BIOI.OOICAL EVAWATION OF THE NUTRITIONAL VAUJE

OF THE PRCYrEINS OF HAITIAN CEREAL-LEGUME BLENDS

by

Koaol Sirinit

Thesis subnitted to Graduate Faculty ot

Virginia Polytechnic Institute

in candidacy for the degree ot

Master of Science

in

Biochemistry

August 1964

Blacksburg, Virginia.

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TABLE OF CONTENTS

List of Tables • • • • • • • • • • • • • • • • • •

List of Figures • • • • • • • • • • • • • • • • •

I. Introduction and Literature Review • • • • • • • • • • •

Page

4

6

7

II. Materials and Methods • • • • • • • • • • • • • • • • • 11

11 a. Processing of Haitian foods • • • • • • • • •

b. Chemical analyses of HaitiAn foods • • • • • • 1. Proximate 81111.lyses • • • • • • • • • • • •

2. Amino Acid analysis • • • • • • • • • • •

11

11

11

c. Preliminary test for Protein Quality of Beans. 11

1. The Effect of Diet&ry Protein Level on PER or Rats .• &'. . . . . . . ..... • • • •

2. Identity of Limiting Amino Acids • • • • •

11

13 J. Ewluation of Prote!h Value of Haitian Beans 14

4. Riboflavin Supplementation of Bean diets • 15

d. Protein Quality Teat of Cereal-Legume Blends • 16

1. Protein Quality Test of Cereal-Legume Blends 16

2. Protein Quality Test of Cerea.1-Legwne Blendis supplemented w.1.th the Limiting Amino Acids 17

III. Results

1. Protein Efficiency Ratio and Weight gA.ins of Rats at the OI_>timum Protein Level (Ta.bles 3,4,5,6 and Figure 1) • • • • • • • . • • • • • • • • • • • 19

2. Composition ot Haitian Foods (Tables 1 and 2). 2l

J. Weight Gain, Feed. Efficiency Md PER after Supple-menting Beans w.1.th Amino Acids for 10-dAy Test Periods (Table 7 and Figure 2) • • • • • • • • 28

IV•

v. VI.

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4. Protein value ot Haitian Beans predicted :rrom--the values of Prote:1n Digestibility, Biologic.tl V&lue, Net Protein Ratio (NPR), Protein Retention Efficiency (PRE), Net Protein Utilization (NPU) and Protein Efficiency Ratio (PER) or rats

Page

(TablEfi 8 and 9 ) • • • • • • • • • • • • • • • • • • • 32

5. PER a.nd Weight Gain of Rats fed Bean diets with Ribotla'fin Supplementation for 4-Week Test Periods (Tables 10, 11 a.nd Figure 3). • • • • • • • • • • • • 35

6. Weight Gain and PER · ot Rate fed Cereal-Legume Blends (Tables 12 and 13, !Uld Figures 4 and 5) • • • • • • • 39

7. - ~ ~"

PER and Weight Gain of Rats fed Cereal-Le~e Blends with 11.miting Amino Acid Supplementation {Tables 14, 15,16,17 ,18,19, 20 and 21, 8.Dd Figurei 6 Md 7 ) ••••

DISCUSSION • • • • • • • •••• • • • • • • • • • • • • • • •

SUMMARY ••• • • • • • • • • • •••••• • • • • • • • •••

• • • • • • • • • • • • • • • • • • ••• • •

VII. BIBLIOORAPHY • • • • • • • • • • • • • • • • • • • • • • • • •

4l:i 63

66 67 6S

71 VIII. vrrA •••• • • • • • • • • • • • • • • • • • • • • • • • • •

Table No.

1

2

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Proximate Analyses of Haitian Foods • • • • • • • • • •

Amino Acid Composition of Haitian Beane !Uld Cereals Compared to Whole Egg Protein • • • • • • • • • • •

3 Effect of Dietary Protein Level on PER of Haitian

4

5

6

7

Foods • • • • • • • • • • • • • • • • • • • • • • •

Effect of Dietary Protein Level on Feed Efficiency of Haitian Beans • • • • • • • • • • • • • • • • •

Effect of Dietary Protein Level on Weight Gain. • • • •

Summarized Results of PER, Feed Efficiency Md Weight Gain for Haitian BeMs at Optimum Protein Level • •

Effect on Weight GA.in, Feed Efficiency and PER of Supplementing Beans with Amino Acids. • • • • • • •

8 Measurerr.ent of Nutritive Value of Protein in Haitian

Page

2J.

22

23

25

26

JO

Beans • • • • • • • • • • • • • • • • • • • • • • • 33

9

10

11

12

13

14

15

16

Comparison of PER, Biologic!il Value and Digestibility of Beans (Phaseolus vulgaria) for rats •••••••

Effect on Weight Gain on Bean Diets with Riboflavin Supplementation for 4-Week Test Periods • • • •

Effect on PER for Rats on Be~m Diet with Riboflavin Supplementation for 4-Week Test Periods • • • •

• •

• • Combinations of Cereal-Legume Blends and Protein Con-

tributions of Ingredients • • • • • • • • • • • ••

Effect of Blend Composition on Weight Ge.in and PER value of Rats fed Cereal-Legume Blends for 2-Week Test Periods. • • • • • • • • • • • • • • • • • • •

The Best Combirm.tions of Cereal-Legume Blends • • • • •

Amino Acid Content in Cerea.1-Legwne 1iets Compired to the Requirement for Growing Rats ••••••••••

Percent LimitL"lg Amino Acids in Cereal-Legume Diets ~s Percent of the Requirement for Growing Rats • • • •

34

J7

40

43

49

50

52

17

18

19

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Amount of Amino Acid added to the Cereal-Legume Diets •

Weight Git.in ~nd PER V'J,'.lue of RA ts fed Cereal-Legume Blends with Amino Acid Supplementation for 2-Week Test Periods ~ • • • • • • • • • • • • • • • • • • •

Percent of the Limiting Amino Acids in Cereal-Legume Blends as Percent ·of the Requirement r or Inf ante • •

Page

53

55

59

20 Comparison of Protein Efficiency I\Atio of Different

21

Sources of Protein with Cereal-Legume Blends lilt. Optimum Protein Levels • • • • • • • • • • • • • • • • • • • 60

Composition of Haitian Blends of good protein qWtlity expressed in tenns of 100 g of blends. • • • • • • • 61

Figure No.

1

2

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LIST OF FIGURES

Etf ect of dietary protein level on PER of Casein !llld Haitian beans (raw PER value) • • • • •• • •• •

Effect of Rm1no Acid supplementation on PER or bean diets • • • • • • • • • • • • • • • • • • • • • • •

Growth or rats as affected by ribonAvin supplementation

Page

31

of bean diets • • • • • • • • • • • • • • • • • • • JS

4 Protein quality test of ceresl-legume blends with vsrying BmOunts of bean showing weight gain tor 2-week test periods • • • • • • • • • • • • • • • • • • • • • • 44

5 Protein quality test of cereal-legume blends with ~rying BmOunts of beAns showing PER value for 2-week test periods • • • • • • • • • • • • • • • • • • • • • • 45

6 Effect ot Amino acid supplementation on PER or rats fed cereal-legtmle blends • • • • • • • • • • • • • • • 57

7 Effect or ~~ino Acid supplementation al weight gain or l"l\ts fed cereal-legume blends • • • • • • • • • • • 58

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I. INTRODUCTID?I AND LlTERATURE REVJ»i

To approach the problem ot meeting the protein requirement by ... - . . -

improving ~he biological value or low quality ~te.17 protein, 8!""ral attempts haft been made by means ot supplementation with essential amino - .

acids. This suggestion is not capable ot implementation on a large -· . ..

scale in the realm ot hlml8ll nutrition. It ia tar more practical to

attempt. to impron the nutritive value ot protein by incorporation ot

other tood stutts into the diet which will supply the deticient amino

acid. In order to use such a tood tor suppl~ting human di~ts, the __ WHO

Protein Advieo1'7 Group (1955 ) has suggested that it ia necess81')" to take

into consideration the following !&ct.ors: (1) the amino acid contri-

but.1.ona ot the indiT.l.dual ingredients and or the final product, (2) the

possible presence or toxic or interfering .tactore, (3) the need tor ob-. . . . -- ..

taining exact specitieations tor each or the components, (4) the necessity - . .. . ..

ot awiding procesa~s that damage the qu&lit!. of protein, (5) the deaira-

bllit1 or using local products, (6) the suitability of the product tor .... -· -- -· feeding waned infants, and (7) the acce~bility ot the product to the

con8\lllere.

The objective of the preeent .eisearch coneiata or detailed

biological evaluation of the nutriti.ansl n.lue ot the proteins ot the

cereal-legume blends formulated from those indigerioua foods ot Haiti

predicted by the chemical data to be moat promieing.

Legumes are potentially an important source or proteins tor ... -·

the people 1n the merging n&tions. Beane proTi.de 20-3Qt ot the pro-

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tein in nvtny such diets. Beans (Phaseolus wlgaris) rAve been ad-

vocated for wide use in the treatment !ind prevention of protein

mAlnutrition (F.A.o. 1956 Nutr. Stud. No. 14). The use of beans has been limited by (1) the presence of

toxic compounds such as trypsin inhibitors (McColl.um ~ al, 1917,

Johns and Finks, 1920 e.nd Liener, 1962) which appear to be the m&jor

cause of the poor utilization of the protein in nw 80'1 bean (Westfall

and Hauge, 1948) e.nd the depression in the growth of re.ts (Klose.~!!.!,

1946, Borchers ~ .!11 1948 and Liener ~ .!J:, 1949) 1md chicles (Westfall

and Hauge, 1948 and Ham~ !l, 1945 ). In general, there is also a lack

or careful assessment of their real nutritive value ptrticularly in

blends with cereals.

Simple means of detoxication of edible beans (Liener, 1962

and Bressani !! ,!!1, 1963) have been etudied, and the nutritional value

of one variety or Phaaeolus vulgaris has been exiunined in some de-

tail by Bressani .!!!: &, (196.3 ). Beans are cooked in order to IIU\ke them

palatable (Eheart and Sholes, 1948) and to increase their digestibility

(Jarri, 1950) as me&sured by' .!n .!!!!? and _Y:! vitro studies (Watenna.n

and Johns, 1921). This improTement in nutritive value is presumably

due 8Jl10ng other things to the destruction of trypsin inhibitors. Proper

cooking increases both weight gain !!lnd Protein Efficiency Ratio in rats

(BresSAni ~ !!,!, 1963 ).

Most of the common vegetable proteins are deficient in me-

thionine and/or lysine. Fortification of food with methionine or lysine

results in ln8rked improvement in the Protein Efficiency Ratio ot these

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proteins. Such legume proteins, when they are fortified with methionine,

show increases in Protein Efficiency Ratio to a marked extent ( 50-lCXlt

according to Richardson (1948),as well as the growth promoting value

of bean diets (Bressani ~ !1, 1963). In these latter studies, further

supplement&tion with other amino acids which are the most limiting

runino acids next to methionine showed little effect.

