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Effect of Histamine on Intestinal Absorption of Gamma Globulin in Newborn Calves
Effect of Histamine on Intestinal Absorption of Gamma Globulin in Newborn Calves Abstract
Newborn calves were isolated from their dams before they were suckled and within 3 hr of birth were fed 1 liter of cow's milk containing either: a) no additives, b) 75.0 g gamma globulin, or c) 75.0 g gamma globulin and 2.0 mg histamine. Additional feedings were at 7 and 15 hr after the initial meal. Serum gamma globulin was determined before and at 1, 3, 5, 7, 11, 15, and 23 hours after the first feeding. Differences in serum gamma globulin between calves that received histaminesupplemented milk and gamma globulin and those that received only added protein in milk were insignificant. I n all calves receiving gamma globulin, serum gamma globulin increased after the first two feedings, but the values plateaued thereafter. The gamma globulin in serum of calves fed only milk remained negligible. Introduction
Previous work (4, 7) has shown that bovine colostrum contains a relatively high concentration of hista,nine. Following parturition, colostral histamine falls rapidly from .9 #g/ml to levels in normal milk, about .3 /~g/ml (7). I t has been postulated: a) that the high concentration of colostral histamine may function in the selective transfer of plasma immune globulins by the nmmmary gland into the colostrum (7), and b) that the high level of this amine prmnotes transfer of ingested globulin across the intestinal epithelium in the newborn calf. The present experiment was designed to test the latter hypothesis. Experimental Procedures
Twelve Holstein calves were isolated from their dams before suckling and were away from the cows throughout the experiment. Each calf was assigned at birth to one of three dietary regimes. A t each feeding, all animals in the first group received 1 liter of cow's milk, calves in the second group consumed 75 g of lyophilized crude gamma globulin in I liter of milk Received for publication on October 1, 1971 as Paper 4059 in the 3ournal Series of the Pennsylvania Agricultural Experiment Station.
and those in the third group received 2 nag of histamine plus 75 g of gamma globulin in 1 liter of milk. Calves were initially fed within 3 hr and usually within 1 hr of parturition. Subsequent feedings were at 7 and 15 h r after the first meal. All calves were fed via a nipple pail. Five milliliters of blood were obtained from the jugular vein before the initial feeding (pre-feed) and at 1, 3, 5, 7, 11, 15, and 23 hr after it; and the sera were analyzed for gamma globulin. Serum gamma globulin was calculated from measurement of total serum protein by the Biuret method and percentage serum gamma globulin by electrophoresis. Biuret determinations were with Uni-Tech Biuret Reagent, and the optical density was read at 540 mtz. Electrophoresis was on cellulose acetate strips (5), and percentage of gamma globulin was determined on a Photovolt Densicord densitometer. Lyophilized crude gamma globulin fed to the calves was prepared from bovine colostrum f r o m normal cows. Colostral casein was removed by adding 4.5 ml of .33% rennin and .5 ml saturated CAC12/150 ml of colostrum. Samples were incubated at 37 C for 20 min to allow the casein to clot~ and subsequently centrifuged for 10 rain. W h e y was poured off and filtered through coarse filter paper. All batches of whey were frozen, then thawed, pooled and refrozen before further treatment. Crude gamma globulin was prepared from this whey by ammonium sulfate fractionation as described by Smith (6), except that whey proteins were precipitated only once. The precipitate was then dialyzed and lyophilized before use. Electrophoresis of the lyophihzed material showed that at least 70% migrated as gamma globulin. Results and Discussion
The results of the experiment, (Fig. 1), revealed no significant differences in serum gamma globulin between calves that received lyophilized bovine gamma globulin and those that consumed gamma globulin and histamine. A p p a r e n t l y supplemental histamine did not increase absorption of gamma globulin by the intestine of these calves even though the concentration of histamine fed to the calves was greater than that in the colostrmn of most dairy cows (4, 7). This suggests that the high 645
646
J O U R N A L OF D A I R Y SCIENCE
Science, The Pennsylvania State University, University Park 16802
