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Mineral Mixture Composition—A Factor in Chick Bioassays of the Protein Quality of Fish Meals
BEHAVIOR AND GONAD IN BLINDED CHICKENS light on the pineal and gonads of the Japanese quail. Arch. Anat. Hist. Embryol. (Paris), SI: 615-626. Underwood, H., and M. Menaker, 1970. Photoperiodically significant photoreceptation in sparrows: Is the retina involved? Science, 167: 298-301. Van Brunt, E. E., M. D. Shepherd, J. R. Wall, W. F. Ganong and M. T. Clegg, 1964. Penetration
1535
of light into the brain of mammals. Ann. New York Acad. Sci. 117: 217-227. Van Tienhoven, A., 1968. Reproductive Physiology of Vertebrates. W. B. Saunders Co., Philadelphia. p. 63. Wetterberg, L., E. Geller and A. Yuwiler, 1970. Harderian gland: An extraretinal photoreceptor influencing the pineal gland in neonatal rats? Science, 167: 884-885.
Mineral Mixture Composition—A Factor in Chick Bioassays of the Protein Quality of Fish Meals MILLER
(Received for publication May 20, 1970)
M
ILLER and Kifer (1970b) showed excess chloride. They showed that such adthat the evaluation of the quality of verse effects also resulted from the addition the protein in fish meal, when the meal was of other chloride and sulfate compounds in fed as the sole source of protein, was influ- diets made up entirely of casein or of soy enced by excesses of lysine present in fish proteins. meal over the concentration needed in the Other possible excess inorganic subdiet and by various anionic components in stances in the diet of chicks could be the the mineral mixture. The excess of lysine high amounts of Ca, P, and Fe contributed was overcome by a dilution of the concen- by fish meals when they are used as the tration of lysine in the diet by the addition only source protein. Some menhaden fish of glutamic acid (free-base form) or was meals containing 6% or more of Ca (Kifer overcome by counteraction from the addi- et al., 1968) will contribute, when fed to tion of arginine. The excess of anionic com- supply 15% protein, about 1.5% Ca to the ponents was overcome by the inclusion in diet. This level of Ca is ordinarily considthe diet of the antiacids Al(OH) 3 and Na- ered as being unnecessarily high. For that HC0 3 . The detrimental anionic effect was reason, a comparative study of a lower evident when the addition of HC1 salt of level of Ca and P was necessary to deterglutamic acid depressed growth. In con- mine if the high levels were detrimental. trast, the free-base form of glutamic acid Similarly, because of the high concentraimproved growth significantly, owing to tions of Fe (438 p.p.m. on the average) in two possibilities: (1) the elimination of the menhaden fish meal (Kifer et al., 1968), a detrimental HC1 portion of this amino acid comparative study of the effect of decreassupplement and (2) the increase of nones- ing the concentration of Fe in the mineral sential amino acid nitrogen (glutamic mixture was necessary. acid). In view of these observations, the comNesheim et al. (1964) also reported that position of the mineral mixture that has glutamic. HC1 was detrimental due to the been used by Grau and Williams (1955),
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DAVID
National Center for Fish Protein Concentrate, Bureau of Commercial Fisheries, College Park, Maryland 20740
1536
D.
MILLER
TABLE 1.—Composition (%) of variousfishmeals Fish meals
Protein
Fat ethyl ether extractable
Ca
P
M T5 TM1 TM2 TM3 TM4 TM5 TM6
65.41 61.89 65.30 65.16 64.92 64.84 64.05 64.24
11.65 13.50 12.93 10.69 12.97 12.33 13.06 13.43
4.45 4.19 4.79 5.05 4.75 4.31 4.25 4.18
2.50 2.33 2.66 2.88 2.72 2.48 2.46 2.44
In Experiment 1, Fish Meal M was studied with eight different mineral supplementations, OM through F. In Experiment 2, five mineral mixtures (OM, A, G, H, and I) were studied with Fish Meal T5. Also the relative growth effect of three different mineral mixtures (OM, A, and I) were comparatively studied with seven different fish meals (T5 and TM1-6). These studies involved a total of 31 treatments.
