ( MR), and the other group received L. acidophilus bacteria added to the MR a t each feeding ( LA). The L. acidophilus culture was cultivated at the Animal Nutrition and Animal Products Institute by culturing the bacteria in Bios 2000 (lactose milk) medium (Laboratorium Wiesby, Niebiill, Germany). Samples of the culture were stored in 1-ml ampules at -18°C until required. Each
ampule contained at least 5 x lo7 viable L. acidophilus cells, and one ampule per feeding was used for each calf in the treatment. All calves received a commercial calf starter pellet (Meadow Feeds, Delmas, South Africa) containing 18% CP and 11.0 M J of metabolizable energykg of DM. Starter pellets were offered for ad libitum intake from 7 d of age. Fecal consistency was scored and recorded daily according to the following definitions: 1 = firm, 2 = normal, 3 = soft but not running, 4 = soft and running, and 5 = watery. Scores of 4 and 5 were considered t o be diarrhea. If diarrhea occurred, milk and starter pellets were removed for 24 h while calves received 1 L of the commercial electrolyte solution three times per day. Milk was then reintroduced at 5% of BW/d for another 24 h before returning to normal feeding. If diarrhea persisted beyond 36 h, calves received two daily oral administrations (1 mV 10 kg of BW per administration) of Diazo1 Doser, a commercial preparation containing 100 mg/ml of furazolidone and 180 mg/ml of sulfamonomethoxine (Elvet, Johannesburg, South Africa) for 3 consecutive d. Calves were weighed weekly, and orts were weighed back at the same time. The experimental period was 6 wk. A completely randomized experimental design was used. A one-way ANOVA was performed on the data (16), except for treatment x time interactions, for which trends over time were tested using polynomial contrasts (27). Data were tested for normality and homogeneity of variances between treatments ( 27 ). Significance was declared at P < 0.05, unless otherwise noted. RESULTS AND DISCUSSION
The BW gain of calves is presented in Figure 1. No significant differences occurred between treatment means in actual BW at any stage during the trial. Trends over the first 3 wk differed ( P < 0.054) between treatments. Total BW gain 113.9kg ( It 0.8 S E ) and 12.9 kg ( k 0.8 SE) for calves on the MR and LA TABLE 1. Composition of commercial electrolyte solution1 used for treatments, respectively] did not differ between treatfirst t w o feedings and for the treatment of diarrhea. ments. However, average daily gain differed and was Sachet B highly significant ( P < 0.01) during the 2nd wk of life Item Sachet A2 . ,-, (Table 21,resulting in a difference in BW gain for the 31.8 Glycerine first 2-wk period. Calves on the MR treatment lost 4% Electrolytes and of initial BW during the first 2 wk, but calves in the citric acid 68.2 ... LA group almost completely maintained BW and lost Dextrose ... 100 only 0.8% of initial BW during the first 2 wk of age 1SmithKline Beecham Animal Health (Johannesburg, South 1). Loss of BW during the 1st or 2nd wk of (Figure Africa). is a phenomenon often observed for calves fed life *Each treatment consisted of two sachets ( A = 19.4 g; B = 44.6 (3,5, 7, 14). Nutrient digestibility was milk replacer g) mixed in 2 L of warm water. Journal of Dairy Science Vol. 79, No. 3, 1996
485
LACTOBAClLLUS SUPPLEMENTATION FOR CALVES
--
,
0
1
2
3
5
4
6
Age IwU
Figure 1. Mean (*SEMI change in BW of calves fed milk replacer (MR; ). or MR with Lactobacillus acidophilus supplementation ( A). No differences ( P > 0.05)occurred between treatment means at any stage.
not measured in this trial; therefore, the apparently beneficial effect of LA cannot be fully explained. Improvement of absorption of nutrients in the lower intestine has been suggested by Sissons (261, and evidence of beneficial effects during the early stages of life in calves supplemented with probiotics has also been reported by others (2, 10, 13, 19, 22, 24). Calves on the LA treatment did not retain the initial advantage from LA, and calves in the MR group appeared to compensate for initial BW loss by tending to grow faster than calves in the LA group ( P < 0.10) during
TABLE 2.Average daily gain of calves fed milk replacer ( M R ) or MR with Lactobacillus acidophilus supplementation (LA).
