Effect of Dietary Manganese on Growth and Manganese Metabolism in Sheep1

Effect of Dietary Manganese on Growth and Manganese Metabolism in Sheep 1 M. I V A N and M . HI D! R O G L O U Animal Research Institute Agriculture Canada Ottawa, Ontario K 1 A 0C6 ABSTRACT

Three groups of five wethers were each fed ad libitum a practical type diet containing 22, 300, or 3000 ppm of manganese for 8 wk. The wethers then were fistulated and killed 13 h after intraruminal dosing with manganese-54. Several tissues were sampled. Average daily gains and feed/gain ratios of sheep fed diets containing 22 and 300 ppm manganese were similar, but gain was lower and the ratio higher for sheep fed the diet containing 3000 ppm manganese than for those fed other diets. Feed intake was approximately the same for all treatments. Increased dietary manganese increased its concentration in soft tissues and bile. There was also a decrease in the uptake and specific activity of manganese-54 in the liver with increasing dietary manganese. Concentration of copper increased and concentration of zinc decreased in liver of sheep with increasing dietary manganese. INTRODUCTION

The essentiality of dietary manganese (Mn) for animals has been documented well but requirements of ruminant animals are not well defined (16). Manganese contents in feedstuffs may vary over a wide range. Thus, the majority of corn silage produced in Ontario contained inadequate amounts of ban (4) whereas concentrations of ban have been excessive in forages in New Zealand (6). Excessive dietary ban intakes by ruminants decreased growth, efficiency of

Received July 9, 1979. 1Contribution No. 857 of the Animal Research Institute. 2Shur-Gain Division, Canada Packers Ltd., Toronto, Ontario. 1980 J Dairy Sei 63:385-390

feed utilization, and feed intake (5, 6, 17), a n d they were associated with decreased absorption of iron (Fe) (7) and with changes in concentrations of Mn, copper (Cu), zinc (Zn), and Fe in some soft tissues (6, 7, 17). In addition, high dietary Mn decreased the relative uptake of s4 Mn by several soft and gastrointestinal tissues indicating Mn homeostasis in ruminants (1, 17). More studies of Mn metabolism in ruminant animals have been with diets containing relatively low (approximately 20 to 40 ppm) and excessive (over 1000 ppm) concentrations of Mn. However the content of ban in most diets of ruminants under practical conditions probably will not exceed 300 ppm. For this reason, our experiment was designed to study effects of low (22 ppm), high (300 ppm), and excessive (3000 ppm) concentrations of dietary ban on growth, feed intake and efficiency and to study some aspects of Mn metabolism in sheep. MATERIALS AND METHODS

Fifteen crossbred wethers were allotted to three groups with equal numbers and fed ad libitum one of three pelleted experimental diets for 8 wk. The control diet consisted o f ground hay (44%), ground barley (50%), soybean meal (5%), Shur salt mix No. 2 without ban 2 (1%), vitamin A (10,000 IU/g) (8 g/100 kg of diet), and vitamin D (1,650 IU/g) (10 g/100 kg of diet). The control diet contained 22 ppm Mn, 8.5 ppm Cu, 31 ppm Zn, and 263 ppm Fe. Two additional experimental diets were formulated by replacement of equal parts of hay with MnSO4"H20 to make final concentrations of 300 and 3000 ppm of Mn. Animals were housed in individual pens in an air-conditioned room and had continuous access to drinking water. Feed consumption was registered daily, and wethers were weighed every 2 wk. After completion of the 8-wk feeding period, each of four wethers from each treatment was equipped with a rumen cannula (8). 385

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IVAN AND HIDIROGLOU

Two weeks later carrier-free SaMn (as MnC12; 99.9% radionuclidic purity) a, diluted with deionized water to give a final concentration of 20 gtCi/ml, was administered to each sheep (10 #Ci/kg body weight) through the rumen fistula 20 min before the morning feeding. Administration of radioisotope was by a modified plastic syringe (50 ml) equipped with a plastic tube (30 cm long). The syringe and plastic tube were rinsed twice with 10 ml of deionized water. All wethers were killed by intravenous injection of T-614 13 h after dosing, and samples of heart, kidney, spleen, liver, pancreas, pituitary, adrenal, bile, rumen (dorsal sac), omasum, abomasum (fundic section), and duodenum were obtained. The samples of rumen, omasum, abomasum, and d u o d e n u m were washed with warm water to remove adhered digesta. The samples then were dried by repeated blotting with paper towels to remove water from the surface. All tissue samples then were homogenized and counted in triplicate in a gamma counter s . Analysis of Mn, Cu, Zn, and Fe in the diet, freeze-dried soft tissues, and bile was on an atomic absorption spectrophotometer 6 after nitric-perchloric acid digestion of samples. Analysis of variance (15) was used for testing differences between treatment means. RESULTS

