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Effect of Autoclaving Field Beans (Phaseolus Vulgaris) and of Supplementing Diets Containing Beans with Amino Acids or Antibiotics on Performance of Young Chicks1,2
Effect of Autoclaving Field Beans {Phaseolus Vulgaris) and of Supplementing Diets Containing Beans with Amino Acids or Antibiotics on Performance of Young Chicks1'2
(Received for publication March 6, 1972)
ABSTRACT Two experiments were conducted with young chicks in which different varieties of dry field beans (Phaseolus vulgaris) were used to furnish approximately 67% of the dietary protein. The beans were used in either raw or cooked form and the diets were supplemented with methionine, a mixture of amino acids or a mixture of antibiotics. The results obtained show that all varieties of beans supported very low growth when fed in raw form and that significant differences in nutritional value were found among varieties. Cooking all varieties of beans by autoclaving gave marked improvements in chick growth. Supplementing diets containing cooked beans with methionine improved growth and feed efficiency to varying degrees, depending upon the variety, but diets containing raw beans were not improved by a methionine supplement. A mixture of four amino acids including methionine was no more effective in stimulating growth of chicks than methionine alone. Adding a mixture of penicillin and streptomycin to diets containing either raw or cooked beans gave a much greater growth response than methionine or the amino acid mixture. All varieties of beans fed in raw form caused marked pancreatic enlargement and cooking the beans reduced this effect but did not completely eliminate the enlargement. POULTSY SCIENCE SI: 1976-1983,
D
RY field beans {Phaseolus vulgaris) constitute a significant source of protein for man in many areas of the world and offer an important potential for increasing protein production to meet increasing protein needs. In spite of the relatively large number of different bean varieties grown, there is relatively little comparative information on nutritional value of the different varieties. The nutritional value of dried Pinto beans, cooked by autoclaving, was shown to be unexpectedly low for chicks (Riley, 1961). These findings suggested that beans contain heat stable growth inhibitors or that they depress growth indirectly in some manner. Studies were undertaken to deter-
1972
mine whether there are differences in nutritional value of some different varieties of dry beans and to observe the effect of different dietary supplements such as amino acids and antibiotics on chicks fed diets containing beans. PROCEDURE
In the two experiments described in this paper each experimental diet was fed to three groups of 12 commercial type broiler chicks from hatching to 3 weeks of age. These chicks were allotted to the experimental groups at random and without sorting by sex. During the experimental period they were kept in electrically heated battery brooders and given feed and water ad libitum. Fourteen hours of light were pro' Scientific Paper No. 3828, College of Agriculvided each day. ture, Washington State University, Pullman. Project 1533. Four varieties of beans (Big Bend, Pinto 2 Supported partially by U.S.D.A. Cooperative 114, Sutter Pink and California Small Agreement 12-14-100-10011 (34). 3 Current address: Food & Drug Administration, White) were used in compounding the experimental diets. Each variety was used in Washington, D.C. 1976
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W. D . GOATCHER3 AND J. McGlNNIS Department of Animal Sciences, Washington State University, Pullman, Washington 99163
FIELD BEANS IN CHICK RATIONS
1977
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TABLE 1.—Percentage composition of control raw form and subjected to an autoclaving and experimental diets cooking treatment (10 minutes at 120°C). Diet The beans were ground in a hammer mill 3 1 2 prior to the cooking treatment and were Item then spread in metal trays in layers apMixed Soybean protein meal Beans proximately 2 cm. deep. They were aircontrol control 10.10 10.10 10.10 dried after autoclaving before mixing in the Corn, ground Corn gluten meal, 41% prot. 3.40 3.40 3.40 feeds. Wheat, ground 27.50 27.50 27.50 Soybean meal, 48.5% prot. 11.56 24.00 — The beans were included in the experi- Meat and bone meal, 50% prot. 4.62 — — Fish meal, herring, 70% prot, 4.62 — • — mental diets to provide approximately 67% Alfalfa1 meal, 20% prot. 2.30 — — Beans — 28.88 — 53.50 29.27 of the total dietary protein which was cal- Cerelose — Cellulose, powdered 4.45 2.44 — 1.00 2.00 1.90 culated to be approximately 17%. The Dicalcium phosphate Calcium carbonate 1.00 0.92 o.so 0.30 0.30 0.30 same level of beans was used in all diets, Salt, iodized 2 Vitamin mix 0.33 0.33 0.33 0.05 0.05 0.05 even though this caused some slight varia- Trace mineral mix8 Fat, animal 2.00 — — tions in protein content of the diets because Protein, %, NX6.25 17.30 17.0-17.6 17.40 of the possibility that some factor in beans 1 Four different varieties of beans were used (Red Mexican, other than protein might influence chick Pinto 114, Sutter's Pink and California Small White). 2 Provided per kg. of diet: vitamin A, 3,300 I.U.; vitamin Da, growth. Two different control diets were 300 I.C.U.; vitamin K, 0.66 mg.; vitamin E, 1.1 I.U.; thiaminNOi_, 2.4 mg.; riboflavin, 4.3 mg.; Ca pantothenate. 13.7 mg.; used, along with the experimental diets niacin, 32.4 mg.; pyridoxine-HCl, 4.4 mg.; folicin, 1.4 mg.; biotin, 0.11 mg.; choline-Cl. 1,560 mg.; vitamin B12, 0.011 mg.; containing different varieties of beans in or- and8 ethoxyquin, 125 mg. per kg. of diet: Mn, 50 mg.; Fe, 50 mg.; Cu. 5 mg.; der to provide data on nutritional value of Zn, 50Provided mg.; 1,1.5 mg.; and Co, 0.5 mg. beans compared with soybean meal and with dietary protein from a wider variety of materials. Composition of the experi- each was removed and weighed to the nearest milligram. mental diets is shown in Table 1. In order to provide data on the effect of Experiment 1. The experimental basal diets containing different varieties of beans feeding diets containing raw beans during a were fed without supplementation and with 3-week period on the response to a change the addition of 0.35% DL-methionine. As in diet, 10 chicks were selected at random indicated earlier, each variety of beans was from each of the unsupplemented diets conused in a raw and cooked form. The sup- taining raw beans and continued for two plementary treatments in outline form are weeks. They were fed Control Diet 1 durshown in Table 2 along with body weight ing this period. Feed consumption and gain so that feed conversion and total pro- body weight gain were determined. Experiment 2. At the conclusion of Extein efficiency (T.P.E.) could be calculated as grams of feed per gram of gain and periment 1, a considerable quantity of each grams of gain per gram of protein con- experimental feed was unconsumed. Some of these experimental diets were modified sumed, respectively. At the end of the 3-week period, when to provide the different treatments for this final body weights and feed consumption experiment. The diets to which DL-methiodata were collected, 10 chicks were selected nine were added in Experiment 1 were adfrom the control treatment fed Diet 1 and ditionally supplemented by adding 0.35% from the treatments containing the differ- DL-threonine, 0.35% L-tyrosine and 0.12% ent varieties of beans in raw and cooked DL-valine. The unsupplemented diets in Exform and weighed individually. They were periment 1 containing the different varithen slaughtered and the pancreas from eties of beans in both raw and heated forms
