Mineral excretion of rats fed on diets containing faba beans (Vicia faba L.) or faba bean fractions.

The effects on faecal mineral excretion of two commercial varieties (local cultivar and Troy cultivar) of raw faba beans (Vicia faba L., minor) meal (VFM) and its fractions have been studied in growing rats. Diets contained local-VFM (dark seed coat) and Troy-VFM (light seed coat) at 474-500 g/kg diet, hull (VFH) from both varieties at 65 g/kg diet, and the insoluble cotyledon residue (VFCR) obtained from the Troy variety at 237 g/kg diet. Rats were pair-fed on diets which had been supplemented with amino acids to target requirements and contained similar amounts of zinc, manganese, iron and copper. With VFM diets the apparent absorption of Zn and Mn was significantly reduced. On the other hand, with hulls the apparent absorption of Fe was reduced while that of Cu slightly increased. As the amounts of calcium, phosphorus and magnesium in VFM and VFH diets were higher than in the controls, the increased intake resulted in a significant increase in both the apparent absorption and the faecal excretion of these minerals. The inclusion of VFCR in the diet had no significant effect on the mineral content of faeces. The relatively low concentrations of phytate in the bean seeds of 7.8 and 6.7 g/kg for the local and Troy cultivars respectively, could not adequately account for the increased mineral excretion. The results suggest that other seed constituents, possibly the soluble non-starch polysaccharides, may be involved in the elevated loss of Zn and Mn in rats fed on diets containing faba bean for extended periods, while some insoluble structural hull components may interfere with the absorption of Fe from the gut.     

Nutritional response of growing rats to faba beans (Vicia faba L., minor) and faba bean fractions.

The effects of raw faba bean (Vicia faba L., minor) meal (VFM) and its fractions on the growth and nitrogen utilization of rats have been determined in two experiments. Two commercial varieties of VFM were tested, local VFM (409-439 g/kg diet) and Troy VFM (439 g/kg diet). The bean fractions tested were V. faba lectin-depleted protein (VFDP), V. faba lectin (VFL) and V. faba cotyledon residue (VFCR). All diets were supplemented with amino acids to target requirements. Body-weight, body N and lipid contents of rats fed on VFM were reduced significantly in comparison with control rats fed on lactalbumin. This was due, in part, to the lower digestibility of the protein, lipid and dry matter (DM) of VFM diets. As a result, net protein utilization (NPU) and biological value (BV) of faba bean proteins were less than expected. Urine and urea-N outputs of the VFM-fed rats were also elevated in both experiments. Increasing the energy content of local VFM diets led to significantly higher dry body-weight, body N and lipid contents, with the result that the NPU and BV values of the protein also increased. However, the NPU values for VFM-fed rats were still significantly lower than those for the controls in both experiments. In contrast, true N, lipid and DM digestibilities in rats given local VFM were not significantly affected by the difference in the energy content of the diets. The replacement of two-thirds of the lactalbumin in the diet with VFDP (65 g/kg) reduced dry body-weight, N and lipid contents, NPU and BV compared with the control rats, even though N, lipid and DM digestibilities were not significantly different. The nutritional performance of rats fed on lactalbumin-based diets containing 7 g VFL/kg was similar to that of the controls. Similarly, the inclusion of the cotyledon residue (237 g VFCR/kg diet) had no appreciable effect on any of the variables studied. As VFL and VFCR had no antinutritional effects in these rats, it appears that the low nutritional value of VFM for rats (NPU 0.66) results not only from the low digestibility of the bean proteins, but also from disturbances in N metabolism.     

Nutritional response of rats to faba beans (Vicia faba L. minor black 'Kalimattar') and its fractions

The investigations on protein quality of raw faba bean seed meal (Vicia faba L. minor) and its fractions were carried out using rats in two experiments. Two varieties of faba seeds were used, locally grown faba bean 'Kalimattar' or 'black' (KSM) (492 g/kg diet) and its exotic counterpart Vicia faba L. minor 'White' (VFW) obtained from International Centre for Agricultural Research in the Dry Areas (ICARDA) (540 g/kg diet). The pulverized cotyledons of 'Kalimattar' (KMC; 438 g/kg diet), hull (KMH; 78 g/kg diet) as well as cotyledons of 'White' faba bean (VFWC; 470 g/kg diet) and hull (VFWH; 82 g/kg diet) fractions were tested. All diets were made isocaloric and isonitrogenous, however, without addition of individual amino acids to the target requirements. Performance of rats fed on both locally grown KSM and exotic FBW meal-fed rats were found to be inferior to those obtained from the rats fed on standard protein diet. The net protein utilization (NPU) of the rats fed on this seed meal was comparatively better than those obtained from the VFW. Replacement of standard protein with cotyledons from the either of the seed (KMC and VFWC) depressed food intake and protein utilization. This was due, in part, to the excretion of comparatively higher amount of nitrogen in faeces. Inclusion of KMH in the diet had no adverse effects on any variable studies as compared to the VFWH, which was found to impose severe restriction on the absorption of standard protein added to this diet. As a result, body weight gain, and NPU value were significantly lower than those obtained from the KMH. We conclude that the locally grown faba bean 'Kalimattar' is better than those of exotic 'White' cultivar and may contain less antinutritional substances. However, this warrants further investigations.     

