Biological Availability of Calcium and Phosphorus in Menhaden Fish Meals

BINDING PROPERTIES OF CHICKEN MEAT

chicken rolls. Chicken rolls and ground chicken meat treated with polyphosphates were found to exhibit increased tear strength of cooked slices. Thus, binding properties or capacity of chicken meat to hold together upon cooking appear to be improved by added polyphosphates. REFERENCES Klose, A. A., A. A. Campbell and H. L. Hanson, 1962. Influence of polyphosphates in chilling water on quality of poultry meat. Poultry Sci. 4 1 : 1655. Klose, A. A., A. A. Campbell and H. L. Hanson, 1963. Influence of polyphosphates in chilling water on quality of poultry meat. Poultry Sci. 42: 743-749. Mahon, J. A., 1962. You can reduce poultry "weep." Poultry Processing and Marketing, 68(8): 16-17, 26. Monk, J. A., G. J. Mountney and I. Prudent, 1964. Effect of phosphate treatment and cook-

1107

ing method on moisture losses of poultry meat. Food Technol. 18 : 104-107. Mountney, G. J., 1962. Some factors influencing the texture of an egg product in sausage casing. Poultry Sci. 4 1 : 1215-1218. Mountney, G. J., and F. C. Organosa, 1962. The effects of polyphosphates on moisture absorption, retention, and cooking losses of broiler carcasses. Poultry Sci. 4 1 : 1668. Schermerhorn, E. P., and W. J. Stadelman, 1962. Effects of polyphosphates on water uptake, moisture retention, and cooking loss in broilers. Poultry Sci. 4 1 : 1680. Snedecor, G. W., 1956. Statistical Methods, Iowa State College Press, Ames, Iowa. Swift, C. E., and R. Ellis, 1956. The action of phosphates in sausage products. I. Factors affecting the water retention of phosphate treated ground meat. Food Technol. 10: 546-552. Swift, C. E., and R. Ellis, 1957. Action of phosphates in sausage products II. Pilot plant studies of the effects of some phosphates on binding and color. Food Technol. 11: 450456.

Biological Availability of Calcium and Phosphorus in Menhaden Fish Meals A. H. SPANDORF AND K. C. LEONG Bureau of Commercial Fisheries Technological Laboratory, College Park, Md. (Received for publication January 25, 1965)

A

LTHOUGH it is generally assumed that the calcium and phosphorus in fish meals are completely available to chickens, this assumption has needed experimental study in view of the variation in availabilities among inorganic mineral supplements (Gillis et al., 1951, 1954; Grau and Zweigert, 1953; Miller and Joukovsky, 1953; Motzok et al., 1956; Nelson and Walker, 1964; and Dilworth and Day, 1964). An erroneous assumption on this point can be especially critical when economic and nutritional considerations indicate the use of high levels of fish meal in poultry diets. If used in substantial amounts, fish

meal can become the sole source of supplemental P and a major source of Ca. This study was therefore conducted to determine experimentally the biological availability of P and Ca in commercially produced menhaden fish meals—the fish meal manufactured in largest quantity in the United States. MATERIALS AND METHODS

Ash analyses of bones taken from chicks previously fed diets supplemented with various levels of minerals, both from inorganic sources and from fish meal, were used as the primary criterion of mineral availability. The general procedures were