1978
W . D . GOATCHER AND J . M c G l N N I S T A B U : 2.—Experiment 1. Effect of autoclaving and supplementing different varieties of beans with methionine on chick growth Av. gain in 3 wks. (gms.) Type of beans
Supplement % Raw beans
Cooked beans 1
None 0.35 DL-methionine
16.0 a2 19.7 a2
139.9 a2 164.4 be3
Pinto 114
None 0.35 DL-methionine
46.5 be2 49.2 c2
144.6 ab 3 174.5 cd3
Sutter Pink
None 0.35 DL-methionine
25.0 abc2 23.5 ab 2
163.0 abc 3 194.6 d3
California Small White
None 0.35 DL-methionine
24.1 ab 2 25.5 abc2
168.5 be' 193.5 d3
Diet 1—Mixed protein control--314.4 Diet 2—Soybean meal control--276.4 1
Beans were spread in trays approximately 2.0 cm. deep and autoclaved 10 minutes at 120°C. Means within each column having different letters and means within each row having different supernumerals are significantly different (P<0.05). 2
were supplemented with a mixture of procaine penicillin G and streptomycin sulfate (0.1% of each). One additional treatment was provided in this experiment by adding a combination of the additional amino acids and antibiotics to the diet from Experiment 1 containing cooked Pinto beans and supplemental methionine. Control Diets 1 and 2 were fed in this trial to give comparative data. Other procedures in this experiment were as described for Experiment 1. RESULTS
Results of the two experiments are expressed primarily on the basis of average gain in weight, total protein efficiency and feed conversion. Total protein efficiency (T.P.E.) is denned as the amount of gain in weight per unit of crude protein consumed. Woodham (1968) showed this to be a reliable measure for comparing relative protein values for chicks. Feed conversion (F/G) represents the amount of feed consumed per unit of gain. Some additional results are given and will be identified as a part of each experiment. Experiment 1. The results on average
chick gain for this experiment are summarized in Table 2. Methionine supplementation of the diets containing raw beans failed to give a significant (P > 0.05) increase in weight gain with all of the different bean varieties. Adding methionine to the diets containing cooked beans gave statistically significant (P < 0.05) increases for all bean varieties and the response to cooking was likewise statistically significant. The two different control diets gave greater gains than any of the experimental diets containing beans. This low weight gain by chicks fed diets containing cooked beans, even after supplementation with methionine, is in agreement with the results obtained by Riley (1961). Results on T.P.E. and F/G are summarized in Table 3. The T.P.E. obtained with diets containing all of the different varieties of beans in raw form was very poor and addition of methionine failed to significantly (P > 0.05) increase T.P.E. The cooking treatment for all varieties of beans significantly increased all T.P.E. values and the improvement by addition of methionine to diets containing cooked beans further sig-
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Small Red
1979
FIELD BEANS IN CHICK RATIONS
not significantly improve F/G for diets containing Pinto or White beans. Autoclaving all of the different varieties of beans gave significant improvements in F/G and methionine supplements of diets containing the different beans further significantly improved F/G. Varietal differences were obtained within the group of diets containing heated beans, with White and Red bean diets giving better F/G than the diets with Red or Pink beans. Adding methionine to the diets containing cooked beans improved F/G for all varieties. The results obtained on pancreas weight of chicks fed diets containing the different varieties of beans, either in raw or cooked form, are summarized in Table 4. Diet containing all of the different varieties of beans in raw form caused pancreatic enlargement and this enlargement was significantly reduced by the cooking treatment. The growth response obtained during a 2-week period following a change from diets containing raw beans to a control diet are summarized in Table 5. These results show a marked increase in rate of gain, in-
TABLE 3.—Experiment 1. Effect of bean variety, cooking treatment and dietary supplements on feed conversion and total protein efficiency of chicks Feed conversion1'2
Total protein emc.1.2
Supplement %
Raw Beans
Cooked Beans
Raw Beans
Cooked Beans
Small Red
None 0.35% DL-meth.
7.49^ 6.82 f
2.65 d 2.45 be
0.786 a 0.845 a
2.22 ab 2.35 be
Pinto 114
None 0.35% DL-meth.
5.15b 4.79 a
2.40 ab 2.21 a
1.13 cd 1.19d
2.42 cd 2.58 d
Sutter Pink
None 0.35% DL-meth.
6.27 e 5.48 c
2.62 cd 2.38 ab
0.902 ab 1.01 be
2.16a 2.33 be
California Small White
None 0.35% DL-meth.
5.85 d 5.71 d
2.52 bed 2.23 a
0.998 be 1.00 be
2.32 abc 2.57 d
Beans
Diet 1 Diet 2 1 2 3
Mixed protein control Soybean meal control
1..77 1 .82
Average of three replicates of 12 chicks fed from hatching to 3 weeks of age. Response on raw beans differs significantly from that on cooked beans in every case. Means within each column having different letters are significantly different (P<0.05).
3.25 3.17
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nificantly increased T.P.E. with the exception of the diet containing Small Red beans. Significant varietal differences were obtained in T.P.E. with both the raw and cooked beans. The T.P.E. value for diets containing raw Pinto beans was significantly greater, either with or without added methionine than that for diets containing either Pink or Small Red beans. This difference, however, was not significant from that for the diet containing raw White beans. Similar differences in T.P.E. values were obtained with diets containing the different varieties of cooked beans. Supplementing the diets containing either cooked Pinto or White beans with methionine gave similar T.P.E. values and these values were signifiantly different from those for Small Red and Pink beans. T.P.E. values for both control diets were much greater than any of the values obtained with the diets containing beans. Feed conversion was improved by adding methionine to the diets containing either Small Red or Pink beans. Methionine did
1980
W . D . GOATCHER AND J .
