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Effect of feeding cellulase enzyme on productive responses of pregnant and lactating ewes and goats
Small Ruminant Research 52 (2004) 137–143
Effect of feeding cellulase enzyme on productive responses of pregnant and lactating ewes and goats H. Titi∗ , W.F. Lubbadeh Department of Animal Production, Faculty of Agriculture, University of Jordan, Amman 11942, Jordan Received 21 August 2002; received in revised form 23 April 2003; accepted 29 June 2003
Abstract An experiment was conducted to study the effect of supplementing the diets of Awassi ewes and Shami goats with exogenous fibrolytic enzyme on birth and weaning weight of lambs and kids and on milk production of dams in the first 60 days of lactation. Two groups of ewes (30 each) and two groups of goats (30 each) were used. One group of each species was supplemented with the fibrolytic enzyme during the last 2 months of pregnancy and the first 60 days of lactation. Birth and weaning weights for lambs and kids were recorded and analyzed, while milk production was measured daily and two milk samples were collected weekly from each dam and analyzed for milk composition. The results showed that treatment had no effect on birth weight, but it increased (P < 0.05) the weaning weight of both Awassi lambs and Shami kids. The treatment resulted in increased (P < 0.05) milk production of both Awassi ewes and Shami goats, with no effect on feed intake. The milk fat and protein percentages in treated Awassi ewes were higher (P < 0.05) than those for untreated ones, while no effect was observed on milk of treated goats. Total solids were higher (P < 0.05) in milk of treated groups of both species than the untreated ones. In conclusion, these results showed that using fibrolytic enzyme treatment improves growth of suckling lambs and kids and improves milk production, though the response may differ between sheep and goats. © 2003 Elsevier B.V. All rights reserved. Keywords: Fibrolytic enzyme; Sheep; Goat; Birth weight; Weaning weight; Milk production; Milk composition
1. Introduction Several studies using feed enzyme supplementation (FES) mixtures have reported that the beneficial effects of such enzymes are mainly due to increased fiber and total tract digestibility in ruminants (Rode et al., 1999; Yang et al., 1999). Earlier studies indicated such effects were most likely due to increased microbial population and microbial colonization of ru-
∗ Corresponding author. Tel.: +962-6-5355511; fax: +962-6-5300077. E-mail address: [email protected] (H. Titi).
men content following enzyme feeding (Beauchemin et al., 1999b, 2000). A great body of evidence showed that fibrolytic FES have also been beneficial when added in to high concentrate diets, and helped to overcome depression in fiber digestion in dairy cows (Yang et al., 1999) and in feedlot cattle (Krause et al., 1998). Addition of fibrolytic FES improved body weight gain of steers fed dry forages (Beauchemin et al., 1995), grazing steers (Feng et al., 1996), and of feedlot cattle (Beauchemin et al., 1999a; McAllister et al., 1999). The effects on dry matter feed intake, feed efficiency and growth rate were variable in feedlot cattle, and this variability was potentially related to the
0921-4488/$ – see front matter © 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0921-4488(03)00254-2
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H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
enzyme level in the supplementation (Beauchemin et al., 1999a, 2000; McAllister et al., 1999). McAllister et al. (1999) reported that feed efficiency in steers was significantly improved, while, it was only numerically improved by 10% due to supplemental enzyme feeding (Beauchemin et al., 1999a). Studies on FES to dairy cows showed that total milk yield, 3.5 and 4% fat corrected milk (FCM) were increased by around 7–15% (Beauchemin et al., 1999b; Rode et al., 1999; Lewis et al., 1999; Yang et al., 1999, 2000; Schingoethe et al., 1999). Such results showed that percentage of organic components in milk either increased due to increased available nutrients as a result of increased digestibility (Yang et al., 1999), remained unchanged (Lewis et al., 1999; Schingoethe et al., 1999; Yang et al., 2000) or dropped due to increased production and/or change in the acetic/propionic acids ratio in the rumen (Rode et al., 1999). FES was most efficient during early lactation or periods of negative energy balance and/or nutritional stress (Schingoethe et al., 1999; Rode et al., 1999; Yang et al., 2000), while no response was found when cows were treated during mid or late lactation stages (Schingoethe et al., 1999; Yang et al., 2000). There is no available literature on the effect of fibrolytic enzymes on the performance of sheep or goats. This experiment was initiated to study the effects of feeding cellulase enzyme mixture to ewes and goats (i) on birth and weaning weight of their lambs and kids and (ii) on milk production and composition of ewes and goats during the first 60 days of lactation.
2. Materials and methods Sixty Awassi ewes and 60 female Shami goats, which had given birth at least once before the experiment, were randomly divided into two treatment groups of equal number from each species. One group from each species was supplemented with enzyme mixture while the other group saved as control. At the beginning of the breeding season, all animals were treated with progesterone impregnated vaginal sponges for 14 days to synchronize estrus. Breeding dates were recorded for each ewe and doe. None returned to estrus for two consecutive cycles and Ultra-
Table 1 Ingredient composition of the concentrate part of the ration Ingredient Barley SBM Wheat bran Lime stone DCP Salt Vitamins and mineralsa Total
% 70 15 13 0.5 1.0 0.4 0.1 100
a
1 g contains: 7.5 MIU Vitamin A; 1.9 MIU Vitamin D3; 4 mg vitamin E (50%); 200 mg magnesium sulfate; 6.4 mg iron sulfate; 1.03 mg cobalt carbonate; 12.96 mg zinc sulfate; 4.84 mg manganese oxide; 0.79 mg potassium iodide; 0.23 mg sodium silinate and 12 mg copper sulfate in addition to antioxidant.
