Relative Bioavailability of Manganese in a Manganese-Methionine Complex for Broiler Chicks1

METABOLISM AND NUTRITION Relative Bioavailability of Manganese in a Manganese-Methionine Complex for Broiler Chicks 1 P. R. HENRY,2 C. B. AMMERMAN,2 and R. D. MILES3 Departments of Animal Science and Poultry Science, University of Florida, Gainesville, Florida 32611 (Received for publication October 19, 1987)

1989 Poultry Science 68:107-112 INTRODUCTION

Biological availability of manganese (Mn) sources is of practical concern when feed ingredients that are low in Mn constitute a large percentage of diets used for poultry. In addition, ingredients including corn, soybean meal, wheat bran, and fish meal have been reported to decrease absorption of Mn (Halpin et al., 1986; Halpin and Baker, 1986). Bioavailability of Mn has become an increasing concern recently because of the extremely rapid growth rate of commercial broiler strains, which puts additional stress on bone structure. Some chelates improve bioavailability of the mineral involved (Kratzer and Vohra, 1986). Fly et al. (1987) suggested that Mn from Mnmethionine (Mn-met) was more available than that from feed grade Mn oxide for chicks; however, Baker and Halpin (1987) reported that Mn from a Mn-protein chelate was similar in value to that from Mn sulfate. The following experi-

'Florida Agricultural Experiment Station Journal Series Number 8440. department of Animal Science. 'Department of Poultry Science.

ment was conducted to determine the relative bioavailability of Mn from reagent grade Mn oxide and feed grade Mn-methionine compared with the standard source for broiler chicks, reagent grade Mn sulfate. MATERIALS AND METHODS

This experiment was conducted with 288 1day-old male, Cobb, feather-sexed chicks assigned to 16 treatment groups. The basal cornsoybean meal diet (Table 1) was formulated to meet the requirements for starting chicks (National Research Council, 1984) and contained 93 ppm Mn on a dry matter basis, by analysis. The basal diet was supplemented with 0, 700, 1,400, or 2,100 ppm Mn (as-fed basis) as either reagent grade Mn sulfate (MnS0 4 -H 2 0), reagent grade Mn monoxide (MnO), or Mn-met complex (35% methionine, 15.7% Mn). In addition, six diets were formulated to contain 700, 1,400, or 2,100 ppm Mn from MnS0 4 -H 2 0 or MnO in combination with .16, .32, or .48% added DL-methionine, respectively, to equalize methionine concentrations in the Mn-met-containing diets. In the event that the high level of added methionine might be detrimental, comparisons of Mn bioavailability could be made

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ABSTRACT The relative bioavailability of Mn from reagent grade Mn monoxide and feed grade Mnmethionine was compared with that from reagent grade Mn sulfate using 288 one-day-old male Cobb chicks. The basal corn-soybean meal diet (93 ppm Mn dry matter basis) was supplemented with 0, 700, 1,400, and 2,100 ppm Mn as Mn sulfate monohydrate, Mn oxide, or Mn-methionine. Additional diets contained 700, 1,400, and 2,100 ppm Mn as sulfate or oxide in combination with .16, .32, or .48% added DLmethionine, respectively, to equalize methionine concentrations in Mn-methionine-containing diets. Diets were fed ad libitum for 3 wk. Tibia and kidney Mn concentrations increased linearly (P<.001) as dietary Mn increased. Addition of methionine to diets containing sulfate and oxide did not influence (P>. 10) tissue Mn concentrations. Based on slope ratios from multiple linear regression of bone and kidney Mn concentrations on added dietary Mn from various sources, the respective relative bioavailability values were 96 and 86% from Mn oxide and 108 and 132% from Mn-methionine compared with 100% from Mn sulfate. Except for the first, all values were significantly different from 100%. Thus, Mn from Mn oxide is significantly less available and Mn from Mn-methionine is significantly more available than that from Mn sulfate monohydrate. (Key words: manganese-methionine, bioavailability, mineral, broiler)

HENRY ETAL.