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Mineral Mixtures. Ten different mineral mixtures (Table 3) were used. Step-wise changes were made to determine the effect of inorganic substances on the growth of the chick when fish meal was the sole source of protein. The OM Mineral Mixture was similar to that used by many investigators (Grau and Williams, 1955; and Ousterhout et al., 1962) except that in the present study the Ca and P were not included in the mineral mixtures but were supplied as variable ingredients dicalcium phosphate and phosphoric acid to maintain a constant dietary level of 1.5 and 1.0%, MATERIALS AND METHODS respectively except in one treatment. In Basal Diets. Table 1 shows the composition Experiment 1, the effect of reducing the Ca of eight menhaden fish meals studied in this concentration to 1.25% and the P to 0.8% investigation, which consisted of two experi- was studied comparatively by decreasing ments. Eight basal diets (Table 2), each the dietary supplements of dicalcium phosone containing a different menhaden fish phate and phosphoric acid in one of the meal, were used to study the effects of dif- treatments containing Mineral Mixture D. ferent mineral mixtures (Table 3) on the At the 5% level the OM Mineral Mixture growth response of the chicks. Diet 1 con- contributed to the diets 0.20% Na and tained 12% protein, all others, 15%. The 0.26% K, which were kept constant in all calorie content of the diet was 1377, which mineral mixtures except in Mix A, which was maintained by adjusting the levels of supplied 0.47% Na (Table 3). The SO* glucose (Cerelose)1 cellulose (Solka Floe),2 content was 0.432% in Mineral Mixtures and corn oil (the lower limit was set at OM, A, B, C and was decreased to 0.032% 2.0%). 3 by the substitution of MgO or MgC0 3 for MgS0 4 in the other mineral mixtures. The 1 Cerelose, Corn Products Company, Argo, Illi- concentration of CI was 0.542% in Mineral nois. 2 Solka Floe, Brown Company, Berlin, New Mixtures OM, A, C, and H and was lowered in the other mineral mixtures to 0.304% Hampshire. 3 The use of trade names is merely to facilitate when KC1 was replaced by K 2 C0 3 or descriptions; no endorsement is implied. KHC0 3 . The concentration of Fe in Min-
Ousterhout and Snyder (1962), Miller et al. (1970), and Miller and Kifer (1970a, b) in diets containing fish meal as the sole source protein was niouincu stepwise, xuis investigation was conducted to determine the effect on chick growth responses from various fish meals as the sole source protein in the diet (1) by decreasing the concentrations of CI, S0 4 , and Fe in the mineral mixtures, (2) by adding antiacids, and (3) by decreasing the concentrations of Ca and P0 4 .
1537
PROTEIN QUALITY BIOASSAYS
TABLE 2.—Composition (%) of diets Diet No.
1
2
3
4
5
6
7
8
Experiment No.
1
2
2
2
2
2
2
2
Fish meal M Fish meal T5 Fish meal TM1 Fish meal TM2 Fish meal TM3 Fish meal TM4 Fish meal TM5 Fish meal T M 6 Cerelose1 Vitamin mix2 Mineral mix3 DiCaP0 4
18.35
—
— —
— — —
— — — —
— — — — —
— — — — — —
— — — — — — —
24.24
— — — — — — —
— — — — — —
23.00
— — — — —
23.02
— — — —
23.09
23.13
— — —
23.41
— —
55.85 5.00 5.00 1.88 0.32 5.51 0.20 2.00
56.46 5.00 5.00 1.54 0.35 6.45 0.20 2.00
57.62 5.00 5.00 1.31 0.32 5.53 0.20 2.00
56.46 5.00 5.00 1.54 0.29 6.40 0.20 2.00
56.80 5.00 5.00 1.92 0.25 5.70 0.20 2.00
56.32 5.00 5.00 1.92 0.22 5.93 0.20 2.00
23.35 56.19 5.00 5.00 2.00 0.22 6.04 0.20 2.00
Protein content (%)
12
15
15
15
15
15
15
15
H3PO4
—
1
Cerelose, Corn Products Company, Argo, Illinois. The vitamin mixture fortified each kilogram of diet with 7 mg. thiamine-HC1, 7 mg. riboflavin, 8 mg. pyridoxineHCl, 60 mg. niacin, 30 mg. calcium pantothenate, 2 mg. folic acid, 2 mg. menadione, 29 jug. vitamin B12, 220 /ig. biotin, 90001.U. vitamin A, 4501.C.U. vitamin fi3, 441.U. dl-alpha tocopherol acetate, and 1.65 gm. choline-chloride. 3 See Table 3 for composition of mineral mixtures. 4 Solka Floe, Brown Company, Berlin, New Hampshire. 6 Ethoxyquin premix ( 1 + 9 cerelose). 2
TABLE 3.—Composition and contribution of various mineral mixtures at a lezel of 5*V0 in the diet Mineral mixture Experiment No.
OM \
A
B
C
D
E
\
1
1
1
1
% of diet Variable portion NaHCOs — 1.0 — — — — Al(OH) 3 — 1.0 — — — — KC1 0.5 0.5 — 0.5 — — K 2 C0 3 — — 0.463 — 0.463 0.463 KHCO3 _ _ _ _ _ _ _ _ MgS0 4 0.5 0.5 0.5 — — — MgO — — — 0.145 0.145 0.145 MgC0 3 _ _ _ _ _ _ _ FeC 6 H 5 0,-3H 2 0 0.08 0.08 0.08 0.08 0.08 0.040 Constant portion* 0.568 in all diets Glucose added to bring total of each mineral mixture to 5 % in each diet. The above mineral mixtures contribute the following to the diets: CI, % 0.542 0.542 0.340 0.542 0.304 0.304 S0 4 , % 0.432 0.432 0.432 0.032 0.032 0.032 CO3, % 0 0.714 0.201 0 0.201 0.201 Na, % 0.200 0.470 0.200 0.200 0.200 0.200 K, % 0.260 0.260 0.260 0.260 0.260 0.260
F *
_
G
H
I
2
2
2
_
0.08
0.5 — — — — o.35 0.08
_ _ — — 0.67 — — 0.35 0.08
0.304 0.432 0.402 0.200 0.260
0.542 0.032 0.249 0.200 0.260
0.304 0.032 0.651 0.200 0.260
_ — 0.463 — 0.145 _ —
0.304 0.032 0.201 0.200 0.260
_ _
— — 0.67 0.5 —
_ _
* Constant portion in all mineral mixtures contains 0 . 5 % NaCl (iodized); 0.03% MnS04-H 2 0; 0.02% ZnS0 4 -7H 2 0; 0 . 0 1 % Al2(S0 4 )3-18H 2 0; 0.004% NaSi0 3 -9H 2 0; 0.002% C u S C v H 2 0 ; 0 . 0 0 1 % Co-(CH 3 COO)2-H2O;0.001%(NH < )2MoO 4 -4H 2 O. Total 0.568%.
