THE MUCOSAL DISACCHARIDASES IN THE SMALL INTESTINE OF THE CALF F. Toofanian, F. W. G. Hill, D. E. Kidder

To cite this version: F. Toofanian, F. W. G. Hill, D. E. Kidder. THE MUCOSAL DISACCHARIDASES IN THE SMALL INTESTINE OF THE CALF. Annales de Recherches V´et´erinaires, INRA Editions, 1973, 4 (1), pp.57-69.

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THE MUCOSAL DISACCHARIDASES IN THE SMALL INTESTINE OF THE CALF F. W. G. HILL

F. TOOFANIAN

D. E. KIDDER

School

of Veterinary S’cicncc, University of liristol, art of l’eterirzavy !llcdiciuo, p De t l n lJl ord, f ford House, LalJg g Lan Bristol (li>ig/fiii«]

SUMMARY The postnatal development and pattern of distribution of the mucosal disaccharidases, lactase, cellobiase, trehalase, maltase, and sucrase have been studied in the young calf, on conventional, or cm high lactose content diets from birth up to six months of age. In the newborn calf, lactase had the highest level of activity, followed by cellobiase, trehalase and maltase activities,

respectively. There was an initial fall in lactase and cellobiasc activities during the first weeks of life. In the ruminant calves, a further decrease in lactase, ccllobiase and trehalase activities occurred at the time of weaning, whereas in the non-ruminants the levels of activity did not significantly change. Maltase activity increased from birth up to 8 weeks and usually remained unchanged thereafter. Lactase, cellobiase and trehalase activities were highest in the proximal jejunum, rather lower in the duodenum, and decreased distally, with a sharp fall in the ileum. Maltase activity showed a very irregular pattern of distribution, with the lowest levels of activity occurring in the duodenum. The non-ruminant calves showed higher levels of lactase, cellobiase and trehalase activities and it is concluded that in the calf, by feeding a high-lactose, low-roughage diet, it is possible to influence and maintain higher disaccharidase levels.

INTRODUCTION

The

dietary carbohydrates, chiefly in the form of polysaccharides (starch and glycogen) and oligosaccharides (mostly disaccharides like lactose and sucrose), must be hydrolyzed to monosaccharides before they can be absorbed and utilized ) New address : 1 ( 1’ahlaki-Shiraz (Iran).

Department

of clinical studies. School of

veterinary

medicine.

University

of

in the body. The pancreatic and the small intestinal mucosal enzymes are responsible for this hydrolysis. In the young pre-ruminant and non-ruminant calves (the term non-ruminant describes an animal in which the ruminal function is artificially prevented), the liquid diet is channelled straight to the abomasum by the closure of the oesophageal groove, making the animal similar to monogastric (simple stomach) species. In the ruminant calf, the principal feature of digestive physiology is that fermentative digestion occurs on a massive scale in the first two compartments of the stomach. Thus, the intestinal disaccharidases have a much smaller role. For this reason the development, pattern of distribution and levels of the intestinal disaccharidases may be expected to differ from that in the non-ruminant calves. It has been found repeatedly that the activity of p-disaccharidases is high in the small intestine of infant mammals, and decreases with age (KoLDovsK9 and HYTII 19 C ,, ). Conversely, the neonate intestine, with the exception of human, is 5 6 relatively deficient in intestinal a.-disaccharidases, as compared with the adult of the same species, and levels of their activity increase with age (DE , rg68). REN The decrease in { 3-disaccharidase (lactase) activity occurs at the time of weaning OLD and CHyTm&dquo; 19 K RETCHMER ig62 ; ovsKÝ K (Dori,r, and , 65), when milk and therefore lactose becomes of lesser importance to the animal. Man is the only known species in which the postweaning decline in lactase activity does not occur (A URIC

al, 1 63). 9 Attempts have been made to prevent the postweaning decline in mucosal lactase by feeding lactose, with variable success. , EKOV H S II ) OEI D L 1951 in rabbits, , ( and HM ETC 6 K R E AS (ig6i) in rats, failed to prevent the LVARE and S A 19 and z ( ) 2 decline in intestinal lactase by increasing the lactose content of the diet. SmDO s V 19 found no marked differences in intestinal lactase activities of ruminant and ( 6S) non-ruminant calves of the same age. On the other hand, GiRARDi;T et al. (rg6 ) and 4 CAIN et al. (io68 and ig6g) have shown induction of lactase to occur in the rat. The postnatal changes in disaccharidase activity in the small intestine of the bovine has been studied by H UB H R EIMKOV ), 1959 E ( 1951 DOLLAR and PORTER ), ( et al. 6 in and Smvows these the studies, 19 ( ) 1 experimental ani(rg68). However, mals were kept basically on a whole milk diet and were not weaned during the expecxio ct

riments. The following investigation was undertaken to study the postnatal development and changes in the pattern of distribution of intestinal disaccharidases from birth up to 6 months of age, and to examine the changes occurring during the weaning period in the calf.