The average daily gain for the 56-day feeding period was lower and feed/gain ratio was higher

3New England Nuclear, Boston, MA. 4 Hoechst Pharmaceuticals, Montreal, Quebec. s Gamma 300 Radiation Counter; Backman Instruments, Inc., lrvine, CA. 6Model 460; Perkin-Elmer, Norwalk, CT.

for lambs fed the diet containing 3000 ppm of Mn than for those fed diets containing 22 or 300 ppm of Mn (Table 1). However, none of the differences was significant (P<.05). Daily feed intakes were similar among treatments. Mean concentrations of Mn, Cu, Zn, and Fe in soft tissues and bile from sheep fed diets with different concentrations of Mn are summarized in Table 2. The increase in dietary Mn, from 22 ppm to 300 ppm and 3000 ppm, increased concentrations of Mn in all measured soft tissues and bile. However, the differences among treatments were not significant (P>.05) for kidney and spleen. With the increased dietary Mn from 22 to 300 ppm, the greatest increase in tissue concentration of Mn was in liver (61%) and the least in adrenal (15%), whereas the increase in dietary Mn from 300 to 3000 ppm resulted in the greatest increase in tissue concentration of Mn in adrenal (234%) and the least in kidney (29%). The concentration of Cu in the liver increased (P<.05) while that of Fe decreased (P<.05) with increasing dietary Mn, but the concentration of Zn was not affected. Differences among treatments in the concentration of Cu, Zn, and Mn in other tissues and bile were not significant (P>.05). The uptake of SaMn as percent of dose per kilogram of fresh liver decreased (P<.05) with increasing concentration of dietary Mn (Table 3). The trend was similar for rumen and duodenal tissues. Increase in 54Mn uptake with increasing dietary Mn was nonsignificant (P>.05) for spleen, pancreas, and adrenal tissues. With the exception of heart, spleen, and pancreas, all other tissues (kidney, liver, pituitary, adrenal) and bile showed decreased specific activity of 54 Mn with increasing dietary Mn (Table 4). However, the only significant differences among treatments were those for

TABLE 1. Performance of sheep fed different concentrations of manganese for 56 days. Dietary Mn (btg/g)a Item Initial weight, kg Gain, g/day Feed intake, g/day Feed/gain ratio

22 24.0 221 1157 5.82

300 24.3 236 1285 5.66

3000 24.1 178 1268 7.64

aFive sheep per treatment; none of the differences between treatment means was significant (P>.05). Journal of Dairy Science Vol. 63, No. 3, 1980

SE 3.47 32.6 157.7 .995

MANGANESE METABOLISM IN SHEEP

387

TABLE 2. C o n c e n t r a t i o n s of manganese, copper, zinc, and iron in selected tissues and bile.