McGlNNIS
significantly greater gains in the absence of supplemental amino acids. Furthermore, gain of chicks in the one treatment fed a Average pancreas weights— mg./g. body wt. 1 diet containing Pinto beans supplemented Diet or bean variety with both the mixtures of amino acids and Raw Cooked beans beans antibiotics significantly increased gain over 2 either supplement alone. These results sug4.8 a Control diet 1 4.8a 8.6 c4 12.7 b 3 Small Red gest that cooked beans exert a depressing 7.1b4 14.6 c3 Pinto 114 4 3 effect on chick growth, possibly through the 7.4 be 12.1b Sutter Pink 7.5 be4 12.2 b 3 California Small White stimulation of an adverse intestinal microflora and that antibiotic supplements stim1 Average of 10 weights. 2 Means within each column having different letulate chick growth by preventing the develters, and means within each row having different opment of such a microflora. supernumerals are significantly different (P<0.05). Diets with cooked Pink and Small White beans supported chick growth when suppledicating that feeding of the raw beans dur- mented with antibiotics that was not signifing the 3-week period from hatching appar- icantly different from that obtained with ently caused no permanent damage. The Control Diet 2. These results are in marked F/G data for this period also show that the contrast to those obtained in Experiment 1 pancreatic enlargement which was evident and those reported by Riley (1961). at 3 weeks of age apparently did not interThe results on F/G and T.P.E. are sumfere with pancreatic enzyme secretion as marized in Table 7. Autoclaving the differthe F/G data compare favorably with previ- ent varieties of beans gave significant imous data with the control diets. provements in F/G as was obtained in ExExperiment 2. The data obtained in Ex- periment 1 and the additional amino acids periment 2 by feeding diets remaining from failed to give results that were significantly Experiment 1, modified as indicated, are better than those obtained in Experiment 1 summarized in Tables 6 and 7. The results with methionine alone. Adding the mixture on chick growth shown in Table 6 indicate of antibiotics significantly improved F/G that the mixture of amino acids failed to for diets containing cooked Pink and White improve growth of chicks fed diets contain- beans. None of the F/G values obtained ing raw beans over the results obtained in with cooked beans supplemented with eiExperiment 1 with methionine alone. In ther amino acids or antibiotics equaled the contrast, the supplement of antibiotics to the F/G values obtained with the control diets diets containing raw beans gave significant (P < 0.0S) increases in gain for all varieties. T A B L E 5.—Response of chicks previously fed diets The Pinto variety fed in raw form gave containing raw beans to a change of diet greater gains, either with the mixture of Av. Av. Av. feed amino acids or the supplement of antibiotRaw bean diet weight gain2 Cons. Feed previously fed1 at 3 wks. 3-5 wks. 3-5 wks. gain ics than other varieties of beans. Even (g.) (g.) (g.) 66.3 261. 8 a' 494.4 1.89 though the supplement of methionine in Small Red Pinto 114 99.5 305.6 b 556.0 1.82 83.0 253.2 a 490.5 1.94 Experiment 1 to diets containing cooked SutterPink California Small White 82.8 293.5 b 573.0 1.95 beans and the addition of the mixture of i The raw bean diets indicated were fed from day of hatch to amino acids in Experiment 2 caused large 3 weeks of age, after which the chicks were switched to the mixed control diet for an additional 2-week period. increases in gain, the addition of the antibi3 Average of 10 chicks. , » Means within the column having different letters are signiotic supplement in this experiment gave ficantly different (P <0.05). T A B L E 4.—Influence of bean variety and cooking of beans on pancreas weight of chicks
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1981
FIELD BEANS IN CHICK RATIONS
TABLE 6.—Experiment 2. Effect of supplementing diets containing different varieties of beans with amino acids or antibiotics and of autoclaving the beans on chick growth Av. gain in 3 wks.fems.)1 J-^JJC U l
beans
Supplement
Small Red
Amino acids 3 Antibiotics4
Pinto 114
Amino acids Antibiotics
Raw beans
Cooked beans 2
30.0 a 167.3 a 61.8 be 220.6 d 56.1 be 190.1 abc 106.6 d 214.1 cd
Amino acids+ antibiotics
—
249.0 e
Sutter Pink Amino acids Antibiotics
38.0 a 80.5 c
184.8 ab 265.7 e
California Amino acids Small White Antibiotics
31.8 ab 78.5 c
205.4 bed 266.1 e
Diet 1 Diet 2
Mixed protein control Soybean meal control
317.8 273.9
1 Means within each column having different letters and all differences between raw and cooked beans are significantly different (P < 0.05). 2 Beans were spread in trays approximately 2.0 cm. deep and autoclaved 10 minutes at 120°C. 3 0.35% DL-methionine, 0.35% DL-threonine, 0.35% L-tyrosine and 0.12% DL-valine. 4 0.1 % procaine penicillin and 0.1% streptomycin sulfate.
TABLE 7.—Experiment 2. Effect of bean variety, cooking treatment and supplements of amino acids and antibiotics on feed conversion and total protein efficiency of chicks Feed conversion1'2 Type of bean
Supplement
%
Raw beans
Total prot. efficiency1'2
Cooked bean's
Raw beans
Cooked beans
Small Red
Amino acids3 Antibiotics4
4.76 d 3.25 be
2.45 c '2.33 be
1.16 a ' 1.81b
2.25 a 2.52 be
Pinto 114
Amino acids Antibiotics Amino acids+antibiotics
4.43 c 3.10 b
2.18 ab 2.08 a 2.16 ab
1.23 a 1.79 b
2.49 be 2.79 d 2.53 be
Sutter Pink
Amino acids Antibiotics
4.40 c 3.11 b
2.30 be 2.15 ab
1.27 a 1.82 b
2.44 b 2.63 cd
California Small White
Amino acids Antibiotics
99 e 82 a
2.17 ab 2.06 a
1.10 a 2.07 c
2.52 be 2.84 d
Diet 1 Diet 2 1
Mixed protein control Soybean meal control
1.77 1.82
Average of 3 replicates of 12 chicks fed from hatching to 3 weeks of age. Response on raw beans differs significantly from that on cooked beans in every case. Mixture of 0.35% DL-methionine, DL-threonine, L-tyrosine and 0.12% DL-valine. 4 0 . 1 % procaine penicillin plus 0 . 1 % streptomycin sulfate. ? Means within each column having,a different letter are significantly different (P<0.05). 2 3
3.25 3.17
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in this experiment. In general, the supplement of antibiotics was more effective in improving F/G than the supplement of amino acids. The T.P.E. values summarized in Table 7 show that all diets containing raw beans gave higher T.P.E. values when antibiotics were added than when the mixture of amino acids was used. The improvements with antibiotic supplements were significant in all cases. T.P.E. values were greatly improved by cooking the beans and again the supplement of antibiotics gave higher T.P.E. values than the supplement of amino acids. The improvements from cooking were significant for all bean varieties and the improvements from the antibiotic supplements were likewise significant. As in Experiment 1, all of the T.P.E. values for diets containing cooked beans and supplemented with either amino acids or antibiotics were better (lower) than the values obtained with the control diets. These differences are closely related to the differences in rates of gain given by the different diets.