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Solka Floe4 Ethoxyquin premix6 Corn oil
60.68 5.00 5.00 2.61 0.22 5.71 0.20 2.87
1538
D. MILLER TABLE 4.—Experiment 1—growth effect of various mineral changes in 12% protein diets containing Menhaden Fish Meal M
Mineral mixtures code
OM A B C D D E F
Chick weights
Substitution of Basic compounds
—
Al(OH) 3 +NaHC0 3 K 2 C0 3 MgO K 2 C0 3 +MgO K 2 C0 3 +MgO K 2 C0 3 +MgO K 2 C0 3 +MgO
For acidic compounds
Decreases
Average 1
Relative growth
Gram
%
169a2
— —
KC1 MgS0 4 KCl+MgSO, KCl+MgS04 KCl+MgS04 KCl+MgSOi
CaHPO,, H3PO4 Fe citrate (J orig.) Fe citrate (complete omission) n)
216b 199b 201 b 235« 234° 216b 203"
1003 128 118 119 139 139 128 120
1
eral Mixture E was reduced to one-half of that in all other mineral mixtures. Chicks. Day-old Arbor Acre cockerels were wingbanded and randomized, 12 chicks per pen, three replicates per treatment. Feed and water were supplied at all times in thermostatic compartmentized chick-starter batteries in a temperature-controlled room. Final individual weights of chicks were obtained on the 19th or 20th day. Only the weights of the heaviest 10 chicks of each pen were included in the final averages. The chick weights were analyzed for significance by Fisher's (1950) modification of Student's t-test. RESULTS AND DISCUSSION The growth effect of various mineral substitutions in a 12% protein diet containing Fish Meal M is shown in Table 4 (Experiment 1). The addition of the antiacids, Al(OH) 3 and NaHC0 3 , to the OM Mineral Mixture produced a 28% greater growth due to two possibilities: they served either to neutralize the effect of acidic components or to supply additional Na to overcome the excessive CI content. This compensatory effect of cations on ex-
cessive anions was substantiated when the concentrations of CI and S0 4 were decreased. For example, substitution of K 2 C0 3 for KC1 yielded an 18% increase in growth, and substituting MgO for MgS04 yielded at 19% increase; combining both K 2 C0 3 and MgO produced a 39% increase. Decreasing the concentration of Ca to 1.25% and that of P to 0.80% did not improve the growth, however, decreasing the concentration of Fe resulted in less growth. These indicate that the two different levels of Ca and P were equally satisfactory but that the addition of 40 p.p.m. Fe was not adequate for optimum growth. The data in Table 5 indicate that, in Experiment 2, the substitution of KHC0 3 for KC1 resulted in a 32% greater growth, which was farther increased by the substitution of MgC0 3 for MgS04. These observations confirm those of the previous experiment that the concentration of CI and S0 4 must be decreased (and as a consequence that of the C0 3 increased) to obtain improved growth from diets containing fish meals as the sole source of protein. Although the antiacids Al(OH) 3 and NaHC0 3 definitely improved Fish Meal M (Table 4) and slightly improved Fish
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Average weight of 19-day old chicks. Average chick weights with different superscripts are significantly different ( P < .01). Relative growth in comparison to that of the 12% protein control diet of fish meal and OM salt mixture wfijch was set arbitrarily at lOOy^. 2
3
PROTEIN QUALITY BIOASSAYS
1539
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TABLE 6.—Experiment 2—comparative effect of Meals T5 and TM6, they did not improve different mineral mixtures on relative growth the other five fish meals (Table 6). In conresponse of various fish meah fed at JS% protein level trast, Mineral Mixture I was definitely beneficial in all seven fish meals studied Mineral mixtures Fish Diet comparatively (Table 6). These results indimeal No. OM A I cate that the concentrations of CI and S0 4 in the diet were important factors in the — Relative growth (%) — 2 100"' 107" 139° T5 growth response, although antiacids may 3 57° 56° 97" TMl d d also play a favorable role in the protein 4 89 84 114b TM2 5 59° 63° 97" TM3 evaluation of certain fish meals, especially 6 96" 97" 127b TM4 inasmuch as the KHC0 3 used in lieu of 7 105" 103" 114b TM5 8 104" 113b 141° TM6 KC1 may also have acted as an antiacid. Nevertheless, Mineral Mixture I, which con1 The growth of the chicks fed the T5 fish meal tained 0.304% CI and 0.032% S0 4 , im- diet containing OM Mineral Mixture is arbitrarily as 100% relative growth. All other growth reproved the growth response considerably, set sponses are comparatively calculated. Relative although not to the same degree for all fish growth percentages with different superscripts are meals (9-40%). The results indicated that significantly different (P<.01). dietary anionic components were hindering According to Leach and Nesheim (1963), the evaluation of fish meal protein quality to its fullest potentiality. This hinderance chicks require about 0.12% CI in the diet. was eliminated by decreasing the concen- These authors also obtained equivalent trations of CI and S0 4 and increasing the growth with 0.30% CI, which is the same concentration of C0 3 in the dietary mineral concentration as was in our experimental Mineral Mixture I. mixture. Leach et al. (1960) depressed the growth TABLE 5.