Dietary Mn (#g/g) 22 Tissue

300

X

SE

X

3000 SE

X

SE

Significance b e t w e e n means

0ag/g dry matter) Heart Mn Cu Zn Fe

2 20 100 239

.2 a 1.6 15.O 2.7

2 21 100 244

.2 a .6 6.5 17.3

3 20 100 252

.3 b 1.0 5.5 11.0

** NS c NS NS

Kidney Mn Cu Zn Fe

5 18 125 288

.6 .6 10.7 24.0

7 19 108 227

.4 .6 2.9 13.3

9 16 108 217

1.9 1.5 11.9 33.9

NS NS NS NS

2 6 141 1940

.1 .2 16.2 462.9

2 5 133 1247

.2 .2 8.4 348.2

3 5 144 1740

.2 .4 14.3 592.9

NS NS NS NS

Liver Mn Cu Zn Fe

8 358 138 326

.6 16.3 15.8 10.7

13 499 139 268

1.1 24.5 13.8 15.5

21 586 120 239

4.2 15.1 16.6 16.2

** * NS *

Pancreas Mn Cu Zn Fe

8 7 108 113

.6 .2 14.7 17.6

10 7 94 84

.6 .5 16.9 2.6

17 8 83 104

2.9 1.8 .9 11.1

** NS NS NS

Pituitary Mn Cu Zn Fe

7 19 107 275

.1 1.0 14.5 38.6

10 23 94 217

3.4 1.7 6.2 44.2

18 23 91 274

3.5 4.1 9.9 60.2

* NS NS NS

Adrenal Mn Cu Zn Fe

5 13 80 331

.4 2.3 8.3 49.1

6 12 88 271

.8 .2 5.3 37.1

20 12 80 308

4.9 .7 4.0 17.2

** NS NS NS

8 12 22 21

2.1 4.6 5.2 2.7

76 8 24 31

13.O 1.8 4.2 11.7

221 8 29 40

15.2 3.0 9.2 21.2

** NS NS NS

Spleen Mn Cu Zn Fe

Bile Mn Cu Zn Fe

aFo ur sheep per t r e a t m e n t . bThree sheep per t r e a t m e n t . cNS, not significant (P>.05). *P<.05. * *P<.O1.

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TABLE 3. Uptake of S4Mn by selected tissues and bile 13 h after intraruminal dosing. Dietary Mn (v.g/g) 22 Tissue

300

.X

SE

.X

3000 SE

.X

SE

Significance between means

(% of dose/kg fresh sample) Heart Kidney Spleen Liver Pancreas Pituitary Adrenal Rumen Omasum Abomasum Duodenum Bile

.013 .067 .033 2.501 .126 .073 .122 21.245 10.515 1.936 1.802 1.216

.0008 a .0107 .0042 .4772 .0473 .0360 .0439 4.9756 2.4579 .3164 .4113 .3564

.021 .082 .045 1.002 .236 .060 .176 19.181 6.109 2.467 1.170 1.048

.0025 a .0249 .0148 .2568 .0953 .0205 .1064 5.4177 .9378 .1880 .2444 .6375

.022 .065 .050 .308 .308 .085 .189 11.863 6.196 2.151 .658 .861

.0105 b .0050 .0060 .0100 .1080 .0650 .0770 3.1855 3.4340 .2510 .0710 .0005

NS c NS NS * NS NS NS NS NS NS NS NS

aFour sheep per treatment. bThree sheep per treatment. cNS, not significant (P>.05). *P<.05.

liver (P<.01) and bile (P<.05). For all soft tissues and bile, the m e a n specific activities of SaMn were lower for the 3000 ppm dietary Mn t r e a t m e n t than for the 22 ppm dietary Mn treatment. F o r conversion of results to fresh or dry m a t t e r basis, means for c o n c e n t r a t i o n o f dry m a t t e r in soft tissues, gastrointestinal tract tissues, and bile are s u m m a r i z e d in Table 5.

DISCUSSION

A l t h o u g h the differences b e t w e e n t r e a t m e n t s for average daily gain and feed/gain ratio were n o t significant, possibly f r o m a short experimental period, results o f the e x p e r i m e n t indicate that sheep fed the diet containing 3000 p p m Mn gained less and utilized more feed per unit of gain than sheep fed the o t h e r diets (22 or 300 p p m Mn). G r o w t h depression was similar in sheep grazing on pasture containing a p p r o x i m a t e l y 160 p p m Mn and supplem e n t e d with 250 or 500 mg Mn per day (6). C u n n i n g h a m and coworkers (5) d e m o n s t r a t e d lower gain and efficiency o f feed utilization by calves fed diets containing over 2000 p p m Mn w h e n feed intake was c o n t r o l l e d ; feed intake Journal of Dairy Science Vol. 63, No. 3, 1980

also was decreased w h e n calves were fed ad libitum. In our e x p e r i m e n t , feed intake was a p p r o x i m a t e l y the same for the t r e a t m e n t s with either 300 or 3000 ppm Mn in the diet, but the efficiency o f feed utilization was 35% lower for the latter treatment. Efficiency of feed utilization was the main factor causing reduced growth rate in sheep fed 3000 p p m Mn. C o n c e n t r a t i o n o f Mn in soft tissues o f animals with increasing dietary Mn intake was increased in our e x p e r i m e n t and also was r e p o r t e d for sheep (6, 12, 17) and cattle (3, 9, 11). Similarly, the significant increase in Cu and a decrease in Fe c o n t e n t o f liver with increasing dietary Mn also has been r e p o r t e d (3, 7, 11, 17). It was suggested (7) that excessive Mn either converts Fe to a f o r m which is physiologically unavailable or antagonizes the e n z y m e systems that oxidize or reduce Fe at the site o f absorption. The significant e f f e c t of high dietary Mn on lowering o f Zn c o n c e n t r a t i o n in the liver o f sheep r e p o r t e d by Watson and coworkers (17) was n o t confirmed. The significant increase in c o n c e n t r a t i o n of stable Mn in liver with increasing dietary Mn was associated with a significant decrease of relative 54Mn uptake and its specific activity.