108 TABLE 1. Composition of basal diet

Ingredient

Amount (as-fed basis)

(%)

Composition 3 Dry matter, % CP, % ME, kcal/kg Manganese, ppm

53.92 37.20 .40 .50

2.50 1.01 1.72 .25

2.50 89.15 23

3,000 93

1 Ingredients supplied per kilogram of diet: vitamin A palmitate, 6,600 IU; vitamin D 3 , 2,200 IU; menadione dimethylpyrimidinol bisulfite, 2.2 mg; riboflavin, 4.4 mg; pantothenic acid, 13 mg; niacin, 40 mg; choline chloride, 500 mg; vitamin B 1 2 , .022 mg; biotin, 1 mg; ethoxyquin, 125 mg; Mn, 60 mg; Fe, 75 mg; Cu, 6 mg; Zn, 36 mg; I, 1.1 mg; Se, .1 mg. 2 Manganese sources and additional DL-methionine added at the expense of equivalent weights of cornstarch. 3 Dry matter basis except for dry matter. Dry matter and Mn were determined by analysis; protein and energy were calculated.

among methionine-containing diets. During the experiment, chicks were housed from 0 to 21 days of age in two electrically heated, thermostatically controlled Petersime batteries with stainless steel fittings (Petersime Incubator Co., Gettysburg, OH). Birds were maintained on a 24-h constant light schedule. Each of 16 diets was fed to three pens of six chicks in a completely randomized design. Feed and tap water were available ad libitum. When mortality occurred, the weight of the remaining feed in that pen was recorded and daily feed intake and feed conversion calculations were adjusted for the dead bird. At the end of the experiment, chicks were weighed individually and feed consumption for each pen was recorded. Chicks were killed by cervical dislocation, and both kidneys and right tibia with associated cartilages were excised and frozen for analysis. Standard reference material (bovine liver-1577 or citrus leaves-1572) from the National Bureau of Standards was analyzed

RESULTS AND DISCUSSION

The reagent grade Mn sulfate monohydrate and Mn oxide and feed grade Mn-met complex contained, by analysis, 32.5, 77.2, and 15.7% Mn (as-fed basis), respectively. The Mn oxide was insoluble (.08%) in water but completely soluble (100%) in .4% HCl, 2% citric acid, and neutral ammonium citrate. The Mn sulfate and Mn-met were completely soluble (100%) in all

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Ground yellow corn Soybean meal (49% CP) Iodized salt Microingredients1 Corn oil Ground limestone Dicaleium phosphate DL-Methionine Cornstarch 2

with all samples of similar matrix. Manganese in diets and tissues was determined by flame atomic absorption spectrophotometry (AAS) on a Perkin-Elmer Model 5000 spectrophotometer (Perkin-Elmer Corporation, Norwalk, CT) with AS-50 autosampler (Anonymous, 1982), following dry ashing and solubilizing in HCl. Manganese sources were refluxed for 4 h on a hotplate in porcelain crucibles with a 1:1 (vol/ vol) mixture of HC1:HN03. The samples were then filtered; Mn was determined by AAS with matrix-matched standards. Relative solubility of a . 1-g sample of Mn sources in 100 mL of water, 2% citric acid, .4% HCl, or neutral ammonium citrate was determined by constant stirring for 1 h at 37 C on a magnetic stir hotplate (Watson etal, 1970). Data were subjected to ANOVA using the general linear models (GLM) procedure in SAS (1982) with a model that included dietary Mn concentration and Mn source as main effects and their interaction. Diets containing sulfate or oxide with and without methionine were considered separate treatments, giving a 3 X 5 factorial arrangement plus a control. For feed intake and feed conversion, pen was the experimental unit; for gain and tissue Mn concentrations, individual chick was the experimental unit. Means were separated by Duncan's multiple range test. After observing that the addition of methionine to the diets containing Mn sulfate or Mn oxide failed to influence tissue Mn concentration, multiple linear regression equations were calculated by least squares using the GLM procedure of SAS (1982). The model used was: y = b 0 + b,x, + b2x2 + b 3 x 3 , where y equals the concentration of Mn in bone or kidney and Xj, x 2 , and x3 equal the quantity of added Mn from Mn sulfate, Mn oxide, and Mn-met, respectively. Slope ratios of b 2 /b! and b3/b[ were calculated to determine the relative amounts of Mn deposited in the tissues from Mn oxide or Mn-met in relation to that from Mn sulfate, following the procedure of Finney (1978).

