Protein quality and digestible energy of selected foods determined in balance trials with rats

Chemical analyses and balance trials with rats of 17 foods showed marked differences in protein quality. According to anin vitro procedure true protein digestibility was high (around 90%). The same was also the case for thein vivo values with the exception of two legumes, beans and chick peas, which were digested to a markedly lower degree when compared to thein vitro values. The assumed reason for this is discussed. Due to extensive differences in the amino acid composition of the 17 samples BV varied considerably with the lowest value (45.5) of rice-wheat gluten breakfast cereal. Energy digestibility was generally high. Amino acid digestibility determinedin vivo followed the pattern of the corresponding protein digestibility, although marked differences occurred from one amino acid to another in the same food.     

Influence of phytate on in vitro digestibility of casein under physiological conditions

The phytic acid content of four different varieties of beans under different processing conditions was estimated. It was highest in red kidney (1.86–2.13%) slightly lower in pigeon (1.86–2.03%), white (1.80–1.96%) and black eyed beans (1.15–1.64%). There was no significant change in phytic acid content of beans after soaking at 25°C for 22 hours. However, both soaking and cooking revealed 26–37% loss of phytic acid in all four varieties of beans.The rate of in vitro casein digestibility with and without phytic acid at concentrations found in legumes was determined at pH 8 and 37°C using multienzyme technique. Addition of 5 mg Na-phytate reduced the casein digestibility up to 20% compared to the control. However, only 25% reduction of casein digestibility was observed in the presence of 25 mg of Na-phytate. Higher concentration of Na-phytate had no significant effect on the rate of casein digestibility. Data strongly suggest the formation of protein phytate complex at alkaline pH of small intestine.Part of this work was presented at the XIII International Congress of Nutrition in Brighton, UK in August 1985.     

Protein quality of some Nigerian traditional diets based on the African yambean (Sphenostylis stenocarpa) and pigeon pea (Cajanus cajan)

The protein quality of Nigerian traditional diets based on the African yambean (AYB) and pigeon pea (PP) were performed by in vivo and in vitro bioassays. The individual foods were processed, cooked, dried and mixed to resemble the traditional diets. Weanling male Wistar rats (45–55 g) were used for the bioassay. The rats were fed ad libitum for 35 days of which 28 days were for growth and 7 days for N-balance. All the eleven diets including casein (control) provided 1.6 g N/100 g diet. The parameters tested include PER, growth, N-balance, BV, NPU and apparent digestibility (AD). The rats fed the AYB diets, except AYB: Agidi (Ag), showed superiority in most of the parameters tested. There were significant positive correlations between PER and NPU (r=0.925;p<0.001); PER and AD (r=0.908;p<0.001); PER and body weight gain (r=0.969;p<0.001). The in vitro protein digestibility of the diets was significantly correlated (r=0.80;p<0.01) with the in vivo apparent digestibility. These parameters were used to rank the various diets. Crayfish protein was a better supplement to legume/cereal or legume starchy staple mixtures than leguminous oil seed. However, supplementation of PP: steamed corn (SC) diet with crayfish did not prove beneficial. The result showed that diets based on the African yambean, an under-exploited legume, are nutritious. The need for its reintroduction into the fare of the populace through increased production and appropriate processing technology is stressed. The results of this study can form a base for the standardization of Nigerian diets based on these legumes.     

SOURCES OF VARIATION IN THE IN VITRO DIGESTIBILITY OF TROPICAL GRASSES

The in vitro DM digestibility of four tropical pasture species, Cenchrus ciliaris, Chloris gayana, Digitaria spp., Setaria spp. and one temperate grass, Lolium perenne , were studied, using the method described by Tilley and Terry (13).
In vitro digestibility was affected by fineness of grinding, sample size, pH of original rumen fluid and size of rumen fluid inoculum. Different relations were found between the in vivo and in vitro results for the five species, with a maximum predicted difference of 3–5 digestibility units.
It was considered that the in vivo digestibility of tropical grasses could be accurately predicted by this method, provided that the procedure was standardized and samples of known in vivo digestibility similar to those being tested are included in each run.     