free histamine in colostrum is not required f o r significant absorption of g a m m a globulin in newborn calves. There were constant increases in serum g a m m a globulin a f t e r the first and second feeding in calves given g a m m a globulin, either with or without additional histamine. I n both cases, the mean values 7 hr a f t e r the initial feeding were significantly higher ( P < . 0 5 ) than the pre-feeding samples~ and likewise, the 11 hr levels were greater ( P < . 0 5 ) than the 7 h r values. However, the serum g a m m a globulin plateaued a f t e r the third feeding, i.e., at approximately 16 hr of age. This agrees with other reports (1, 2) t h a t the ability of calves to absorb maeromolecules declines within several hours of birth. A low gamma globulin was in the serum sample of all calves before feeding. I n those receiving only milk, the concentration remained low throughout the experimental period, t t y p o gammaglobulinemia in newborn colostrumdeprived calves has been well established (1,
References (1) Brambe]l, 1~. W. R. 1969. The transmission of immune globulins from the mother to the foetal and newborn young. Proe. Nutrition Sot., 28: 35. (2) Butler, J. ]~. 1969. Bovine inmmnoglobulins: a review. E. Dairy Sei., 52: 1895. (3) Fey, H. 1971. Immunology of the newborn calf : its relationship to colisepticemia. Ann. New York Aead. Sei., 176: 49. (4) X~uquay, J. W., E. M. Kesler, and A. Zarkower. 1969. Effect of prepartum and postpartum diet on histamine metabolism of young Holstein cows. g. Dairy Sci., 52: 1781. (5) Gelman Procedures, Techniques and Apparatus for Eleetrophoresis. 1968. Gelman Instrument Co., Ann Arbor, ]~ichlgan. (6) Smith, E. L. 1948. Isolation and properties of the immune proteins of bovine milk and eolostrum and their role in immunity: a review. J. Dairy Sci., 31: 127. (7) Zarkower, A. 1967. Histamine in the cow: pre- an4 postparturition histamine concentrations in plasma, milk and tissue. Amer. J. Vet. Res., 28: 1751.
2, 3). J. A. PATT, Jr., A. ZARKOWER, and R. J. EBERHART, Department of Veterinary
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FIG. 1. Serum gamma globulin in newborn calves after receiving 1 liter cow's milk per feed ( A ) , 1 liter cow's milk plus 75 g ]yophilized bovine gamma globulin per feed ( O ) , or 1 liter cow's milk, plus 75 g ]yophi]ized bovine gamma globulin, plus 2 mg histamine per feed ( [ ] ) . Values are means ~__ standard error. JOURlqAL OF DAIRY SCIENCE VOL. 55, NO. 5
Newborn calves were isolated from their dams before they were suckled and within 3 hr of birth were fed 1 liter of cow's milk containing either: a) no additives, b) 75.0 g gamma globulin, or c) 75.0 g gamma globulin and 2.0 mg histamine. Additional feedings were at 7 and 15 hr after the initial meal. Serum gamma globulin was determined before and at 1, 3, 5, 7, 11, 15, and 23 hours after the first feeding. Differences in serum gamma globulin between calves that received histaminesupplemented milk and gamma globulin and those that received only added protein in milk were insignificant. I n all calves receiving gamma globulin, serum gamma globulin increased after the first two feedings, but the values plateaued thereafter. The gamma globulin in serum of calves fed only milk remained negligible. Introduction
Previous work (4, 7) has shown that bovine colostrum contains a relatively high concentration of hista,nine. Following parturition, colostral histamine falls rapidly from .9 #g/ml to levels in normal milk, about .3 /~g/ml (7). I t has been postulated: a) that the high concentration of colostral histamine may function in the selective transfer of plasma immune globulins by the nmmmary gland into the colostrum (7), and b) that the high level of this amine prmnotes transfer of ingested globulin across the intestinal epithelium in the newborn calf. The present experiment was designed to test the latter hypothesis. Experimental Procedures
Twelve Holstein calves were isolated from their dams before suckling and were away from the cows throughout the experiment. Each calf was assigned at birth to one of three dietary regimes. A t each feeding, all animals in the first group received 1 liter of cow's milk, calves in the second group consumed 75 g of lyophilized crude gamma globulin in I liter of milk Received for publication on October 1, 1971 as Paper 4059 in the 3ournal Series of the Pennsylvania Agricultural Experiment Station.
