Development of Intestinal Disaccharidase Activities in Nursing Calves

Size: px

Start display at page:

Download "Development of Intestinal Disaccharidase Activities in Nursing Calves"

Homer Booker
6 years ago
Views:

1 Development of Intestinal Disaccharidase Activities in Nursing Calves Toshikazu MIYASHIGE and Shigefusa YAHATA Chugoku National Agricultural Experiment Station, Oda-shi (Received July 5,1979) Abstract 1. The disaccharidase activities of mucosal homogenates from the small intestine, caecum and colon of cattle were observed. 2. In nursing calves, lactase, cellobiase and trehalase activities were located mainly in the upper part of the small intestine, whereas maltase and isomaltase activities were more uniformly distributed along the small intestine and were also found in the caecum and colon. Intestinal cellobiase, trehalase, maltase and isomaltase activities were very low compared with lactase activity and intestinal sucrase activity was not detected. 3. Intestinal lactase, cellobiase and trehalase activities were the highest at birth and decreased rapidly with age. These activities increased during the late fetal period. On the other hand, intestinal maltase and isomaltase activities showed no increase during the late fetal period and nursing period of 6 months, 4. Relatively low ph values of intestinal contents of , which were close to optimum ph of intestinal disaccharidases, were observed in the upper part of the small intestine. 5. In adult cattle, intestinal lactase and cellobiase activities were found but only with negligible levels. Intestinal trehalase activity was also found with low level. Intestinal maltase and isomaltase activities were found to be similar levels to those found in calves. Intestinal sucrase activity was not detected. 6. Based on the results of disaccharidase activities and tissue weights, overall utilizable quantities of lactose and maltose by small intestinal mucosa in nursing calves were estimated, Jap. J. Zootech. Sci., 51(1):58-68,1980 It has been known that intestinal lactase activity of calves is high in the early weeks of life but decreases with age1-3). And it has been also shown that intestinal maltase activity is very low and shows no increase with age, and intestinal activity is absent1,2,4). From these observations, sucrase it is seemed that young calves are able to utilize lactose but possess poor ability to utilize maltose and starch, and are unable to utilize sucrose. However these activities reported are expressed either as units per mg protein of mucosa or as units per g dry tissue, and therefore do not necessarily imply the digestive capacity of these carbohydrates by intestinal mucosa. The capacity seems to also depend largely on increase of tissue weight and on change of distribution of activity along the small intestine, and may be influenced by change of luminal ph. However little detailed observation including these factors has been reported of development of disaccharidase activities in calves. In the present study, we investigated the sequential development of mucosal disaccharidase activities and of patterns of their distributions in the small intestine, caecum and colon of nursing calves with intestinal tissue weights and content ph's, Jap. J. Zootech. Sci., 51(1):

2 Intestinal Disaccharidase of Nursing Calf and estimated mucosa. the overall utilizable quantities of lactose and maltose by small intestinal Materials and Methods Thirteen calves and three fetuses of Japanese Black Cattle were used. The calves were kept with their dams and nursed voluntarily. They were given concentrates and hay ad libitum from 1-4 weeks of age. And two or three calves were killed at 4,6-10,12 and 26 weeks of age by bleeding out from the carotid artery after local infusion of procaine hydrochloride solution. Two newborn calves were also killed within 24 hours after birth. Of these calves, one nursed about a day, but the other removed from its dam immediately after birth. The fetuses were removed from their dams which were normally fed and killed by electrical stunning and bleeding at 5.5 and 7 months after conception. The mean body weights of experimental animals at slaughter are shown in Table 1. Two adult cows and two fattenning steers were included in this study as the reference of adult levels of disaccharidase activities. The cows, weighed 332kg, were fed only rice straw for three months before slaughter and the steers, weighed 531kg, were fed concentrates ad libitum. They were killed by electrical stunning and bleeding. Immediately after slaughter, the small and large intestines were removed as soon as possible and weighed with their contents. In calves, samples of intestines for enzyme assay and of contents were taken from seven sites along the small intestine at These sites were decided arbitrarily and were referred to as duodenum, proximal jejunum, middle jejunum, distal jejunum, proximal ileum, middle ileum and distalileum, respectively. Samples were also taken from the caecum and sigmoid colon. In fetuses, samples for enzyme assay were taken from four sites along the small intestine arbi- tine from the pylorus and were referred to as duodenum, proximal jejuno-ileum, middle jejuno-ileum and distal jejuno-ileum, respectively. Weights of the empty intestines were measured. Weights of the reticulo-rumen and abomasum with and without their contents were also measured. The intestine samples were chilled with ice and transported to the laboratory. The intestinal contents from calves were preserved with a saturated solution of HgCl2. And then the ph of these contents were measured by Beckman Zeromatic ph Meter (Beckman-Toshiba Ltd, Tokyo, Japan). Each intestine sample was cut open, washed in ice-cold solution of 0.15M-NaCl and gently blotted with a piece of filter paper. The mucosa was scraped off with a glass slide and weighed. A known quantity of the mucosa was homogenized with a given quantity of ice-cold 0.15M-NaCl solution in Universal Homogenizer (Nihon Seiki Ltd, Tokyo, Japan) at top speed for 3 minutes. Then the homogenate was centrifuged at 3000rpm for 10 minutes in order to remove nuclei and larger cell debris, and the supernatant was