1108

A. H. SPANDORF AND K. C. LEONG TABLE 1.—Proximate analysis, Ca, P, and Ca'.P ratios of 12 commercial menhaden fish meals

Fish meal

Protein

No. 1 2 3 4 5 6 7 8 9 10 11 12

64.1 61.7 62.8 60.7 62.4 63.0 63.2 60.9 61.0 59.1 63.8 63.1

6.2 6.7 7.5 7.6 7.8 8.7 7.0 7.4 7.0 7.7 6.9 5.7

10.3 11.6 13.1 10.5 13.4 9.1 11.9 10.1 12.6 9.7 11.5 9.7

/o 15.8 18.0 16.0 19.4 15.2 19.1 16.7 20.0 19.2 22.4 17.2 20.9

2.90 2.88 2.39 3.10 2.58 3.09 2.84 3.28 3.18 3.66 2.58 3.36

5.61 5.44 3.94 5.79 4.80 5.85 5.18 6.06 5.85 6.80 4.72 5.96

1.93:1 1.89 1.65 1.87 1.86 1.89 1.82 1.85 1.84 1.86 1.83 1.77

62 .1

7.2

11 .1

18.3

2.99

5..49

1.84:1

0.37

0 .72

0.07

Average Standard deviation

Ether extract

H20

%

%

/o

1 .48

0.79

1 .40

Ash 0/

2.22

Ca

%

%

Ca:P ratio

similar to those suggested by Nelson and Walker (1964), who recently reviewed methods for the biological evaluation of compounds containing P. Modifications were necessary in order to factor out the protein supplemental value of the fish meals from their mineral contributions.

termination was by the Fiske and SubbaRow method as described in Practical Physiological Chemistry, 13th edition, 1954, by P. B. Hawk et al. These values agree closely with those previously published (N.R.C., 1956; Snyder et al., 1962).

Fish Meals. Twelve commercially manufactured menhaden {Brevoortia tyrannus) fish meals were used in this study. They were produced between September 1963 and January 1964. These meals were collected from 11 fish-meal manufacturers on the East and Gulf coasts of the United States and were numbered from 1 to 12. The proximate analysis, Ca, P, and ratios of Ca to P of these meals are shown in Table 1. Proximate and Ca analyses were by the A.O.A.C. method. Phosphorus de-

Dietary Treatments. Using the analyzed results for the fish meals and for commercial sources of dicalcium phosphate and limestone, we formulated the following diets (Table 2).

TABLE 2 . --Plan of study: sources and levels Of added P Added P levels (%)

Source Dicalcium phosphate

0.0> 0.1

0.2

0.3 0.4

Menhaden F.M. 1-12

0.2

0.4

Fish-meal ash2

0.2

0.4

0.5

i Analysis of basal diet =0.24% P, 0.20% Ca. * Fish Meal #6 ashed at 650°C. for 2 hours.

0.6 0.7

1. Eight control diets supplemented with sufficient discalcium phosphate to provide 0 to 0.7% P to the basal diet. Limestone was used to maintain the Ca:P ratio at 1.84:1, the average ratio of the 12 fish meals under test. 2. Diets in which each of the 12 fish meals was substituted into the basal diet to provide both 0.2 and 0.4% P. (No attempt was made to control the precise Ca levels in these diets, since the chemical analyses had indicated only small variations in the Ca:P ratio among the fish meals.) 3. Two diets were formulated as added controls in which the ash from one

1109

AVAILABILITY OF CA AND P IN FISH MEALS

of the fish meals ( # 6 ) under test was used as the supplemental source of 0.2 and 0.4% P to the basal diet. (These control diets were included in order to compare the possible effects of the addition of trace minerals, which might be present in the diets containing fish meals but not in those supplemented with inorganic P sources. Fish meal # 6 was chosen as the source of the ash because it most nearly represented the average proximate analyses of the 12 fish meals.) Basal Diet. Table 3 shows the composition of the basal diet. In all cases, the diet was modified by various alterations in such a manner as to maintain the protein level at 20% and the metabolizable energy (M.E.) at 3025 kilocalories per kilo. In the diets containing fish meals, isonitrogenous amounts of fish protein replaced soybean meal (S.B.M.) supplemented with 0.57% (level based on methionine + cystine requirement equal to 0.53%/megacalorie; M.E.) methionine hydroxy analogue (M.H.A.). Previous studies in this laboratory (unpublished) have shown about equal chick-growth potential from methionine-supplemented S.B.M. and from unsupplemented fish-meal protein when each was used as a sole source of protein in otherwise nutritionally adequate diets. Levels of cerelose and corn oil were adjusted to maintain the diets isocaloric. Chicks. The 34 experimental diets were fed ad libitum in two replicate pens of 10 1-day-old Arbor Acre male chicks housed in electrically heated wire brooders. Body weight and feed consumption for each bird were recorded weekly. On the 21st day of the experiment, all surviving chicks were sacrificed, and their tibiae removed and prepared for bone ash