MANGANESE METABOLISM IN SHEEP

389

TABLE 4. Specific activity of s4 Mn in selected tissues and bile 13 h after intraruminal dosing. Dietary Mn (~g/g) 22 Tissue

300

X

SE

3000

X

SE

X

Significance between means

SE

(DPM//~g Mn) Heart Kidney Spleen Liver Pancreas Pituitary Adrenal Bile

278 523 719 6631 456 328 882 11191

84.5 a 130.6

383 387 762 1790 479 196 826 1387

227.2

1158.5 126.8 118.9 472.9 3444.6

69.3 a 103.2 255.5 383.2 164.1 26.2 439.8 164.4

196 168 485 236 255 96 211 210

93.0 b 4.0

NS c NS NS ** NS NS NS *

62.5

12.5 43.0 67.0 63.0 11.5

aFour sheep per treatment. bThree sheep per treatment. cNS, not significant (P>.05). *P<.05. * *P<.O1.

i n t e s t i n a l t i s s u e s . Ivan ( 1 0 ) f o u n d t h a t t h e S4Mn u p t a k e b y t h e r e t i c u l u m a n d r u m e n t i s s u e s was e v e n h i g h e r t h a n t h a t o f 6 S Z n b u t t h a t t h e S4Mn u p t a k e was m u c h l o w e r b y o t h e r g a s t r o i n t e s t i n a l t i s s u e s . Miller et al. ( 1 4 ) sugg e s t e d t h a t m u c h o f t h e Mn t a k e n u p b y

R e l a t i o n s h i p w a s also similar for bile. In addit i o n , t h e relative u p t a k e o f SaMn b y r u m e n a n d d u o d e n a l tissues decreased with increasing d i e t a r y Mn. A r o r a et al. (2) s h o w e d t h a t r u m e n t i s s u e h a s t h e a b i l i t y to a b s o r b a n a p p r e c i a b l e a m o u n t o f Zn in c o m p a r i s o n w i t h o t h e r g a s t r o -

TABLE 5. Dry matter of selected tissues and bile. Dietary Mn (~g/g)a 32 Tissue

300

X

SE

X

21.7 22.1 22.2 32.0 24.7 22.9 22.7 17.5 18.7 18.0 16.7 14.2

.99b .62 .25 .64 .28 .61 .94 .85 1.12 3.28 .85 3.02

20.7 21.7 21.7 30.5 24.8 22.7 22.1 20.6 15.2 22.5 14.8 12.5

3000 SE

X

SE

.36b .68 .13 1.17 .64 .96 .58 3.15 .28 .96 1.10 3.21

21.1 22.2 22.4 31.2 24.6 24.2 22.0 19.3 17.3 22.3 16.5 9.1

.57c .35 .70 .18 .62 .32 .49 .59 1.86 1.19 1.28 .66

(%) Heart Kidney Spleen Liver Pancreas Pituitary Adrenal Rumen Omasum Abomasum Duodenum Bile

aNone of the differences between treatment means was significant (P>.O5). hFour sheep per treatment. CThree sheep per treatment. Journal of Dairy Science Vol. 63, No. 3, 1980