One prsctical method of providing a low-cost nutritiorullly

balanced protein supplement Msed on cereal-legume blends utilizing

the available indigenous foods consists, for example, of com which

is deficient in lysine and tryptophM (Sauberllch ~ !J:, 1953 and

Bressani ~ ,!!, 1958) supplemented with beans which contain a consider-

able amomt of methionine, lysine &nd tryptophan (FAO, 1953). These two

staple foods should complement each other efficiently. It has been re-

ported that on blending beans with rice or corn a synergistic effect

was obtained in the apparent nutritive nlue or the mixed protein

(Breesani ~ !,!, 1962 a, b ). The available data including the dat11

of the present study suggests that in lIVU1Y ruitions considere.ble public

health progress might be made by developing cereal-legume blends cap-

italizing on this type or nutritional synergism.

This report deals primarily with a program for improving the

nutritional status of the population ot the Republic of Haiti. Mal-

nutrition is generally recognized to be one of the most important health

problems in this llrea (Jellitfe and Jelliffe, 1960 and Sebrell ~ !!, 1959). Important advances can therefore be expected by improvement of

the nutritional status associated with medic~l improvement, education

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and economic developnent. (Jelli!fe and Jellif!e, 1961).

In order to underat&nd the Haiti&n situation &s a whole,

specific into:nnation can be obtained from the following references:

1. Jelllffe, D.B. and E.F. Patricia Jellitte. 1961. The Nutritional Status or Haitian Children. Acta Tropica ];§, 1.

2. King, K.w., Foucauld, J ., Fougere, w., and Severingha.us, E.L. 196.3. Height and Weight or Haitian Children. Am. J • Clin. Nutrition .!J, 106.

J. King, K.w., Sebrell, W.H., Severinghaus, E.L. And Storrlck, w.o., 196.3. Lysine Fortification of Wheat Bread Fed to Haitian School Children. Am. J. Clin. Nutrition ~' 36.

4. Sebrell, W.H., Jr., Smith, s.c., Severinghaus, E.L., Delva, H., Reid, B.L., Olcott, H.s., Beniadotte, J., Fougere, w., Barron, G.P., Nicolas, G., King, K.w., Brinkman, G.L., and French, C.E. 1959. Appraisal of Nutritinn in Haiti. Am. J. Clin. Nutrition 1, 1.

5. Jellitre, D.B. and E.F. Patricia. Jellltte. 1960. Pre-valence of protein-.ca.lorie malnutrition in Haitian pre-school children. Am. J. of Public HeRlth. 2.Q,1355.

6. Grant, F.w. and Groom, D., 1958. A diet&ry study in Haiti. J. Amer. Diet. Aeaoc. J!t, 708.

7. Olcott, Harold s. 1959. Food Technology in H&iti. Food TechJaol. ,2, 10.

8. Holly, M.A. 1955. Agriculture in Haiti. Vantage Presa, Inc., New York.

9. George, D.A. 1959. Educ&tion in Republic or H&iti. Bull. No. 20. U.S. Department of Health, Education and Welf&.re. United States Governnent Printing Office, Washington.

10. F.A.o. 1958 • Report No. 721, Rome.

11. Ruth, w.o. 1957. Here is Haiti. Phlloaophic&l Library, New York.

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II. MATERIALS AND METHODS

A. Procesaing of Haitian Food• . ..

Beans and ce~als used in all ot t~ experiments were re-

ceived from variou1 localities 1n Haiti and ha.Te been described in de-

W.l prerloualy (King, 1964). .. . -~

Betna: To ea~h kilogram ot be&ns W_I! added two liters ~r

water and the mixtura was cooked in the autoclave tor 1.5 minutes at 12J.0c., . --

then the beana were dried at scPc in a forced-draft oven tor 18 hours.

They wre ground in a 40-mesh Wiley Mill and stored in a retrigerator

rocm until used.

Cereals: These were ground in a 40-meah Wiley Mill &nd stored

in a refrigerator rocn until use4.

B. Chemical Analyses

l. Proximate Anal.188a

Proximate a.nalyaea were m&de on a dried bssis tor crwle . . . --protein by the Kjeldahl method and for a.ah, moisture, ether extract and

crude fiber. (A.O.A.c., 1960 a,b,c,d,and Whitehouse,!!:.!;!, 1945).

2. Amino Acid Analysis

The Amino Acid comp:>sition of Haitian foods vas ob-

tained from the data of King (1964).

C. Prel.iminarr Teat for Protein Quality of Beana

l. The Effect of Dietary Protein LeTel on Protein Ettioienc:y

Ratio (PER) of rats

Experimental

An initial test for protein quality of beans 1""8 conducted to

detennine the optimuxn diet1U7 level of be8ll protein by meaBUring the .. .. . ..

protein etticieney ratio (PER), teed e.ttioieno;y and weight gain of rats.

Diets; ~re formulated to __ contain 10,12,15,18 and 2~ protein

(N x 6.25 ). The rat, moisture, ash and tiber contents or the n.rious

dieta were equalized, 1rid'lere poasible at S,6.2, 5.0, and 6.o, rea- .':-

pectively, by the addit~ of appropriate amounts, ~aed upton the prox-

imate compoeition of U8 sample, of cottoneeed oil, water, mineral mix-

ture and cellulose (A.O.A.c., 1960 e ). All diets were supplemented with

aalt mixture USP XVI (A.O.A.c., 1960 t) and vitamin mixture l g per 100

g of diet (Ca-panthothenate, 2.0 g; niacin, 1.0 g; inositol, l.O gs thiamine HCl, 0.5 g; menadione, 0.4 g; ribo!lATin 0.3 g; pyridoxine

HCl, 0.3 g; tolic acid, 0.02 g; biotin, 0.01 g; Vitamin B12 , O.OOl g;

and sucrose tanmakB 1000 g of the mixture). To each kilogram of di•t,

5 ml or 20' choline chloride solution was added. Two drops ot Vitamins

A, D and E (1qt; of vitamin A and D a.s Percomorph oil with lo,( of •' m o r

vitamin E acet&te &t the expe11ae or cottonseed oil) llere given to m• ••

each rat wee~. Rats of all the experiments wre kept, on stock diet

(8 g cottonseed oil, 5 g a&lt mixture USP XVI, 2.4 g solkafloc, l g . .. . -.. vitamin mixture, 5 g water, 15 g vitamin-tree casein and 6J.6 g ot

sucrose) tor 7 days before .feedirg a test diet.

Rats: The rate used were males of the Sprague-Dawle1 strain

lfeighing 40 ! 5 g. At the beginning s.nd the end of the experimentu - .

period, the animals were starved for 24 hours to clear their intestinal . --

tract a ot !ood. Then the animals were divided into 20 groups of 6 each

on a basis such that the :individual and average -weights o! each group

- 13 -

were as similar as possible. The mean initial weight ot the animals

was between 58 Md 67 g and the mean weight among the groupe was

62 :!: 1 g. The animals were kept in individual wire-screen cages. Food

;tnd 'W8ter were given fil! libitum. All experiments lasted 10 days; gains

in weight and food intake were recorded e.t the end of the experimental

period a rter a 2J+ hour fast.

PKRs ( g gain/ g protein intake ) were calculated in addition to

feed efficienc7 (g gain/g food intake). The PER values obtained are

given in Table 3, Feed Efficiencies in Table 4 and weight gains in Table

5. Summarized results a.re seen in Table 6 and growth cUrYea in Figure 1.

2. Identity of Limiting Amino Acids

Experimental

This experiment vas conducted to evaluate the magnitude of

the eftect of amino acid supplementation of the Haitian beans at

optimum protein level aa determined from the previous experiment by

measuring PER and weight gairul.

Diets: These were tonnula.ted in e. similar wa:r to those or the

previous experiments, to contain l~ protein level for pois rouge and

pois noir and 15% for pois buerre e.nd pois cmigo diets. The diets wre

supplemented with methionine,itoleucine, valine, leucine or tryptophan

aa seen in Table 7. These amino acids were indicated from aruU.y'tical

data to be most likely to be limiting.

~: The rats used 1i1ere males of the Sprague-Dawley strain

weighing between 51 and 70 g, and the mean weight among the groups waa

61 ± l g. The rats were divided into 19 groups of 6 rats per group.

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The experimental procedure was simil&r to the prerloua one. The

experimental period lasted 10 days. Weight gains were recoroed and

PER vw.lues wre c&lculated. The results are seen in Table 7.

J. Eftl.uation ot Protein Value ot Haitian Beans

Experimental

Thia experiment va.s co~ducted to evaluate the protein vw.lue

ot the more promising Haitian beans by determining the protein digest-

ibili ty, biological nlue, net protein r&tio (NPR), protein retention

efficiency (PRE), net protein utilization (NPU) and protein efficieney

rstio (PER) for rats (Mitchell, 1924). Diets; were formulated as described in the previous experi-

ment, tor pois rouge, pois noir and pois buerre. For the pois congo

diet, and the mixture or pois rouge and pois congo diets were tor--

mulated to contain 1S% protein. The pois rouge and pois congo diet

had a ratio 50:50 by weight And the protein rstio was 7.74:7.'Jh (%).

For the protein-tree diet the composition was the same as the others

but protein va.s replaced by sucrose.

~: Male ~ts of the Spr&gu~Da:wlq strain were di Tided

into 7 grQUps with 6 rate per group. The initial weight or the

animals was between 61 and 70 g, and the mean weight ot each group

was 64.0 g.

Food and water were given J!S libitum. The food wss changed

daily, and the intake was recoroed. The animal' a weights were recorded

at the beginning and at the end ot the 7-day experiment.Al period. Feces

were collected daily and frozen. Urine was collected in HCl under toluene.

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The teces and urine were analyzed tor tot&l nitrogen. The w.lue

of protein digestibility, biological v8lue, NPR, PRE, NPU, and

PER obtained a.re seen in Table 8.

4. Ribofiavin Supplementation ot Bean Diets

Experimental

This experiment was conducted in order to see whether the

tull level of riboflavin ( o. 3 mg/100 g ot diet) or halt that level

( 0.15 mg/100 g of diet) of ribofiavin supplementation would affect the

growth of rats fed tean diets as sole source of riboflavin. Bec~uae

riboflavin deficiency is the most frequently encountered B-vitamin de-

ficiency in Haiti by growth response it was important to know 'Whether

the cooked beAns vould contribute significantly to ribofiavin nutriture

in addition to improving protein intake.

Diets: Ten groups of diets without ribofiavin were formulated

to contain 1$ protein, for pois rouge an:l pois noir diets and 15'% pro-

tein which was optimum for pois buerre. From each bean diet sublots

were prepR.red into which three levels of riboflavin wre then incorporated,

(the normal level, 5Qt ot the normal level and no ribofi~vin ). The e.x-

periJnent lasted 4 weeks. The groups with 5<$ of' the nonnal riboflavin

supplementation were carried for 2 weeks. At the end of the 3rd week,

the .full level of riboflavin was added to the riboflavin-free diets and

feeding wsa continued tor one more week.

Rats: Male rats of the Sprague-Dawley strRin were divided

into 10 groups with 6 rats per group. The initial weight of the 4 ~ - ' ~

animals was between 55 and 65 g and the mean weight or each group 'tmS

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+ 5S - 1 g.

Food &nd water were given !,S! libitum, Food was changed

daily. The animal's weights Md food intake were recorded weekly.