TABLE 3.—Composition of basal diet Ingredient Soybean meal with MHA Cerelose Corn oil Cellulose Vitamin mix2 Trace mineral mix3

Parts/100 1

40.23 37.77 7.00 5.00 5.00 5.00

1 50% protein soybean meal blended with 0.568% M.H.A. to provide minimum sulfur amino acid requirement in 20% protein, 3025 kilocalories M.E. per kilo diet. 2 Provides the following amounts per kg. diet: thiamine hydrochloride, 7.0 mg.; riboflavin, 7.0 mg.; pyridoxine, 8.0 mg.; niacin, 60.0 mg.; calcium pantothenate, 30.0 mg.; folic acid, 4.0 mg.; menadione, 2.0 mg.; B12, 2.9 meg.; biotin, 0.22 mg.; vitamin A, 9,000 I.U.; vitamin D3, 1,500 I.C.U.; dl-atocopherol acetate, 44.0 mg.; choline chloride, 2.00 g.; and cerelose to 50 g. 3 Provides the following per kg. diet: iodized salt (NaCl), 5 g.; potassium chloride, 5.0 g.; magnesium sulfate, 5.0 g.; ferric sulfate, 800 mg.; manganese sulfate, 300 mg.; zinc sulfate, 200 mg.; sodium silicate, 40 mg.; cupric sulfate, 20 mg.; cobaltous acetate, 10 mg.; ammonium molybdate, 10 mg.; and cerelose to 50 g.

analysis according to the procedure described by the A.O.A.C. (1960) in the vitamin D assay. All left tibia obtained from the surviving chicks in each pen were divided into two replicate composite groups of five or less bones; the bone ash results, therefore, are an average of four determinations per dietary treatment. RESULTS AND DISCUSSION Control Diets. The results obtained with the control diets are shown in Table 4. Chemical analyses of the mixed diets indicate close agreement between the actual P values found and those calculated. Except for the diet supplemented with 0.7% P from dicalcium phosphate, the variation in values obtained is within that normally expected (±0.03% with the analytical procedures used). Body weights and cumulative feed conversions are shown also. These results indicate good growth of all groups of chicks fed diets supplemented with 0.2% or more P from dicalcium phosphate.

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A. H. SPANDORF AND K. C. LEONG

TABLE 4.-

-Body weight, feed utilization, and bone ash response of chicks fed diets varying in P levels and sources1 Level P added

Source

%

Total analyzed P in diet

Av. weight

%

Feed/gain

Total feed/chick

Tibia bone ash (S.E.)

%

Dicalcium phosphate

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7

0.24 2 0.32 0.44 0.51 0.65 0.71 0.82 0.86

g126 318 385 377 398 387 384 391

2.18 1.66 1.52 1.56 1.53 1.58 1.54 1.54

g197 471 530 535 556 555 538 547

22.0 (.79) 34.0 (.49) 41.1(.64) 44.2 (.51) 45.9 (.10) 45.6 (.43) 47.2 (.56) 46.6 (.66)

Fish meal ash 3

0.2 0.4

0.47 0.68

375 371

1.58 1.54

537 526

41.3 (.54) 44.8 (.47)

Average 12 fish meals

0.2 0.4

0.39(0.013)" 0.55(0.017) 4

368 375

1.62 1.64

542 554

41.0(.315) 46.2 (.336)

1

Period of study from 1 to 21 days of age. Basal diet analyzed 0.20% Ca., other diets supplemented with dicalcium phosphate used limestone to adjust Ca:P ratio to 1.84:1. 3 Fish meal §6 ashed at 650° C. for 2 hours. 4 Figures in parentheses are S.D. Theoretically these diets should analyze 0.39 and 0.55% P, assuming that 8.5 and 17% S.B.M. are removed to maintain isonitrogenous diets when fish meals are used to supply 0.2 and 0.4% P, respectively. 2