BIOAVAILABILITY OF MANGANESE

TABLE 2. Effect of manganese source on chick performance

Source

1

Daily feed intake 2

Daily gain3

Feed conversion2

29.8 27.8 27.4 28.2 27.7 27.6

(g/g) 1.52a 1.52 a 1.55 a 1.50 a 1.54 a 1.44 b

(g) Control Sulfate Oxide Sulfate + met 4 Oxide + met 4 Mn-methionine Pooled SE

42.1a 38.9bc 39.3bc 39.2 b e 39.6 b 36.8 C .69

.46

.02

Means in columns with no common superscripts differ significantly (P<.05). 1 Basal diet contained 93 ppm Mn (dry matter basis) by analysis. 2 Each value represents the mean of three pens of six chicks fed from 0 to 21 days of age. 3 Each value represents the mean of 18 chicks fed from 0 to 21 days of age.

"met = DL-Methionine; added to the diet at the concentration provided by a similar dietary addition of Mn-methionine.

approximately .38% methionine in feed ingredients. Griminger and Fisher (1967) reported a growth depression in chicks fed 1% added DLmethionine for 3 wk in a diet that was calculated to contain .39% methionine from other ingredients. There were no differences in feed intake or gain among treatments when DL-methionine was added to diets (.54% methionine) for broilers for 4 wk at 1%, but 2% supplemental methionine decreased feed intake and weight gain (Nam et al., 1984). Dietary Mn concentration did not influence (P>. 10) daily feed intake or daily gain, which is in agreement with results of Black et al. (1984a), based on feeding of as much as 4,000 ppm Mn for 26 days. Bone Mn concentration increased (P<.01) from a mean of 5.5 to 26.1, 47.2, and 65.7 ppm on an ash weight basis as added dietary Mn concentration from all sources increased from 0 to 700, 1,400, and 2,100 ppm, respectively (Table 3). Mean bone Mn concentrations for birds fed Mn sulfate, Mn oxide (with and without methionine), and Mn-met were 46.1, 44.7, and 49.9 ppm on an ash weight basis, respectively. There was no effect (P>.10) of methionine addition to sulfate or oxide diets, so data were pooled within source for regression analysis (Table 4). Multiple regression of bone Mn concentration on quantity of added dietary Mn from the three sources resulted in an equation with an adequate fit to the linear model (R2 = .81; n = 285). The slope for Mn-met was greater (P<.01) than those for Mn sulfate or Mn oxide. Kidney Mn concentration increased (P<.01) as dietary Mn concentration increased; 8.7, 15.6, 20.2, and 24.2 ppm on a dry matter basis, respectively (Table 3). Kidney Mn concentration was also greater (P<.01) in birds fed Mnmet than those fed sulfate or oxide with and without methionine (22.6, 19.8, and 18.9 ppm, respectively). Methionine addition to sulfate or oxide diets had no effect on kidney Mn concentration, so data were pooled within source for regression analyses. Kidney was less sensitive to dietary Mn than bone, as indicated by the regression coefficients (Table 4). Kidney Mn did not fit the linear model as well as bone (R2 = .70; n = 285). The slope representing birds fed Mn-met complex was greater (P<.01) than slopes representing birds fed sulfate or oxide (Table 4). Relative bioavailability estimates of Mn sources and confidence limits based on multiple linear regression slopes for bone and kidney Mn

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four solvents. Low solubility of Mn oxides in water has been reported previously (Watson et al., 1970; Black et al, 1984a; Henry et al., 1987). Mortality averaged 1.0% and was not related to treatment. Birds fed diets containing Mn-met ate less feed (P<.05) than those fed the unsupplemented control diet or those fed the diet containing MnO with methionine, but differences among intakes of other treatments were not significant (Table 2). There was an interaction (P<.05) between Mn source and level on feed intake: birds fed diets containing 2,100 ppm Mn from Mn-met ate less feed than birds fed the same dietary level from other sources. There was no effect (P>. 10) of Mn source on average daily gain; however, birds fed diets with Mn-met had more efficient feed conversion rates than birds fed other diets. There was no main effect (P>. 10) of Mn level on performance of chicks. There was no effect (P>.10) of additional methionine in birds fed Mn sulfate or oxide, so the reduction in feed intake was unlikely to be due to excess methionine. Diets with 2,100 ppm Mn contained a total of .72% added DL-methionine, and there was

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HENRY ET AL.

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TABLE 3. Effect of manganese source and dietary manganese concentration on tissue manganese concentration of chicks

Source

Added Mn 1

Bone Mn (ash basis) 2 ' 3

Control Sulfate Sulfate Sulfate Oxide Oxide Oxide Sulfate + met" Sulfate + met 4 Sulfate + met" Oxide + met 4 Oxide + met 4 Oxide + met 4 Mn-methionine Mn-methionine Mn-methionine

0 700 1,400 2,100 700 1,400 2,100 700 1,400 2,100 700 1,400 2,100 700 1,400 2,100

5.5 25.4 47.2 67.6 25.8 46.5 58.3 26.8 44.8 65.0 24.1 46.6 67.3 28.7 51.1 69.8

Kidney Mn (dry basis) 2 ' 3

(ppm)

1.25

.335

1

Basal diet contained 93 ppm Mn (dry matter basis) by analysis.