The digestibility in piglets of faba bean (Vicia faba L.) as affected by breeding towards the absence of condensed tannins.

Seed samples from two near-isogenic faba bean (Vicia faba L.) lines were examined for the levels of so-called anti-nutritional factors (ANF). From the ANF known to be present in faba beans, trypsin inhibitor activity, functional lectins, condensed tannins and pyrimidine glycosides were analysed. It was concluded that the lines differed only in the content of condensed tannins being < 0.5 g/kg and 5.2 (SD 0.2) g/kg for the low (LT)- and high (HT)-tannin lines respectively. In addition, the level of pyrimidine glycosides in the LT line was slightly higher than that in the HT line. The LT line showed a reduced proportion of the seed coat (105 v. 119 g/kg) and a lower seed weight (0.85 v. 1.01 g). The apparent ileal and faecal digestibility values of dry matter and nitrogen from the HT and LT line were determined for piglets which were fed on diets containing chromium oxide as a marker. The mean apparent ileal and faecal digestibility values for dry matter for the LT line were 0.694 and 0.889 and for N 0.828 and 0.879 respectively. For the HT line, these values were approximately 0.05 and 0.10 lower (P > 0.05). A multienzyme technique was used to predict the in vitro protein digestibility (IVPD) of the two lines. The IVPD of the LT line was 0.965 and about 0.05 higher than the HT line, confirming the difference in digestibility as measured in vivo. The present study shows the positive effects on digestibility of removal of condensed tannins in faba beans which was achieved by plant breeding.     

Oxidative stress and DNA damage in broad bean (Vicia faba L.) seedlings induced by thallium

Thallium (Tl) is a metal of great toxicological concern because it is highly toxic to all living organisms through mechanisms that are yet poorly understood. Since Tl is accumulated by important crops, the present study aimed to analyze the biological effects induced by bioaccumulation of Tl in broad bean (Vicia faba L.) as well as the plant's antioxidative defense mechanisms usually activated by heavy metals. Thallium toxicity was related to production of reactive oxygen species in leaves and roots of broad bean seedlings following short-term (72 h) exposure to thallium (I) acetate (0, 0.5, 1, 5, and 10 mg/L) by evaluating DNA damage and oxidative stress parameters as well as antioxidative response. The possible antagonistic effect of potassium (K) was tested by combined treatment with 5 mg/L of Tl (Tl+) and 10 mg/L of potassium (K+) acetate. Accumulation of Tl+ in roots was 50 to 250 times higher than in broad bean shoots and was accompanied by increase in dry weight and proline. Despite responsive antioxidative defense (increased activities of superoxide dismutase, ascorbate peroxidase, and pyrogallol peroxidase), Tl+ caused oxidative damage to lipids and proteins as evaluated by malondialdehyde and carbonyl group levels, and induced DNA strand breaks. Combined treatment caused no oxidative alternations to lipids and proteins though it induced DNA damage. The difference in Tl-induced genotoxicity following both acellular and cellular exposure implies indirect DNA damage. Results obtained indicate that oxidative stress is involved in the mechanism of Tl toxicity and that the tolerance of broad bean to Tl is achieved, at least in part, through the increased activity of antioxidant enzymes.     

Carbohydrates digestibility and faecal nitrogen excretion in rats fed raw or germinated faba bean (Vicia faba)- and chickpea (Cicer arietinum)-based diets