1982
W. D. GOATCHER AND J. McGlNNIS DISCUSSION
and to a correcting of a limiting deficiency of total sulfur amino acids or to a combination of these factors. Further amino acid supplementation (methionine plus threonine, valine and tyrosine) did not generally improve bean nutritive value over the improvement obtained with methionine alone. This finding is in contrast to the results obtained with soybean meal where supplementing with these amino acids increased its value (Booth et al, 1960). Wagh et al. (1963), working with chicks, equated the essential amino acid composition of a red kidney bean diet with that of a control ration and failed to obtain equal growth of chicks fed the two diets. They suggested that the failure may have been due to such factors as a thermostable toxin in beans, poor availability of other amino acids, or a dietary energy deficit. Antibiotic supplementation of bean diets resulted in about a 200% increase in gain when the beans were raw and about a 56% increase with autoclaved beans. Other studies in this laboratory have shown that large antibiotic responses are also obtained with diets containing either dry peas or lentils (Goatcher and McGinnis, 1972) and with diets containing rye (MacAuliffe and McGinnis, 1971). The chick growth response which is usually obtained with antibiotic supplements is about 5-10% (Matterson and Singsen, 1951; Heuser and Norris, 1952). The addition of an antibiotic supplement to the mixed protein control diet 1 used in these studies produced data showing that antibiotic supplementation only gave a 6% improvement in growth. Thus, it appears that the antibiotic growth response is markedly influenced by ration composition. The data of Kakade and Evans (1964, 1966) indicate that antibiotics are less effective in promoting growth of rats fed diets containing beans than they are in pro-
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Raw beans of all varieties supported poor chick growth. This may have been due to generally poor digestibility of protein and/or energy or due to the presence of toxic factors or some combination of these. Riley (1961) observed an increase in metabolizable energy of cull field beans from 449 to 877 Kcal./kg. when these beans were autoclaved and included in chick diets at a level of 40%. Autoclaving the beans resulted in an increase in 3-week gain of about 500% and an increase in T.P.E. values and feed consumption of about 150%. However, performance was still below that of chicks fed control diets. The increase in nutritive value of beans with autoclaving is probably due to increases in digestibility as well as inactivation of some toxic factors known to be present in many legumes. Liener (1953) demonstrated that soybean hemagglutinin was growth-inhibitory to rats. Honavar et al. (1962) showed that Phaseolus vulgaris hemagglutinin depressed chick growth. Trypsin inhibitors do not appear to account completely for the poor chick growth obtained with Phaseolus vulgaris (Riley, 1961) or Glycine max (Saxena, 1964). The data indicate that the varieties of beans studied differed in nutritive value. Generally speaking, the White and Pink beans were of higher nutritional quality than the Pinto and Red beans. Kakade and Evans (1965), using rats as the experimental animal, did not detect a difference in nutritional value of 5 varieties of navy beans. Methionine supplementation of heated bean diets improved performance characteristics. Supplementing the raw bean diets with methionine failed to improve bean nutritive value. The response to methionine additions to diets containing heated beans could be related to decreased growth depression by heat labile growth inhibitors
1983
FIELD BEANS IN CHICK RATIONS
moting chick growth. In their work, however, dietary bean level was about 41.6% whereas in the present study it was 53.6%.
1. 2. 3. 4. 5. 6.
7. 8. °. 10. 11.
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REFERENCES Booth, A. N., D. J. Robbins, W. E. Ribelin and F. DeEds, 1960. Effect of raw soybean meal and amino acids on pancreatic hypertrophy in rats. Proc. Soc. Exp. Biol. Med. 104: 681-683. Goatcher, W. D., and J. McGinnis, 1972. Influence of beans, peas and lentils as a dietary component and growth response of chicks to antibiotics and methionine supplementation of the diet. Poultry Sci. 51: 440-443. Heuser, G. F., and L. C. Norris, 1952. Some results of feeding antibiotics to chickens. Poultry Sci. 3 1 : 857-862. Honavar, P. M., C. Shih and I. E. Liener, 1962. Inhibition of the growth of rats by purified hemagglutinin fractions isolated from Phaseolus vulgaris. J. Nutrition, 77: 109-114. Kakade, M. L., and R. J. Evans, 1964. Effect of methionine, vitamin B12 and antibiotics supplementation on protein nutritive value of navy beans. Proc. Soc. Exp. Biol. Med. 115: 890-893. Kakade, M. L., and R. J. Evans, 1965. Nutritive value of different varieties of navy beans. Michigan State Univ. Agr. Expt. Sta. Quart. Bui. 48: 89-93.
Kakade, M. L., and R. J. Evans, 1966. Growth inhibition of rats fed raw navy beans (Phaseolus vulgaris). J. Nutrition, 90: 191-198. Liener, I. E., 1953. Soyin, a toxic protein from the soybean. I. Inhibition of rat growth. J. Nutrition, 49: 527-539. MacAuliffe, T., and J. McGinnis, 1971. Effect of antibiotic supplements to diets containing rye on chick growth. Poultry Sci. 50: 1130-1134. Matterson, L. D., and E. P. Singsen, 1951. A comparison of several antibiotics as growth stimulants in practical chick-starting rations. Storrs (Connecticut) Agr. Exp. Sta. Bui. 275. Riley, D. J., 1961. Studies on the nutritional value of some little-used protein feedstuffs for poultry. M.S. Thesis, Washington State University, Pullman, WA. Saxena, H. C , 1964. Investigations on the mechanism of growth depression and pancreatic hypertrophy by raw soybean meal in the chick. Ph.D. Thesis, Washington State University, Pullman, WA. Wagh, P. V., D. F. Klaustermeier, P. E. Waibel and I. E. Liener, 1963. Nutritive value of red kidney beans (Phaseolus vulgaris) for chicks. J. Nutrition, 80: 191-195. Woodham, A. A., 1968. A chick growth test for the evaluation of protein quality in cereal-based diets. 1. Development of the method. British Poultry Sci. 9 : 53-63.