—Experiment 2—effect of different mineral of chicks by adding 1.2% S0 4 in a diet conl substitutions on protein bioassay of fish meal T5 taining casein supplemented with methionine, glycine, and arginine. Our experiments Mineral mixtures Average Relative Code indicate that decreasing the concentration 2 growth Additions or substitutions weight of S0 4 in the mineral mixture from 0.432% Gram % to 0.032% was just as beneficial as was lowering that of CI. However, the combined 242«3 1004 OM — decrease in S0 4 and CI resulted in signifi107 A +AIfOH) 3 258" cantly greater improvement than a decrease +NaHC03 in either one alone. These decreases were 132 320^ G KHC0 3 for KCl accompanied by increases in the C0 3 . 284b H MgC0 3 for MgSO, 117 Nesheim et al. (1964) also obtained a depression of growth in chicks due to exces139 335° I K H C 0 3 + M g C 0 3 for KCl+MgS04 ses of CI or S0 4 in casein diets and in 1 isolated soy-protein diets. They indicated Sole source protein to supply 15% protein. 2 Average of 30 chicks (three replicate peris of 10 that a relation existed between dietary levels chicks each). Age/of chicks: 20 days when experiof the anions. CI and S0 4 and the cations ment, terminated. 3 Average chick weights with different super- Na and K. The.concentrations of these anscripts are significantly different ( P < .01). 4 The growth obtained with reference control diet ions in their studies extended over a wide (containing OM mineral mixture) is arbitrarily set range-r-in some cases the concentrations at 100 percent and all other growth responses are were as high as 2.5%. In contrast, in our relative to the control diet growth response.
1S40
D. MILLER
investigations, these anions were studied at comparatively low levels in diets containing fish meal as the sole source of protein. Our diets required modified salt mixtures supplying less CI and S0 4 . Evidently, to assume that all mineral mixtures are interchangeable and proper for all experimental diets is erroneous. SUMMARY
AllLiaUUJ
VV . *-•L l>
1U U l i u
IU
Ut
uvuCuviui
supplements for some fish meals. They may play a role in counteracting acidic components. However, decreasing the mineralmixture dietary contribution of CI from 0.542 to 0.304% and S0 4 from 0.432 to 0.032% (and simultaneously increasing the C0 3 content) improved the growth response by 9 to 40% for various fish meals fed to supply the entire 15% dietary protein. The 1.25% Ca and 0.8% P levels used in the diets in these experiments resulted in the same growth response as the 1.50% Ca and 1.0% P levels. The addition of 40 ppm Fe was not adequate for optimum growth.
NEWS AND NOTES (Continued from page 1500) Grow Out Manager of the Pillsbury Company's poultry operations in Alabama. Currently a member of the Executive Board of the Industrial Veterinarians Association, Dr. Dillehay is also a member of the American Veterinary Medical Association, the American Association of Avian Pathologists, the North Alabama Poultry Health Association, and the U.S. Sanitary Livestock Association.
CANADA BRANCH W.P.S.A. The following officers and executive committee have been elected: Chairman—A. D. Davey; Secretary-Treasurer—W. H. Pope; Executive Committee: D. F. Archibald, J. Biely, C. H. Bigland, J. R. Cavers, D. R. Clandinin, R. D. Crawford, J. H. Hare, P. A. Kondra, N. Nikolaiczuk and J. D. Summers. A. D. Davey and J. D. Summers have been
(Continued on page 1S54)
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The evaluation of the quality of the protein in fish meals fed to chicks as a sole source protein is influenced by the composition of the mineral mixture included in the test diet.
REFERENCES Fisher, R. A., 1950. Statistical Methods for Research Workers, 11th edition, Hafner Publishing Company, New York, N.Y. Grau, C. R., and M. A. Williams, 1955. Fish meals as amino acid sources in chick rations. Poultry Sci. 34: 810-817. Kifer, R. R., W. L. Payne, D. Miller and M. E. Ambrose, 1968. The nutritive content of menhaden {Brevoortia tyrannus and patronus) fish meal evaluated by chemical methods. Feedstuffs, 40(20): 36-37. Leach, R. M., and M. C. Nesheim, 1963. Studies on chloride deficiency in chicks. J. Nutr. 8 1 : 193-199. Leach, R. M., T. R. Zeigler and L. C. Norris, 1960. The effect of dietary sulfate on the growth rate of chicks fed a purified diet. Poultry Sci. 39: 1577-1578. Miller, D., and R. R. Kifer, 1970a. Factors affecting protein evaluation of fish meal by chick bioassay. Poultry Sci. 49: 999-1004. Miller, D., and R. R. Kifer, 1970b. Effect of glutamic acid and antiacids on chick bioassay of protein quality of fish meals. Poultry Sci. 49 : 1327-1334. Miller, D., R. R. Kifer and M. E. Ambrose, 1970. Effect of storage on fish meal quality as evaluated by chemical indices and chick bioassay. Poultry Sci. 49 : 1005-1010. Nesheim, M. C , R. M. Leach, Jr., T. R. Zeigler and J. A. Serafin, 1964. Interrelationships between dietary levels of sodium, chlorine and potassium. J. Nutr. 84: 361-366. Ousterhout, L. E., and D. G. Snyder, 1962. Nutritional evaluation of fish meals using four shortterm chick tests. Poultry Sci. 4 1 : 1753-1757.