Chicks fed the basal diet with no P or with only 0.1% P did not grow normally. Gross symptoms of rickets were observed, starting at 7 days of age, in chicks fed the basal diet. Analysis of variance of the 3-week body-weight data indicates no statistically significant difference among the

~55

Si

53

53

53

55 6S~ 0.7

0.8

0.9

TOTAL /MAILABLE P IN DIET (%) 18 28 .38 .48 LOG OF % TOTAL AVKILABLE P IN DIET

5 8 .68 .78

FIG. 1. Standard bone-ash response curve from the addition of 0.07% P from a commercial dicalcium phosphate source to a basal diet containing 0.24% total P from soybean meal assumed to be one-third available.

chicks fed diets supplemented with 0.2 to 0.7% P from dicalcium phosphate. Supplementation of the basal diet with fish meal ash containing 0.2 and 0.4% P resulted in growth which did not differ significantly from that obtained with control rations containing dicalcium phosphate sources or from the growth obtained with diets supplemented with an equivalent amount of P derived from the intact fish meal from which the fish meal ash was made. Feedintake data indicate that total average consumption did not vary greatly among the groups of chicks fed diets adequate in P. The normal growth rate and constant feed intakes among the groups of chicks fed the various P levels and sources is in contrast to many of the studies previously cited, in which chicks that grow slowly, such as Leghorns, have often been used. Bone ash values obtained from the control diets are given in Table 4 and are graphically illustrated in Figure 1. The response to mineral supplementation of the basal diet as measured by bone ash fol-

1111

AVAILABILITY OF CA AND P IN FISH MEALS

TABLE 5.—Results offeeding diets supplemented with 0.2 and 0.4% P from various fish-meal and control sources

% added P

Number of surviving chicks1 ,

21-day body weight g.

Fee d/unit g<*m

Tibia bone ash (S.E.) 2

0.2

0.4

0.2

0.4

0.2

0.4

0.2

0.4

P source Dicalcium phosphate 20 Fish meal ash4 20

20 19

385 375

398»3 371" b

1.52 1.58

1.53 1.54

41.1 (.64) 41.3 (.55)

45.9 (.10) 44.8 (.47)

Fish meal #1 2 3 4 5 6 7 8 9 10 11 12

20 18 19 20 18 19 20 18 19 18 20 19

383 384 372 366 379 356 358 382 361 361 348 370

396«b 406" 368abo 383»b 391 ab 381»b 383 ab 397»b 347b° 390ab 328= 36 7abc

1.59 1.71 1.62 1.64 1.56 1.60 1.59 1.65 1.68 1.58 1.65 1.61

1.56 1.57 1.62 1.60 1.54 1.63 1.56 1.63 1.77 1.65 1.89 1.61

42.2 (.38) 41.9 (.26) 39.5 (.84) 41.6(.62) 41.0 (.63) 40.8 (.45) 42.0(.95) 41.4(.54) 41.1 (.39) 41.2 (.39) 40.8 (.54) 40.1(.33)

46.2 (.64) 47.3 (.41) 45.9(1.33) 46.3 (.33) 46.7 (.71) 45.9 (.55) 46.5 (.28) 46.7(.66) 44.6(.62) 47.0 (.51) 45.6 (.66) 45.9 (.27)

370

379

1.61

1.62

41.1(.32)

46.1 (.34)