PERs (g gain/g protein intake) were c&lculAted. The reeults ot weight

gains and PER value are seen. in T&ble 10 a.nd 11 and growth cunea in

Figure 3.

5. Protein Quality Test of Cereal-Legume Blends

Experimental

This experiment was conducted to detezmi.ne the weight gain

and PER tor rats on several cereal-legume blends in order to find out

which combination gave better growth.

Diets: Fourty diets were formulAted to contain 8,l( protein

for com-bean diets, 6.4% protein for rice-bean diets Md lOC protein

for the vitamin-free casein diet. The combinations of the cereal-

legume diets and the ratios in terme of % ot the protein contributed

by cereals and by legumes are seen in Table 12. Ea.ch diet aeries in-

cluded the be.sic cereal alone and supplemented with 10,20, or )Qt ot

each bean tested.

~: Ma.le rats or Sprague-Dawley strain were divided into

40 groups with 6 nts per group. The initiAl weight or the anim&ls

was between 52 a.nd 7S g and the mean wight of each group was 65 :! 1 g,

Food and water were given !,S! libitum. Food was chAnged daily.

The animal's weights and food intake were recorded weekly. The ex-

periment lasted 2 weeks. PER's (g gain/g protein intake) were c&l-

culAted. PER values and weight gaine &re seen in Table 13. Growth

- 17 -

and PER curves appear in Figures 4 and 5.

6. Protein Quality Test or Cereal-Legume Blends Supplemented with the Limiting Amino Acids

Experimental

This experiment wss conducted to determine the PER md the

growth response of rats fed cereal-legume diets o! the more promising

combinations Rs determined from the earlier experiments when supple-

mented with the limiting amino acids as indicated by calculations using

the data on amino acid composition or Haitian foods by King (1964)

A.ssoeiated with the data on the requirement for growing rats by Rama

Rao .21 !l (1964). The MJino acids l\re added 1n order of the most to

the least limiting.

Diets: Thirty-five diets of the better combinations wre

formulated to contain 10' protein for corn-bean and sorghum-bean diets

and 9% protein tor rice-bean diets. The combinations or cereal-legume

diets and the percentage of protein contributed by cereals and legumes

are seen in Table U.. These diets were supplemented with single or

with combinations of amino acids which were indicated by calculation to

be the limiting amino acids as lieted :in Ta.b1e 16. The amotm.t of amino

acids present in the diets and the smount added to the diets are seen

in Table• 15 and 17. Supplementation was made with chromatographical.ly

pure, al.lo-tree, L-amino acids in amounts to bring the total level to

10~ ot the growing rat's requirement.

Rats: Male rats of Sprague-Dawley strain were divided into

36 groups with 6 rats per group. The initial weight of the animals

- 18 -

+ was between 52 and 78 g. The mean wight or each group was 66 - 1 g.

The experimental procedures were the same ss deecribed pre-

viously. The experiment lasted 2 weeks.

PERs were ctlcul.ated. PER values and weight gains are seen

in Table 18. Growth and PER curves sppe&r in Figures 6 and 7.

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III. RESULTS

Experiment 1

Effect of Dietary Protein Level on PER or R&ts

Table 1 shows the composition ot the Haitian .foods obtained

!'ran the proximate analysis. The figures or tl"Ue protein represent the

sum or amino acid plus NH • 3

Table 2 shows the amino acid composition or Haitian foods

compBred to whole egg protein. The essential amino acid indices are

70.8, 69.5, and 72.<Jf> for pois rouge, pois noir and pois 'Werre and

58.J, 56.0 and 68.Ji .for com, sorghum and rice respectively. The most

limiting &mi.no acids are methionine, for beans, tJ7Pt.ophan, for com,

lysine, for sorghum and threonine, for rice. The chemical scores are

20' for pois rouge and pois buerre 1~ tor pois noir, 'JJ.d, for com,

21% tor sorghum and 47% for rice. (King, 1964 ).

Table 3 shows the effect of dietary protein level on PER of

Haitian beans for a 10-day experimental period. Optimum protein level

judged b;y the ms.ximt.nn PER was obtained at l~ protein for pois rouge And

pois no1r and 15% for pois buerre, the PER& being 1.16, 1.17 and 1.34

respectively.

Table 4 shows the effect of dietary protein level on feed

efficiency. The three beans were very similar, maximum values being

o.25, o.26 and 0.25 for p:>is rouge, pois noir and pois buerre res-

pectively.

Table 5 shows the effect of dietary protein level on weight

gain of rats fed Haitian beans as sole source of protein. Maximum

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values were found to be 33, 32 and Jl g per 10 days tor pois rouge,

pois noir snd pois buerre respectively.

Table 6 showe the summarized results of PER, teed efficiency

and weight gain for Haitian beans a.t opt.imwn protein level.

Figure 1 show the eftect ot dietsry protein level on PER or casein (fortified with o.2' eystine) and Haitian be&nS. The PER nlues

given in this figure are nw PER vslues (not adjusted to PER for the

standard casein • 2.50).

Table 1 Proximate analysis of Haiti.an Food.a (dey basis)

Expressed as Percentage

True Crude Ash Moisture Ether Food protein2 protein Ex:t.ract

Beans pois rouge(Phaseolus !Ulga.ris) 20.2 23.2 3.4 6.7 1.6

pois noir~ wl.garis) 21.7 25.5 3.5 6.0 2.7

pois bl&nc (.f!. ~ria) 21.6 2.4.3 3.5 5.1 1.0

pois buerre{.f.t. yulgaris) 20.9 a.a 3.5 6.2 2.5

pois congo( Ca.janus indicus) 20.2 21.9 2.8 5.9 2.5

Com (~ mAYS) 9.5 ll.1 1.2 ll.9 5.3

Sorghum (Sorghum yaj..gare) 10.4 10.6 o.6 12.0 1.3

Rice (Qriza. sati va) 7.9 8.1 0.3 11.2 0.7

Crude Nitrogen Fiber free Ex- Total

tmse!i1

6.8 58.3 100

5.6 56.7 100

7.7 5s.4 100

6.) 56.7 100

a.1 58.8 100

1.8 68.7 100

0.2 75.3 100

0.3 79.4 100

1 Nitrogen tree extra.ct obtained by subtracting the crude protein, ash, moisture, ether extract, and crude fiber percentages trom one hundred.

2 True protein = sum of Amino Acids plus NH3 , Figure obtained trom King, K.w. (1964) Amino Acid Composition of Haiti:m Foods. Ecananic Botany (in press).

~

Table 2 Amino Acid Compositicnof Haitian beans and cereals compared to whole egg protein

Expressed ae g. Mlino acid/100 g. crude protein

Beans Cereals Amino Acid pois rouge poi.a noir pois buerre Com Sorghum Rice ' Phaseolua Pha.aeolus {Pha.seolus (Zea (Sor~ (Oriza Whole Egg

vul~ria) vulgaria wlgtiriO) umys) vul&!re) §!!tin)

lg sine 8.06 8.24 8.(J7 2.99 1.51 3.76 7.9 Histidine 2.92 2.92 2.97 3.12 2.02 2.20 2.7 Anmonia 1.87 1.93 1.87 2.37 2.92 2.28 Arginine 4.76 5.36 5.15 3.45 2.m. 5.58 6.7 Teyptophan 1.62 1.58 1.62 0.52 1.25 1.47 1.42 Aspartic acid ll.42 11.41 11.97 5.69 5.75 8.44 10.5 Threonine 3.61 3.37 J.65 2.56 2.39 2.28 4.5 Serine 5.20 5.12 5.19 3.46 J.00 2.95 6.1 ~ Glut~mic acid 14.16 13.77 14.30 17.13 21.71 15.02 13.2 Pro line 3.44 3.50 3.63 8.26 7.85 J.57 4.1 Glycine J.60 J.42 3.65 3.00 2.21.. 3.46 2.J Al~nine 4.02 3.79 4.02 7.17 9.25 4.89 6.9 Cystine 0.79 o.66 0.74 1.78 2.Ci> 2.19 2.42 V~line 4.36 4.05 4.35 3.91 4.36 5.'Zl 7.1 Methionine 0.53 0.52 0.58 l.JO o.86 1.72 3.0 lsoleucine J.74 3.69 J.85 2.99 J.60 3.58 6.o Leucine 7 .Cf) 6.97 7.21 10.92 13.16 6.72 9.1 Tyrosine 2.61 2.54 2.78 2.25 1.98 2.20 3.1 Phen:yl~lanine 4.95 4.87 5.04 4.20 4.30 4.53 5.8

TGrAL 80.65 87.71 90.66 87 .(JJ 92.22 82.lJ 102.9

Beans and cereals: King, K.w. (1964) Amino Acid composition or Haitian foods. Economic Botany (in press) 'Lepore, P.D.(1962) Ph.D. Thesis, Virginia Po]Jt.echnic Institute, Blacksburg,

Virginia 2 Block,R.J. end Bolling, D. (1956) Amino Acid Handbook. Springfield, Ill.

c.c. ThorMe Publisher

- 23 -

Table 3

Effect of dietaey protein level on PER of Haitian beans

Diet

pois rouge

pois noir

pois buerre

casein (fortified with o.~ cystine)

10

1 0 69 2 ( • . 0.84)

1.00 (1.22)

0.90 (1.09)

(4.19)

Protein level (% )

12 15

0.83 1.14 (1.01) (l.JS)

o.83 1.06 (1.00) (1.28)

0.96 1 • .34 (1.16) (l.6J)

(3.64) (2.97)

18 20

1.16 0.92 (1.41) (1.12)

1.17 1.09 (1.42) (1.32)

0.98 1.02 (1.19) (1.24)

(2.54) (2.20)

1The PER values obtained were corrected by a factor such th.qt the value for the casein st~mdArd is 2. 50, correct sample PER • 2. 50 x sAlllple PER

2Raw PER values are in parenthesis Re.t"erence casein PER

- 21+ -

Table 4

Effect of dietary protein level on Feed Efficiency1or Haiti~n BeMis

Diet Protein level (% )

10 12 15 18 20

pois rouge o.oa 0.12 0.21 0.25 0.23

pois noir 0.13 0.12 0.19 o.26 o.zr pois buerre o.n 0.14 0.25 0.21 0.25

casein 0.42 0.44 0.45 0.46 0.44 (fortified with

o.Z( cystine

1 gram gam/ gram food ea ten ·

- 25 -

Table 5

Effect of dietary protein level on Weight gain (g/lOdays)

Diet Protein level (% )

10 12 15 18 20

pois rouge 8 14 a 33 29

pois noir 13 12 24 32 33 pois buerre ll 14 31 Zl 32

casein 63 66 67 67 6.3 (fortified with

o. zt; cystine )

,

- 26 -

Table 6

SU!!IMrized results ot PER, Feed Efficiency and Weight gain tor Haitian beans at optimum protein level

Diet PER Feed Weight gain Efficiency (g/10 days)

pois rouge (l~ )1 1.16 0.25 33 pois noir ( 1~ ) 1.17 o.26 32

pois buerre (15%) 1 • .34 0.25 31

casein (lQt) 4.19 o.46 63 (fortified vi.th

cyatine)

Protein level

- 'Zl -

Figure l. Effect ot dietary protein levels on PER (ni.w PER VBlue) of Cssein and Haitbn beans.