Protein evaluation of four oat (Avena sativa L.) cultivars adapted for cultivation in the south of Brazil

Four oat cultivars adapted for soil and climate conditions in the southern region of Brazil were evaluated for protein nutritive value. Evaluations were done both in vitro and in vivo. In vitro evaluation was done by essential amino acid profile, available lysine, amino acid scoring, and protein digestibility corrected amino acid-scoring (PDCAAS). Nitrogen balance indices and PER were determined in vivo with rats. In all four cultivars (UFP-15, UFP-16, CTC-03, UFRGS-14), lysine was the most limiting amino acid. Available lysine, amino acid score and PDCAAS were highest for cultivar UFRGS-14 and lowest for CTC-03. When compared to casein, only nitrogen retention for UFRGS-14 did not differ statistically (p>0.05); all other indices of protein quality were inferior to casein for the oat cultivars. The oat cultivars tended to be identical among themselves, except for apparent protein digestibility which was significantly higher in the UFRGS-14 and CTC-03 cultivars. On average, the PER values of the oat cultivars were 82% of casein; the net protein utilization was 88% of casein as determined in vivo and 49% by the estimation in vitro (PDCAAS).     

Effect of heat processing on in vitro protein digestibility and some chemical properties of African breadfruit (Treculia africana Decne) seeds

The effects of dry heat (roasting) and moist heat (boiling) on in vitro protein digestibility, protein fractions and other chemical properties of African breadfruit (Treculia africana Decne) seed that affect their utilization as a source of human food were investigated. Chemical analyses showed that the crude protein and fat contents of the unprocessed (raw) seeds were 20.1% and 13.7%, respectively. The level of phytic acid in the raw seed (1.19 mg/g) was lower than the levels found in some commonly consumed pulses in Nigeria. Albumin and globulin protein fractions were found to be the major seed proteins of African breadfruit seed, constituting 67.8% of the total protein of the raw seed. There were no significant (p<0.05) differences between crude protein, ash and fat contents of the raw and heat processed samples. Boiling proved more effective than roasting for improving protein digestibility and for reducing the levels of trypsin inhibitor, phytic acid and polyphenols of the samples. The complete removal of these antinutrients, however, would require a more severe heat treatment of the seed, which in turn would profoundly reduce the nutritional value and availability of proteins, as demonstrated by the low values obtained for in vitro protein digestibility, protein fractions and protein extractability.     

Digestibility of protein and amino acids in selected foods as determined by a rat balance method

Values (%) for true digestibility of crude protein and individual amino acids in 20 selected foods were determined by the rat balance (fecal) method. The products were fed as the sole source of protein in diets containing 8% crude protein (N × 6.25). Lowest true protein digestibility values (79–84) were obtained for pinto beans, kidney beans and lentils; intermediate values (89–92) were obtained for chick peas, beef stew, skim milk (over heated), rolled oats, whole wheat cereal, and pea protein concentrate; and highest values (94–100) were obtained for sausage, macaroni-cheese, rice-wheat gluten cereal, skim milk, tuna, soy isolate, peanut butter, chicken frankfurters, beef salami, casein and casein + methionine. In animal foods, peanut butter and soy isolate, the differences between true digestibility of crude protein and most individual amino acids were less than 5%. However, the values for true digestibility of methionine and cystine were up to 44% lower than those of crude protein in pinto beans, kidney beans, lentils, chick peas and pea concentrate. In these legumes, digestibility of crude protein was not a good predictor of digestibility of the limiting amino acids.     

Effect of processing on the in vitro digestibility of proteins and carbohydrates in some Indian legumes

The effect of different types of processing such as boiling, pressure cooking, puffing, frying, germination, and germination followed by cooking on the protein and carbohydrate digestibility of chickpea, horsegram and cowpea were studied in in vitro systems. In the case of chickpea, the protein digestibility was not significantly improved by any of the treatments. However, for horsegram and cowpea, improvement in protein digestibility was observed after some of the different processing treatments. Frying decreased the protein digestibility in all of the pulses. All of the treatments, except germination, caused a marked increase in in vitro carbohydrate digestibility.     