sound Doppler Pregnancy Scanner (Medata, Medata systems LTD., West Sussex, England) were used for pregnancy diagnosis. Animals were fed a concentrate mixture at a level of 1.35 kg per head per day dry matter and alfalfa hay ad lib, which should provide the nutritional requirements according to the NRC recommendations for sheep (National Research Council, 1985) and goats (National Research Council, 1989). Hay consumed for each group was recorded daily on individual basis and average total feed consumption of dry matter per head was calculated for each group. Percentage composition and proximate analysis of the concentrate ration are presented in Tables 1 and 2. A commercial dried powdered cellulase enzyme derived from Trichoderma group was offered at the level of 150 g enzyme/t of alfalfa hay consumed (Maxicel 200L® , George A. Jeffreys Company Inc., Salem, VA, USA). Treatment started from the beginning of the third month of pregnancy through the first 60 days of lactation. Amount of enzyme required for 1 t concentrate mix was diluted in Table 2 Proximate analysis (%) of ration fed (DM) Dry matter Crude protein Crude fiber NDF ADF Ash ME (MJ/Kg DM)
90.53 15.70 6.63 15.87 5.78 2.81 11.99
H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
a 500 g of ground corn, mixed with the micronutrient part of the ration and added to the concentrate mixture. Feed samples were collected at mixing time and analyzed according to AOAC (1984). Crude protein was determined by the macro Kjeldahl method for N analysis (N × 6.25) utilizing a 1031 Kjaltec analyzer unit (Tecator, Tecator AB, Hoganas, Sweden). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined according to Georing and Van Soest (1970). Birth weights and sex of lambs and kids were recorded. The weaning weight at 60 days old for lambs and kids were also recorded. The lambs and kids were left to suckle their dams until the date of weaning. Milk production for the first 60 days of lactation was estimated by measuring the production of 1 day once a week for each dam. Milk samples from every dam were collected twice a week, preserved with 2–3 drops of potassium dichromate and stored at −4 ◦ C for analysis. Samples were analyzed for composition of crude protein (Kjeldahl), fat (Gerber), SNF, and TS (Richardson, 1985). Protein values were determined by multiplying the nitrogen results by 6.38. Depended variables were analyzed using the General Linear model of SAS (1988). ANOVA was performed and treatment means were compared by using the LSD test. The model for birth and weaning weights analysis included the effects of treatment with species, birth and weaning weight, sex, number of kids per lambs, and all possible interactions. Yijkl = Si + Tj + Nk + Gl + Nk ∗ Si + Tj ∗ Gl +Si ∗ Tj + Nk ∗ Gl + Nk ∗ Tj + Si ∗ Gl +Si ∗ Tj ∗ Gl + Si ∗ Nk ∗ Gl + Si ∗ Nk ∗ Tj +Tj ∗ Nk ∗ Gl + Si ∗ Tj ∗ Nk ∗ Gl +residual error, where S = species, T = treatment, N = number of kids per lambs and G = gender of kids per lambs. Meanwhile, the model for dry matter intake and milk analysis was designed to determine the effect of the treatment on species, sex, and type of birth, and their interactions. Furthermore, week effect was introduced in the model in a repeated measure design to study treatment effect throughout the experiment. However, non significant effects were removed by stepwise methodology with the rep(TRT*SPP) was
139
used as an error term for testing. Yijk = Si + Tj + Si ∗ Tj + rep(Si ∗ Tj ) + Wk +Tj ∗ Wk + Si ∗ Wk + Sj ∗ Tj ∗ Wk +residual error, where S = species, T = treatment, W = week. 3. Results and discussion Mean values for birth and weaning weights of lambs and kids are presented in Table 3. There were no significant differences in average birth weight between treated and untreated groups for both lambs and kids. We are not aware of any results regarding the effect of fibrolytic FES for sheep and goats on milk production, birth or weaning weights for lambs and kids to compare our results. However, several studies reported that such enzymes had a beneficial effect in dairy cows during periods of nutritional stress (Lewis et al., 1999; Yang et al., 2000). Such conditions are prevailing during the late stages of pregnancy and suckling period in sheep and goats, especially with dams carrying twins. In the present experiment, however, the lack of significant differences in birth weight might be due to the feeding regime followed during the late pregnancy period. Animals were fed a diet of 15.70% CP and 11.99 MJ/Kg ME, in addition to free access to alfalfa hay (Table 2), which seems that the nutritional requirements were all met which might have masked the enzyme effect on the treated groups. Moreover, feeding easily digestible materials like alfalfa hay or the concentrate part (Table 1) could have resulted in no effect of the enzyme added. Yang et al. (2000) found no effect of fibrolytic FES when applied with highly digestible diets components to lactating dairy cows. Weaning weights of lambs from sheep fed FES were higher (P < 0.05) than those of the control group, while no differences were observed between treated and control groups of in goats. Weaning weight of males was also not different while those of treated single and twin male lambs and treated single male kids were higher (P < 0.05) than untreated ones. Meanwhile, female kids, single female lambs, and twin female kids of the treated groups had higher (P < 0.05) weaning weight than untreated ones.