1535
of light into the brain of mammals. Ann. New York Acad. Sci. 117: 217-227. Van Tienhoven, A., 1968. Reproductive Physiology of Vertebrates. W. B. Saunders Co., Philadelphia. p. 63. Wetterberg, L., E. Geller and A. Yuwiler, 1970. Harderian gland: An extraretinal photoreceptor influencing the pineal gland in neonatal rats? Science, 167: 884-885.
Mineral Mixture Composition—A Factor in Chick Bioassays of the Protein Quality of Fish Meals MILLER
(Received for publication May 20, 1970)
M
ILLER and Kifer (1970b) showed excess chloride. They showed that such adthat the evaluation of the quality of verse effects also resulted from the addition the protein in fish meal, when the meal was of other chloride and sulfate compounds in fed as the sole source of protein, was influ- diets made up entirely of casein or of soy enced by excesses of lysine present in fish proteins. meal over the concentration needed in the Other possible excess inorganic subdiet and by various anionic components in stances in the diet of chicks could be the the mineral mixture. The excess of lysine high amounts of Ca, P, and Fe contributed was overcome by a dilution of the concen- by fish meals when they are used as the tration of lysine in the diet by the addition only source protein. Some menhaden fish of glutamic acid (free-base form) or was meals containing 6% or more of Ca (Kifer overcome by counteraction from the addi- et al., 1968) will contribute, when fed to tion of arginine. The excess of anionic com- supply 15% protein, about 1.5% Ca to the ponents was overcome by the inclusion in diet. This level of Ca is ordinarily considthe diet of the antiacids Al(OH) 3 and Na- ered as being unnecessarily high. For that HC0 3 . The detrimental anionic effect was reason, a comparative study of a lower evident when the addition of HC1 salt of level of Ca and P was necessary to deterglutamic acid depressed growth. In con- mine if the high levels were detrimental. trast, the free-base form of glutamic acid Similarly, because of the high concentraimproved growth significantly, owing to tions of Fe (438 p.p.m. on the average) in two possibilities: (1) the elimination of the menhaden fish meal (Kifer et al., 1968), a detrimental HC1 portion of this amino acid comparative study of the effect of decreassupplement and (2) the increase of nones- ing the concentration of Fe in the mineral sential amino acid nitrogen (glutamic mixture was necessary. acid). In view of these observations, the comNesheim et al. (1964) also reported that position of the mineral mixture that has glutamic. HC1 was detrimental due to the been used by Grau and Williams (1955),
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
DAVID
National Center for Fish Protein Concentrate, Bureau of Commercial Fisheries, College Park, Maryland 20740
1536
D.
MILLER
TABLE 1.—Composition (%) of variousfishmeals Fish meals
Protein
Fat ethyl ether extractable
Ca
P
M T5 TM1 TM2 TM3 TM4 TM5 TM6
65.41 61.89 65.30 65.16 64.92 64.84 64.05 64.24
11.65 13.50 12.93 10.69 12.97 12.33 13.06 13.43
4.45 4.19 4.79 5.05 4.75 4.31 4.25 4.18
2.50 2.33 2.66 2.88 2.72 2.48 2.46 2.44
In Experiment 1, Fish Meal M was studied with eight different mineral supplementations, OM through F. In Experiment 2, five mineral mixtures (OM, A, G, H, and I) were studied with Fish Meal T5. Also the relative growth effect of three different mineral mixtures (OM, A, and I) were comparatively studied with seven different fish meals (T5 and TM1-6). These studies involved a total of 31 treatments.
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
Mineral Mixtures. Ten different mineral mixtures (Table 3) were used. Step-wise changes were made to determine the effect of inorganic substances on the growth of the chick when fish meal was the sole source of protein. The OM Mineral Mixture was similar to that used by many investigators (Grau and Williams, 1955; and Ousterhout et al., 1962) except that in the present study the Ca and P were not included in the mineral mixtures but were supplied as variable ingredients dicalcium phosphate and phosphoric acid to maintain a constant dietary level of 1.5 and 1.0%, MATERIALS AND METHODS respectively except in one treatment. In Basal Diets. Table 1 shows the composition Experiment 1, the effect of reducing the Ca of eight menhaden fish meals studied in this concentration to 1.25% and the P to 0.8% investigation, which consisted of two experi- was studied comparatively by decreasing ments. Eight basal diets (Table 2), each the dietary supplements of dicalcium phosone containing a different menhaden fish phate and phosphoric acid in one of the meal, were used to study the effects of dif- treatments containing Mineral Mixture D. ferent mineral mixtures (Table 3) on the At the 5% level the OM Mineral Mixture growth response of the chicks. Diet 1 con- contributed to the diets 0.20% Na and tained 12% protein, all others, 15%. The 0.26% K, which were kept constant in all calorie content of the diet was 1377, which mineral mixtures except in Mix A, which was maintained by adjusting the levels of supplied 0.47% Na (Table 3). The SO* glucose (Cerelose)1 cellulose (Solka Floe),2 content was 0.432% in Mineral Mixtures and corn oil (the lower limit was set at OM, A, B, C and was decreased to 0.032% 2.0%). 3 by the substitution of MgO or MgC0 3 for MgS0 4 in the other mineral mixtures. The 1 Cerelose, Corn Products Company, Argo, Illi- concentration of CI was 0.542% in Mineral nois. 2 Solka Floe, Brown Company, Berlin, New Mixtures OM, A, C, and H and was lowered in the other mineral mixtures to 0.304% Hampshire. 3 The use of trade names is merely to facilitate when KC1 was replaced by K 2 C0 3 or descriptions; no endorsement is implied. KHC0 3 . The concentration of Fe in Min-
Ousterhout and Snyder (1962), Miller et al. (1970), and Miller and Kifer (1970a, b) in diets containing fish meal as the sole source protein was niouincu stepwise, xuis investigation was conducted to determine the effect on chick growth responses from various fish meals as the sole source protein in the diet (1) by decreasing the concentrations of CI, S0 4 , and Fe in the mineral mixtures, (2) by adding antiacids, and (3) by decreasing the concentrations of Ca and P0 4 .