2

Each value represents the mean of individual samples from 18 chicks fed from 0 to 21 days of age.

3

Significant main effects of source and dietary Mn level (P<.01).

4

met = DL-Methionine; added to the diet at the concentration provided by a similar dietary addition of Mn-methionine.

TABLE 4. Relative value of manganese sources1

Tissue

Mn source

Regression coefficient (slope ± SE)

Relative value

Confidence limits

Bone

Sulfate2 Oxide 2 Mn-methionine

.0284 b .0273 b .0308 a

± .00091 ± .00091 ± .00129

100 96 108

94 to 99 102 to 115

Kidney

Sulfate2 Oxide 2 Mn-methionine

.00660 b ± .00030 .00565 b ± .00030 .00871 a ± .00043

100 86 132

84 to 87 123 to 141

(,u)

a ' b Slopes differ (P<.01) within a tissue. 'Calculated from multiple regression: y t = 6.42 + .0284 x, + .0273 x 2 + .0308 x 3 ; (R 2 = .81; SD = 9.12; n = 285) and y 2 = 10.59 + .00660 x, + .00568 x 2 + .00873 x 3 ; (R2 = .70; SD = 3.02; n = 285), where y, equals parts per million bone Mn (ash weight basis), y2 equals parts per million kidney Mn (dry basis), and x , , x 2 , and x 3 equal parts per million added dietary Mn from Mn sulfate, Mn oxide, and Mn-methionine, respectively. 2

Data pooled from treatment groups with and without added methionine.

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Pooled SE

8.7 14.6 20.2 24.1 15.8 19.2 20.4 15.8 19.9 24.4 15.8 19.1 22.9 16.0 22.8 29.1

BIOAVAILABILITY OF MANGANESE

Aranda etal., 1983). The kinetics of the absorption process were compatible with a high-affinity, low-capacity, active transport mechanism. Thus, there may be a limited role-for small molecular weight ligands associated with both diffusion and active transport (Kratzer and Vohra, 1986). Baker and Halpin( 1987) reported that Mn in an Mn-proteinate was similar to that of Mn sulfate. Fly et al. (1987) also reported that Mn-met was more available than MnO in purified diets for chicks, especially with the inclusion of 10% corn-soybean meal in the diet. The phytate, fiber, or both in corn and soybean meal have been shown to decrease Mn availability from supplemental Mn sources (Halpin et al, 1986; Halpin and Baker, 1986). All or part of the amino acid-complex Mn in Mn-met may have been protected from binding by ligands in corn or soybean meal or both, which would have rendered this source more available than Mn sulfate. ACKNOWLEDGMENT

The authors wish to acknowledge Zinpro Corp., Bloomington, MN for funds in support of this research and for supplying the Mn-met complex. REFERENCES Anonymous, 1982. Analytical Methods for Atomic Absorption Spectrophotometry. The Perkin-Elmer Corp., Norwalk, CT. Baker, D. H., and K. M. Halpin, 1987. Efficacy of a manganese-protein chelate compared with that of manganese sulfate for chicks. Poultry Sci. 66:1561-1563. Black, J. R., C. B. Ammerman, P. R. Henry, and R. D. Miles, 1984a. Biological availability of manganese sources and effects of high dietary manganese on tissue mineral composition of broiler-type chicks. Poultry Sci. 63:1999-2006. Black, J. R., C. B. Ammerman, P. R. Henry, and R. D. Miles, 1984b. Tissue manganese uptake as a measure of manganese bioavailability. Nutr. Rep. Int. 29:807814. Finney, D. J., 1978. Statistical Method in Biological Assay. 3rd ed. Charles Griffin & Co., London, UK. Fly, A. D., O. A. Izquierdo, K. L. Lowry, and D. H. Baker, 1987. Manganese (Mn) bioavailability in a Mnmethionine chelate. Fed. Proc. 46:911. (Abstr.) Garcia-Aranda, J. A., R. A. Wapnir, and F. Lifshitz, 1983. In vivo intestinal absorption of manganese in the rat. J. Nutr. 113:2601-2607. Griminger, P., and H. Fisher, 1967. Methionine excess and chick growth. Poultry Sci. 47:1271-1273. Halpin, K. M., and D. H. Baker, 1986. Manganese utilization in the chick: effects of corn, soybean meal, fish meal, wheat bran, and rice bran on tissue uptake of manganese. Poultry Sci. 65:995-1003.