0 t-c( a::

4-0

>- 3·0 0 z I.LI -(J

LI.. LI.. w2·0 z w t-o

-·---· 0---<0

-·--·-· a.A---o

casein fortified with cyatine

pois rouge

pois noir

pois buerre

g: I ·O ~::a....c::::::;~?

0 .._~.___.___.._ ___ ___. __ ___. ______ ___. ______ __

10 II 12 13 14 I~ 16 17 18 19 20

PR 0 TE I N LEVE L (0/o)

- 28 -

Experiment 2

Identity of Limiting Amino Acid in Haitian Beans

The data on the amino acid composition of Haitian foods (King,

1964) suggests that methionine (with cyetine) is the most limiting amino

acid in this variety or beans (Phaaeolµs YUlgaris ). Leucine, iaoleucine,

va.line or tryptopha.n also were indicated trom analytical data to be

somewhat limiting. For this reaean, Haitian beans supplemented with

these amino &cids, were tested for nutritional response to BUpplementa-

tion. In Table 7, the effect on weight gain, faed efficiency and PER of

supplementing beans vi.th amino acids for 10-day test periods is seen.

At optimum protein level, the addition of O.:?% methionine to

the bean diets, resulted in increases in both weight gain and PER. Either

isoleucine or the combination of isoleucine and methionine added to

the diet caused no improvement in either weight gain Bnd PER beyond the

diet supplemented with methionine alone except in the pois buerre diet

where the combination of both amino acids resulted in improved weight

gain and PER over that of methionine added alone. The growth depression

by iaoleucine in pois rouge, poie buerre, and pois congo 1-e anomalous.

For tryptophan supplemented pois congo diets improvement in weight gain

and PER was seen. The other combinations of amino 11.cids tested seemed

not to affect animal performance greatly unless tryptophan and methionine

had already been added to the diet.

The maximum PER and weight ga.in of the amino !\Cid supplemented

bean diets (10 days test period) were as follows:

- 29 -

11. pois rouge + methionine, 1.63 for PER e.nd 46 g for weight gal. n

b. pois noir + methionine, 1.67 for FER and 45 g for weight gain

c. pois buerre + methionine, + iaoleucine, 1.83 tor PER and 44 for weight gain

d. pois congo + tryptoph&n + ieoleucine + valine + methionine 2.00 for PER and 52 g tor weight g8.in

Figure 2 shows the PER of bean diets with smino scids

supplementation.

Table 7

Effect on Weight Gain, Feed Efficiency and PER of Supplementing Beans with Amino Acids

Diet Feed Gsin No. PER Efficiency (g I 10 days)

1 pois rouge (Phaseolus vulmris) 0.98 ~1.19 r 0.21 23 2 (1«tt )2 + methi0nine1 1.63 1.98 0.36 46 3 + isoleueine o.82 (o. 99) 0.18 20 4 + methionine + isoleucine 1.51 (1.83) 0.33 45 5 rois noir (Phaseolus vulgaris) 0.94 (1.14) 0.12 2.4 6 {l~) + methionine 1.67 (2.02) 0.36 45 7 + isoleueine I.CY} (1.29) 0.23 28 8 + methionine + isoleucine 1.49 (1.81) 0.33 42 9 his buerre(Pha.seolus vulga.ris) 0.91 (1.10) 0.17 19 \...>

0 10 15% ) + :oethionine 1.54 (1.m) 0.28 J6 11 + ieoleucine l.ll (1.35) 0.20 2J+ 12 + met~ionine + isoleucine 1.83 (2.22) 0.34 44 13 his congo (Ca._jMus indicus) 1.47 (1.78) O.Zl 36 14 15% ) + tryptophlUl 1.53 (1.85) o.28 J9 15 + tryptophan + isoleucine 1.34 (1.63) 0.24 33 16 + tryptoplmn + mline 1.67 (2.02) 0.30 41 17 + tryptophl\l'l + isoleucine +

valine + methionine 2.00 (2.43) 0.36 52 18 + isoleucine + mline + methio-

ni..'1.e 1.45 (1.76) O.Z? 34 19 Casein (fortified with cystine) (3. 76) 0.39 57

1 Amino acids supplemented at O. of diet except methionine added at o. for pois congo

2Protein level 3~w PER value in parenthesis, other values adjusted to casein • 2.50

- 31 -

Figure 2. Effect ot amino acid supplementation on PER of bean diets.

Protein levels: lat for JX>is rouge Md JX>is noir

15% for pois buerre and pois congo

Shaded column represents wisupplemented bean.

0 N . .,, 0 01 0 01 0 rri .,, :::D

0 -(/) n

> MET :::D Ul

0 "' c ISO z Gl ITI MET, ISO > ...

0 .,, ';/! 0 u

N en MET di z -Q ISO

::u MET, ISO

.,, 0 -VJ m MET c ITI ISO ::u ::u MET,150 1'11

.,, 0 VJ 0 " , ISO 0 z " ,VAL G') 0 " ,150,VAL,MET

ISO, VAL,MET

- 32 -

Experiment 3

Evaluation of Protein Value of H8itian Beans

In Table S me11surementa of nutritive VA.lue of protein in

Haitian beans shov that the biological V!tlue of p:>ia buerre ie a.

little better than the other tw beans 11lthough quite similar to p:>is

noir. Digestibility (true digestibility) of pois buerre, is Tery

little better than the other tlift) beans (between 68-7CJI, ). The Mlino

acid ba.lB.nce or digested protein from pois buerre seemed superior to

that of the other two beans as indiceted by the N.P.U. e.nd B.V. data.

PER is slightly less for pois buerre than for pois noir but better

thAn for pois rouge.

Table 9 shOW'S the biological value and digestibility, com-

pared to the data of Breesani et!.!• (1963) and Powrie et~· (1964).

Table 8

Mea.surements ot Nutritive Va.lue of protein in Haitian Beans

Casein pois pois pois pois rouge:pois Protein- ( + C7etein ) rouge noir buerre congo 50: 50 by tree lat iag 18% 1$% weight

1

N. intAke (g) 0 1.:33 2.14 2.34 1.94 1.97 Fectl. N (~) o.06 0.14 0.74 o.ao o.63 o.69 Urine N (g 0.10 0.25 0.77 0.79 o.68 0.61 Weight gain ( g) -8 33 17 21 18 18 PER (casein • 2.50) (3.95) 1.06 1.17 1.15 1.16

{l.29 )1 (1.42) (1.39) (1.40) Feed Efficiency 0.40 0.23 O.Zl 0.21 0.21 True Digestibility 93.98 68.22 68.)8 70.62 68.02 NPR 4.88 1.87 1.99 2.15 2.11 PRE 78.08 'Z}.92 31.81+ 31+.40 33.76 B.V. 88 54 57 58 62 NPU 0.83 0.']7 0.39 0.41 0.42

1PEIC(Protein efficiency ratio)• weight gain (g) Feed Efficiency = weight gs.in Protein consumed (g) food intake ~g

True Digestibility (D) = Absorbed N =- Food N - (fecal N-meta.bolic N) x 100 Food N Food N

NPR (Net Protein Ratio) • Weight gain on test protein + weight loss on non-protein group Weight of test protein conSlllned

PRE (Protein Retention Efficiency) a NPR x 16 . B.V. (Biological value)• Retained N • N intake - fecal N-metabolie N - Urine N- o enc>Us N )x 100

Absorbed N N intake - fecal N-metabolic N NPU (Net Protein Utilization) • B.V. x D

1Rav PER value in parenthesis. All fonnulas used in this experiment (Anonyrm:>us 1963, Enluation ot Protein Quality NAS-NRC

Publication 1100)

'-'> VJ I

- 34 -

Table 9

Complrieon of PER, Biologicn.l value and Digestibility of beans (Phaseolus wlgaris) for rats

Protein Biological True digestibility Efficiency Ratio Vfllue

Present study l.'1>,1.17,1.15 54,57,58 68.22,68.38,70.62

Bresse.ni et al (1963) - - i.09,1.20,1.14, 56,61,76,64 62,65,69,65

1.00 Powrie et al

C1964T- 1.06-1.53 33-37 63-69'

1Apparent digestibility

- .35 -

Experiment 4

Riboflavin Supplementation of Bean Diets

In Table 10 the results show clearly that the ribofiaTin

in beans was sufficient to support growth of rats. Arter a .3-week

test period, adding ribofiavin at the full level to the control gl'Qup

showed no effect. After the 3-week test period on the beM diet without

rlbofiaVin supplementation, no sign of riboflavin deficiency appeared.

PER values measured at the end of the third week as seen in

Table 11 show slight differences between diets with and without ribo-

flavin supplementation. To test these results further at the end of the

third week, ribofiavin was added to the control diets and feeding was

continued one more week. PER measured at the end of the fourth week

showed no distinct respaa to ribofi1'vin addition. Growth curves show-

ing the response of rats to ribofhvin supplementa.tion appeAr in

Figure J.

- 36 -

Table 10

Effect on Weight gain on bean diets •dth riboflavin supplementation for 4 week test periods

Weight gain (g) Diet let 2Dd 3rd 4th

week week 'f9ek week

pois rouge + full ribofia.vin 36 65 86 11.3

+ half ribofliwin 31 60

+ Nonel 30 53 77 1<1>

pois noir + full riboflavin 28 59 75 104

+ half ribofia.vin .32 61

+ None JO 59 85 114 pois buerre+ full riboflavin 28 55 74 100

+ half rilx>flavin 21 52

+ None 2, 11 61 82

Casein (fortified with o.zt (lat:) eyatine) 35 62 92 132

1 At the end of the 3rd week, the full level of riboflA.ru was added to the baalll diet and feeding WRB continued for one more week.