1537
PROTEIN QUALITY BIOASSAYS
TABLE 2.—Composition (%) of diets Diet No.
1
2
3
4
5
6
7
8
Experiment No.
1
2
2
2
2
2
2
2
Fish meal M Fish meal T5 Fish meal TM1 Fish meal TM2 Fish meal TM3 Fish meal TM4 Fish meal TM5 Fish meal T M 6 Cerelose1 Vitamin mix2 Mineral mix3 DiCaP0 4
18.35
—
— —
— — —
— — — —
— — — — —
— — — — — —
— — — — — — —
24.24
— — — — — — —
— — — — — —
23.00
— — — — —
23.02
— — — —
23.09
23.13
— — —
23.41
— —
55.85 5.00 5.00 1.88 0.32 5.51 0.20 2.00
56.46 5.00 5.00 1.54 0.35 6.45 0.20 2.00
57.62 5.00 5.00 1.31 0.32 5.53 0.20 2.00
56.46 5.00 5.00 1.54 0.29 6.40 0.20 2.00
56.80 5.00 5.00 1.92 0.25 5.70 0.20 2.00
56.32 5.00 5.00 1.92 0.22 5.93 0.20 2.00
23.35 56.19 5.00 5.00 2.00 0.22 6.04 0.20 2.00
Protein content (%)
12
15
15
15
15
15
15
15
H3PO4
—
1
Cerelose, Corn Products Company, Argo, Illinois. The vitamin mixture fortified each kilogram of diet with 7 mg. thiamine-HC1, 7 mg. riboflavin, 8 mg. pyridoxineHCl, 60 mg. niacin, 30 mg. calcium pantothenate, 2 mg. folic acid, 2 mg. menadione, 29 jug. vitamin B12, 220 /ig. biotin, 90001.U. vitamin A, 4501.C.U. vitamin fi3, 441.U. dl-alpha tocopherol acetate, and 1.65 gm. choline-chloride. 3 See Table 3 for composition of mineral mixtures. 4 Solka Floe, Brown Company, Berlin, New Hampshire. 6 Ethoxyquin premix ( 1 + 9 cerelose). 2
TABLE 3.—Composition and contribution of various mineral mixtures at a lezel of 5*V0 in the diet Mineral mixture Experiment No.
OM \
A
B
C
D
E
\
1
1
1
1
% of diet Variable portion NaHCOs — 1.0 — — — — Al(OH) 3 — 1.0 — — — — KC1 0.5 0.5 — 0.5 — — K 2 C0 3 — — 0.463 — 0.463 0.463 KHCO3 _ _ _ _ _ _ _ _ MgS0 4 0.5 0.5 0.5 — — — MgO — — — 0.145 0.145 0.145 MgC0 3 _ _ _ _ _ _ _ FeC 6 H 5 0,-3H 2 0 0.08 0.08 0.08 0.08 0.08 0.040 Constant portion* 0.568 in all diets Glucose added to bring total of each mineral mixture to 5 % in each diet. The above mineral mixtures contribute the following to the diets: CI, % 0.542 0.542 0.340 0.542 0.304 0.304 S0 4 , % 0.432 0.432 0.432 0.032 0.032 0.032 CO3, % 0 0.714 0.201 0 0.201 0.201 Na, % 0.200 0.470 0.200 0.200 0.200 0.200 K, % 0.260 0.260 0.260 0.260 0.260 0.260
F *
_
G
H
I
2
2
2
_
0.08
0.5 — — — — o.35 0.08
_ _ — — 0.67 — — 0.35 0.08
0.304 0.432 0.402 0.200 0.260
0.542 0.032 0.249 0.200 0.260
0.304 0.032 0.651 0.200 0.260
_ — 0.463 — 0.145 _ —
0.304 0.032 0.201 0.200 0.260
_ _
— — 0.67 0.5 —
_ _
* Constant portion in all mineral mixtures contains 0 . 5 % NaCl (iodized); 0.03% MnS04-H 2 0; 0.02% ZnS0 4 -7H 2 0; 0 . 0 1 % Al2(S0 4 )3-18H 2 0; 0.004% NaSi0 3 -9H 2 0; 0.002% C u S C v H 2 0 ; 0 . 0 0 1 % Co-(CH 3 COO)2-H2O;0.001%(NH < )2MoO 4 -4H 2 O. Total 0.568%.