(Received for publication March 6, 1972)
ABSTRACT Two experiments were conducted with young chicks in which different varieties of dry field beans (Phaseolus vulgaris) were used to furnish approximately 67% of the dietary protein. The beans were used in either raw or cooked form and the diets were supplemented with methionine, a mixture of amino acids or a mixture of antibiotics. The results obtained show that all varieties of beans supported very low growth when fed in raw form and that significant differences in nutritional value were found among varieties. Cooking all varieties of beans by autoclaving gave marked improvements in chick growth. Supplementing diets containing cooked beans with methionine improved growth and feed efficiency to varying degrees, depending upon the variety, but diets containing raw beans were not improved by a methionine supplement. A mixture of four amino acids including methionine was no more effective in stimulating growth of chicks than methionine alone. Adding a mixture of penicillin and streptomycin to diets containing either raw or cooked beans gave a much greater growth response than methionine or the amino acid mixture. All varieties of beans fed in raw form caused marked pancreatic enlargement and cooking the beans reduced this effect but did not completely eliminate the enlargement. POULTSY SCIENCE SI: 1976-1983,
D
RY field beans {Phaseolus vulgaris) constitute a significant source of protein for man in many areas of the world and offer an important potential for increasing protein production to meet increasing protein needs. In spite of the relatively large number of different bean varieties grown, there is relatively little comparative information on nutritional value of the different varieties. The nutritional value of dried Pinto beans, cooked by autoclaving, was shown to be unexpectedly low for chicks (Riley, 1961). These findings suggested that beans contain heat stable growth inhibitors or that they depress growth indirectly in some manner. Studies were undertaken to deter-
1972
mine whether there are differences in nutritional value of some different varieties of dry beans and to observe the effect of different dietary supplements such as amino acids and antibiotics on chicks fed diets containing beans. PROCEDURE
In the two experiments described in this paper each experimental diet was fed to three groups of 12 commercial type broiler chicks from hatching to 3 weeks of age. These chicks were allotted to the experimental groups at random and without sorting by sex. During the experimental period they were kept in electrically heated battery brooders and given feed and water ad libitum. Fourteen hours of light were pro' Scientific Paper No. 3828, College of Agriculvided each day. ture, Washington State University, Pullman. Project 1533. Four varieties of beans (Big Bend, Pinto 2 Supported partially by U.S.D.A. Cooperative 114, Sutter Pink and California Small Agreement 12-14-100-10011 (34). 3 Current address: Food & Drug Administration, White) were used in compounding the experimental diets. Each variety was used in Washington, D.C. 1976
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W. D . GOATCHER3 AND J. McGlNNIS Department of Animal Sciences, Washington State University, Pullman, Washington 99163
FIELD BEANS IN CHICK RATIONS
1977
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TABLE 1.—Percentage composition of control raw form and subjected to an autoclaving and experimental diets cooking treatment (10 minutes at 120°C). Diet The beans were ground in a hammer mill 3 1 2 prior to the cooking treatment and were Item then spread in metal trays in layers apMixed Soybean protein meal Beans proximately 2 cm. deep. They were aircontrol control 10.10 10.10 10.10 dried after autoclaving before mixing in the Corn, ground Corn gluten meal, 41% prot. 3.40 3.40 3.40 feeds. Wheat, ground 27.50 27.50 27.50 Soybean meal, 48.5% prot. 11.56 24.00 — The beans were included in the experi- Meat and bone meal, 50% prot. 4.62 — — Fish meal, herring, 70% prot, 4.62 — • — mental diets to provide approximately 67% Alfalfa1 meal, 20% prot. 2.30 — — Beans — 28.88 — 53.50 29.27 of the total dietary protein which was cal- Cerelose — Cellulose, powdered 4.45 2.44 — 1.00 2.00 1.90 culated to be approximately 17%. The Dicalcium phosphate Calcium carbonate 1.00 0.92 o.so 0.30 0.30 0.30 same level of beans was used in all diets, Salt, iodized 2 Vitamin mix 0.33 0.33 0.33 0.05 0.05 0.05 even though this caused some slight varia- Trace mineral mix8 Fat, animal 2.00 — — tions in protein content of the diets because Protein, %, NX6.25 17.30 17.0-17.6 17.40 of the possibility that some factor in beans 1 Four different varieties of beans were used (Red Mexican, other than protein might influence chick Pinto 114, Sutter's Pink and California Small White). 2 Provided per kg. of diet: vitamin A, 3,300 I.U.; vitamin Da, growth. Two different control diets were 300 I.C.U.; vitamin K, 0.66 mg.; vitamin E, 1.1 I.U.; thiaminNOi_, 2.4 mg.; riboflavin, 4.3 mg.; Ca pantothenate. 13.7 mg.; used, along with the experimental diets niacin, 32.4 mg.; pyridoxine-HCl, 4.4 mg.; folicin, 1.4 mg.; biotin, 0.11 mg.; choline-Cl. 1,560 mg.; vitamin B12, 0.011 mg.; containing different varieties of beans in or- and8 ethoxyquin, 125 mg. per kg. of diet: Mn, 50 mg.; Fe, 50 mg.; Cu. 5 mg.; der to provide data on nutritional value of Zn, 50Provided mg.; 1,1.5 mg.; and Co, 0.5 mg. beans compared with soybean meal and with dietary protein from a wider variety of materials. Composition of the experi- each was removed and weighed to the nearest milligram. mental diets is shown in Table 1. In order to provide data on the effect of Experiment 1. The experimental basal diets containing different varieties of beans feeding diets containing raw beans during a were fed without supplementation and with 3-week period on the response to a change the addition of 0.35% DL-methionine. As in diet, 10 chicks were selected at random indicated earlier, each variety of beans was from each of the unsupplemented diets conused in a raw and cooked form. The sup- taining raw beans and continued for two plementary treatments in outline form are weeks. They were fed Control Diet 1 durshown in Table 2 along with body weight ing this period. Feed consumption and gain so that feed conversion and total pro- body weight gain were determined. Experiment 2. At the conclusion of Extein efficiency (T.P.E.) could be calculated as grams of feed per gram of gain and periment 1, a considerable quantity of each grams of gain per gram of protein con- experimental feed was unconsumed. Some of these experimental diets were modified sumed, respectively. At the end of the 3-week period, when to provide the different treatments for this final body weights and feed consumption experiment. The diets to which DL-methiodata were collected, 10 chicks were selected nine were added in Experiment 1 were adfrom the control treatment fed Diet 1 and ditionally supplemented by adding 0.35% from the treatments containing the differ- DL-threonine, 0.35% L-tyrosine and 0.12% ent varieties of beans in raw and cooked DL-valine. The unsupplemented diets in Exform and weighed individually. They were periment 1 containing the different varithen slaughtered and the pancreas from eties of beans in both raw and heated forms