Nutritional improvement of lentils,chick pea,rice and wheat by natural fermentation

Effect of natural fermentation process (4 days) on the non-protein nitrogen, crude and true protein, amino acids content and in vitro digestibility of two kinds of legumes (lentils and chick pea) and two kinds of cereals (rice and wheat) was investigated. Non-protein nitrogen increased significantly (p<0.001) in the fermented products. Little increase has occured in the crude protein while no significant change was observed in the fermented true protein samples. It was observed that methionine and cystine which are considered the limiting amino acids in legume seeds were close to those of FAO/WHO patterns. Also, lysine content (the first limiting amino acid in cereals was higher in fermented rice than that of FAO/WHO pattern. Moreover, fermentation process improved significantly, the in vitro digestibility of both legume and cereal products.     

THE USE OF THE IN VITRO TECHNIQUE IN THE DETERMINATION OF THE DIGESTIBILITY OF GRASS/LEGUME MIXTURES

The in vitro dry-matter digestibility was determined of two samples of Digitaria decumbens , one sample of Medicago sativa , and one sample of Trifolium repens and nine mixtures prepared from these samples.
When the results were compared with in vivo digestibilities the standard errors of the regressions for the three combinations of legume, grass and legume/grass mixtures were ±0.6, ±0.6 and ±1.5 percentage units, respectively.
It is considered that in vivo digestibility of mixtures of grasses and legumes can be estimated by the in vitro technique, provided that suitable grass and legume standards are included.     

Compositional,functional and storage properties of flours from raw and heat processed African breadfruit (Treculia africana Decne) seeds

African breadfruit (Treculia africana Decne) seeds were either boiled or roasted and then milled into flour. Chemical composition, functional properties and storage characteristics of raw and treated flours and the effect of partial proteolysis on selected functional properties of the raw flour were determined. Raw flour contained 20.1% crude protein, 2.5% total ash and 13.7% fat. Heat processing significantly (p<0.05) improved in vitro protein digestibility, and water and fat absorption capacities but decreased bulk density, nitrogen solubility, emulsion and foaming properties, trypsin inhibitor, and phytic acid and polyphenol contents of the samples. Boiling proved more effective than roasting for improving protein digestibility, emulsion capacity and foam stability and reducing antinutritional factor levels. Partial proteolysis increased nitrogen solubility, bulk density and water and fat absorption capacities but decreased foam capacity at hydrolysis levels greater than 35%. Fatty acid and peroxide values of the samples increased during storage. Compared to raw samples, heat processed samples had significantly (p<0.05) lower and more acceptable peroxide values and free fatty acid contents and higher and more stable water (3.0 g/g sample) and fat (2.4 g/g sample) absorption capacities.     

Intake and digestibility of components of forage rape (Brassica napus) by sheep

Herbage from forage rape ( Brassica napus cv. Lair), harvested in the autumn, was separated into different morphological components and offered ad libitum to weaned lambs and adult sheep in two indoor pen-feeding experiments. In vivo. digestibility and the voluntary intake of herbage components were measured and related to chemical composition and in vitro digestibility. The organic matter digestibility of all plant components, especially of petiole, was high; lamina, 0·847: petiole, 0·892; upper stem. 0·865; lower stem, 0·771. Lambs and adults digested 'leaf' (0·83) vs 0·846) or 'stem' (0·782 vs 0·789) equally well. Both lambs and adults ate much less of the components or of whole crop than would be predicted with grass crops of similar digestibility or fibre content. With the exception of lamina, intake was closely related positively to digestibility and negatively lo fibre content. Low intakes of lamina were associated with high concentrations of glucosinolates. In vitro digestibility values obtained on small samples of forage show that the process is adequate for the prediction of in vivo digestibility.     

A comparison of physical properties, oxalate-oxalic acid soluble substances, protein content, and in vitro protein digestibility of the blue-green algaNostoc commune Vauch. from the Philippines and Japan

The physical properties, amount of oxalate-oxalic acid soluble substances (OOSS), protein and in vitro protein digestibility ofNostoc commune from the Philippines and Japan were determined. The samples were the spherical type from the Philippines (PS), the dry field discoid type from the Philippines (PD), and the dry field discoid type from Japan (JD). The discoid types, PD and JD, regardless of origin showed higher settling volume in water (SV) and bulk density than PS. The OOSS values obtained for all samples were directly related to SV and bulk density. The WHC level was highest in PS. This desirable characteristic makes PS a technologically potential sample for large-scale food production applications in the future. Protein and in vitro protein digestibility varied significantly among all samples. In general, the algal samples examined posses desirable physical properties which are expected to be related to physiological functions.     