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Solka Floe4 Ethoxyquin premix6 Corn oil
60.68 5.00 5.00 2.61 0.22 5.71 0.20 2.87
1538
D. MILLER TABLE 4.—Experiment 1—growth effect of various mineral changes in 12% protein diets containing Menhaden Fish Meal M
Mineral mixtures code
OM A B C D D E F
Chick weights
Substitution of Basic compounds
—
Al(OH) 3 +NaHC0 3 K 2 C0 3 MgO K 2 C0 3 +MgO K 2 C0 3 +MgO K 2 C0 3 +MgO K 2 C0 3 +MgO
For acidic compounds
Decreases
Average 1
Relative growth
Gram
%
169a2
— —
KC1 MgS0 4 KCl+MgSO, KCl+MgS04 KCl+MgS04 KCl+MgSOi
CaHPO,, H3PO4 Fe citrate (J orig.) Fe citrate (complete omission) n)
216b 199b 201 b 235« 234° 216b 203"
1003 128 118 119 139 139 128 120
1
eral Mixture E was reduced to one-half of that in all other mineral mixtures. Chicks. Day-old Arbor Acre cockerels were wingbanded and randomized, 12 chicks per pen, three replicates per treatment. Feed and water were supplied at all times in thermostatic compartmentized chick-starter batteries in a temperature-controlled room. Final individual weights of chicks were obtained on the 19th or 20th day. Only the weights of the heaviest 10 chicks of each pen were included in the final averages. The chick weights were analyzed for significance by Fisher's (1950) modification of Student's t-test. RESULTS AND DISCUSSION The growth effect of various mineral substitutions in a 12% protein diet containing Fish Meal M is shown in Table 4 (Experiment 1). The addition of the antiacids, Al(OH) 3 and NaHC0 3 , to the OM Mineral Mixture produced a 28% greater growth due to two possibilities: they served either to neutralize the effect of acidic components or to supply additional Na to overcome the excessive CI content. This compensatory effect of cations on ex-
cessive anions was substantiated when the concentrations of CI and S0 4 were decreased. For example, substitution of K 2 C0 3 for KC1 yielded an 18% increase in growth, and substituting MgO for MgS04 yielded at 19% increase; combining both K 2 C0 3 and MgO produced a 39% increase. Decreasing the concentration of Ca to 1.25% and that of P to 0.80% did not improve the growth, however, decreasing the concentration of Fe resulted in less growth. These indicate that the two different levels of Ca and P were equally satisfactory but that the addition of 40 p.p.m. Fe was not adequate for optimum growth. The data in Table 5 indicate that, in Experiment 2, the substitution of KHC0 3 for KC1 resulted in a 32% greater growth, which was farther increased by the substitution of MgC0 3 for MgS04. These observations confirm those of the previous experiment that the concentration of CI and S0 4 must be decreased (and as a consequence that of the C0 3 increased) to obtain improved growth from diets containing fish meals as the sole source of protein. Although the antiacids Al(OH) 3 and NaHC0 3 definitely improved Fish Meal M (Table 4) and slightly improved Fish
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Average weight of 19-day old chicks. Average chick weights with different superscripts are significantly different ( P < .01). Relative growth in comparison to that of the 12% protein control diet of fish meal and OM salt mixture wfijch was set arbitrarily at lOOy^. 2
3
PROTEIN QUALITY BIOASSAYS
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TABLE 6.—Experiment 2—comparative effect of Meals T5 and TM6, they did not improve different mineral mixtures on relative growth the other five fish meals (Table 6). In conresponse of various fish meah fed at JS% protein level trast, Mineral Mixture I was definitely beneficial in all seven fish meals studied Mineral mixtures Fish Diet comparatively (Table 6). These results indimeal No. OM A I cate that the concentrations of CI and S0 4 in the diet were important factors in the — Relative growth (%) — 2 100"' 107" 139° T5 growth response, although antiacids may 3 57° 56° 97" TMl d d also play a favorable role in the protein 4 89 84 114b TM2 5 59° 63° 97" TM3 evaluation of certain fish meals, especially 6 96" 97" 127b TM4 inasmuch as the KHC0 3 used in lieu of 7 105" 103" 114b TM5 8 104" 113b 141° TM6 KC1 may also have acted as an antiacid. Nevertheless, Mineral Mixture I, which con1 The growth of the chicks fed the T5 fish meal tained 0.304% CI and 0.032% S0 4 , im- diet containing OM Mineral Mixture is arbitrarily as 100% relative growth. All other growth reproved the growth response considerably, set sponses are comparatively calculated. Relative although not to the same degree for all fish growth percentages with different superscripts are meals (9-40%). The results indicated that significantly different (P<.01). dietary anionic components were hindering According to Leach and Nesheim (1963), the evaluation of fish meal protein quality to its fullest potentiality. This hinderance chicks require about 0.12% CI in the diet. was eliminated by decreasing the concen- These authors also obtained equivalent trations of CI and S0 4 and increasing the growth with 0.30% CI, which is the same concentration of C0 3 in the dietary mineral concentration as was in our experimental Mineral Mixture I. mixture. Leach et al. (1960) depressed the growth TABLE 5.—Experiment 2—effect of different mineral of chicks by adding 1.2% S0 4 in a diet conl substitutions on protein bioassay of fish meal T5 taining casein supplemented with methionine, glycine, and arginine. Our experiments Mineral mixtures Average Relative Code indicate that decreasing the concentration 2 growth Additions or substitutions weight of S0 4 in the mineral mixture from 0.432% Gram % to 0.032% was just as beneficial as was lowering that of CI. However, the combined 242«3 1004 OM — decrease in S0 4 and CI resulted in signifi107 A +AIfOH) 3 258" cantly greater improvement than a decrease +NaHC03 in either one alone. These decreases were 132 320^ G KHC0 3 for KCl accompanied by increases in the C0 3 . 284b H MgC0 3 for MgSO, 117 Nesheim et al. (1964) also obtained a depression of growth in chicks due to exces139 335° I K H C 0 3 + M g C 0 3 for KCl+MgS04 ses of CI or S0 4 in casein diets and in 1 isolated soy-protein diets. They indicated Sole source protein to supply 15% protein. 2 Average of 30 chicks (three replicate peris of 10 that a relation existed between dietary levels chicks each). Age/of chicks: 20 days when experiof the anions. CI and S0 4 and the cations ment, terminated. 3 Average chick weights with different super- Na and K. The.concentrations of these anscripts are significantly different ( P < .01). 4 The growth obtained with reference control diet ions in their studies extended over a wide (containing OM mineral mixture) is arbitrarily set range-r-in some cases the concentrations at 100 percent and all other growth responses are were as high as 2.5%. In contrast, in our relative to the control diet growth response.