and those in the third group received 2 nag of histamine plus 75 g of gamma globulin in 1 liter of milk. Calves were initially fed within 3 hr and usually within 1 hr of parturition. Subsequent feedings were at 7 and 15 h r after the first meal. All calves were fed via a nipple pail. Five milliliters of blood were obtained from the jugular vein before the initial feeding (pre-feed) and at 1, 3, 5, 7, 11, 15, and 23 hr after it; and the sera were analyzed for gamma globulin. Serum gamma globulin was calculated from measurement of total serum protein by the Biuret method and percentage serum gamma globulin by electrophoresis. Biuret determinations were with Uni-Tech Biuret Reagent, and the optical density was read at 540 mtz. Electrophoresis was on cellulose acetate strips (5), and percentage of gamma globulin was determined on a Photovolt Densicord densitometer. Lyophilized crude gamma globulin fed to the calves was prepared from bovine colostrum f r o m normal cows. Colostral casein was removed by adding 4.5 ml of .33% rennin and .5 ml saturated CAC12/150 ml of colostrum. Samples were incubated at 37 C for 20 min to allow the casein to clot~ and subsequently centrifuged for 10 rain. W h e y was poured off and filtered through coarse filter paper. All batches of whey were frozen, then thawed, pooled and refrozen before further treatment. Crude gamma globulin was prepared from this whey by ammonium sulfate fractionation as described by Smith (6), except that whey proteins were precipitated only once. The precipitate was then dialyzed and lyophilized before use. Electrophoresis of the lyophihzed material showed that at least 70% migrated as gamma globulin. Results and Discussion
The results of the experiment, (Fig. 1), revealed no significant differences in serum gamma globulin between calves that received lyophilized bovine gamma globulin and those that consumed gamma globulin and histamine. A p p a r e n t l y supplemental histamine did not increase absorption of gamma globulin by the intestine of these calves even though the concentration of histamine fed to the calves was greater than that in the colostrmn of most dairy cows (4, 7). This suggests that the high 645
646
J O U R N A L OF D A I R Y SCIENCE
Science, The Pennsylvania State University, University Park 16802
free histamine in colostrum is not required f o r significant absorption of g a m m a globulin in newborn calves. There were constant increases in serum g a m m a globulin a f t e r the first and second feeding in calves given g a m m a globulin, either with or without additional histamine. I n both cases, the mean values 7 hr a f t e r the initial feeding were significantly higher ( P < . 0 5 ) than the pre-feeding samples~ and likewise, the 11 hr levels were greater ( P < . 0 5 ) than the 7 h r values. However, the serum g a m m a globulin plateaued a f t e r the third feeding, i.e., at approximately 16 hr of age. This agrees with other reports (1, 2) t h a t the ability of calves to absorb maeromolecules declines within several hours of birth. A low gamma globulin was in the serum sample of all calves before feeding. I n those receiving only milk, the concentration remained low throughout the experimental period, t t y p o gammaglobulinemia in newborn colostrumdeprived calves has been well established (1,
References (1) Brambe]l, 1~. W. R. 1969. The transmission of immune globulins from the mother to the foetal and newborn young. Proe. Nutrition Sot., 28: 35. (2) Butler, J. ]~. 1969. Bovine inmmnoglobulins: a review. E. Dairy Sei., 52: 1895. (3) Fey, H. 1971. Immunology of the newborn calf : its relationship to colisepticemia. Ann. New York Aead. Sei., 176: 49. (4) X~uquay, J. W., E. M. Kesler, and A. Zarkower. 1969. Effect of prepartum and postpartum diet on histamine metabolism of young Holstein cows. g. Dairy Sci., 52: 1781. (5) Gelman Procedures, Techniques and Apparatus for Eleetrophoresis. 1968. Gelman Instrument Co., Ann Arbor, ]~ichlgan. (6) Smith, E. L. 1948. Isolation and properties of the immune proteins of bovine milk and eolostrum and their role in immunity: a review. J. Dairy Sci., 31: 127. (7) Zarkower, A. 1967. Histamine in the cow: pre- an4 postparturition histamine concentrations in plasma, milk and tissue. Amer. J. Vet. Res., 28: 1751.
2, 3). J. A. PATT, Jr., A. ZARKOWER, and R. J. EBERHART, Department of Veterinary
;
$
FEEDING
I
1,5
z .J rn O ..J (.9
~E
1.O
0.5
o~ w
L~/L
I
I
I
Q 0
5
I0
15
"' IJ. W
TIME AFTER INITIAL FEEDING
I 20
I 25
(hours)
t~
n
FIG. 1. Serum gamma globulin in newborn calves after receiving 1 liter cow's milk per feed ( A ) , 1 liter cow's milk plus 75 g ]yophilized bovine gamma globulin per feed ( O ) , or 1 liter cow's milk, plus 75 g ]yophi]ized bovine gamma globulin, plus 2 mg histamine per feed ( [ ] ) . Values are means ~__ standard error. JOURlqAL OF DAIRY SCIENCE VOL. 55, NO. 5