Effect of a Bangladeshi home-cooking procedure on the amino acid content,trypsin inhibitor activity and in vitro digestibility of some legume seeds

Raw and cooked samples of cultivars ofLens esculenta (Lentils),Pisum sativum (peas),Phaseolus vulgaris (beans),Phaseolus aureus (navy beans)Cicer arietnum (gram), andLathyrus sativus (dhal) as well as precooked commercial products were analysed for amino acids, trypsin inhibitor activity and in vitro protein digestibility. Of the fifteen samples used in the study one lentil sample, one pea sample, two gram samples and one sample of khesari dhal were from Bangladesh, one gram sample was from Sri Lanka. The other samples were obtained either in shops in Norway or from an industrial firm. The latter were also obtained precooked and dried. The two samples obtained in shops were used with hull and dehulled.Neither cooking by a Bangladeshi household procedure nor industrial precooking and drying had any effect on the amino acid contents of the samples.Cooking and precooking reduced the trypsin inhibitor activity of the raw samples more when the original activity was high then when it was low. The inhibitor activity was similar between samples after cooking.Cooking and precooking and drying increased the in vitro protein digestibility in all samples except in the lentils in which the digestibility was reduced.In the raw samples protein digestibility was negatively correlated with the trypsin inhibitor activity.     

A TWO-STAGE TECHNIQUE FOR THE IN VITRO DIGESTION OF FORAGE CROPS

A simple technique for the determination in vitro of the dry- or organic-matter digestibility of small (0·5 g) samples of dried forages is described. It involves incubation first with rumen liquor and then with acid pepsin. Using 146 samples of grass, clover and lucerne of known in vivo digestibility ( Y ), the regression equation Y = 0·99 X– 1·01 (S.E. ± 2·31) has been calculated, where X = in vitro dry-matter digestibility. This technique has been used for the study of the digestibilities of plant breeder's material, of the leaf and stem fractions of herbage and of herbage consumed by animals.     

The prediction of the in vivodigestibility of the diet of sheep and cattle grazing indigenous hill plant communities by in vitrodigestion, faecal nitrogen concentration or indigestible' acid-detergent fibre

Herbage and faeces samples were retained from an intake and in vivo digestibility trial using material harvested at three different stages from five indigenous hill plant communities (those dominated by Agrostis-Festuca., Nardus stricta., Molinia caerulea., Eriophorum vaginatum and Tri-chophorum caespitosum) and from sown swards of ryegrass or white clover. Samples of the herbages and of their separate components, together with extrusa samples of the same herbages recovered from pen-fed sheep and cattle oesophageal fistulates, were digested in vitro using rumen liquor. Measurements were made also of faecal nitrogen concentration (FN) and of indigestible acid-detergent fibre (IADF) using the samples from the original trial.
The range of in vitro disappearance values of the herbages (IVOMD; 0809-0278) was slightly wider than those of in vivo digestibility (OMD; 0-796-0-37I). Differences between OMD and IVOMD were greatest at low quality and relationships were best described by two separate linear regressions of OMD on IVOMD for (a) ryegrass, white clover, Agrostis-Festuca and Nardus (RSD; 0 0185) and, (b) Molinia, Tricho-phorum and Eriophorum (RSD; 00246). In vitro disappearance values of extrusa were higher than those of herbages offered by 00503 and 00156 units for sheep and cattle respectively, partly because of greater levels of ensalivation, especially in the sheep. The relationships of OMD to IADF and to FN were poor, though inclusion of faeces output and fitting parallel lines for communities in the regression of OMD on FN reduced the RSD to 0020. We conclude that in vitro digestion of samples of extrusa, using appropriate standards, is the best method of diet digestibility prediction for sheep and cattle grazing these communities.     

Immunomodulatory activities of non-prolamin proteins in wheat germ and gluten

Albumin (Alb), globulin (Glo), glutelin (Gll) and glutenin (Gln) were separately extracted from wheat germ and wheat gluten. Amino acisd composition, molecular weight distribution, solubility, in vitro digestibility, and immunomodulatory activities were all analyzed. Gll and Gln have similar molecular weight distributions, which differed from those of Alb and Glo. Alb showed the highest solubility at various pH values (except pH 4.0), whereas Glo showed the highest in vitro digestibility. Glo and Gll have the highest proportion of essential to total amino acids, while Alb and Gll have the highest protein digestibility-corrected amino acid scores. Gll had the strongest immunomodulatory effects in terms of stimulation of RAW 264.7 cells to produce IL-6, TNF-α, and IL-10, and good stimulatory effects on splenocyte proliferation, production of IL-2, phagocytosis, and secretion of nitric oxide in RAW 264.7 cells. Gll can be considered a good protein source for use in health foods.     