Average all sources

20 20 20 19 20 19 18 18 19 20 17 17

1

Two replicate pens of 10 chicks each started. Lett tibia removed from surviving chicks divided into four composite groups per treatment. s Significant mean differences subjected to Duncan's multiple range test, 5% probability level. Treatments with common letters are not significantly different from each other. 4 See Note 3, Table 4. 2

lowed the law of diminishing returns. With supplementation of from 0 to 0.4% P, the plot of percent bone ash to log of percent total available P is a straight line. This meets the criterion required for a valid P availability assay using the parallel-line assay technique comparing an unknown to a known source. As in previous studies, maximum bone ash required greater levels than did maximum body weight. Fish Meal Diets. Body weights, feed utilization, and bone ash analyses of chicks previously fed diets containing 0.2 and 0.4% P from various sources are shown in Table 5. Analysis of variance of the data on the individual body weights indicates no statistically significant differences among the groups fed the various sources of 0.2% P. Significant differences occurred in the mean weights among the groups fed the 0.4% P supplemental diets. These values were analyzed according to Duncan's

multiple range test (Duncan, 1955) and indicate that growth of the chicks fed diets supplemented with Fish Meals #9 and # 1 1 were significantly lower than those obtained from chicks fed the control diets. These differences are apparently an effect of variability in the quality of the proteins in the meals, since at the levels of supplementation employed, fish meals provided between one-third and one-half of the dietary protein. Variations in feed utilization among the groups appeared to be related to their respective growth rates. The bone ash data shown in Table 5 indicate that the average values were 41.1 and 46.1% for chicks fed diets supplemented with 0.2 and 0.4% P, respectively. At the 0.2% supplementation level, the values ranged from 39.5% for chicks fed Fish Meal # 3 to 42.2% from those fed Fish Meal # 1 . Statistically, these values are no different from those obtained when the standard dicalcium phosphate source

1112

A. H. SPANDORF AND K. C. LEONG

was fed (41.1%). Only the difference between the lowest and highest bone ash values are significant at the 5% level of probability when evaluated by Student's "t" test. In the results obtained from supplementation with 0.4% P, the bone ash values from Fish Meals #9 and # 2 were 44.6 and 47.3%, respectively. Statistically, only the bone ash values obtained from feeding the fish meal ash source (44.8%) differed significantly from the controls at the 5% level of probability, but not at the 1%. On the average, the bone ash values obtained from feeding the intact fish meal sources were 100.1 and 100.7% as great as those obtained from the standard sources when used at 0.2 and 0.4% levels of supplementation, respectively. In studies concerned with biological availability of inorganic phosphate sources, Gillis et al. (1954) calculated the amount of P from an unknown source that would be required to achieve the same percentage of bone ash as resulted from supplementation by a standard beta-tricalcium phosphate source, which by definition is assumed to have a biological value (B.V.) equal to 100. A critical study of this method by Nelson and Walker (1964) showed that, as an example, when a single phosphate sample had a B.V. of 95 its standard deviation was 4.3 B.V. units in 23 repeated determinations. Calculations of B.V. values for fish meals in this study did not appear to be justified, since no statistically significant differences in bone ash values between unknown and the standard source were revealed. The mean and standard error of bone ash values from chicks fed 0.2 and 0.4%, P were 41.1 (.315) and 46.1 (.336), respectively. Thus, no variation in P availability existed among the 12 menhaden fish meals tested; as sources of available Ca and P, they were equal to the standard to which they were compared.

SUMMARY

Twelve menhaden fish meals (F.M.) of normal commercial production were assayed to determine the biological availability of their calcium and phosphorus contents. Growth, feed efficiency, and bone ash values of broiler-type male chicks fed the experimental diets until 3 weeks of age were compared. A semi-purified diet with 20% protein, 3025 kilocalories M.E. per kilo with M.H.A. supplemented S.B.M. as sole source of protein was used. Three series of experimental diets were formulated: (1) a standard assay series in which dicalcium phosphate was added to supplement the diet with 0 to 0.7% P by 0.1% increments and limestone to maintain a constant Ca:P ratio of 1.84:1 (the average value found in the 12 F.M.); (2) a series of 24 assay diets made by substituting each of the 12 meals under test into the basal diet to provide 0.2 and 0.4% P. No additional Ca other than that obtained from the F.M. was added; and (3) additional control diets in which a low-temperature ash made from one of the F.M. under test was used as a source of 0.2 and 0.4% P. The diets were kept isonitrogenous and isocaloric. Chicks fed the basal diet showed signs of severe rickets after 7 days. Supplementation of this diet with 0.2% or more P from all sources resulted in average body weights at 21 days of 375 g. with a feed efficiency of 1.62. Body weights were not a sensitive criterion of mineral availability. Bone ash results from the chicks fed the control series of diets indicated a log dose response curve between 0 and 0.4% levels of P addition. Comparison of the ash values from diets supplemented with fish meal sources indicates that the biological availability of the Ca and P in fish meals averaged 100.4% and ranged from 96.1 to