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H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
Table 3 Means for birth weights and weaning weights of Awassi lambs and Shami goat kids of dams fed cellulase enzyme Awassi ewe Treated Birth weight (Kg) Male Single Twin
± ± ± ±
n
Untreated ± ± ± ±
n
Treated ± ± ± ±
n
Untreated ± ± ± ±
n
0.13 0.20 0.23 0.32
38 21 12 9
4.41 4.28 4.47 4.41
0.28 0.21 0.23 0.33
36 16 10 6
4.11 4.37 4.54 4.10
0.14 0.16 0.25 0.26
40 25 15 10
3.88 4.05 4.10 4.00
0.14 0.16 0.28 0.16
46 22 9 13
Female Single Twin
4.13 ± 0.19 4.35 ± 0.28 3.92 ± 0.25
17 10 7
4.11 ± 0.19 4.43 ± 0.18 3.87 ± 0.30
20 14 6
3.79 ± 0.23 3.70 ± 0.40 3.69 ± 0.32
15 5 10
3.72 ± 0.23 3.77 ± 0.33 3.67 ± 0.33
24 9 15
Single Twin
4.30 ± 0.18 4.17 ± 0.21
22 16
4.45 ± 0.15 4.15 ± 0.23
24 12
4.12 ± 0.24 4.04 ± 0.17
20 20
4.45 ± 0.15 3.83 ± 0.18
18 28
± ± ± ±
0.50 ac 0.52 c 0.58 ac 0.50ac
38 21 12 9
14.20 14.45 14.50 14.33
± ± ± ±
36 16 10 6
13.96 14.59 14.70 14.46
± ± ± ±
0.75 d 0.41d 0.23 ad 0.30 d
40 25 15 10
13.68 14.35 14.47 14.21
± ± ± ±
46 22 9 13
Female Single Twin
14.53 ± 0.47 c 15.67 ± 0.47 ac 13.40 ± 0.63
17 10 7
13.95 ± 0.36 c 14.87 ± 0.71 bc 13.12 ± 0.49
20 14 6
13.33 ± 0.57 ad 13.50 ± 0.82 d 13.16 ± 0.82 a
15 5 10
12.90 ± 0.59 bd 13.20 ± 1.00 d 12.80 ± 0.63 b
24 9 15
Single Twin
15.79 ± 0.37 ac 14.45 ± 0.51 ac
22 16
14.69 ± 0.45 b 13.78 ± 0.44 b
24 12
14.10 ± 0.54 d 13.83 ± 0.45 ad
20 20
13.85 ± 0.59 13.51 ± 0.41 b
18 28
Weaning weight (Kg) Male Single Twin
4.20 4.34 4.25 4.43
Shami goat
15.14 15.70 15.91 15.50
0.34 b 0.58 0.51 b 0.46 b
0.64 0.71 0.10 b 0.24
Means with different letters (a and b) in the same line within the same species are different (P < 0.05). Means with different letters (c and d) in the same line within the same treatment are different (P < 0.05).
Total single lambs from treated sheep had higher (P < 0.05) weaning weight than those of the control, while no differences were obtained between treated and untreated goat singles. Weaning weights of twins from treated groups were higher (P < 0.05) than controls for both sheep and goats. Results for dry matter intake, milk production, and milk composition are presented in Table 4. No differences in dry matter intake throughout the experiment
were observed between treated and untreated groups in both species. Intake was not affected by exogenous FES. The lack of effect of enzyme supplementation obtained was consistent with previous studies which found no effect of adding different fibrolytic FES on feed intake of lactating and feedlot cattle fed forage, concentrate or total mixed rations (Beauchemin et al., 1999a,b; Rode et al., 1999; Yang et al., 2000; Zheng et al., 2000).
Table 4 Dry matter intake, milk production and milk composition of Awassi ewes and Shami goats fed cellulase enzymes Awassi ewes Treated ± 0.10
Dry matter intake (Kg per head per day) Concentrate mix Milk production (Kg) Fat (%) Protein (%)
2.28 1.35 50.21 7.33 6.11
Total solids (%)
18.33 ± 0.54 ac
± 3.1 ad ± 0.39 ac ± 0.26 ac
Shami goats Untreated 2.30 1.35 45.76 6.49 5.44
± 0.11 ± 3.09 bd ± 0.40 bc ± 0.34 bc
16.91 ± 0.59 bc
Treated 2.19 1.35 61.23 4.70 4.61
± 0.09 ± 3.3 ac ± 3.80 d ± 0.36 d
13.40 ± 0.43 ad
Untreated 2.23 1.35 54.49 4.09 3.98
± 0.12 ± 3.10 bc ± 0.39 d ± 0.39 d
12.53 ± 0.55 bd
Means with different letters (a and b) in the same line within the same species are different (P < 0.05). Means with different letters (c and d) in the same line within the same treatment are different (P < 0.05).