wk 3 and 4, but weaning BW was not higher for calves on the MR treatment at 6 wk. Calves are vulnerable and quite susceptible to diseases during the first few weeks of life; therefore, the apparent beneficial effect of LA during wk 1 and 2 of this trial would support recommendation of the prophylactic use of probiotics in calves 52 wk of age. Feed intake and occurrence of diarrhea data are presented in Table 3. The MR DMI was determined by initial BW and, therefore, could not be affected by treatment. Treatment had no effect on DMI from starter pellets or on total DMI. This result is in contrast with those reported by Higginbotham and Bath ( 13) and by Ruppert et al. ( 23 ) . According to Ruppert et al. (231, when the diet was supplemented with a probiotic and when calves were kept under stressful conditions, feed intake of calves ( 2 to 28 d ) was higher than intake for calves in the negative control group. The probiotic in that experiment contained a combination of L. acidophilus, Streptococcus cervisiae, and Streptococcus faecium. The higher feed intake observed by those researchers also resulted in faster BW gains than those of the control group. Morrill et al. (171, however, reported lower starter intakes for calves receiving cultured LA than for calves receiving pasteurized milk. The lack of response to LA in our study was probably because the calves were not stressed. Because total BW gain and total DMI did not differ between treatment means, feed efficiency was the same for both treatments. Feed efficiency in the current study corresponded with reported data on MR fed at the same level (4, 5 , 15) but was lower than that reported for MR fed at higher levels ( 6 ) or for whole milk (4, 5). Rose11 ( 2 2 ) reported improved feed efficiency in calves fed LA, S. faecium, and a yeast culture. Treatment had no effect on the occurrence of diarrhea. Also, there was no relationship between occur-
Treatment Item Calves, no. Average gain, g/d wk 1 wk 2 wk 1 and 2 wk 3 wk 4 wk 3 and 4 wk 5 wk 6 wk 5 and 6 Total period, g/d
LA 20 -43.9 -1.3 -22.6 265.7 421.5 343.7 555.0 650.7 602.9 306.9
MR
SEM
20 -57.9 -165.1** -111.5* 381.6 517.8 449.7t 659.3 645.8 652.5 330.2
*Means within a row tended to differ ( P < 0.10). *Means within a row differ ( P < 0.05). **Means within a row differ ( P < 0.01).
34.9 30.8 21.6 39.8 38.6 29.8 34.4 40.4 27.0 13.4
TABLE 3. Feed intake, feed efficiency, and occurrence of diarrhea in calves fed milk replacer ( M R ) or MR with Lactobacillus acidophilus supplementation (LA).1 Treatment
LA
Item
-~
Calves, no. Milk replacer intake, kg of DM Starter intake, kg of DM Total intake, kg of DM Feed efficiency, kg of DMYkg of gain Diarrhea, mean days ~
MR
SEM ~
~
20
20
21.0 20.2 41.2
20.7 17.4 38.0
0.4 1.0 1.3
3.1 2.2
3.1 2.3
0.1 0.3
~
'Differences were not significant ( P > 0.05) Journal of Dairy Science Vol. 79, No. 3, 1996
486
CRUYWAGEN ET AL.
rence or severity of diarrhea and BW changes. The occurrence of diarrhea was low and at an acceptable level; LA, therefore, did not have any additional effect. Contrasting reports (2, 8, 15, 17, 18, 22, 24) on the effect of probiotics on the occurrence of diarrhea have been documented. A significant reduction in mortality rate has been reported by Rosell ( 22 ). The effects of feeding regimen and stress appeared to have complicated the observation of absolute effects of probiotics on calf performance, and, according to Fuller (9 1, probiotics are only effective when animals are stressed by the presence of a microbial population that depresses growth. Management and feeding conditions in our study were good, and LA might have had different effects under suboptimal conditions. CONCLUSIONS
In our study, LA had no effect on weaning BW, feed efficiency, or general health of calves. There was no difference between groups in general appearance or in response to treatment for diarrhea. However, LA calves almost maintained initial BW during the first 2 wk of life, but the MR group lost 4% of BW. Calves are vulnerable and susceptible to diseases during the first few weeks of life, and maintenance of BW during that stage could improve resistance against diseases; therefore, the prophylactic use of probiotics during the first 2 wk of life is recommended, especially for calves fed MR or for calves reared under suboptimal management conditions. ACKNOWLEDGMENTS