AVAILABILITY OF CA AND P IN FISH MEALS

102.7% of the values obtained with corresponding mineral levels from dicalcium phosphate and limestone. Average bone ash values for the diets supplemented with 0.2 and 0.4% P from dicalcium phosphate and limestone were 41.1 and 45.9%, respectively, compared with 40.1 and 46.2% from fish-meal supplementation. Since none of the bone ash values obtained from chicks fed the fish-meal sources differed statistically from those obtained from the standard sources, their biological value as mineral sources can be assumed to be equal. ACKNOWLEDGMENT

V. Scarborough and Preston Smith, Jr., provided the bone ash and phosphorus analyses, respectively. REFERENCES Association of Official Agricultural Chemists, 1960. Official Methods of Analysis, 9th Ed., Washington, D.C. Dilworth, B. C , and E. J. Day, 1964. Phosphorus availability studies with feed grade phosphates. Poultry Sci. 43 : 1039-1044.

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Duncan, D. B., 1955. Multiple range and multiple " F " tests. Biometrics, 11: 1-12. Gillis, M. B., L. C. Norris and G. F. Heuser, 1951. Biological availability of inorganic phosphates. Poultry Sci. 30: 914. Gillis, M. B., L. C. Norris and G. F. Heuser, 1954. Studies on the biological value of inorganic phosphates. J. Nutr. 52: 115-126. Grau, C. R., and P. A. Zweigert, 1953. Phosphatic clay as a phosphorus source for chicks. Poultry Sci. 32: 500-503. Hawk, P. B., B. L. Oser and W. H. Summerson, 1954. Practical Physiological Chemistry, 13th Ed., pp. 630-632. Motzok, I., D. Arthur and H. D. Branion, 1956. Utilization of phosphorus from various phosphate supplements by chicks. Poultry Sci. 35: 627-649. Miller, M. W., and V. V. Joukovsky, 1953. Availability of phosphorus from various phosphate materials for chicks. Poultry Sci. 32: 78-81. National Research Council, 1956. Nutrient requirements for domestic animals. No. 1. Nutrient requirements for poultry. Nelson, T. S., and A. C. Walker, 1964 The biological evaluation of phosphorus compounds. Poultry Sci. 43 : 94-98. Snyder, D. G., L. E. Ousterhout, H. W. Titus, K. Morgareidge and S. Kellenbarger, 1962. The evaluation of the nutritive content of fish meals by chemical methods. Poultry Sci. 41 : 1736-1740.

Relationship of Protein Level, Age and Ambient Temperature to Laying Hen Performance B. L. REID, A. A. KUENICK 1 AND B. J. HULETT Poultry Science Department, University of Arizona, Tucson, Arizona (Received for publication January 25, 1965)

T

HE requirement of laying hens for protein has been the subject of numerous investigations, and a wide range of protein levels has been found to support reasonable rates of production. Reports 1 Present address: The Ray Ewing Co., Div. of Hoffman-LaRoche Inc., Pasadena, California. Arizona Agricultural Experiment Station, Journal Article #914.

have indicated the protein needs of the laying hen to be from 11 to 18% (Heywang et al., 1955; Miller et al., 1956; Frank and Waibel, 1959; Griminger and Fisher, 1959; Thornton et al, 1957; Quisenberry and Williams, 1952; Milton and Ingram, 1957; Creek and Combs, 1958). Based on these reports, a number of factors would seem to be involved in an