1

Table 11

Effect on PER for ra.ts on be8.Il diet with Ribofiavin supplemenUl.tion for 4 week test periods

Diet lat week 2nd week 3rd week

pois rouge + full riboflavin 2.08 (2.52)1 1.58 (1.91) l.J2 (1.60)

+ half riboflavin 1.86 (2.26) 1.56 (1.89)

+ None 1.93 (2.34) 1.50 (1.82) 1.36 (1.65)

pois noir + full riooflavin 1.80 (2.18) 1.49 (1.81) 1.20 (1.46)

+ half riboflavin 1.77 (2.15) 1. 52 (1.87)

+ None 1.86 (2.26) 1.52 (1.84) 1.33 (1.61)

pois buerre+ full ribof'l.avin 1.93 (2.34) 1.63 (1.98) 1.36 (l.65)

+ half riboflavin 2.16 (2.62) 1.66 (2.01)

+ None 1.85 (2.24) 1.46 (1.77) 1.39 {1.68)

Casein (fortified with eystine) (4.34) (3.55) (J.20)

Figures in parenthesis are rn.w PER values

4th week

1.25 (1.52)

1.28 (1.55)

1.19 (1.44) \..,) -.J

1.25 (1.52)

1.33 (1.61)

1.33 (1.61)

(J.20)

- 38 -

Figure 3. Growth of rats as affected bf ribofiArln supplement&tinn ot bean diets.

o~--o

• • -----full level of ribofi&vin added to the diets

no ribofiavin added to the diets

full leTel ot ribofiavin added to the diets at the end of the third week

Vertical bar represents the range ot the animal's weights. WE r G H T GAIN ( G)

~ ~ g ~

N

"'O g en z 0 -

.,, 0 en m c: "' ::u ::u "'

- .39 -

Experiment 5

Protein Quality Teet of Cereal-Legume Blends

Table 12 showe the canbinatimus of cereal-legume blends

in terms of their ratios and the protein contributed by the cere&l

and the legume. T11ble 13 shows the etfect of blend compoeition on

weight gain &nd PER value of rats &fter 2-week test periods. The

beet combinlltions considered from growth response blending besne with

com, sorghum and rice were as followe:

for pois rouge: 20-30,20-.30 and 2oi respectively

for pois noir: 20, 30 and 2Qt respectively

for pois blanc: .30, 30 and 20'-3~ reepectively

for pois buerre: 20, 30 and 20' respectively

Two-week weight gains corresponding to the blended series or the best combinations were as follows:

pois rouge .30, 30 and 41 g respectively

pois noir .'.36, 4.3 and 39 g respectively

pois blanc 31, J6 and 39 g respectively

pois buerre 36, .38 and 42 g respectivelJr

PERs measured as follows:

pois rouge 2.05, 2.13 and J.03 at 20, .30 and 2Ql respectively

pois noir 2.30, 2.45 and 2.95 at 20, 30 ani 2Qt respectively

pois blanc 2.00, 2.30 and 2.98 at 30, 30 and JO,;( respectively

pois buerre 2.28, 2.45 &nd 2.90 at 20, 30 and 20,:( respectively

Growth patterns ehow.i.ng wight gains Md PER values &re seen

in Figures 4 and 5.

- 40 -

Table 12

Combinations of Ce~l-Legume Blends and Protein Cootributions of Ingredients

Diet Cereal- Ratio in weight Weight per Protein distri-No. legume Cereal:legume 100 g diet but ion

Diet C~H~al:l~~e CS!real:le~

l' corn alone 100 : 0 84.7 : 0 s.30 : 0

2 com-pois rouge 90 : 10 68.0 : 7.6 6.66 : 1.64

3 corn-pois rouge 80 : 20 54.4 : 13.6 5.33 : 2.95

4 corn-pois rouge 70 : 30 43.3 : 18.6 4.24 : 4.04

5 corn-pois noir 90 : 10 66.7 7.4 6.54 : 1.78

6 corn-pois noir 80 : 20 52.7 13.2 5.16 3.17

7 coin-pois noir 70 : JO 41.2 : 17.7 4.04 4.'JS

8 com-pois blanc 90 : 10 66.2 : 7.4 6.49 : 1.80

9 com-pois blanc 80 20 52.3 : lJ.l 5.13 : 3.18

10 corn-poie blanc 70 30 41.0 : 17.6 4.02 : 4.28

11 com-pois buerre 90 10 66.7 : 7.4 6.54 : 1.72

12 com-pois buerre so : 20 53.1 : lJ.3 5.20 : 3.10

13 com-pois buerre 70 : 30 41.s : 17.9 4.10 : 4.17

14 sorghum alone 100 : 0 89.2 : 0 s.30 : o 15 sorghum-pois rouge 90 : 10 71.1 : 7.9 6.61 : 1.71

16 sorghum-pois rouge 80 : 20 56.2 : 14.1 5.23 : 3.Ci>

17 sorghum-pois rouge 70 : 30 44.7 : 19.l 4.16 : 4.14

f Continued) · · corn-bean and sorghu.•·n-bean diets (Diet Nos. 1-26) contained 8. )%protein

- 41 -

Table 12 (Continued)

Combinations of Cereal-Legume Blends and Protein Contributions of Ingredients

Diet No.

Cerea.1-legume Diet

Ratio in· wight Weight per Protein dietri-Cereal: legume 100 g diet bution

18

19

20

21

22

23

24

25

26

sorghum-pois noir 90 : 10

sorghum-pois noir 80 : 20

sorghum-pois noir 70 : 30

sorghum-pois blanc 90 : 10

sorghum-pois blanc 80 20

sorghtnn-pois blanc 70 30

sorghum-pois buerre 90 : 10

sorghum-pois buerre 80 : 20

sorghw-pois buerre 70 30

Z{2 rice alone 100 : 0

28 rice-pois rouge

~

30

.31

32

33

34

rice-pois rouge

rice-pois rouge

rice-pois noir

rice-pois noir

rice-pois noir

rice-pois blanc

(Continued)

90 : 10

80 : 20

70 : 30

90 : 10

80 : 20

70 : 30

90 : 10

cere~l:legume cereal:legume

69.2 : 7.7

54.4 : 13.6

42.4 : 18.2

69.2 : 7.7

54.0 : 13.5

42.1 : 18.0

69.8 : 7.8

54.9 : 13.7 43.1 18.5

88.9 : 0

66.2 : 7.4

50.7 12.7

38.6 16.6

64.7 : 7.2

48.J : 12.1

36.8 : 15.8

64.7 : 7.2

6.44 : 1.85

5.06 : 3.26

3.94 : 4.Y?

6.44 : 1.87

5.02 : 3.24 3.92 : 4.'37

6.49 : 1.82

;.n : 3.19

4.01 : 4.31

6.40 : 0

4.77 : 1.61

J.65 : 2.76

2.78 : 3.60

4.66 : 1.73

3.4a : 2.90

2.65 : 3.79

4.66 : 1.75

2rice-bean diets (Diets No. 'Z/-39) cont.Ained 6.4% protein.

Diet No.

35

.36 YI

38

39

40

- 42 -

Table 12 (Continued)

Combinations ot Cereal-LeP,Unle Blend~ and Protein Contributions of Ingredients

Cereal- Ratio in weight Weight per legume cereal: legume 100 g diet Diet cereal: legume

rice-pois blanc 80 20 4s.3 12.1

rice-pois blanc 70 30 36.4 15.6

rice-pois buerre 90 : 10 65.4 . 7.3 . rice-pois blerre 80 : 20 49.2 : 12.3

rice-pois buerre 70 : 30 '37.3 : 16.o

casein u.o

Protein distri-but ion cereal: legume

3.48 : 2.94

2.62 : 3.79

4.71 : 1.70

3.54 : 2.f!"/

2.69 : 3.73

10

Ta.ble 13

Effect of Blend Composition on Weight Ga.in {g) and PER '981.ue or nta ted Cereal-Legume Blends for 2- Week Test Periods

Cmn-be&n Diet 1 SC>l"&hum-bean Diet 1 Rice-bean Diet2 Weight PER Weight PER Weight PER ~in~g2 ~8in~g2 6~1n~~~

Cereal alone 17 1.40 (1.80)3 6 0.53 (0.69) Zl 2.38 (J.ct>)

Cereal: pois rouge(90:1ot 2b 1.78 (2.28) 17 1.22 (1.57) 34 2.60 (3.36) p>is rouge (80:20) JO 2.05 (2.63) JO 1.83 (2.'37 ~ 41 3.0J (J.90) p>is rouge (?0:30) 30 1.98 (2.55) 30 2.13 (2.74 36 2.90 (J.76)

p>ia noir ( 90:10) Zl 1.78 (2.28) 23 1.65 (2.J.1,.) JS 2.83 (J.64) pois noir (80:20) 36 2.30 ~2.98} 36 2.13 ~2.75) 'J9 2.95 (J.80) poie noir (70:.30) 32 2.10 2.70 43 2.45 3.15) 36 2.75 (3.54)

pois blanc (90:10) 23 1.68 (2.18) 21 1.50 ~1.95) 36 2.65 (J.42) pois blanc (00:20) 28 1.83 (2.36) 35 2.25 2.91) 'J9 2.80 (J.63) pois blanc {70:30) 31 2.00 (2.60) 36 2.30 (2.98) YJ 2.98 (J.85)

:sx>is buerre (90:10) 22 1.53 (1.98) 20 1.43 (1.84) 41 2. 78: (J.60) pois buerre ~ 80: 20) 36 2.29 (2.94) 33 1.98 (2.54) 42 2.90 (3. 76) pois buerre 70:30) 35 2.13 (2.74) 38 2.45 (3.15) 30 2.35 (3.04)

Casein l~ 64 (3.23)

t com and sorghum diets contained s.'!t crude protein 2rice diets contained 6.1/J, crude protein 3 "Vnlues in parenthesis are raw PER values ~ratios Of Ce1"93.l:legume w/w

I ~ \...> I

- 44 -Figure 4. Protein quality test of cere'll-legume blends with v1u7ing

amounts of beans showing weight gains {g) tor 2-week teat periods.

(Weight gains of rats fed cereal diets alone are shown by shaded a.rea ).

A. corn diets, protein level of all diets ""'-S 8.1' B. sorghum diets, protein level o! ill diets ""'-S 8.]t

C. rice diets, protein level of all diets ""'-S 6.4% WEIGHT IAIN ( 8) 40

40

0 10 2.030 POIS ROUGE

lO 2030 POISNOIR

10 20 30 % BEAM POIS BUERRE

102030 102030 0/0 BEAN POIS8LANC POISBUERRE

A

B

c

- 45 -Firuge 5. Protein quality test of cereAl-legume blends with VRrying

levels of beans showing PER value for 2-weeka test period.

PER nlue of rats fed cereal diets done are show by shaded "'rea.).

A. corn diete, protein level of all diets ~ s 8 • .3%

B. sorghum diets, proteir: level of Ali diets waes s • .3%

C. rice diets, protein level of all diets was 6.4%

PER (CASEIN AT 10%=2·5} 2-5

2-0

2·5

2'0

1·5

1-0

0·5

0 10 2030 POIS ROUGE

3·0

2·5

1·5

l·O

0·5

0 102030 POIS ROUGE

102030 POISNOI~

102030 POIS NOIR

10 2030 POIS NOIR

10 2030 10 2030 % BEAN POIS BLANC POISBUERRE

10 2030 10 2030 %BEAN POIS BLANC POIS BUERRE

10 2030 10 2'030 %BEAN POIS BLANC POfS BUE.RR£

A

B

c

- 46 -

Experiment 6

Protein Quality Test ot Cereal-Legume Blends Supplemented with the Limiting Amino Acids

Table 14 shows tor the best ccm.binations ot cereal-legume

blends tor ratios ot cereal to legume, the actual amounts in the teat

diets and the percentage contributions ot each tood to the total pro-

tein.

Table 15 shows the amino acid content ot the best combinations

of cereal-1•«=• blends ccapared to the requirement for growing rats

(Rama Rao, 1964).

Table 16 shows the limiting amino acids in the cereal-legume

blends ccpreaaed as percent of the requirement tor growing rate listed

in order trcm the most to the least limiting.

Table 17 shows the amount of amino acids added to the cereal-

legume diets.

Table l.S shows the response ot weight gain and PER value ot

rats fed cereal-legume blends with amino acid supplementation tor 2-

week test periods.

The ettecta on weight gain ot amino acid supplementation ot

cereal-legume blends were aa follows:

a. com-pois rouge supplmented with the combination of methio-

nine, lysine and threonine (Diet Ho. 4) gives better grovth than the

other ccmbinations or single amino acid aupplmentatiQn.

b. sorghum-pois noir diet supplemented vi th the combination

ot methionine, lysine and threonine (Diet No. 10) is again the best mixture.