MATERIALS AND METHODS Experimental

calves and their

management

The conventional (ruminant) animals were offered hay and water ad libitum, and concenwere weaned at 5 -6 weeks of age. The non-ruminant animals were kept on a commercial milk substitute diet containing 37 8 p. ioo dry matter lactose, without having any access to 3 roughage and concentrates.

trates, and

Expevin2ental design The 4 , 3 , 2 , 1 6, 20 and 8 calves studied were assigned to age groups of newborn, 1 , 4 , 8, 12 24 weeks of age, 3 calves in each group (in the age group of 4 weeks, 6 calves were assigned). Intestinal

preparations

Calves were slaughtered in the usual way, the entire gastro-intestinal tract was removed from the abdominal cavity, and quickly carried to a cold room. The small intestine was separated from the rest of the tract, and the total length from the pylorus to the ileocaecal valve measured. 0 8 0 and 9j per cent of the Next, the small intestine was sampled at six definite sites at 5 , 40 , 20 , 6 total length of the small intestine. These sites were decided arbitrarily and represent duodenum, proximal jejunum, middle jejunum, distal jejunum, proximal ileum and distal ileum, respectively. The samples consisted of approximately 3 cm segments of the intestine which were cut open, gently rinsed in ice cold distilled water, wrapped separately and frozen rapidly by plunging into a flask of liquid nitrogen (― igj!C).

Ho2nogenate preparations The frozen samples were thawed, the mucosa scraped off with a metal spatula, mixed with ice cold distilled water and homogenized with an ultrasonic probe for 20 seconds. The homogenates were kept at °C until analyzed. 20 -

Disacchavidase determination The frozen homogenates were thawed and their disaccharidase activities were assayed by the classical method of D AHLQVIST 6 1, ( ) 4 9 lactase, cellobiase and trehalase at pH 5 4 and maltase .

at pH j.8. Pvotein determination The

using

a

protein content of the homogenate was assayed by homogenate, and bovine plasma albumin was

fresh

Calculations and statistical

the method of LowRY et al. used to prepare a standard

ji), 9 (i curve.

analysis

The enzyme activity was calculated as e international units o (IU) per gram of protein. One IU is one ,mole disaccharide hydrolyzed per minute at 37 LORKIN C under optimum conditions (F 0 and , TOTZ j). S 96 r A multifactorial analysis of variance was conducted via a BMD 02 V computer programme. A negative exponential curve was calculated, using the mean values, and fitted for each enzyme.

RESULTS Table I gives the mean specific values for disaccharidase activities along the entire length of the small intestine of the experimental calves and figures i-g illustrate the pattern of development, and the distribution of the disaccharidases studied. Postn.atal

development of

disaccharidases

The postnatal development of the disaccharidases is described and discussed in three separate periods : i) from birth up to 4 weeks ; 2 ) from4 to 8 weeks, during this period the ruminant calves were completely weaned onto solid food ; 3 ) from

8 to 24 weeks. Since the ruminant calves did not consume much hay and concentrates while they were still on milk, the calves in the I to 4 weeks age groups were considered non-ruminants as well. Lactase mean specific activity decreased significantly (P < ) 05 within the . 0 first 4 weeks of life, with the sharpest fall occurring during the first week. The ruminant calves were weaned during the 5 to 6 weeks, and there was a decrease of 3 6 p. ioo between the values recorded at 4 weeks and at 8 weeks, which is highly significant (P < o.or), whereas in the non-ruminant calves the lactase mean specific activity did not change markedly from 4 to 8 weeks. In both ruminant and non-ruminant calves, from 8 to 24 weeks of age, although the levels of activity fluctuated, there was no significant change in lactase levels. However, the total mean specific activity of lactase, over this latter period (from 8 to 24 weeks of age), was 123 p. 100 more in non-ruminant calves than in comparable ruminants (fig. I et 2 ). Cellobiase activity behaved similarly to that of lactase, a significant decrease .8 p. 100 fall during the during the first 4 weeks (P < ), 05 and in ruminants a 34 . 0 < while in non-ruminants it remained unchanged. In the weaning period (P 05 . 0 ), from 8 to the non-ruminant calves showed a weeks, 45 period 24 p. ioo more cellobiase activity than the corresponding ruminants (fig. 3 et 4 ). The overall change in trehalase activity within the first 4 weeks was not significant. A 47 p. ioo decrease occurred during the weaning period (P < o.oi), while in non-ruminants the changes were not significant. The non-ruminant calves showed 7 p. 100 more trehalase activity than the ruminant ones (fig. 5 et 6). 6 There was a gradual increase in maltase activity during the first 4 weeks after birth (P < ). 05 In the ruminant calves, the increase during the weaning pe. 0 riod, in maltase activity, was significant (P < o.o ), while in the non-ruminants, 5 it not it increased was statistically significant. Thereafter, in both rumialthough nant and non-ruminant calves, the changes were insignificant (fig. 7 et 8). Sucrase was detected and then with levels of activity rarely only activity. negligible Pattern The