H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
In contrast, others have reported increased feed intake of dairy cows and feedlot cattle supplemented with cellulase enzymes (Lewis et al., 1999; McAllister et al., 1999). Meanwhile, Beauchemin et al. (1999a), Beauchemin et al. (2000) and Yang et al. (2000) reported that the effects of exogenous fibrolytic enzymes on dry matter intake appear to differ among enzyme products, and some, but not all enzyme mixtures may increase feed intake. Results for milk production for both species are presented in Table 4. ANOVA results showed that, in both species, milk production was increased (P < 0.05) by 10–12% affected by the FES treatment with no effect of sex, species or any of their interaction. The rate of response varied between the two species. Awassi ewes fed the FES diet produced higher (P < 0.05) milk than those of the unsupplemented ewes. Similarly, enzyme fed goats produced higher (P < 0.05) milk than control goats. The rate of increase in milk production reached around 10 and 12% compared to the control ewes and goats, respectively. We are not aware of any published results concerning the effect of fibrolytic FES on lactating ewes or goats. However, nearly all of the available published literature revealed increased milk production of dairy cows fed enzyme treated feeds especially in early lactation periods when cows were in negative energy balance (Beauchemin et al., 1999b; Lewis et al., 1999; Rode et al., 1999; Schingoethe et al., 1999; Yang et al., 1999, 2000; Zheng et al., 2000). As no difference were obtained in feed intake between treated and untreated groups, these results indicate that the effect of exogenous fibrolytic enzymes was related to factors other than feed intake. Although not measured in this study, improved milk production in both species could be attributed to increased total tract digestibility of the dry matter fed. These findings are in line with other studies which reported that the improvement in milk production of FES dairy cows fed alfalfa hay or total mixed ration was a direct result of increased feed digestibility rather than to increased feed intake (Beauchemin et al., 1999a,b; Yang et al., 2000), which lead to increased energy available for milk production (Lewis et al., 1999; Yang et al., 1999). Response in milk composition was variable (Table 4). Treated Awassi ewes had higher (P < 0.05) milk fat and protein when compared with the control ewes, while, no differences were found between
141
treated and untreated goat groups. However, treated goats had numerically higher values for fat and protein percentages than the control group. In both species, mean values for total solids were higher (P < 0.05) in the treated groups compared to untreated groups for sheep and goats. The improvement in sheep is consistent with similar studies with milk of dairy cows fed fibrolytic FES (Lewis et al., 1999; Schingoethe et al., 1999; Zheng et al., 2000), though, Beauchemin et al. (2000) reported improvement in milk protein percent only while Rode et al. (1999) found a drop in milk fat and protein composition. The higher milk components of FES treated animals than controls were most likely due to more nutrients that were available as a result of improved digestibility following enzyme treatment (Yang et al., 1999). In contrast, some studies reported a drop in milk fat and protein composition of dairy cows fed fibrolytic enzyme treated diet (Rode et al., 1999). The response in goats was similar to that in sheep though it was significant only for total solid. Results for goats are in line with those of other studies which reported no effect of feeding enzyme treated forages to dairy cows on the concentration of all or some of the milk components (Lewis et al., 1999; Beauchemin et al., 2000). However, content of total solids was significant due to the cumulative effect of cellulase treatment on the fat and protein concentrations as both were numerically higher for the treated group compared to the control group. Likewise, Zheng et al. (2000) reported that total solids in milk of fibrolytic enzyme treated dairy cows tended to be higher reflecting the higher fat and protein yields. On the other hand, the lack of differences in percentage milk composition between treated and untreated goats, compared to those of sheep, might be due to that the level of increase in milk production of treated goats was higher than that of treated ewes. Due to the negative relationship between milk yield and composition, it seems that as the level of milk production increased the effect of FES on milk composition tended to be low. Some studies in diary cows fed fibrolytic FES showed inverse relation between milk yield and milk components (Rode et al., 1999). Thus, the differences in milk yield between the sheep and goat may explain the differences in milk composition.
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Fig. 1. Treatment week interaction of Awassi ewes and Shami goats fed cellulase enzyme. The symbol (∗) indicates means within the same species are different (P < 0.05).
Fig. 1 shows the interaction of cellulase treatment by week for both species. Analysis of variance showed a significant (P < 0.05) interaction of treatment by week. No differences in weekly milk production were observed during the first 2 weeks (Fig. 1). However, treated groups of in both species produced higher (P < 0.05) milk during weeks 3–5 of lactation when animals were in at peak lactation and disappeared towards the end of the treatment period (weeks 6–8, Fig. 1). It seems that FES treatment is effective if enzymes were added during periods of early lactation when dams are in negative energy balance, while no or very little response was obtained if enzymes were added in mid or late lactation. Similar results were obtained by Rode et al. (1999), Schingoethe et al. (1999), and Beauchemin et al. (2000) who reported that best benefits obtained when FES was given to early lactating cows while treating cows past their peak production was ineffective. In conclusions, FES treatment may be used to improve weaning weight of lambs or kids and the improvement may be attributed to improvement in milk quantity and quality. This response was particularly evident in twins. The lack of differences between treated and untreated groups in the overall weaning
weight of goat kids or in the goat singles may be related to higher milk production in the goats, which probably fulfilled the kids’ requirements. Improved milk production occurred without apparent changes in feed intake, which might suggest that improvement was through improved feed utilization.
References Association of Official Analytical Chemists, 1984. Official Methods of Analysis, 13th ed. AOAC, Washington, DC. Beauchemin, K.A., Rode, L.M., Sewalt, V.J., 1995. Fibrolytic enzymes increase fiber digestibility and growth rate of steers fed dry forages. Can. J. Anim. Sci. 75, 641. Beauchemin, K.A., Yang, W.Z., Rode, L.M., 1999a. Effects og grain source and enzyme additive on site and extent of nutrient digestion in dairy cows. J. Dairy Sci. 82, 378. Beauchemin, K.A., Rode, L.M., Karren, D., 1999b. Use of feed enzymes in feedlot finishing diets. Can. J. Anim. Sci. 79, 243. Beauchemin, K.A., Rode, L.M., Macaw, M., Morgan, DP, Kampent, R., 2000. Evaluation of a nonstarch polysaccharidase feed enzyme in dairy cow diets. J. Dairy Sci. 83, 543. Feng, P., Hunt, C.W., Pritchard, G.T., Julien, W.E., 1996. Effect of enzyme preparations and in vitro degradation and in vitro digestive characteristics of mature cool-season grass forage in beef steers. J. Anim. Sci. 74, 1349.
H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143 Georing, H.K., Van Soest, P.J., 1970. Forage Fiber Analysis. Agriculture Handbook No. 379. Agriculture Research Service, USDA, Washington, DC. Krause, M., Beauchemin, K.A., Rode, L.M., Farr, B.I., Norgaard, P., 1998. Fibrolytic enzyme treatment of barley grain and source of forage in high-grain diets fed to growing cattle. J. Anim. Sci. 76, 2912. Lewis, G.E., Sanchez, W.K., Hunt, C.W., Guy, M.A., Pritchard, G.T., Swanson, B.I., Treacher, R.J., 1999. Effect of direct-fed fibrolytic enzymes on lactational performance of dairy cows. J. Dairy Sci. 82, 611. McAllister, T.A., Oosting, S.J., Popp, J.D., Mir, Z., Yanke, L.J., Hristov, A.N., Treacher, R.J., Cheng, K.J., 1999. Effect of exogenous enzyme on digestibility of barley silage and growth performance of feedlot cattle. Can. J. Anim. Sci. 79, 353. National Research Council, 1985. Nutrient Requirements of Domestic Animals: Nutrient Requirements of Sheep. Nutritional Academy of Science, National Academy Presses, Washington, DC. National Research Council, 1989. Nutrient Requirements of Domestic Animals: Nutrient Requirements of Goats. Nutritional Academy of Science, National Academy Presses, Washington, DC.