- 47 -

c. rice-pois rouge diet supplemented with the combination

of methionine, threonine, lysine, isoleucine, and t;rrosine (Diet No. 111

and Diet No. 2.0 with nllne added) gives better growth.

d. com-poie buerre diet supplemented with the combination

ot methionine, lysine, threonine, isoleucine, nline,tyroeine and

t?7]Jtophan (Diet No. 25) gives the best growth.

e. sorghum-pois buerre diet supplemented with the combi.n&-

tion of methionine and lysine (Diet No. 28), gives the best growth and

all ot the increase is attribut&ble to the lysine.

r. rice-pois buerre diet supplemented with the combirultion

of methionine, threonine and lysine (Diet No. 34) gives Teey close

results in weight gain with the above combinatian plus isoleucine, ty-

rosine, leueine, Te.line and histidine {Diet No. 35). These give better

growth than the others in the same group. Again the major response or

this diet is attributable to lysine.

PER values correspond to the weight gains except in those groups

which were eqU&l in weight gain but more complete amino acid supple-

mentation, give better PER values. These IU"e diets No. 20, and 19, and

35 and 34. Weight gains and PER values of the best diets (Rs seen in

Table 18) are listed as follows:

Diet No. 4, gain in weight 66 g and PER 2.69

Diet No. 10 gain in weight 77 g and PER 3.29

Diet No. 20 gain in weight 70 g and PER 3.74

- 48 -

Diet No. 25 gain in weight 65 g and PER J.09

Diet No. 28 gain in weight 77 g Md PER J.27

Diet No • .35 gain in weight 72 g Md PER J.82

Figure 6 shows the eftect on PER of rats fed cereal-legume

blends with amino &cid supplementAtion.

Figure 7 shows the effect on weight gti:i.n of rats fed cereal-

legume blends with amino acid supplementation.

Table 19 shows the limiting amino acid in cereal-legume blends

of the best combination of the limiting Amino acids being expressed as

percent of the requirement for infants.

In several instR.11ces the predictions from analytic~ data re-

garding the Vl'llue of runino acid supplementation A.re not bome out in

animal performance. This e.nomaly is seen in the case of lysine !Uld iso-

leucine in Diets 1-6, of threonine And isoleucine in Diets 14-20, of

methionine and lysine in Diets 21-25 and of methionine and threonine in

Diets 26-JO. Presumably these anoJM.lies results prinmrily from varying

biological aVlll.ilability of the various proteins since the true digest-

ibility among the beans, for eXAmple, are in the rRnge of only 68.I"laf,.

Table 21 shows the compositicn of Haitian blends of good

protein quality compsred to the new recommended dietary &lloW811ces.

TA.ble 14

The Best Combinations of Cereal-Legume Blends

Diet Protein RAtio by Weight in % Protein % Protein Nos. level Cerea.1-legume weight diet in diet in

tested cereAl:legume cereAl:legume from mixture % cereAl:legwne alone

-1-6 10 corn-pois rouge 70 : 30 52.2 : 22.4 5.J 2 : 4.86 13.4

7-13 10 sorghurn-pois noir 70 : JO 51.1 : 21.9 4.75 : 5.27 12.8 I

14-20 9 rice-pois rouge 80 : 20 ?1.J : 17.8 5.13 : J.86 10.1 .r:-"'

21-25 10 com-pois buerre 80 : 20 64.0 : 16.0 6.27 : J.73 12.5 I

2£-30 10 sorghtm1.-pois buerre 70 : JO 51.9 : 22.2 4.83 : 5.17 13.5

31-35 9 rice-pois buerre 80 : 20 69.2 : 17.J 4.98 : 4.03 10.4

T!lble 15

Amino Acid Content in Cereal-Legume Diets Compired to the Requirement for Growing Rats

Expressed as g/100 g of diet

Diet

com-pois rouge ratio 70:30 by wt.

sorghum-pois noir ratio 70:30 by wt.

rice-pois rouge ratio 80: 20 by wt.

com-pois buerre rotio 80: 20 by wt.

sorghum-pois buerre ntio 70:)0 by wt.

rice-pois buerre rRtio 80:20 by wt.

TAble 15 (Continued)

Amino Acid Content in CereAl-Legume Diets Com:pi.red to the Requirement for Growing Rats

Expressed AB g/100 g of diet

Methionine Isoleucine Leucine

0.089 0 • .34 0.90

o.066 0.36 0.99

C.11 o.JJ 0.62

0.10 O.JJ 0.95

0.(!72 O.Y/ 1.00

0.11 0.34 o.62

Requirement for growing rrlts (Ra.ma RAo et al, 1964) 0.16 -- 0.55 0.70

Tyrosine Phenylal8nine

0.24 0.46

0.23 0.46

V"I 0.21 0.42 I-'

0.25 0.45

0.24 0.47

0.23 0.43

O.JO 0.42

Diet

corn-pois rouge 70:30

sorghum-pois noir 70:30

rice-pois rouge 80:20

com-pois buerre 80:20

sorghtm1-pois buerre 70:JO

rice-pois buerre 80:20

Table 16

Percent Limiting Amino Acids in Cerel!ll-Legume Diets :qs Percent of the Requirement for Growing IU\ts

Expressed as percent of the Requirement

% Limiting Amino Acid

cystine1, methionine 44, lysine 40, threonine 38, isoleucine JS, valine 25, a.nd tyrosine 20. (Excess in Jiienylalanine 10, histidine 20 and leucine 29, equal in tryptoprum ).

cystine 62, methionine 59, lysine /J+, threonine 42, isoleucine 35, valine 24, tyrosine 23, and histidine 4. (Excess in }'.ilenylalrmine 10, tryptophan Z7 and leucine 41 ).

cyetine 59, threonine 48, lysine 43, isoleucine 40, methionine 31, tyrosine JO, valine 22, histidine 12, and leucine 11. (Excess in tryptophan 'Zl and equal in Jbenylalanine ).

cyet:i.ne 59, lysine 46, threonine 40, isoleucine 40, methionine Tl, valine Z7, tyrosine 17 1md tryptophan 14. (Excess in phenylal~n:i.ne 7, histidine 20 and leucine 36 ).

cystine 59, methionine 55, lysine 46, threonine 40, isoleucine 33, valine 20, and tyrosine 20. (Excess in Jflenylal'Uline 12, tryptophan 'Zl and leucine 43, equal in histidine).

cystine 59, threonine 48, lysine 43, isoleucine 38, methionine Jl, tyrosine 23, leucine 11, 'V'lline 11, and histidine 8. (Excess in phenylalanine and trypto-ph,i.n 'Zl ) •

1Cyst:i.ne replaced by methionine at equal a.mount.

Vt N

Diet No.

1 2 3 4 5 6 7 8 9

10 11 12 13

14 15 16 17 18 19 20

Table 17

Amount of Amino Acids added to the Cere111l-Legume Diets Expressed as g/100 g of diet

None Methionine 0.28 Methionine 0.28 and lysine 0.36

Amino Acid Added

Methionine 0.28, lysine 0.36, and threonine 0.19 Methionine 0.28, lysine 0.36, threonine 0.19, &nd isoleucine 0.21 Methionine 0.281 lysine 0.36, threonine 0.191 isoleucine, 0.21 and valine 0.14 None Methionine 0.30 Methionine 0.30, lysine 0.40 Methionine 0.30, lysine 0.40, Md threonine 0.21 Methionine 0.30, lysine 0.40, threonine 0.21, and isoleucine 0.21 Methionine O.JO, lysine 0.40, threonine 0.21, isoleucine 0.19, and V7\line O.lJ Methionine 0.30, lysine 0.40, threonine 0.21, isoleucine 0.19, valine O.lJ, tyrosine O.Cfl,

and histidine 0.01 None Methionine 0.25 Methimine 0.25 and threonine 0.24 Methionine 0.25, threonine 0.24, Mld lysine 0.41 Methionine 0.25, threonine 0.24, lysine 0.41, and isoleucine o.22 Methionine 0.25, threonine 0.24., lysine 0.41, isoleucine 0.22 and tyrosine O.C1] Methionine 0.25, threonine 0.24, lysine 0.41, isoleucine 0~22, tyrosine O.Cf), valine 0.12,

+ Leucine 0.08 and histidine (O.OJ).

Continued

VI w

Diet No.

Zl 22 23 24 25

26 'Z1 28 29 JO 31 32 33 34 35

Table 17 (Continued)

Amount of Amino Acids added to the Cereal-Legume Diets Expressed as g/100 g ot diet

Amino Acid Added

None Methionine, 0.26 Methionine 0.26, and lysine 0.41 Methionine 0.26, lysine 0.41, threonine 0.20, A.nd isoleucine 0.22 Methionine 0.26, lys:ine 0.41, threonine 0.20, isoleucine 0.22, valine 0.15, tyrosine 0.05,

A.nd tryptophan 0.02. None Methionine 0.29 Methionine 0.2$, lyeine 0.41, Methionine 0.29, lysine 0.41, And threonine 0.20 Methionine 0.29, lysine 0.41., threonine 0.20, isoleucine 0.18, va.line 0.11, and tyrosine O.C6 None Methionine 0.25 Methionine 0.25, nnd threonine 0.24 Methionine 0.25, threonine 0.2.4, and lysine 0.39 Methionine 0.25, threonine 0.24, lysine 0.39, isoleucine 0.21, tyrosine O.fJl, leucine o.oo.

valine o.06, and histidine 0.02.

VI ~

1 2 3 4 5 6

7 8 9

10 11 12

13

14 15 16 17 18 19

20

Diet2

Table lS

Weight Gain ( g) and PER value of nts fed Cereal-Legume Blends with Amino Acid Supplementation for 2-Weeks Test Period

Amino Acid Added Gain in weight

___ t~L

com-pois rouge None 47 Methionine 58 Methionine and lysine 42 Methionine, lysine, and threonine 66 Methionine, lysine, threonine, Md isoleucine 43 Methionine, lysine, threonine, isoleucine A.Ild

VRline 43 sorghum-pois noir None 56

Methionine 58 Methionine and 1.ysi.~e 64 Methionine,lysine and threonine 77 Methionine, lysine, threonine and ieoleucine 67 Methionine,lysine, threonine, ieoleucine and

wline 69 Methionine, lysine, threonine, iaolrucine, vallne,

tyrosine and histidine 72 rice-pois rouge None 57

Methionine 62 Methionine and threonine 57 Methionme, threonine and l;ysine 67 Methionine, threonine, lysine and isoleucine 63 Methionine, threonine, lysine, iaoleucine and

tyrosine 70 Methionine, threonine, lysine, iaoleucine, tyrosine

and Wlline 70

rFigures 1.n~i:srenthesis~are Raw rm--ruues.

PER

2.41 (2.92) 2.60 (J.15) 2.35 (2.85) 2.69 (J.26) 2.38 (2.87)

2.45 (2.97) 2.54 (3.08) 2.61 (J.16) 3.05 (3.70) 3.29 (J.99) 3.16 (J.83)

3.33 (4.04)

3.41 (4.13) 2.85 (3.46) 2.98 (3.61) 2.93 (3.55) 3.20 (J.88) 3.69 (4.47)

J.68 (4.4b)

3.74 (4.53)

'

2Diet No. 1-6, 7-13, and 26-JO, cereal bean ratio = 70:30 by weight and all diets contain lat pro-tein. Diet No. 14-20, 21-25 Rnd 31-35 cereal bean ratio "" 80:20 by weight a.nd protein level ,.. 9, 1 O and ~ respecti vel.y.