Nutritional and Antinutritional Characteristics of Seven South Indian Wild Legumes

Seeds from seven species of wild legumes of the South India were analyzed for proximate composition, mineral profiles, amino acid profiles of total seed protein, in vitro protein digestibility, and certain antinutritional factors to assess their potential as alternative sources of protein crops. The major findings of the study were as follows: crude protein ranged from 20.3 to 35.0%, crude lipid 3.1–9.6%, crude fiber 5.9–12.1%, ash 2.7–5.1%, and carbohydrates 49.2–61.8%. Minerals viz., sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc, and manganese occurred in the range 42.9–135.9, 556.2–1639.5, 304.5–572.2, 174.9–686.7, 98.4–947.8, 3.6–16.4, 0.2–1.2, 2.0–30.0, and 1.0–3.9 mg/100 g seed flour, respectively. Profiles of amino acids of total seed proteins detected in the present study revealed that levels of valine, phenylalanine, tyrosine, isoleucine, and histidine of all the seven wild legume seed samples, threonine of Canavalia ensiformis and C. gladiata, leucine of Mucuna pruriens var. pruriens, and lysine of Cassia floribunda and C. obtusifolia were found to be higher than FAO/WHO (1991) requirement pattern. The in vitro protein digestibility of the legumes under study ranged from 63.39 to 83.32%. Antinutritional factors such as total free phenolics ranged from 0.41 to 5.96%, tannins from 0.04 to 0.60%, L-DOPA from 1.34 to 8.37% and trypsin inhibitor activity from 13.48 to 65.43 TIU/mg protein. The detected antinutritional factors probably have little nutritional significance if the seeds are properly processed.     

Processing of commercial peanut cake into food-grade meal and its utilization in preparation of cookies

The commercial cake produced during expeller pressing of peanuts was extracted with n-hexane, and 80% ethanol followed by sieving through 80 mesh, to remove residual oil, pigments, bitter taste and fibrous material. The processed meal exhibited comparable composition with defatted peanut flour prepared in the laboratory by solvent extraction of full-fat peanut meal. However, the processed cake meal exhibited low methionine content and in vitro protein digestibility as compared with defatted peanut flour. The processed cake meal can be blended with wheat flour to the extent of 10% (w/w) to prepare acceptable cookies with improved protein and mineral contents.     

Relationship between amino acid scores and protein quality indices based on rat growth

Protein efficiency ratio (PER), relative PER (RPER), net protein ratio (NPR) and relative NPR (RNPR) values, and amino acid scores were calculated for 20 food products (casein, casein + Met, beef salami, skim milk, tuna, chicken frankfurters, sausage, heated skim milk, peanut butter, rolled oats, soy isolate, chick peas, pea concentrate, kidney beans, wheat cereal, pinto bean, lentils, rice-wheat gluten cereal, macaroni-cheese, and beef stew). In most cases, PER, RPER, NPR or RNPR ranked the products in the same order and positive correlations among the protein quality methods were highly significant (r=0.98–0.99). Amino acid scores (based on the first limiting amino acid, Lys-Met-Cys, Lys-Met-Cys-Trp or lys-Met-Cys-Trp-Thr) were positively correlated to the PER, RPER, NPR or RNPR data (r=0.61–0.75). Inclusion of the correction for true digestibility of protein improved the correlations between amino acid scores and the indices based on rat growth. The correlations were especially high between Lys-Met-Cys scores (corrected for true digestibility of protein) and PER, RPER, NPR or RNPR (r=0.86–0.91). Inclusion of the correction for true digestibility of individual amino acids did not result in further improvements of the correlations in most cases. It is concluded that adjusting amino acid scores for true digestibility of protein would be sufficient and further correction for digestibility of amino acids would be unnecessary in mixed diets.This paper was presented in part at the 71st annual FASAB meeting, Washington, DC, March 29–April 2, 1987.