of

disaccharidase distribution

along

the snaall 1:ntestine

pattern of disaccharidase distribution along the small intestine of rumi-

was similar. The highest lactase and cellobiase activities occurred in the proximaljejunum, relatively lower in the duodenum, and decreased distally towards the ileo-caecal valve. This decrease was more gradual in younger calves, whereas in older ones a sharp fall occurred in distal jejunum, and in many cases the ileum was actually devoid of any lactase and cellobiase activity. The distribution of trehalase activity was similar to those of lactase and cellobiase, with the exceptions that highest activities were recorded in proximal and middle jejunum, and the distal decrease was more gradual and less pronounced. Maltase activity was more uniformly distributed along the length of the small intestine with highest activities occurring in distal jejunum and proximal ileum ; however, maximum levels also occurred in proximaljejunum. Maltase activity decreased distally towards the ileo-caecal valve, which nevertheless was higher than that of the duodenum

nant and non-ruminant calves

(fig. 9 ).

DISCUSSION AND CONCLUSIONS

In several mammalian species, the small intestine in the newborn has

high

lactase

ST, HLQVI activity, and very low activity of other disaccharidases (BAILEY et al., zg56; DA , 19 R E HM 4 b ; DOELI, and KRETC , rg6 2 6 ). The activity of p-disaccharidases 3 , rg6 1 6 10 decreases with age, occurring at the time of weaning, (DOE E; R K T , and CHMER I OVSK and ’rm, D KoL Y HV 19 C 65), whereas the levels of a-disaccharidases activity increase with age (D , ig68). The present findings in the calf agree well with EREN this developmental pattern and correspond with the physiological need for lactase during the first period of extrauterine life, when milk is the most important nutrient. EILSKOY ), O D L AR I and PORTER ), Although the previous reports of H 1951 , ( 1959 ( in and SmDOVS showed the same developUBER et al. 6 H terms, 19 ( ) 1 (ig68), general mental pattern, these never substantiated the precise period at which the changes

occurred. Moreover, in those studies, the experimental calves were not weaned, and thus the effects of weaning on the levels of intestinal disaccharidases was unclear. U H R E et B al. (r 6q) found a 3 9 -times higher total lactase activity in the intestinal mucosa of calves receiving varying amounts of lactose than those receiving a control hay-grain ration. Although this report shows the possibility of lactase adaptation in the calf, it is not clear from the text if the weaning decline in lactase activity was IDDONS 68) prevented. S 19 found no marked differences in lactase activities in ( 4 month old calves fed solely on milk, and calves of the same age that had been weaned when 6 weeks old. Nevertheless, the present study clearly demonstrates that, in the calf, by feeding a high-lactase, low-roughage diet, it is possible to prevent the decrease in lactase activity occurring during the weaning period, and to maintain higher levels of certain disaccharidases. Higher levels of lactase activity, by feeding lactose, might be expected. However, the appreciably lower lactase activity in the ruminant calves could be a consequence of either the absence of exposure of the intestinal mucosa to lactose, or of other changes consequent upon ruminal functional development. The increase in cellobiase activity is not unexpected, as the lactase and cellobiase activities are both believed to be exerted by the same enzyme , QVIST 19 AHI (D 4 a). Since 6 trehalase appears to have absolute specificity for its substrate unlike the other intestinal disaccharidases (D , 19 T IS ALHQV 4 a), the prevention of its postweaning decline, and 6 its higher activity observed in milk-fed calves, is presumably not a case of substrate induction, and the exact nature of this phenomenon is not clear. R EDDY et al. (ig6s) also found an increase in trehalase activity in maltose-fed rats. In conclusion, in the calf, there is a postnatal physiological fall in #-disaccharidase activity and an increase in x-disaccharidase activity, during the first4 weeks of life, which could not be manipulated by dietary substances and is probably regulated by genetical and hormonal factors, but the alterations occurring during the weaning period could be prevented by substrate feeding, and moreover, higher levels of activity could be maintained if the substrate feeding is continued.

ACKNOWLEDGEMENTS The authors would like to express their sincere thanks to Professor C. S. G. R G UNSELL for his sustained interest in all aspects of the work. This work was supported by a grant from the University of Pahlavi, Shiraz, Iran, which is gratefully acknowledged. Thanks are also due to Miss M. McAvoy for technical assistance.

RÉSUMÉ LES DISACCHARIDASES DANS LA Le

développement

lase, maltase de l’intestin

MUQUEUSE DE L’INTESTIN GRÊLE

et la distribution des disaccharidases mucosales :

et sucrase, ont été étudiées dans six

grêle.

DU VEAU

lactase, cellobiase, trehasegments, également disposés sur la longueur

Le niveau d’activité des enzymes décroît

rapidement après

la naissance du

veau, mais s’il se

maintenir

reçoit une nourriture pendant six mois.

riche

en

lactose

un

niveau très élevé de

ces

enzymes

peut

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