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Richardson, G.H., 1985. Standard Methods for the Examination of Dairy Products, 15th ed. American Public Health Association, Washington, DC. Rode, L.M., Yang, W.Z., Beauchemin, K.A., 1999. Fibrolytic enzyme supplements for dairy cows in early lactation. J. Dairy Sci. 82, 2121. SAS/STAT User’s Guide, 1988. Release 6.03 ed. SAS Institute Inc., Cary, NC, USA. Schingoethe, D.J., Stegeman, G.A., Treacher, R.J., 1999. Response of lactating dairy cows to a cellulase and xylanase enzyme mixture applied to forages at the time of feeding. J. Dairy Sci. 82, 996. Yang, W.Z., Beauchemin, K.A., Rode, L.M., 1999. Effects of an enzyme feed additive on extent of digestion and milk production of lactating dairy cows. J. Dairy Sci. 82, 391. Yang, W.Z., Beauchemin, K.A., Rode, L.M., 2000. A comparison of methods of adding fibrolytic enzymes to lactating cow diets. J. Dairy Sci. 83, 2512. Zheng, W., Schingoethe, D.J., Stegeman, G.A., Hippen, A.R., Treachert, R.J., 2000. Determination of when during the lactation cycle to start feeding a cellulase and xylanase enzyme mixture to dairy cows. J. Dairy Sci. 83, 2319.
Effect of feeding cellulase enzyme on productive responses of pregnant and lactating ewes and goats H. Titi∗ , W.F. Lubbadeh Department of Animal Production, Faculty of Agriculture, University of Jordan, Amman 11942, Jordan Received 21 August 2002; received in revised form 23 April 2003; accepted 29 June 2003
Abstract An experiment was conducted to study the effect of supplementing the diets of Awassi ewes and Shami goats with exogenous fibrolytic enzyme on birth and weaning weight of lambs and kids and on milk production of dams in the first 60 days of lactation. Two groups of ewes (30 each) and two groups of goats (30 each) were used. One group of each species was supplemented with the fibrolytic enzyme during the last 2 months of pregnancy and the first 60 days of lactation. Birth and weaning weights for lambs and kids were recorded and analyzed, while milk production was measured daily and two milk samples were collected weekly from each dam and analyzed for milk composition. The results showed that treatment had no effect on birth weight, but it increased (P < 0.05) the weaning weight of both Awassi lambs and Shami kids. The treatment resulted in increased (P < 0.05) milk production of both Awassi ewes and Shami goats, with no effect on feed intake. The milk fat and protein percentages in treated Awassi ewes were higher (P < 0.05) than those for untreated ones, while no effect was observed on milk of treated goats. Total solids were higher (P < 0.05) in milk of treated groups of both species than the untreated ones. In conclusion, these results showed that using fibrolytic enzyme treatment improves growth of suckling lambs and kids and improves milk production, though the response may differ between sheep and goats. © 2003 Elsevier B.V. All rights reserved. Keywords: Fibrolytic enzyme; Sheep; Goat; Birth weight; Weaning weight; Milk production; Milk composition
1. Introduction Several studies using feed enzyme supplementation (FES) mixtures have reported that the beneficial effects of such enzymes are mainly due to increased fiber and total tract digestibility in ruminants (Rode et al., 1999; Yang et al., 1999). Earlier studies indicated such effects were most likely due to increased microbial population and microbial colonization of ru-
∗ Corresponding author. Tel.: +962-6-5355511; fax: +962-6-5300077. E-mail address: [email protected] (H. Titi).
men content following enzyme feeding (Beauchemin et al., 1999b, 2000). A great body of evidence showed that fibrolytic FES have also been beneficial when added in to high concentrate diets, and helped to overcome depression in fiber digestion in dairy cows (Yang et al., 1999) and in feedlot cattle (Krause et al., 1998). Addition of fibrolytic FES improved body weight gain of steers fed dry forages (Beauchemin et al., 1995), grazing steers (Feng et al., 1996), and of feedlot cattle (Beauchemin et al., 1999a; McAllister et al., 1999). The effects on dry matter feed intake, feed efficiency and growth rate were variable in feedlot cattle, and this variability was potentially related to the
0921-4488/$ – see front matter © 2003 Elsevier B.V. All rights reserved. doi:10.1016/S0921-4488(03)00254-2
138
H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
enzyme level in the supplementation (Beauchemin et al., 1999a, 2000; McAllister et al., 1999). McAllister et al. (1999) reported that feed efficiency in steers was significantly improved, while, it was only numerically improved by 10% due to supplemental enzyme feeding (Beauchemin et al., 1999a). Studies on FES to dairy cows showed that total milk yield, 3.5 and 4% fat corrected milk (FCM) were increased by around 7–15% (Beauchemin et al., 1999b; Rode et al., 1999; Lewis et al., 1999; Yang et al., 1999, 2000; Schingoethe et al., 1999). Such results showed that percentage of organic components in milk either increased due to increased available nutrients as a result of increased digestibility (Yang et al., 1999), remained unchanged (Lewis et al., 1999; Schingoethe et al., 1999; Yang et al., 2000) or dropped due to increased production and/or change in the acetic/propionic acids ratio in the rumen (Rode et al., 1999). FES was most efficient during early lactation or periods of negative energy balance and/or nutritional stress (Schingoethe et al., 1999; Rode et al., 1999; Yang et al., 2000), while no response was found when cows were treated during mid or late lactation stages (Schingoethe et al., 1999; Yang et al., 2000). There is no available literature on the effect of fibrolytic enzymes on the performance of sheep or goats. This experiment was initiated to study the effects of feeding cellulase enzyme mixture to ewes and goats (i) on birth and weaning weight of their lambs and kids and (ii) on milk production and composition of ewes and goats during the first 60 days of lactation.