1978
W . D . GOATCHER AND J . M c G l N N I S T A B U : 2.—Experiment 1. Effect of autoclaving and supplementing different varieties of beans with methionine on chick growth Av. gain in 3 wks. (gms.) Type of beans
Supplement % Raw beans
Cooked beans 1
None 0.35 DL-methionine
16.0 a2 19.7 a2
139.9 a2 164.4 be3
Pinto 114
None 0.35 DL-methionine
46.5 be2 49.2 c2
144.6 ab 3 174.5 cd3
Sutter Pink
None 0.35 DL-methionine
25.0 abc2 23.5 ab 2
163.0 abc 3 194.6 d3
California Small White
None 0.35 DL-methionine
24.1 ab 2 25.5 abc2
168.5 be' 193.5 d3
Diet 1—Mixed protein control--314.4 Diet 2—Soybean meal control--276.4 1
Beans were spread in trays approximately 2.0 cm. deep and autoclaved 10 minutes at 120°C. Means within each column having different letters and means within each row having different supernumerals are significantly different (P<0.05). 2
were supplemented with a mixture of procaine penicillin G and streptomycin sulfate (0.1% of each). One additional treatment was provided in this experiment by adding a combination of the additional amino acids and antibiotics to the diet from Experiment 1 containing cooked Pinto beans and supplemental methionine. Control Diets 1 and 2 were fed in this trial to give comparative data. Other procedures in this experiment were as described for Experiment 1. RESULTS
Results of the two experiments are expressed primarily on the basis of average gain in weight, total protein efficiency and feed conversion. Total protein efficiency (T.P.E.) is denned as the amount of gain in weight per unit of crude protein consumed. Woodham (1968) showed this to be a reliable measure for comparing relative protein values for chicks. Feed conversion (F/G) represents the amount of feed consumed per unit of gain. Some additional results are given and will be identified as a part of each experiment. Experiment 1. The results on average
chick gain for this experiment are summarized in Table 2. Methionine supplementation of the diets containing raw beans failed to give a significant (P > 0.05) increase in weight gain with all of the different bean varieties. Adding methionine to the diets containing cooked beans gave statistically significant (P < 0.05) increases for all bean varieties and the response to cooking was likewise statistically significant. The two different control diets gave greater gains than any of the experimental diets containing beans. This low weight gain by chicks fed diets containing cooked beans, even after supplementation with methionine, is in agreement with the results obtained by Riley (1961). Results on T.P.E. and F/G are summarized in Table 3. The T.P.E. obtained with diets containing all of the different varieties of beans in raw form was very poor and addition of methionine failed to significantly (P > 0.05) increase T.P.E. The cooking treatment for all varieties of beans significantly increased all T.P.E. values and the improvement by addition of methionine to diets containing cooked beans further sig-
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Small Red
1979
FIELD BEANS IN CHICK RATIONS
not significantly improve F/G for diets containing Pinto or White beans. Autoclaving all of the different varieties of beans gave significant improvements in F/G and methionine supplements of diets containing the different beans further significantly improved F/G. Varietal differences were obtained within the group of diets containing heated beans, with White and Red bean diets giving better F/G than the diets with Red or Pink beans. Adding methionine to the diets containing cooked beans improved F/G for all varieties. The results obtained on pancreas weight of chicks fed diets containing the different varieties of beans, either in raw or cooked form, are summarized in Table 4. Diet containing all of the different varieties of beans in raw form caused pancreatic enlargement and this enlargement was significantly reduced by the cooking treatment. The growth response obtained during a 2-week period following a change from diets containing raw beans to a control diet are summarized in Table 5. These results show a marked increase in rate of gain, in-
TABLE 3.—Experiment 1. Effect of bean variety, cooking treatment and dietary supplements on feed conversion and total protein efficiency of chicks Feed conversion1'2
Total protein emc.1.2
Supplement %
Raw Beans
Cooked Beans
Raw Beans
Cooked Beans
Small Red
None 0.35% DL-meth.
7.49^ 6.82 f
2.65 d 2.45 be
0.786 a 0.845 a
2.22 ab 2.35 be
Pinto 114
None 0.35% DL-meth.
5.15b 4.79 a
2.40 ab 2.21 a
1.13 cd 1.19d
2.42 cd 2.58 d
Sutter Pink
None 0.35% DL-meth.
6.27 e 5.48 c
2.62 cd 2.38 ab
0.902 ab 1.01 be
2.16a 2.33 be
California Small White
None 0.35% DL-meth.
5.85 d 5.71 d
2.52 bed 2.23 a
0.998 be 1.00 be
2.32 abc 2.57 d
Beans
Diet 1 Diet 2 1 2 3
Mixed protein control Soybean meal control
1..77 1 .82
Average of three replicates of 12 chicks fed from hatching to 3 weeks of age. Response on raw beans differs significantly from that on cooked beans in every case. Means within each column having different letters are significantly different (P<0.05).
3.25 3.17
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nificantly increased T.P.E. with the exception of the diet containing Small Red beans. Significant varietal differences were obtained in T.P.E. with both the raw and cooked beans. The T.P.E. value for diets containing raw Pinto beans was significantly greater, either with or without added methionine than that for diets containing either Pink or Small Red beans. This difference, however, was not significant from that for the diet containing raw White beans. Similar differences in T.P.E. values were obtained with diets containing the different varieties of cooked beans. Supplementing the diets containing either cooked Pinto or White beans with methionine gave similar T.P.E. values and these values were signifiantly different from those for Small Red and Pink beans. T.P.E. values for both control diets were much greater than any of the values obtained with the diets containing beans. Feed conversion was improved by adding methionine to the diets containing either Small Red or Pink beans. Methionine did
1980
W . D . GOATCHER AND J .