3 MIYASHIGE and YAHATA all the activity of each disaccharidase of the whole homogenate, it was referred to as the homogenate hereafter. For preparation of, homogenates of mucosa in adult cattle, samples were similarly taken from the small intestine at seven sites and from the caecum and sigmoid colon. Determination of disaccharidase activities of the homogenate was carried out within two weeks after preparation. Under these conditions, the lactase activity of the homogenate, which was stored for 2 months after preparation, that of the fresh homogenate. was found to be 97% of The frozen homogenates were allowed to thaw and their disaccharidase activities were measured by the method of DAHLQVIST5), using substrates made up in 0.1M-sodium 28mM maleate, ph 5.6 for lactase, cellobiase and trehalase and ph 5.8 for maltase, isomaltase and sucrase. One unit of disaccharidase acti- The protein content of homogenate was assayed by the method of LOWRY et al.6). Results Development of intestinal tissue weights Table 1 shows tissue weights of the reticulo-rumen, abomasum, small intestine and large intestine, and content weights of the total stomach and intestines. The relative tissue weight of the small intestine to the body weight decreased rapidly after 4 weeks Table 1. Body weights, tissue weights of the reticulo-rumen, abomasum, small intestine_??_and large intestine, which are expressed as an actual weight and as a weight per kilogram body weight, and content weights of the total stomach and intestines of fetuses and calves. 60

4 Intestinal Disaccharidase of Nursing Calf of age and that. of the large intestine decreased slightly, though the actual tissue weights of both intestines increased greatly. Changes of ph of intestinal contents Influence of ph on disaccharidase activities in homogenate of calf small intestinal mucosa is shown in Fig. 1 and changes of ph of contents from different sites of the small intestine and from the caecum and colon of calves are shown in Fig. 2. Relati- Fig. 1. PH-activity curves for disaccharidase activities of homogenate of calf small intestinal mucosa. Enzyme activities are expressed as percentage of the activity at optimum ph. The buffers used were: ph ,0.1M-sodium acetate; ph , 0.1M-sodium maleate; ph ,0.1 M-sodium phosphate. Fig. 2. Changes of ph of contents from different sites of the small intestine and from the caecum and colon of calves. No contents in the colon were obtained in newborn- and 4 weeks old-calves. 61

5 MIYASHIGE and YAHATA vely low ph values of the contents of , which were close to optimum ph of intestinal disaccharidases, were observed all over the sampling sites of the small intestine in newborn calves. At 4 and 26 weeks of age, relatively low ph values of the. contents were observed only in the upper part of the small intestine. Development of intestinal disaccharidase activities Pre- and post-natal development of mean disaccharidase activities of small intestinal mucosa is shown in Fig. 3. Apparently high activity of lactase and distinctly low activities of cellobiase, trehalase, maltase and isomaltase were observed in fetuses and_ calves. Sucrase activity was rarely detected but only with negligible level in fetuses and calves. Lactase activity was the highest at birth and decreased rapidly with age. Similar tendency with age was observed in cellobiase and trehalase. At birth, of the, Fig. 3. Pre-and post-natal development of disaccharidase activities of homogenates of small i ntestinal mucosa. Each point is expressed as the weighted average of activiti es of 7 sites in _ calves or 4 sites in fet uses for each corresponding site of the small intestine. 62

6 Intestinal Disaccharidase of Nursing Calf two newborn calves slaughtered, the unsuckled one showed apparently higher activity of lactase than the suckled one. In fetuses after 5.5 months of fetal age, the activities of these disaccharidases increased rapidly with age. On the other hand, maltase and isomaltase activities did not show any definite trend with age before and after birth. The disaccharidase activities of mucosa from different sites of the small intestine and from the caecum and colon of newborn, 4 weeks old- and 26 weeks old-calves are shown in Table 2. At birth, lactase activity was found all over the sampling sites of the small intestine and was found even in the caecum and colon though it was very low. The activity was the highest in the proximal jejunum, slightly lower in the duodenum and decreased in the distal sampling sites. In older calves, the pattern of distribution of lactase activity was unchanged, though overall activity decreased with age. The pattern of distribution of cellobiase activity was very similar to that of lactase activity at any age of calves. Trehalase activity was also found mainly in the upper part of the small intestine. Maltase and isomaltase activities were low but nearly constant along the small intestine at birth and 4 weeks of age. At 26 weeks of age, apparently higher activities of these disaccharidases were observed in the lower part of the small intestine than in the upper part. Some maltase activity was found in the caecum and colon as well as in the small intestine. Table 2. Disaccharidase activities of mucosa from different sites of the small intestine and from the caecum and colon of calves (mu/mg protein). 63