V'I Vl

21. 22 23 24 25

26 'Zl 28 29 30

31 32 33 34 35

36

Table 18 (Continued)

Weight Gain (g) And PER V8.l.ue of rats fed Cereal-Legume Blends with Amino Acid Supplementation for 2-Weeka Test Period

Diet Amino Acid Added Gain in ,,,~t

conl-pois buerre None 52 Methionine 51 Methionine and lysine 4h Methionine, lysine, threonine and isoleucine 49 Methionine,lysine, threonine, isoleucine, VA.line,

and tyrosine a.nd tryptopian 65 eorghum-pois buerre None 50

Methionine 50 Methionine And ~ ysine 77 Methionine, lysine and threonine 65 Methionine, lysine, threonine, isoleucine, VA.line,

And tyrosine 66 rice-pois buerre None 62

Methionine 63 Methionine 8lld threonine 66 Methionine, threonine and lysine TJ Methionine, threonine, lysine, isoleucine, tyrosine,

leucine, valine and histidine 72 Casein (1<$) 50

PER

2.50 (3.03) 2.56 (J.10) 2.49 ~3.02~ 2.94 3.56

J.09 (3.74) 2.41.. ( 2. 96 ) 2.47 (2.99) "' °' 3.'Zl (J.9f.) I 3.18 (3.86)

3.28 (3.98) 3.00 (3.73) 3.13 (3. 79) 3.17 (J.84) J.66 (4.44)

3.82 (4.63) J.OJ

- 57 -Figure 6. Effeet of amino acid supplementation on PER or no.ts fed

cere~l-legume blends.

PER values for cereal-legume diets alone are shown in shaded column •

Composition (ratio by weight) of cereals t:md legumes and protein.

Levels (%)as follows:

com-pois rouge 70: 30, iqg

sorghum-pois no1r 70:30, left

rice-pois rouge 80:20, % corn-pois buerre 80:20, lqg

sorghum-pois buerre ?O:JO, iqg

rice-pois buerre 80:20, 9f,

PER (CASEIN AT 10% = 2·5)

4-0

3·5

3·0 r _J I

2·5

2·0

1·5

1·0

05

~

-.._

"' I- ~-w : :I.

>-er ,_ I~-I-

i _J

~-1g :

1;;;- -:i: . ,__ -: :

- -: :

I I I

0 ':?

er fl) :r : >-I- .J

cr -"' :i: • ::;_ :

I- I- I- -UJ . : = I UJ . : :I. 2

I-

"' :r .J

~ :::> ':'j a: >-,_ 0 '::! :

·~ • 1

CORN-P.ROUGE SORGHUM-P,NOIR RICE-P.ROUGE CORN-P.BUERRE SORG.HUM- RICE~P.BUERRE

P. BUERRE

- .58 -

Figure 7. Effect ot amino acid supplementation on weight gain of rats fed cereal-legume blends.

Weight gains or rats fed cereal-legume diets alone are shown in shaded column.

Composition (ratio by weight) of cereals and legmnes snd protein levels (% ) are as follows:

com-pois rouge 70:.30, l°"

sorghum-pois noir 70: .30, l°"

rice-pois rouge 80:20, <!$

com-pois buerre 80:20, l°"

sorghum-pois buerre 70:30, 10'

rice-pois buerre 80:20, 'II>

WEIGHT GAIN(G} 80

70

60

50

40

30

20

10

0

0 (II

a: :--. :c • .... _

. .

.... (II :c :::> w _J

CORN-P.ROUGE SORGHUM-P.NOIR. RICE-P. ROUGE CORN-P.BUERRE SORGHUM- RICE-P.BUERRE P.BUERRE

Table 19

Percent ot the Limiting Amino Acids in Cereal-legume Blends as Percent or the Requirement tor Infants'

Expressed SS percent or the Requirement for Infants

Diet

com-pois rouge rrltio 70:30 by wt.

sorghum-pois noir J"S\tio 70:.30 by wt.

rice-pois rouge rntio 80:20 by wt.

sorghum-pois buerre rntio 70:.30 by wt.

com-pois buerre ratio 80:20 by wt.

rice-pois buerre ratio 80:20

% Limiting Amino Acid

sulfur &mi.no llCid 65, isoleucine 48, 'VB.line 40, tryptophan 17, threonine .30, phenylA.lanine .30, lysine 2£, and leucine 17. (Excess in histidine 21.)

sulfur smino acid 68, isoleucine 44, valine 17, threonine 34, lysine 32~ pheny1A.lanine .30, trypto}il1\n 12 -..nd leucine 9. (Excess in histidine 4J

sulfur amino acid 55, isoleucine JJ+, leucine 17, threonine 34, phenyla-lanine ?!1, nline Z7, ~md lysine a. (Excess in histidine 4 8Jld equal in tryptophan)

sul.f'ur amino acid 66, isoleucine 44, wline 34, lysine 34, threonine 32, phenylalanine ~, tryptopuui 12, and leucine 7. (Excess in histidine 4)

sulfur amino acid 61, i90leucine 50, wline ~. tryptophan '37, lysine 34, threonine 32, Jiienylalanine 32 and leucine 13. (Excess in histidine 25)

sulfur Mlino t11cid 55, isoleucine 42, leucine Y/, threonine 34, phenyla-lrurlne Zl, lysine 2.3, valine 18 and trypt.ophtln 6. (Excess in histidine 8)

1 E'V'llustion ot protein quality. NAS-NRC (Publication 1100, PP• 14)

\J'I -.a

- (IJ -

Table 20

Comparisons or Protein Efficiency Ratio or Rats on Dirf'erent sources o! Protein with Cereal-Legume Blends at Optimum Protein Levels

Source of protein

'Dried def'atted egg Meat meal Dried milk Fish meal

2Rice-black bean Lime-treated com-black bean

3Rioe-beans {SO :20 )'t Com-beans (80:20) Sorghum-beans (70:30)

1Bender and Doeli {1958) 2Anonymoua (1961) NAS-NRC Publlcati.Dn 843 3Present study ~Figures in parenthesis are ratio w/w

Protein Efficiency Ratio (Cuein • 2,50)

2.65 2.19

2,9-3.0 2.0-2.J 2.1-2.5

Table 21

Composition of Haitian Blends of good protein quality Expressed in terme or 100 g of blends

Total Food Moisture Protein Fat carbo- Fiber

Energy hydrate

1 Cal % Blends ~ gm gm f:1Il

Rice-bean 358.6 12.0 10.16 0.80 75.92 1.34 80:20

Sorghum-bean .340.5 10.6 12.76 2.72 71.65 2.90 70:30

Com-bean 356.2 10.88 11.92 J.76 71.68 2.30 80:20 -

Recanmended2 Infants to 1 yr. kg xll5 --- kgx2.5 Children 3-6 yrs. 1600 --- 40 Boys 12-15 yrs. 3000 -- 75 Girls 12-15 yrs. 2500 --- 62 Men 18-35 yrs. 2900 -- 70 Women 18-35 yrs. 2100 ----- 58 Women, lact.R.ting +1000 --- +40

Ash

gm

1.12

2.Z7

1.76

Ca

gm

.024

.039

.024

.7

.8 1.4 1.3 .8 .8

+ .5

•Calculations based on the data of Composition of foods cOIIIDOnly used in l.Atin Amerlc1U1 Countries. Food Ccmposition Table for use in !Atin America. June, 1961 DJCAP-ICNND, ?JID, Bethesda, Maryland, U .s .A.

2a. w. Engel (1964) 1963 Recommended Dietaey Allowances J. Am. Diet. Assoc • .!JA:93 (Continued)

°' ~ I

T&ble 21 (Continued)

Composition of Haitian Blends of good protein quallt7 Expressed in terms Of 100 g Of blends

p Fe Vit. Thia- Ribofia- Nia- Ascor-A mine T.in cin bic

acid

Blends1 mg mg 1.u. mg mg mg mg

ruce-bean 1.32.6 2.56 1.6 0.172 o.C62 1.70 o.6 80:20

Sorghum-bean 283.4 4.frl 14 0.449 0.141 2.303 0.9 °' Z\) 70:30 I

Com·-beM 281.4 J.52 95 0.452 o.ns J.92 o.6 00:20

*Recommended Wants to 1 yr. ---- kgxl.O 1500 .4 .6 6 .30 Children J-6 yrs. -- 10 2500 .6 1.0 11 50 Boys 12-15 yrs. --- 15 5000 1.2 1.8 20 80 Girls 12-15 ;yrs. ---- 15 5000 1.0 1.5 17 80 Men 18-.35 yrs. ----- 10 5000 1.2 1.7 19 70 Women 18-35 yrs. -- 15 5000 .8 1.3 14 70 Women, lact&ting --- +5 +3000 + .4 + .6 +7 + 30

--·· -·--·-~ -----

1 Ibid 2Ibid, 9.3

- 63 -

IV. DISCUSSION

The high incidence of protein malnutrition in Haiti is

particularly evident in the low consumption of good quality protein

during the critical age of 1-3 yeers (Jelliffe and Jelliffe, 1961).

If a good combination of cerea.1-legume in the diet could be developed

to feed during this period, its widespread use should help in prevention

of protein malnutrition.

From the preliminary study, the optiruum dietary protein level

for ea.ch of several beans alone wss foWld to be 15-lat, results which

a.re similAr to those reported for a black variety of Phluseolue wJ.garie

by Breesani (1963).

In the studies on supplementA.tion of beans with a.mi.no acids,

methionine was fom1d to be the most limiting rurlno ncid as has been de-

monstrated by several investigators, (Jaff~, 1949, Schneider and ¥J.ller,

1954; Russell~~. 1946 and Sherwood ~ .!!J., 1954 ). Isoleucine, valine

and tryptophAn were predicted to be the other l:iJniting amino acids by -- '.

the R!lAlytical data on the a.mi.no acid composition of Haitian Foods

(King, 1964 ). The supplementation of both methionine and isoleucine

caueed improvement in nutritive value particuln.~ly in the ve.rietiee of . . . . .

Phaseolus vulgnria beyond the nlue obtained fron methionine alone. In

Cajanis indicus, one of the less promieing beRns, the nutritive value was

improved quite markedly when supplemented with the combination of tryptophan,

ieoleucine, valine and methionine, but not tryptopum a.lone or the other

- 64 -

combinR.tions. A response to supplementation with ieoleucine wn.e

expected because of its deficiency in beans as indicated by ana]Jrtieal

datl\ previousJ.7 mentioned; however, the results shawed no benefit from

supplementing with this amino acid !\lone.

In the protein evaluation study, the values of true digest-

ibility are somewhat low in the three most promising beAns (6S-7~ ).

Thia suggests that the amino acids in beans are only pRrtially ava.ilabla

to the animals. The biological values are somewh.":!.t low ttlso due to the

poor balance of digested protein. Both ot these ve.lues and the PER ue

compared to the data of other workers for this variety of bean in Table 9.