2. Materials and methods Sixty Awassi ewes and 60 female Shami goats, which had given birth at least once before the experiment, were randomly divided into two treatment groups of equal number from each species. One group from each species was supplemented with enzyme mixture while the other group saved as control. At the beginning of the breeding season, all animals were treated with progesterone impregnated vaginal sponges for 14 days to synchronize estrus. Breeding dates were recorded for each ewe and doe. None returned to estrus for two consecutive cycles and Ultra-
Table 1 Ingredient composition of the concentrate part of the ration Ingredient Barley SBM Wheat bran Lime stone DCP Salt Vitamins and mineralsa Total
% 70 15 13 0.5 1.0 0.4 0.1 100
a
1 g contains: 7.5 MIU Vitamin A; 1.9 MIU Vitamin D3; 4 mg vitamin E (50%); 200 mg magnesium sulfate; 6.4 mg iron sulfate; 1.03 mg cobalt carbonate; 12.96 mg zinc sulfate; 4.84 mg manganese oxide; 0.79 mg potassium iodide; 0.23 mg sodium silinate and 12 mg copper sulfate in addition to antioxidant.
sound Doppler Pregnancy Scanner (Medata, Medata systems LTD., West Sussex, England) were used for pregnancy diagnosis. Animals were fed a concentrate mixture at a level of 1.35 kg per head per day dry matter and alfalfa hay ad lib, which should provide the nutritional requirements according to the NRC recommendations for sheep (National Research Council, 1985) and goats (National Research Council, 1989). Hay consumed for each group was recorded daily on individual basis and average total feed consumption of dry matter per head was calculated for each group. Percentage composition and proximate analysis of the concentrate ration are presented in Tables 1 and 2. A commercial dried powdered cellulase enzyme derived from Trichoderma group was offered at the level of 150 g enzyme/t of alfalfa hay consumed (Maxicel 200L® , George A. Jeffreys Company Inc., Salem, VA, USA). Treatment started from the beginning of the third month of pregnancy through the first 60 days of lactation. Amount of enzyme required for 1 t concentrate mix was diluted in Table 2 Proximate analysis (%) of ration fed (DM) Dry matter Crude protein Crude fiber NDF ADF Ash ME (MJ/Kg DM)
90.53 15.70 6.63 15.87 5.78 2.81 11.99
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a 500 g of ground corn, mixed with the micronutrient part of the ration and added to the concentrate mixture. Feed samples were collected at mixing time and analyzed according to AOAC (1984). Crude protein was determined by the macro Kjeldahl method for N analysis (N × 6.25) utilizing a 1031 Kjaltec analyzer unit (Tecator, Tecator AB, Hoganas, Sweden). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined according to Georing and Van Soest (1970). Birth weights and sex of lambs and kids were recorded. The weaning weight at 60 days old for lambs and kids were also recorded. The lambs and kids were left to suckle their dams until the date of weaning. Milk production for the first 60 days of lactation was estimated by measuring the production of 1 day once a week for each dam. Milk samples from every dam were collected twice a week, preserved with 2–3 drops of potassium dichromate and stored at −4 ◦ C for analysis. Samples were analyzed for composition of crude protein (Kjeldahl), fat (Gerber), SNF, and TS (Richardson, 1985). Protein values were determined by multiplying the nitrogen results by 6.38. Depended variables were analyzed using the General Linear model of SAS (1988). ANOVA was performed and treatment means were compared by using the LSD test. The model for birth and weaning weights analysis included the effects of treatment with species, birth and weaning weight, sex, number of kids per lambs, and all possible interactions. Yijkl = Si + Tj + Nk + Gl + Nk ∗ Si + Tj ∗ Gl +Si ∗ Tj + Nk ∗ Gl + Nk ∗ Tj + Si ∗ Gl +Si ∗ Tj ∗ Gl + Si ∗ Nk ∗ Gl + Si ∗ Nk ∗ Tj +Tj ∗ Nk ∗ Gl + Si ∗ Tj ∗ Nk ∗ Gl +residual error, where S = species, T = treatment, N = number of kids per lambs and G = gender of kids per lambs. Meanwhile, the model for dry matter intake and milk analysis was designed to determine the effect of the treatment on species, sex, and type of birth, and their interactions. Furthermore, week effect was introduced in the model in a repeated measure design to study treatment effect throughout the experiment. However, non significant effects were removed by stepwise methodology with the rep(TRT*SPP) was
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used as an error term for testing. Yijk = Si + Tj + Si ∗ Tj + rep(Si ∗ Tj ) + Wk +Tj ∗ Wk + Si ∗ Wk + Sj ∗ Tj ∗ Wk +residual error, where S = species, T = treatment, W = week. 3. Results and discussion Mean values for birth and weaning weights of lambs and kids are presented in Table 3. There were no significant differences in average birth weight between treated and untreated groups for both lambs and kids. We are not aware of any results regarding the effect of fibrolytic FES for sheep and goats on milk production, birth or weaning weights for lambs and kids to compare our results. However, several studies reported that such enzymes had a beneficial effect in dairy cows during periods of nutritional stress (Lewis et al., 1999; Yang et al., 2000). Such conditions are prevailing during the late stages of pregnancy and suckling period in sheep and goats, especially with dams carrying twins. In the present experiment, however, the lack of significant differences in birth weight might be due to the feeding regime followed during the late pregnancy period. Animals were fed a diet of 15.70% CP and 11.99 MJ/Kg ME, in addition to free access to alfalfa hay (Table 2), which seems that the nutritional requirements were all met which might have masked the enzyme effect on the treated groups. Moreover, feeding easily digestible materials like alfalfa hay or the concentrate part (Table 1) could have resulted in no effect of the enzyme added. Yang et al. (2000) found no effect of fibrolytic FES when applied with highly digestible diets components to lactating dairy cows. Weaning weights of lambs from sheep fed FES were higher (P < 0.05) than those of the control group, while no differences were observed between treated and control groups of in goats. Weaning weight of males was also not different while those of treated single and twin male lambs and treated single male kids were higher (P < 0.05) than untreated ones. Meanwhile, female kids, single female lambs, and twin female kids of the treated groups had higher (P < 0.05) weaning weight than untreated ones.