McGlNNIS
significantly greater gains in the absence of supplemental amino acids. Furthermore, gain of chicks in the one treatment fed a Average pancreas weights— mg./g. body wt. 1 diet containing Pinto beans supplemented Diet or bean variety with both the mixtures of amino acids and Raw Cooked beans beans antibiotics significantly increased gain over 2 either supplement alone. These results sug4.8 a Control diet 1 4.8a 8.6 c4 12.7 b 3 Small Red gest that cooked beans exert a depressing 7.1b4 14.6 c3 Pinto 114 4 3 effect on chick growth, possibly through the 7.4 be 12.1b Sutter Pink 7.5 be4 12.2 b 3 California Small White stimulation of an adverse intestinal microflora and that antibiotic supplements stim1 Average of 10 weights. 2 Means within each column having different letulate chick growth by preventing the develters, and means within each row having different opment of such a microflora. supernumerals are significantly different (P<0.05). Diets with cooked Pink and Small White beans supported chick growth when suppledicating that feeding of the raw beans dur- mented with antibiotics that was not signifing the 3-week period from hatching appar- icantly different from that obtained with ently caused no permanent damage. The Control Diet 2. These results are in marked F/G data for this period also show that the contrast to those obtained in Experiment 1 pancreatic enlargement which was evident and those reported by Riley (1961). at 3 weeks of age apparently did not interThe results on F/G and T.P.E. are sumfere with pancreatic enzyme secretion as marized in Table 7. Autoclaving the differthe F/G data compare favorably with previ- ent varieties of beans gave significant imous data with the control diets. provements in F/G as was obtained in ExExperiment 2. The data obtained in Ex- periment 1 and the additional amino acids periment 2 by feeding diets remaining from failed to give results that were significantly Experiment 1, modified as indicated, are better than those obtained in Experiment 1 summarized in Tables 6 and 7. The results with methionine alone. Adding the mixture on chick growth shown in Table 6 indicate of antibiotics significantly improved F/G that the mixture of amino acids failed to for diets containing cooked Pink and White improve growth of chicks fed diets contain- beans. None of the F/G values obtained ing raw beans over the results obtained in with cooked beans supplemented with eiExperiment 1 with methionine alone. In ther amino acids or antibiotics equaled the contrast, the supplement of antibiotics to the F/G values obtained with the control diets diets containing raw beans gave significant (P < 0.0S) increases in gain for all varieties. T A B L E 5.—Response of chicks previously fed diets The Pinto variety fed in raw form gave containing raw beans to a change of diet greater gains, either with the mixture of Av. Av. Av. feed amino acids or the supplement of antibiotRaw bean diet weight gain2 Cons. Feed previously fed1 at 3 wks. 3-5 wks. 3-5 wks. gain ics than other varieties of beans. Even (g.) (g.) (g.) 66.3 261. 8 a' 494.4 1.89 though the supplement of methionine in Small Red Pinto 114 99.5 305.6 b 556.0 1.82 83.0 253.2 a 490.5 1.94 Experiment 1 to diets containing cooked SutterPink California Small White 82.8 293.5 b 573.0 1.95 beans and the addition of the mixture of i The raw bean diets indicated were fed from day of hatch to amino acids in Experiment 2 caused large 3 weeks of age, after which the chicks were switched to the mixed control diet for an additional 2-week period. increases in gain, the addition of the antibi3 Average of 10 chicks. , » Means within the column having different letters are signiotic supplement in this experiment gave ficantly different (P <0.05). T A B L E 4.—Influence of bean variety and cooking of beans on pancreas weight of chicks
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1981
FIELD BEANS IN CHICK RATIONS
TABLE 6.—Experiment 2. Effect of supplementing diets containing different varieties of beans with amino acids or antibiotics and of autoclaving the beans on chick growth Av. gain in 3 wks.fems.)1 J-^JJC U l
beans
Supplement
Small Red
Amino acids 3 Antibiotics4
Pinto 114
Amino acids Antibiotics
Raw beans
Cooked beans 2
30.0 a 167.3 a 61.8 be 220.6 d 56.1 be 190.1 abc 106.6 d 214.1 cd
Amino acids+ antibiotics
—
249.0 e
Sutter Pink Amino acids Antibiotics
38.0 a 80.5 c
184.8 ab 265.7 e
California Amino acids Small White Antibiotics
31.8 ab 78.5 c
205.4 bed 266.1 e
Diet 1 Diet 2
Mixed protein control Soybean meal control
317.8 273.9
1 Means within each column having different letters and all differences between raw and cooked beans are significantly different (P < 0.05). 2 Beans were spread in trays approximately 2.0 cm. deep and autoclaved 10 minutes at 120°C. 3 0.35% DL-methionine, 0.35% DL-threonine, 0.35% L-tyrosine and 0.12% DL-valine. 4 0.1 % procaine penicillin and 0.1% streptomycin sulfate.
TABLE 7.—Experiment 2. Effect of bean variety, cooking treatment and supplements of amino acids and antibiotics on feed conversion and total protein efficiency of chicks Feed conversion1'2 Type of bean
Supplement
%
Raw beans
Total prot. efficiency1'2
Cooked bean's
Raw beans
Cooked beans
Small Red
Amino acids3 Antibiotics4
4.76 d 3.25 be
2.45 c '2.33 be
1.16 a ' 1.81b
2.25 a 2.52 be
Pinto 114
Amino acids Antibiotics Amino acids+antibiotics
4.43 c 3.10 b
2.18 ab 2.08 a 2.16 ab
1.23 a 1.79 b
2.49 be 2.79 d 2.53 be
Sutter Pink
Amino acids Antibiotics
4.40 c 3.11 b
2.30 be 2.15 ab
1.27 a 1.82 b
2.44 b 2.63 cd
California Small White
Amino acids Antibiotics
99 e 82 a
2.17 ab 2.06 a
1.10 a 2.07 c
2.52 be 2.84 d
Diet 1 Diet 2 1
Mixed protein control Soybean meal control
1.77 1.82
Average of 3 replicates of 12 chicks fed from hatching to 3 weeks of age. Response on raw beans differs significantly from that on cooked beans in every case. Mixture of 0.35% DL-methionine, DL-threonine, L-tyrosine and 0.12% DL-valine. 4 0 . 1 % procaine penicillin plus 0 . 1 % streptomycin sulfate. ? Means within each column having,a different letter are significantly different (P<0.05). 2 3
3.25 3.17
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in this experiment. In general, the supplement of antibiotics was more effective in improving F/G than the supplement of amino acids. The T.P.E. values summarized in Table 7 show that all diets containing raw beans gave higher T.P.E. values when antibiotics were added than when the mixture of amino acids was used. The improvements with antibiotic supplements were significant in all cases. T.P.E. values were greatly improved by cooking the beans and again the supplement of antibiotics gave higher T.P.E. values than the supplement of amino acids. The improvements from cooking were significant for all bean varieties and the improvements from the antibiotic supplements were likewise significant. As in Experiment 1, all of the T.P.E. values for diets containing cooked beans and supplemented with either amino acids or antibiotics were better (lower) than the values obtained with the control diets. These differences are closely related to the differences in rates of gain given by the different diets.