7 MIYASHIGE and YAHATA The pattern of distribution of disaccharidase activities of fetuses, shown in Table 3, was similar to those of newborn calves. The pattern of distribution of disaccharidase activities of cows and steers is shown in Table 4. Lactase and cellobiase activities were found in adult cattle but with neg- Table 3. Disaccharidase activities of mucosa from different sites of the small intestine and from the caecum of fetuses (mu/mg protein). Table 4. Disaccharidase activities of mucosa from different sites of the small intestine and from the caeaum and colon of adult cattle (mu/mg protein). 64

8 Intestinal Disaccharidase of Nursing Calf ligible levels. Trehalase activity was also found at similar level to that of calves at 26 weeks of age. Maltase and isomaltase activities also showed similar levels to those of calves, though much higher activities occurred in the middle part of the small intestine. In cows, extremely high levels of these activities were found only in the distal jejunum, whereas in steers, high levels of these activities were found both in the jejunum and in the ileum. Sucrase activity was not detected even in adult cattle as well as in calves. Discussion Up to this time, using various criteria such as growth, digestion data and postprandial blood sugar levels, the digestive capacity of carbohydrates in calves has been discussed. The enzyme levels in pancreatic and intestinal tissue have been also investigated for this purpose. And it is known that lactose is well utilized by young calves, and this is correlated with the high activity of intestinal lactase7,8). Our results also support this. The finding of the decrease in activity of lactase with age is similar to observations reported by HUBER et al.1), SIDDONS2) and TOOFANIAN et al.3) in calves fed solely on milk and in calves weaned at 5-6 weeks of age, in which the activity decreased with age up to 3 to 8 weeks but changed little thereafter. Of course, change of the activity expressed as units per mg protein of mucosa does not correspond to that of the amount of overall activity in the small intestine. But it is seemed that in addition to the decrease of the relative tissue weight of the small intestine to the body weight, the decrease of the specific activity of lactase with age indicates the decline of nutritional contribution of intestinal lactase to the calf. And adult cattle showed almost no activity of lactase, as reported by SIDDONS2). This post-natal change of lactase activity agree with the observation in blood sugar level reported by HUBER et al.9), in which an elevation of blood sugar level after lactose feeding decreased with advance of age from 22 to 600 days. It is seemed that intestinal lactase activity of fetuses increases their fetal period, as reported rapidly throughout by BRITT et al.10). But after birth, the activity decreases sharply in the first few weeks, as shown by earlier workers and by us. The cause of the sharp decrease of activity is not known. TOOFANIAN et al.3) reported that the decrease of activity was especially drastic in the first week. The fact that of the two newborn calves, the suckled one showed apparently lower activity than the unsuckled one suggests that the nursing and the subsequent flow of chyme along the small intestine in newborn calves might probably result in physical damage of intestinal mucosa that was developed during the fetal period, and therefore in decrease of mucosal lactase activity. Our result of intestinal maltase activity is similar to those reported by HUBER et al.1), SIDDONS2), and COOMBE and SIDDONS4) in milk fed calves and even in early weaned calves, and also agree with observations maltose feeding of a small elevation of blood sugar level after by HUBER et al.9), VELU et al.11) and SIDDONS et al.12). From the distribution of maltase activity, it is expected that maltose entering the duodenum would 65

9 MIYASHIGE and YAHATA be digested little by little throughout the small intestine, but the digestive capacity of maltose would be only a limited extent. The low activities of intestinal maltase and isomaltase of calves also suggest utilization of dietary starch in the small intestine. In the early weeks of life of calves, testinal mucosa. But it is predicted that the activity may be due to an adsorption of cur largely in the lumen of the small intestine. Therefore, it is conceivable that the region where maltose is produced and digested is limited to much lower part of the small intestine. In nursing calves, intake of concentrates increased rapidly after 12 weeks of age16), and therefore it is expected that apparently higher activity of maltase in the lower part of the small intestine at 26 weeks of age would be correlated with the increase of quantity of this substrate in the luminal contents of this region. MAYES and ORSKOV17 found in sheep infused a solution of gelled maize starch via the abomasum that considerable quantities of glucose, maltose and maltotriose were found in ileal digesta. Our result of high maltase activity in the middle and lower part of the amylase secreted. The fact that a marked difference in pattern of distribution of maltase activity along the small intestine between the cows and the steers suggests occurrence of dietary effect. The digestive capacity of various substrates poor by small intestinal mucosa seemed to depend on the amount of overall activity of corresponding enzyme. We calculated the quantities of lactose and maltose which are able to be hydrolyzed by mucosal lactase and maltase of the overall small intestine, respectively, and showed in Table 5. Table 5. Calculation of utilizable quantities of lactose and maltose by overall small intestinal mucosa in calves. Mucosa weight is postulated as 50% of tissue weight after the result of BRITT and HUBER18). Disaccharidase activity is the weighted average of 7 sites along the small intestine. 66