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Table 3 Means for birth weights and weaning weights of Awassi lambs and Shami goat kids of dams fed cellulase enzyme Awassi ewe Treated Birth weight (Kg) Male Single Twin
± ± ± ±
n
Untreated ± ± ± ±
n
Treated ± ± ± ±
n
Untreated ± ± ± ±
n
0.13 0.20 0.23 0.32
38 21 12 9
4.41 4.28 4.47 4.41
0.28 0.21 0.23 0.33
36 16 10 6
4.11 4.37 4.54 4.10
0.14 0.16 0.25 0.26
40 25 15 10
3.88 4.05 4.10 4.00
0.14 0.16 0.28 0.16
46 22 9 13
Female Single Twin
4.13 ± 0.19 4.35 ± 0.28 3.92 ± 0.25
17 10 7
4.11 ± 0.19 4.43 ± 0.18 3.87 ± 0.30
20 14 6
3.79 ± 0.23 3.70 ± 0.40 3.69 ± 0.32
15 5 10
3.72 ± 0.23 3.77 ± 0.33 3.67 ± 0.33
24 9 15
Single Twin
4.30 ± 0.18 4.17 ± 0.21
22 16
4.45 ± 0.15 4.15 ± 0.23
24 12
4.12 ± 0.24 4.04 ± 0.17
20 20
4.45 ± 0.15 3.83 ± 0.18
18 28
± ± ± ±
0.50 ac 0.52 c 0.58 ac 0.50ac
38 21 12 9
14.20 14.45 14.50 14.33
± ± ± ±
36 16 10 6
13.96 14.59 14.70 14.46
± ± ± ±
0.75 d 0.41d 0.23 ad 0.30 d
40 25 15 10
13.68 14.35 14.47 14.21
± ± ± ±
46 22 9 13
Female Single Twin
14.53 ± 0.47 c 15.67 ± 0.47 ac 13.40 ± 0.63
17 10 7
13.95 ± 0.36 c 14.87 ± 0.71 bc 13.12 ± 0.49
20 14 6
13.33 ± 0.57 ad 13.50 ± 0.82 d 13.16 ± 0.82 a
15 5 10
12.90 ± 0.59 bd 13.20 ± 1.00 d 12.80 ± 0.63 b
24 9 15
Single Twin
15.79 ± 0.37 ac 14.45 ± 0.51 ac
22 16
14.69 ± 0.45 b 13.78 ± 0.44 b
24 12
14.10 ± 0.54 d 13.83 ± 0.45 ad
20 20
13.85 ± 0.59 13.51 ± 0.41 b
18 28
Weaning weight (Kg) Male Single Twin
4.20 4.34 4.25 4.43
Shami goat
15.14 15.70 15.91 15.50
0.34 b 0.58 0.51 b 0.46 b
0.64 0.71 0.10 b 0.24
Means with different letters (a and b) in the same line within the same species are different (P < 0.05). Means with different letters (c and d) in the same line within the same treatment are different (P < 0.05).
Total single lambs from treated sheep had higher (P < 0.05) weaning weight than those of the control, while no differences were obtained between treated and untreated goat singles. Weaning weights of twins from treated groups were higher (P < 0.05) than controls for both sheep and goats. Results for dry matter intake, milk production, and milk composition are presented in Table 4. No differences in dry matter intake throughout the experiment
were observed between treated and untreated groups in both species. Intake was not affected by exogenous FES. The lack of effect of enzyme supplementation obtained was consistent with previous studies which found no effect of adding different fibrolytic FES on feed intake of lactating and feedlot cattle fed forage, concentrate or total mixed rations (Beauchemin et al., 1999a,b; Rode et al., 1999; Yang et al., 2000; Zheng et al., 2000).
Table 4 Dry matter intake, milk production and milk composition of Awassi ewes and Shami goats fed cellulase enzymes Awassi ewes Treated ± 0.10
Dry matter intake (Kg per head per day) Concentrate mix Milk production (Kg) Fat (%) Protein (%)
2.28 1.35 50.21 7.33 6.11
Total solids (%)
18.33 ± 0.54 ac
± 3.1 ad ± 0.39 ac ± 0.26 ac
Shami goats Untreated 2.30 1.35 45.76 6.49 5.44
± 0.11 ± 3.09 bd ± 0.40 bc ± 0.34 bc
16.91 ± 0.59 bc
Treated 2.19 1.35 61.23 4.70 4.61
± 0.09 ± 3.3 ac ± 3.80 d ± 0.36 d
13.40 ± 0.43 ad
Untreated 2.23 1.35 54.49 4.09 3.98
± 0.12 ± 3.10 bc ± 0.39 d ± 0.39 d
12.53 ± 0.55 bd
Means with different letters (a and b) in the same line within the same species are different (P < 0.05). Means with different letters (c and d) in the same line within the same treatment are different (P < 0.05).