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concentrations are found in Table 4. Availability of Mn from Mn oxide and Mn-met was 96 and 108% that from Mn sulfate, based on bone, and 86 and 132%, based on kidney. When these values were averaged and sulfate was set at 100%, relative values were 91 and 120% for Mn oxide and Mn-met, respectively. The Mnmet was 133% available compared with reagent grade oxide. The average estimate of 91% availability for reagent grade MnO in the present study was greater than other values reported for the compound. Linear and multiple linear regression slopes and increase in bone and liver Mn concentrations (Black et al., 1984a) and bone and kidney Mn concentrations (Wong-Valle et al., 1988) estimated relative Mn bioavailability values for reagent grade MnO at 71 and 85% of reagent grade Mn sulfate, respectively. Liver Mn concentration did not respond as readily to dietary Mn as did kidney Mn (Black et al., 1984b), which may account for the lower availability estimate reported by Black etal.(\984a). Wong-Valle et al. (1988) estimated bioavailability of Mn from reagent grade Mn oxide and three feed grade Mn oxides to be 85, 62, 64, and 82% compared with Mn sulfate. The Mn in Mn-met could therefore probably be given a value, compared with feed grade Mn oxides, higher than the 133% calculated for reagent grade Mn oxide in the present experiment. The high relative bioavailability of Mn-met may be related partially to its high relative solubility and small particle size. Fly et al. (1987) suggested that Mn from Mn-met was more available than that from feed grade oxide, but they did not provide a numerical value. Southern and Baker (1983) used multiple regression of bile Mn concentration on Mn intake at 0, 3,000, and 4,000 ppm to give an equation that determined a value for Mn from MnCl 2 , 4H 2 0, another highly soluble source of Mn, at 102% of sulfate. Many mineral-amino acid chelates, especially those containing Cu, Zn, and Fe have been shown to have generally greater bioavailability than inorganic sources of the mineral (Kratzer and Vohra, 1986). Copper, Zn, and Fe, unlike Mn, must be bound in the intestinal lumen prior to absorption (Underwood, 1977); therefore, their increased bioavailability when chelated with an amino acid can be explained. Absorption of Mn from ileum or jejunum in rats was more rapid when L-histidine or citrate was included in the perfusion solution (Garcia-

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Halpin, K. M., D. G. Chausow, and D. H. Baker, 1986. Efficiency of manganese absorption in chicks fed cornsoy and casein diets. J. Nutr. 116:1747-1751. Henry, P. R., C. B. Ammerman, and R. D. Miles, 1987. Bioavailability of manganese monoxide and manganese dioxide for broiler chicks. Nutr. Rep. Int. 36:425-433. Kratzer, F. H., and P. Vohra, 1986. Chelates in Nutrition. CRC Press, Inc., Boca Raton, FL. Nam, D. S., I. K. Han, and J. D. Kim, 1984. Effects of supplemental copper and excess dietary methionine on the growing performance of broiler chicks. Korean J. Anim. Sci. 26:621-630. National Research Council, 1984. Nutrient Requirements of Domestic Animals. No. 1. Nutrient Requirements

ofPoultry. 8thed. Natl. Acad. Sci., Washington, DC. Southern, L. L., and D. H. Baker, 1983. Excess manganese ingestion in the chick. Poultry Sci. 62:642-646. SAS, 1982. SAS User's Guide: Statistics. SAS Inst. Inc., Cary, NC. Underwood, E. J., 1977. Trace Elements in Human and Animal Nutrition. 4th ed. Academic Press, New York, NY. Watson, L. T., C. B. Ammerman, S. M. Miller, and R. H. Harms, 1970. Biological assay of inorganic manganese for chicks. Poultry Sci. 49:1548-1554. Wong-Valle, J., C. B. Ammerman, P. R. Henry, and R. D. Miles, 1988. Bioavailability of Mn as feed grade Mn oxides for chicks. Poultry Sci. 67(Suppl. 1):41. (Abstr.) Downloaded from http://ps.oxfordjournals.org/ at University of Strathclyde on April 9, 2015