The authors gratefully acknowledge the assistance of Amos Shabangu and Jerry Shongwe in the care of the animals. Denkavit SA ( P t y . ) Ltd. is also thanked for supplying the milk replacer. The assistance of Cobus Brayshaw with the statistical analyses is also appreciated. REFERENCES 1Atherton, D., and S. Robbins. 1987. Probiotics-European perspective. Page 167 in Biotechnology in the Feed Industry. T. P. Lyons, ed. Alltech Tech. Publ., Nicholasville, KY. 2 Bechman, T. J., J. V. Chambers, and M. D. Cunningham. 1977. Influence of Lactobacillus acidophilus on the performance of young calves. J. Dairy Sci. Gl(Supp1. 1):74.(Abstr.) 3 Cruywagen, C. W. 1982. Optimizing soybean flour, whey powder and colostrum ratios for rearing dairy calves. S. Afr. J. h i m . Sci. 12:103. 4Cruywageq C. W., G. J. Brisson, G. F. Tremblay, and H. H. Meissner. 1990. Effect of curd suppression in a milk replacer on physiological parameters in calves. I. Digestibility of nutrients and body mass gain. S. Afr. J. Anim. Sci. 20~234. 5 Cruywagen, C. W., and J. G. Horn. 1985. Pre-weaning growth and feed intake of dairy calves receiving different combinations Journal of Dairy Science Vol. 79, No. 3, 19%
of soybean flour, whey powder and colostrum. S. Afr. J. Anim. Sci. 15:ll. 6 Cruywagen, C. W., and J. G. Horn-Quass. 1991. Effect of curd suppression in a calf milk replacer fed a t increasing levels on nutrient digestibility and body mass gain. S. Afr. J. Anim. Sci. 21:153. 7Downes, T.E.H., C. W. Cruywagen, G. A. Smith, and J. C. Pelster. 1982. The use of whey, whey protein concentrate and spray dried blood powders in the manufacture of milk replacers for calves. S. A&. J . Dairy Technol. 14:81. 8 Ellinger, D. K, L. D. Muller, and P. J. Glantz. 1978. Influence of feeding fermented colostrum and Latobacillus acidophilus on fecal flora and selected blood parameters of young dairy calves. J. Dairy Sci. 61(Suppl. 1):126.(Abstr.) 9 Fuller, R. 1989. Probiotics in man and animals. A review. J. Appl. Bacteriol. 66365. 10 Gilliland, S. E., B. B. Bruce, L. J. Bush, and T. E. Staley. 1980. Comparison of two strains of Lactobacillus acidophilus as dietary adjuncts for young calves. J. Dairy Sci. 63:964. 11Hatch, R. C., R. 0. Thomas, and W. V. Thayne. 1973. Effect of adding Bacillus acidophilus to milk fed to baby calves. J. Dairy Sci. 56682. 12 Hertrampf, J . 1979. Wachstumsforderer der Zukunft. Mdhle und Mischfuttertechnik 116:611.(Abstr.) 13Higginbotham, G. E., and D. L. Bath. 1993. Evaluation of Lactobacillus fermentation cultures in calf feeding systems. J. Dairy Sci. 76:615. 14Jaster, E. H., G. C. McCoy, and R. L. Fernando. 1990. Dietary fat in milk or milk replacers for dairy calves raised during the winter. J. Dairy Sci. 73:1843. 15Jenny, B. F., H. J. Vandijk, and J. A. Collins. 1991. Performance and fecal flora of calves fed a Bacillus subtilis concentrate. J. Dairy Sci. 74:1968. 16Little, T. M., and F. J. Hills. 1975. Statistical Methods in Agricultural Research. Univ. California, Davis. 17 Morrill, J. L., A. D. Dayton, and R. Mickelsen. 1977. Cultured milk and antibiotics for young calves. J. Dairy Sci. 60:1105. laOwen, F. G., and L. L. Larsen. 1984. Effect of Probiocin and starter preparations on calf performance. J. Dairy Sci. 61(Suppl. l):139.(Abstr.) 19 Quintero-Gonzalez, C. I., J. W. Comerford, G. A. Varga, and T. W. Cassidy. 1994. Effects of direct-fed microbials on productivity and blood parameters in young - calves. J . Dairy Sci. 77(Suppl. 1):89.(Abstr.) 20Radulovic. D.. P. Krdzalic. M. Marinac. D. Bresianac. and S. Stojicevic.’1976. The effect’of Lactobacilius on thd development of intestinal flora of the healthy and affected calves. Vet. Glas. 30:911.(Abstr.) 21 Raibaud, P., and J. P. Raynaud. 1991. Experimental data on the modes of action of probiotics. Page 269 in New Trends in Veal Calf Production. Proc. Int. Symp. Veal Calf Prod., Wageningen, The Netherlands. Eur. Assoc. h i m . Prod. Publ. No. 52. J.H.M. Metz and C. M. Groenestein, ed. Pudoc, Wageningen, The Netherlands. 22 Rosell, V. 1987. Acidification and probiotics in Spanish pig and calf rearing. Page 177 in Biotechnology in the Feed Industry. T. P. Lyons, ed. Alltech Tech. Publ., Nicholasville, KY. 23Ruppert, L. D., G. C. McCoy, and M. F. Hutjens. 1994. Feeding of probiotics to calves. J. Dairy Sci. 77(Suppl. 1):296.(Abstr.) 24Schwab, C. G., J. J. Moore, 111, P. M. Hoyt, and J. L. Prentice. 1980. Performance and fecal flora of calves fed a nonviable Lactobacillus bulgaricus fermentation product. J. Dairy Sci. 63:1412. 25 Sharpe, M. E. 1986. Identification of lactic acid bacteria. Page 1218 in Bergey’s Manual of Systematic Bacteriology. Vol. 11. Williams and Wilkins, London, England. 26 Sissons, J. W. 1989. Potential of probiotic organisms tu prevent diarrhea and promote digestion in farm animals-a review. J . Sci. Food Agric. 49:l. 27 Snedecor, G. W., and W. G. Cochran. 1980. Statistical Methods. 7th ed. Iowa State Univ. Press, Ames.