1S40
D. MILLER
investigations, these anions were studied at comparatively low levels in diets containing fish meal as the sole source of protein. Our diets required modified salt mixtures supplying less CI and S0 4 . Evidently, to assume that all mineral mixtures are interchangeable and proper for all experimental diets is erroneous. SUMMARY
AllLiaUUJ
VV . *-•L l>
1U U l i u
IU
Ut
uvuCuviui
supplements for some fish meals. They may play a role in counteracting acidic components. However, decreasing the mineralmixture dietary contribution of CI from 0.542 to 0.304% and S0 4 from 0.432 to 0.032% (and simultaneously increasing the C0 3 content) improved the growth response by 9 to 40% for various fish meals fed to supply the entire 15% dietary protein. The 1.25% Ca and 0.8% P levels used in the diets in these experiments resulted in the same growth response as the 1.50% Ca and 1.0% P levels. The addition of 40 ppm Fe was not adequate for optimum growth.
NEWS AND NOTES (Continued from page 1500) Grow Out Manager of the Pillsbury Company's poultry operations in Alabama. Currently a member of the Executive Board of the Industrial Veterinarians Association, Dr. Dillehay is also a member of the American Veterinary Medical Association, the American Association of Avian Pathologists, the North Alabama Poultry Health Association, and the U.S. Sanitary Livestock Association.
CANADA BRANCH W.P.S.A. The following officers and executive committee have been elected: Chairman—A. D. Davey; Secretary-Treasurer—W. H. Pope; Executive Committee: D. F. Archibald, J. Biely, C. H. Bigland, J. R. Cavers, D. R. Clandinin, R. D. Crawford, J. H. Hare, P. A. Kondra, N. Nikolaiczuk and J. D. Summers. A. D. Davey and J. D. Summers have been
(Continued on page 1S54)
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The evaluation of the quality of the protein in fish meals fed to chicks as a sole source protein is influenced by the composition of the mineral mixture included in the test diet.
REFERENCES Fisher, R. A., 1950. Statistical Methods for Research Workers, 11th edition, Hafner Publishing Company, New York, N.Y. Grau, C. R., and M. A. Williams, 1955. Fish meals as amino acid sources in chick rations. Poultry Sci. 34: 810-817. Kifer, R. R., W. L. Payne, D. Miller and M. E. Ambrose, 1968. The nutritive content of menhaden {Brevoortia tyrannus and patronus) fish meal evaluated by chemical methods. Feedstuffs, 40(20): 36-37. Leach, R. M., and M. C. Nesheim, 1963. Studies on chloride deficiency in chicks. J. Nutr. 8 1 : 193-199. Leach, R. M., T. R. Zeigler and L. C. Norris, 1960. The effect of dietary sulfate on the growth rate of chicks fed a purified diet. Poultry Sci. 39: 1577-1578. Miller, D., and R. R. Kifer, 1970a. Factors affecting protein evaluation of fish meal by chick bioassay. Poultry Sci. 49: 999-1004. Miller, D., and R. R. Kifer, 1970b. Effect of glutamic acid and antiacids on chick bioassay of protein quality of fish meals. Poultry Sci. 49 : 1327-1334. Miller, D., R. R. Kifer and M. E. Ambrose, 1970. Effect of storage on fish meal quality as evaluated by chemical indices and chick bioassay. Poultry Sci. 49 : 1005-1010. Nesheim, M. C , R. M. Leach, Jr., T. R. Zeigler and J. A. Serafin, 1964. Interrelationships between dietary levels of sodium, chlorine and potassium. J. Nutr. 84: 361-366. Ousterhout, L. E., and D. G. Snyder, 1962. Nutritional evaluation of fish meals using four shortterm chick tests. Poultry Sci. 4 1 : 1753-1757.