1982
W. D. GOATCHER AND J. McGlNNIS DISCUSSION
and to a correcting of a limiting deficiency of total sulfur amino acids or to a combination of these factors. Further amino acid supplementation (methionine plus threonine, valine and tyrosine) did not generally improve bean nutritive value over the improvement obtained with methionine alone. This finding is in contrast to the results obtained with soybean meal where supplementing with these amino acids increased its value (Booth et al, 1960). Wagh et al. (1963), working with chicks, equated the essential amino acid composition of a red kidney bean diet with that of a control ration and failed to obtain equal growth of chicks fed the two diets. They suggested that the failure may have been due to such factors as a thermostable toxin in beans, poor availability of other amino acids, or a dietary energy deficit. Antibiotic supplementation of bean diets resulted in about a 200% increase in gain when the beans were raw and about a 56% increase with autoclaved beans. Other studies in this laboratory have shown that large antibiotic responses are also obtained with diets containing either dry peas or lentils (Goatcher and McGinnis, 1972) and with diets containing rye (MacAuliffe and McGinnis, 1971). The chick growth response which is usually obtained with antibiotic supplements is about 5-10% (Matterson and Singsen, 1951; Heuser and Norris, 1952). The addition of an antibiotic supplement to the mixed protein control diet 1 used in these studies produced data showing that antibiotic supplementation only gave a 6% improvement in growth. Thus, it appears that the antibiotic growth response is markedly influenced by ration composition. The data of Kakade and Evans (1964, 1966) indicate that antibiotics are less effective in promoting growth of rats fed diets containing beans than they are in pro-
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Raw beans of all varieties supported poor chick growth. This may have been due to generally poor digestibility of protein and/or energy or due to the presence of toxic factors or some combination of these. Riley (1961) observed an increase in metabolizable energy of cull field beans from 449 to 877 Kcal./kg. when these beans were autoclaved and included in chick diets at a level of 40%. Autoclaving the beans resulted in an increase in 3-week gain of about 500% and an increase in T.P.E. values and feed consumption of about 150%. However, performance was still below that of chicks fed control diets. The increase in nutritive value of beans with autoclaving is probably due to increases in digestibility as well as inactivation of some toxic factors known to be present in many legumes. Liener (1953) demonstrated that soybean hemagglutinin was growth-inhibitory to rats. Honavar et al. (1962) showed that Phaseolus vulgaris hemagglutinin depressed chick growth. Trypsin inhibitors do not appear to account completely for the poor chick growth obtained with Phaseolus vulgaris (Riley, 1961) or Glycine max (Saxena, 1964). The data indicate that the varieties of beans studied differed in nutritive value. Generally speaking, the White and Pink beans were of higher nutritional quality than the Pinto and Red beans. Kakade and Evans (1965), using rats as the experimental animal, did not detect a difference in nutritional value of 5 varieties of navy beans. Methionine supplementation of heated bean diets improved performance characteristics. Supplementing the raw bean diets with methionine failed to improve bean nutritive value. The response to methionine additions to diets containing heated beans could be related to decreased growth depression by heat labile growth inhibitors
1983
FIELD BEANS IN CHICK RATIONS
moting chick growth. In their work, however, dietary bean level was about 41.6% whereas in the present study it was 53.6%.
1. 2. 3. 4. 5. 6.
7. 8. °. 10. 11.
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REFERENCES Booth, A. N., D. J. Robbins, W. E. Ribelin and F. DeEds, 1960. Effect of raw soybean meal and amino acids on pancreatic hypertrophy in rats. Proc. Soc. Exp. Biol. Med. 104: 681-683. Goatcher, W. D., and J. McGinnis, 1972. Influence of beans, peas and lentils as a dietary component and growth response of chicks to antibiotics and methionine supplementation of the diet. Poultry Sci. 51: 440-443. Heuser, G. F., and L. C. Norris, 1952. Some results of feeding antibiotics to chickens. Poultry Sci. 3 1 : 857-862. Honavar, P. M., C. Shih and I. E. Liener, 1962. Inhibition of the growth of rats by purified hemagglutinin fractions isolated from Phaseolus vulgaris. J. Nutrition, 77: 109-114. Kakade, M. L., and R. J. Evans, 1964. Effect of methionine, vitamin B12 and antibiotics supplementation on protein nutritive value of navy beans. Proc. Soc. Exp. Biol. Med. 115: 890-893. Kakade, M. L., and R. J. Evans, 1965. Nutritive value of different varieties of navy beans. Michigan State Univ. Agr. Expt. Sta. Quart. Bui. 48: 89-93.
Kakade, M. L., and R. J. Evans, 1966. Growth inhibition of rats fed raw navy beans (Phaseolus vulgaris). J. Nutrition, 90: 191-198. Liener, I. E., 1953. Soyin, a toxic protein from the soybean. I. Inhibition of rat growth. J. Nutrition, 49: 527-539. MacAuliffe, T., and J. McGinnis, 1971. Effect of antibiotic supplements to diets containing rye on chick growth. Poultry Sci. 50: 1130-1134. Matterson, L. D., and E. P. Singsen, 1951. A comparison of several antibiotics as growth stimulants in practical chick-starting rations. Storrs (Connecticut) Agr. Exp. Sta. Bui. 275. Riley, D. J., 1961. Studies on the nutritional value of some little-used protein feedstuffs for poultry. M.S. Thesis, Washington State University, Pullman, WA. Saxena, H. C , 1964. Investigations on the mechanism of growth depression and pancreatic hypertrophy by raw soybean meal in the chick. Ph.D. Thesis, Washington State University, Pullman, WA. Wagh, P. V., D. F. Klaustermeier, P. E. Waibel and I. E. Liener, 1963. Nutritive value of red kidney beans (Phaseolus vulgaris) for chicks. J. Nutrition, 80: 191-195. Woodham, A. A., 1968. A chick growth test for the evaluation of protein quality in cereal-based diets. 1. Development of the method. British Poultry Sci. 9 : 53-63.