10 Intestinal Disaccharidase of Nursing Calf The calculation was based on the enzyme activities and tissue weights of the small intestine. As shown in this table, the utilizable quantity of lactose a day decreased from birth up to 4 weeks of age, then was assumed to be fairly stable up to 12 weeks of age and increased at 26 weeks of age. The utilizable quantity of maltose showed a steady increase with age, but remained still small compared with lactose. The milk intake of nursing calves of Japanese Black Cattle at 1,3,11 and 25 weeks of age were 7.00,7.26,5.67 and 3.51kg a day, respectively, according to YOSHIDA19). And the average lactose content in milk of Japanese Black cows was 4.42%, according to HABU and UESAKA20). Therefore, lactose intake can be estimated at 309, 321, 251 and 155g a day, respectively. These intake will be well within the hydrolysing capacity of lactose. Table 5 also shows the utilizable quantities of lactose and maltose per kg body weight. These values indicate the decrease of nutritional importance of lactose and a very little contribution of maltose to energy demand of calves, as described above. In addition to disaccharidase activities, the activity of microbes present in the small intestine may have an important role in intestinal carbohydrate utilization. Relatively high ph environment in the lower part of the small intestine after 4 weeks of age may be rather favorable to microbial activity. The quantitative importance of microbial utilization in the small intestine is further to be elucidated. References 1) HUBER, J.T., N.L. JACOBSON, R.S. ALLEN and P.A. HARTMAN, J. Dairy Sci., 44: ) SIDDONS, R.C., Biochem. J., 108: ) TOOFANIAN, F., F.W.G. HILL and D.E. KIDDER, Ann. Rech. veter., 4: ) COOMBE, N.B. and R.C. SIDDONS, Br. J. Nutr., 30: ) DAHLQVIST, A., Anal. Biochem., 7: ) LOWRY, O.H., N.J. ROSEBROUGH, A.L. FARR and R.J. RANDALL, J. Biol. Chem., 193: ) HUBER, J.T., J. Dairy Sci., 52: ) RADOSTITS, O.M. and J.M. BELL, Can. J. Anim. Sci., 50: ) HUBER, J.T., N.L. JACOASON, A.D. MCGILLIARD and R.S. ALLEN, J. Dairy Sci., 44: ) BRITT, D.G., W. OXENDER, F.B. STIFEL and J.T. HUBER, J. Dairy Sci., 57: ) VELU, J.G., K.A. KENDALL and K.E. GARDNER, J. Dairy Sci., 43: ) SIDDONS, R.C., R.H. SMITH, M.J. HENSCHEL, W.B. HILL and J.W.G. PORTER, Br. J. Nutr., 23: ) MORRILL, J.L., W.E. STEWART, R.J. MCCORMICK and H.C. FRYER, J. Dairy Sci., 53: ) TERNOUTH, J.H., J.H.B. Roy and S.M. SHOTTON, Br. J. Nutr., 36: ) TOOFANIAN, F., D.E. KIDDER and F.W.G. HILL, Br. vet. J., 130: ) TERADA, T., S. WATANABE, T. MIYASHIGE and S. OZAWA, Proc. Jap. Soc. Anim. Nutr. Meta., 17: ) MAYES, R.W. and E.R. ORSKOV, Br. J. Nutr., 32: ) BRITT, D.G. and J.T. HUBER, J. Dairy Sci., 57: ) YOSHIDA, S., in Studies on Japanese Feeding Standard for Beef Cattle. (Agriculture, Forestry and Fisherys Research Council ed.) Norinkosaikai. Tokyo ) HABU, Y. and S. UESAKA, Jap. J. Zootech. Sci., 17:

11 MIYASHIGE and YAHATA

Calf Notes.com. Calf Note #155 Day 2. Introduction

Calf Notes.com. Calf Note #155 Day 2. Introduction Calf Notes.com Calf Note #155 Day 2 Introduction Calf nutrition and management seems to be divided into two distinct periods namely, the first day of life and everything after. We all know of the importance