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intestinal m u c o s a o f calves is n o t t r a n s p o r t e d to t h e b l o o d b u t in s o m e w a y r e e n t e r s t h e intestinal c o n t e n t s s o o n after u p t a k e , t h u s n o t c o m pleting t h e a b s o r p t i o n process. It w o u l d be i n t e r e s t i n g t o k n o w if similar process takes place in t h e r u m e n . Since higher d i e t a r y Mn d e c r e a s e d t h e relative S4Mn u p t a k e b y s o m e g a s t r o i n t e s t i n a l and s o f t tissues and bile, d i e t a r y Mn a p p e a r s t o a f f e c t a b s o r p t i o n and secret i o n o f Mn. This agrees with t h e suggestion o f Miller et al. (13) and A b r a m s et al. (1) t h a t tissue h o m e o s t a s i s o f Mn m a y be c o n t r o l l e d largely t h r o u g h the rate o f a b s o r p t i o n f r o m the g a s t r o i n t e s t i n a l t r a c t and e n d o g e n o u s fecal excretion. A l t h o u g h s o m e tissue c o n c e n t r a t i o n s o f Mn, Cu, and Fe change, t h e g r o w t h rate o f s h e e p and e f f i c i e n c y o f feed utilization are n o t a f f e c t e d b y d i e t a r y Mn up to 300 p p m . ACKNOWLEDGMENTS

A p p r e c i a t i o n is e x p r e s s e d to P. Jui for statistical analysis o f the data, to M. K. Bryan, H. Mode, and J. S h a c k l e t o n for t e c h n i c a l assistance, and to F. Fisher for care o f e x p e r i m e n t a l animals. REFERENCES

1 Abrams, E., J. W. Lassiter, W. J. Miller, M. W. Neathery, R. P. Gentry, and D. M. Blackmon. 1977. Effect of normal and high manganese diets on the role of bile in manganese metabolism of calves. J. Anim. Sci. 45:1108. 2 Arora, S. P., E. E. Hatfield, D. S. Garrigus, T. G. Lohman, and B. B. Doane. 1969. Zinc-65 uptake by rumen tissue. J. Nutr. 97:25. 3 Boogaerdt, J. 1961. Harmful effects from too high amounts of manganese for cows. Tijdschrift Voor Dierengeneeskunde 86:956. 4 Buchanan-Smith, J. G., E. Evans, and S. O. Poluch. 1974. Mineral analyses of corn silage produced in Ontario. Can. J. Anim. Sci. 54:253.

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5 Cunningham, G. N., M. B. Wise, and E. R. Barrich. 1966. Effect of high dietary levels of manganese on the performance and blood constituents of calves. J. Anim. Sci. 25:532. 6 Grace, N. D. 1973. Effect of high dietary Mn levels on the growth rate and the level of mineral elements in the plasma and soft tissues of sheep. New Zealand J. Agric. Res. 16:177. 7 Hartman, R. H., G. Matrone, and G. H. Wise. 1955. Effect of high dietary manganese on hemoglobin formation. J. Nutr. 57:429. 8 Hecker, J. F. 1969. A simple rapid method for inserting rumen cannulae in sheep. Australian Vet. J. 45:293. 9 Howes, A. D., and I. A. Dyer. 1971. Diet and supplemental mineral effects on manganese metabolism in newborn calves. J. Anim. Sci. 32:141. 10 Ivan, M. 1979. Metabolism of radiomanganese and radiozinc in sheep - effects of intraruminal dosing with nitrilotriacetic acid. Can. J. Anim. Sci. 59:283. 11 Ivan, M., and C. M. Grieve. 1975. Effects of zinc, copper, and manganese supplementation of highconcentrate ration on digestibility, growth, and tissue content of Holstein calves. J. Dairy Sci. 58:410. 12 Lassiter, J. W., and J. D. Morton. 1968. Effects of a low manganese diet on certain ovine characteristics. J. Anim. Sci. 27:776. 13 Miller, W. J., M. W. Neathery, R. P. Gentry, D. M. Blackmon, and J. W. Lassiter. 1973. Fecal excretion, tissue accumulation and turnover of manganese-54 after intravenous dosing in Holstein calves fed a practical-type diet. J. Anim. Sci. 37:827. 14 Miller, W. J., M. W. Neathery, R. P. Gentry, D. M. Blackmon, J. W. Lassiter, and F. M. Pate. 1972. Distribution and turnover rates of radioactive manganese in various tissues after duodenal dosing in Holstein calves fed a practical-type diet. J. Anita. Sci. 34:460. 15 Steel, R.G.D., and J. H. Torrie. 1960. Principles and procedures of statistics. McGraw-Hill, New York, NY. 16 Underwood, E. J. 1971. Trace elements in human and animal nutrition. 3rd ed. Academic Press, New York, NY. 17 Watson, L. T., C. B. Ammerman, J. P. Feaster, and C. E. Roessler. 1973. Influence of manganese intake on metabolism of manganese and other minerals in sheep. J. Anim. Sci. 36:131.