H. Titi, W.F. Lubbadeh / Small Ruminant Research 52 (2004) 137–143
In contrast, others have reported increased feed intake of dairy cows and feedlot cattle supplemented with cellulase enzymes (Lewis et al., 1999; McAllister et al., 1999). Meanwhile, Beauchemin et al. (1999a), Beauchemin et al. (2000) and Yang et al. (2000) reported that the effects of exogenous fibrolytic enzymes on dry matter intake appear to differ among enzyme products, and some, but not all enzyme mixtures may increase feed intake. Results for milk production for both species are presented in Table 4. ANOVA results showed that, in both species, milk production was increased (P < 0.05) by 10–12% affected by the FES treatment with no effect of sex, species or any of their interaction. The rate of response varied between the two species. Awassi ewes fed the FES diet produced higher (P < 0.05) milk than those of the unsupplemented ewes. Similarly, enzyme fed goats produced higher (P < 0.05) milk than control goats. The rate of increase in milk production reached around 10 and 12% compared to the control ewes and goats, respectively. We are not aware of any published results concerning the effect of fibrolytic FES on lactating ewes or goats. However, nearly all of the available published literature revealed increased milk production of dairy cows fed enzyme treated feeds especially in early lactation periods when cows were in negative energy balance (Beauchemin et al., 1999b; Lewis et al., 1999; Rode et al., 1999; Schingoethe et al., 1999; Yang et al., 1999, 2000; Zheng et al., 2000). As no difference were obtained in feed intake between treated and untreated groups, these results indicate that the effect of exogenous fibrolytic enzymes was related to factors other than feed intake. Although not measured in this study, improved milk production in both species could be attributed to increased total tract digestibility of the dry matter fed. These findings are in line with other studies which reported that the improvement in milk production of FES dairy cows fed alfalfa hay or total mixed ration was a direct result of increased feed digestibility rather than to increased feed intake (Beauchemin et al., 1999a,b; Yang et al., 2000), which lead to increased energy available for milk production (Lewis et al., 1999; Yang et al., 1999). Response in milk composition was variable (Table 4). Treated Awassi ewes had higher (P < 0.05) milk fat and protein when compared with the control ewes, while, no differences were found between
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treated and untreated goat groups. However, treated goats had numerically higher values for fat and protein percentages than the control group. In both species, mean values for total solids were higher (P < 0.05) in the treated groups compared to untreated groups for sheep and goats. The improvement in sheep is consistent with similar studies with milk of dairy cows fed fibrolytic FES (Lewis et al., 1999; Schingoethe et al., 1999; Zheng et al., 2000), though, Beauchemin et al. (2000) reported improvement in milk protein percent only while Rode et al. (1999) found a drop in milk fat and protein composition. The higher milk components of FES treated animals than controls were most likely due to more nutrients that were available as a result of improved digestibility following enzyme treatment (Yang et al., 1999). In contrast, some studies reported a drop in milk fat and protein composition of dairy cows fed fibrolytic enzyme treated diet (Rode et al., 1999). The response in goats was similar to that in sheep though it was significant only for total solid. Results for goats are in line with those of other studies which reported no effect of feeding enzyme treated forages to dairy cows on the concentration of all or some of the milk components (Lewis et al., 1999; Beauchemin et al., 2000). However, content of total solids was significant due to the cumulative effect of cellulase treatment on the fat and protein concentrations as both were numerically higher for the treated group compared to the control group. Likewise, Zheng et al. (2000) reported that total solids in milk of fibrolytic enzyme treated dairy cows tended to be higher reflecting the higher fat and protein yields. On the other hand, the lack of differences in percentage milk composition between treated and untreated goats, compared to those of sheep, might be due to that the level of increase in milk production of treated goats was higher than that of treated ewes. Due to the negative relationship between milk yield and composition, it seems that as the level of milk production increased the effect of FES on milk composition tended to be low. Some studies in diary cows fed fibrolytic FES showed inverse relation between milk yield and milk components (Rode et al., 1999). Thus, the differences in milk yield between the sheep and goat may explain the differences in milk composition.
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Fig. 1. Treatment week interaction of Awassi ewes and Shami goats fed cellulase enzyme. The symbol (∗) indicates means within the same species are different (P < 0.05).
Fig. 1 shows the interaction of cellulase treatment by week for both species. Analysis of variance showed a significant (P < 0.05) interaction of treatment by week. No differences in weekly milk production were observed during the first 2 weeks (Fig. 1). However, treated groups of in both species produced higher (P < 0.05) milk during weeks 3–5 of lactation when animals were in at peak lactation and disappeared towards the end of the treatment period (weeks 6–8, Fig. 1). It seems that FES treatment is effective if enzymes were added during periods of early lactation when dams are in negative energy balance, while no or very little response was obtained if enzymes were added in mid or late lactation. Similar results were obtained by Rode et al. (1999), Schingoethe et al. (1999), and Beauchemin et al. (2000) who reported that best benefits obtained when FES was given to early lactating cows while treating cows past their peak production was ineffective. In conclusions, FES treatment may be used to improve weaning weight of lambs or kids and the improvement may be attributed to improvement in milk quantity and quality. This response was particularly evident in twins. The lack of differences between treated and untreated groups in the overall weaning
weight of goat kids or in the goat singles may be related to higher milk production in the goats, which probably fulfilled the kids’ requirements. Improved milk production occurred without apparent changes in feed intake, which might suggest that improvement was through improved feed utilization.
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