There was evidence that on nddit.i.nn of methionine the biological "f'llue ... ~ ~ ~ -

of beans was significantly increased, without an increase in digest-

ibility (BressAni ~ M• 1963 ).

In view of the limitation of Haitian meals, riboflavin con-

tent is apt to be veey low in the Haitian dietary (Sebrell ~ _tl, 1959 ).

From the riboflavin supplementation study, no evidence of riboflavin

deficiency appeared lthen the three most promising beans sened as sole

sources of protein. In optimum combinations of cereals a.nd legumes

studied here 64-4J1, of the protein ctune from cereals and 37-5~ from

beans. PERa ranged from 2.90 to J.OJ. All these mixtures were of

fairly good protein quality as judged by the PER (See Table 20 ). The

PERe were highest with rice-bean mixtures followed by sorghum beAn and

corn-bean mixtures in that order. Thie experiment indicAted that the

mixtures were pala.~ And gave good growth and efficient feed utilization.

- 65 -

The results of amino acid supple~.entation of cereal-

legume diets, showed no marked improvement in nutritive value in

general. The results indicate that in these mixtures, cereal and

legumes were complementing each other very well, there being only snail

value to supplementing turthel" with amino acids. It is noted that by

supplementation of the Umiting B.ID.ino acids in this study, some amino

acids Wiich wre limiting by calculation .f'rom analytical dat~, pre-

viously mentioned, did not show up in the biologic~ study.

- 66 -

V. StMiARY

Biological evaluation of the nutritional value ot the protein

of cereal-legune blends was studied using the three most promising beans

of Haiti including confirmatinn of the identity of the limiting runino

acids by supplementation and also Vitamin B content by depletion-2

repletion exper:bnents and .tina.lly by eupplementation of the limiting

Amino acids in the blends as indicated by calculation. Procedures of

protein evaluation in this study were based on NAS-NRC Publication 1100

(1963) and Nutrition Document R.G. Add. 1 (1963), P.A.G. of WHO-FAO-

UNICEF. Haitian beans were teated to detennine protein quality at

different protein levels. Optimum diet8.ry levels wre found Flt 15-18% •

.&.t opt,imum protein levels, biological value and true d~gestibility were

aomewhat low, but rilx>fiavin content was au!ficient as tested independ-

ently by using beans as a sole source of ribofiavin in the diet. Beans

supplemented with the lli.iting amino acid, showed marked improvement i.'1

nutritive value. Cere~l-legume blends were tested for protein quality

at the ratio of cereals to beans found optimum for growth. At these

levels protein qui.lity wns fairly high, indic:iting Unt the ~ino acids

were fairly well balanced aa confirmed by d~or..stration of only slight

improvement when these blends were supplemented with :i."Tlino acids. Blends

of good protein qutlity were obtained using the following mixtures:

rice-pois rouge at a ratio of 80:20 w/w sort;huin-pois blanc-ratio~ rice-pois noir " tt " " 80:20 " sorghum-pois buerre " rice-pois blanc " " " " 70: 30 " corn-pois rouge " sorghum-pois buerre " 11 " 80: 20 " com-pois noir " sorghlml-pois rouge " " " ?O:JO " corn-pois blMc n SDrghlml-pois noir " " " 70:30 " com-pois buerre "

70:30 v/w 70:30 " 80:20 " 80:20 t1

70:30 " 80:20 tt

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VI. ACKNCWLEIGME1ITS

The author Vi.shes to express his sincere appreciation to

Dr. K.w. King, tor inspiring guidance and experience; to Dr. R.w. Engel

tor his suggestions and patience. He is also gratetul to Mr. J.M. Albert,

Mr. M. Nunn, Mrs. B.W. Hall and Mrs. Z.R. Poe, tor their invaluable

technical aid.

The author is indeed thankful to Dr. A.L. Forbes ot ICNND,

NIH who made arrangements tor financial support and the United States

Arrrry Military Assistance Training Program (MAP) tor its financial

support during this study.

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vn. BJBLIOORAPHY

1. A.O.A.C. Ot!icial Method ot Analysis, 9t.h Ed., 1960 , .. , Sec. 2.o.31+,2.035 .th: Sec. 28.003 •••• sec. 4.003 ~ .. Sec. 22.033, 22.034 , .. ' Sec. 39.113 ~t~ :r •. · Seo. .39.ll4 2

2. Ano~ue (196.3) Evaluation ot Protein Quality, NAS-NRC, Publication llOO, PP• 74 •

.). Anonymous August 196.3 Nutrition Docmnent R. 6/Add. 1, P.A.G. Meeting

4. Anoeymous (1961) Meeting Protein Needs of Infants a.nd Children NAS-NRC Publication 843, PP• 35-48.

5. Borchers, R. Ackerson, c.w., Muesehl, F.E., and Moehl, A. 1948 -· Trypsin inhibitor VID Growth inhibiting properties of a soybean Teypsin inhibitor. Arch. Biochem. ,12: 317

6. Bresse.ni, R. Elia.s, L.G. and Narnrete, D .A. 1962(a) All vegetable mixture for human feedings. VI. The value of comMnation or lime-treated corn and cooked black bean. J. Food Sci. 1J.: .394

?. Bressani, R.N., Scrimshaw, N.s., Behar, M. and Viteri, F. 1958 Supplementation of cereal protein with amino acid II. Effect o! amino acid of com masa at intemediate level of protein intake on the nitrogen retention of young children. J. Nutr. ~: 501

8. Bender, A. E. and Doell, B.H. 1958 · Biological evaluation of pro-teins, a new aspect. Brit. J. Nutr. 11: 140

9. Bress:mi, R., Md A. Valiente 1962. (b) All vegetable protein mix-tures for human feeding. VII. Protein complementation between polished rice a.nd cooked bl~ck beans. J. Food Sci. gj_: 401

10. Bresaani, L.G., Elias Md Ana Terresen Valiente 196) Effect of cooking md am:ino acid 8UJ.?plementation on nutritive value or black bea.ns (Phaseolue vulgAris) Brit. J. Nutr. 17: 69

11., Ehe~, M.S. and Sholes, M.L. 1948 Effect of old fashioned and modern methods of cooking on retention ot nutrients in vegetables. II. Snapbesn Food Rea • .!J: 'JZ7

\

- l:9 -

12. F.A.o. (1953) Nutr. Stud. No. 9

lJ. F.A.o. (1956) Nutr. Stud. No. 14

14. Joint F.A.O.~O expert. committee on nutrition 1955. WHO Tech-nical report. aeries No. 97

15. Ham, W.E., Sandstedt, R.M. and Mussehl, F.E. 1945. The proteolytic inhibiting substMee in the e.xtntct rran unheated soybean meal and its effect upon growth in chicks. J. Biol. Chem. ill: 6J5.

16. Johns, c.o., and Finks, A.J. 1920 Studied in Nutrition II. The role of eystine in nutrition as exemplified by nutrition experiments with the proteins of the navy bean, Phaseolus vu.lg_1iris. J. Biol. Chem. J.t!: 719

17. Jaff~, W .G. 1949 Limiting essentitl amino acids for some legmne seeds Proc. Soc. Exper. Biol. and Med. _ll: 398

18. Jaff~, W.G. 1950 Protein digestibility and trypsin inhibitor activity of legume seeds. Proc. ~c. Exper. Biol. and Med • .z2: 219

19. Jellitf'e, D.B. and E.F. Patrici1l Jelliffe 1960 Prenlence of pro-tein-calorie malnutrition in Haiti.An preschool children. Am. J. ot Public Health 2Q:1355

20. Jelli!fe, D.B. and E.F. Patricia Jelliffe 1961 The Nutritional Status of Haitian Children Act1i Tropicn .,lg: 1

21. Ia.ose, A.A., Hill, B. and Fe"«Jld, H.L. 1946 Presence of a growth inhibiting substance in raw soybeans. Proc. Soc. Exper. Biol. and Med. g: 10

22. King, K.w. 1%4 Amino acid composition of Haitian foods. Econroic Botany (In press)

23. Liener, I.E., Deuel, H.J.,Jr., and Fevold, H.L. 1949 The Effect of supplementation methionine on the nutritive TI..l.ue of diets containing concentrates of the soybe1m trypein inhibitor. J. Nutr • .J2: 325

24. Liener, I.E. 1962 Toxic fA.ctors in edible legumes and their elimination Am. J. of Clin. Nutr. ]l: 282

25. Mitchell, H.H. 1924 A method of determ.ining the biological nlue of protein J. Biol. Chem. ~: ff73

- 70 -

26. McCollum, E.V., Simona, N. and Pitz, w. 1917 The dietary de-ficiencies ot the white bean, Phaaeolua yyl..garie, J. Biol. Chem. Z2: 521

Z7. Powrie, W.D. end Laml::ert, E.L. 1964 Nutritive -n.lue ot proteins in canned Il8Ty beans Food Technol. ll: 112

28. Richardson, L.R. 1948 Southem peas and other legume eeeda as a source ot protein for growth of rate. J. Nutr • .J2: 451

z:t. Russell, w.c., Taylor, M.w. Mehrhot, H.rr~ and Hirsch, R.R. 1946 The nutritive value of the protein ot nrieties legumes and the e!tect of methionine eupplementAtion. J. Nutr. 32: 31.3 - 0-

30. Rama RAo, P.B. Norton, H.w. and Connor, J.B. 1964 Amino acid composition and niitritive value of proteins V. Amino acid re-quirements as a pattern tor protein evaluation. J. Nutr. ~: 88

:u. Sehnei<ier, B.H., and Miller, D.F. 1954 The biologicAl nlue of Alaska pea proteins J. Nutr. _8: 581

32. Sherwood, F.w., Weldon, v. and Petera0n, W.J. 1954 Effect ot cooking and of methionine BUpplementation on the growth promoting ot cow pea {Vigna sienaia) protein. J. Nutr. £: 199

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.34. Sebrell, W.H.,Jr., Smith, S.C., Se-.eringhaus, E.L., Delva~ H., Reid, B.L., Olacott, H.s., Bernadotte,J., Fougere, w., Barron, G.P., Nicol.As, G., King, K,W., Brmkman, G.L, and French, C.E. 1959 Appraisal of nutrition in Haiti Afl. J. Clin. Nutr. 1: 1

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J?. Whitehouse, K., Zero, A. and Shey, H. 1945 Rapid Method of determining crude tiber in distillers dried grain. J.A.o.A.c. !§.:147

The vita has been removed from the scanned document

ABSTRACT

The nutritive vnlue of Haitian cereal-legume blends in optimum . . -

combinations was fairly high UidicA.ting a reasonably good OOlance of

amino acids. No i:nru-ked mprovement in nutritive value of the blends

resulted when they are supplemented with the amino acids indicated to

be limiting by calculatior:. Beans a.lone aro poor in nutritive value afl . . -

shown by low PER values, low digestibility and low biologietl value even -- --

when teated at opt:imura protein level. Nutritive w.lue of the beans was ~ . " . ~ -.

improved by supplementation with the limiting runirlO acids a.s indicated

by rma.lytica.l data. No evidence of ribollivin deficiency appeared when

beAns were used as a sole eource of ribo.fltlVill in the diet.