Raw or germinated faba bean (Vicia faba minor var. Alameda) and chickpea (Cicer arietinum kabuli var. Athenas) seed meals were incorporated in essential amino acid-supplemented and energy-equalized diets for growing (65 (SD 1) g) male Wistar rats as the only sources of dietary protein. A lactalbumin-based diet was used as the control. Faecal dry weight and N excretion of animals fed legume-containing diets were greater (P<0.01) and DM digestibility lower (P<0.01) than controls. Apparent faecal digestibilities of amino acids were found to be not different or lower (P<0.01) than controls in rats fed diets containing legume seeds, either germinated or not. Total diaminopimelic acid (DAPA) and purine bases excretion were significantly (P<0.01) higher than controls in rats fed both legume seed meals. Faecal bacterial N calculated according to DAPA or purine bases values was similar and significantly higher (P<0.01) than controls in rats fed legume seed meals. Bacterial N accounted for 50 to 80 % of total faecal N in rats fed legume diets. Apparent faecal N digestibility values (53-65 %) of rats fed legume-based diets were lower (P<0.01) than controls, but became substantially higher (85-92 %) when calculated taking into account bacterial N values. Faecal NSP digestibility values for legume diets were 40-57 g/100 g ingested. Germination decreased starch faecal excretion (P<0.05) and increased (P<0.05) faecal starch and NSP digestibilities of faba bean seeds.     

The utilization of raw and autoclaved faba beans (Vicia faba L., var. minor) and faba bean fractions in diets for growing boiler chickens

The effects of the inclusion of raw and autoclaved whole faba beans (Vicia faba; RFB and AFB respectively) or faba bean fractions (cotyledons and hulls) in diets for growing broiler chickens (0-4 weeks of age) on performance, intestinal physiology and jejunal histological structure have been studied in three experiments. Significant decreases in body-weight as well as lower food consumption and higher food intake:weight gain ratio were observed in those animals fed on diets containing 250, 350 and 500 RFB/kg in the diet. Birds fed on AFB diets (500 g/kg) had significantly greater body-weights than chicks fed on RFB or raw faba bean cotyledons (RC). Significant increases in the relative lengths of duodenum, jejunum, ileum and caeca, pancreas relative weight, and intestinal transit time of birds fed on diets containing 250, 350 and 500 g RFB/kg compared with control birds were observed. Including AFB (500 g/kg) in the diet significantly increased body-weight and significantly decreased pancreas weight compared with RFB (500 g/kg)-fed birds. The inclusion of RFB hulls had no effect on these variables. Dehulling or autoclaving of faba beans, or both, proved to have no significant effect on relative lengths of duodenum, jejunum, ileum and caeca, nor on caecal volatile fatty acid concentration in birds fed on 500 g faba beans/kg diet. Electron microscopy of the jejunal mucosa revealed discrete hyperplasia of polysomes and mitochondrial swelling in those animals fed on AFB (500 g/kg) or AC (426.4 g/kg). Pronounced strangulations were also observed along the microvilli, whose length was similar to that of control birds. The inclusion of RFB hulls, either autoclaved or raw, led to no ultrastructural changes in the enterocytes, as detected by electron microscopy. Birds fed on diets containing the cotyledons of RFB (RC, 426.4 g/kg) rather than whole RFB showed the same ultrastructural disorders as RFB (500 g/kg)-fed birds. The present study shows that factors other than those usually claimed, i.e. protease inhibitors, phytates, tannins and lectins, may be contributing to the low nutritional value of V. faba seeds for growing chickens.     

Nutritional outcome and immunocompetence in mice fed on a diet containing raw field beans (Vicia faba, var. minor) as the source of protein.

Feeding growth mice on diets containing raw field beans (Vicia faba var. minor) as the only source of protein brought about an impairment in growth, muscle mass and liver weight. No changes in food consumption were observed, but the food intake:weight gain ratio was increased in those animals. Plasma protein, triacylglycerols and cholesterol values were not affected by the dietary treatment although serum glucose and zinc levels fell after legume intake as well as the number of circulating erythrocytes. The relative enlargement of thymus and spleen in the legume-fed mice was apparently accompanied by a reduction in the cell number and an increase in cell size, while the protein synthesis capacity followed differentiated patterns in both tissues when assessed through protein, DNA and RNA determinations. The haemagglutination titres and the number of rosette-forming cells were lower in those animals fed on the field bean diet as well as the splenic lymphocyte responses to phytohaemagglutinin, Concanavalin A or lipopolysaccharide mitogens used to evaluate the functional status of T and B lymphocytes. The present study describes, apparently for the first time in mice, the involvement of field bean intake in some immunological disturbances affecting both humoral- and cell-mediated aspects of the immune response.     

Hemoglobin repletion in anemic rats fed a new bean cultivar (Phaseolus vulgaris L.) as source of iron.

Utilization of iron from a new cultivar of dry bean (Carioca 80 SH) was estimated using anemic rat by two methods: the slope ratio and the hemoglobin regeneration methods. The bean was incorporated into the diet in the form of whole bean flour (WBF) 7.45 mg Fe/100 g or as partially decorticated bean flour (DBF), 8.57 mg Fe/100 g. Hemoglobin concentration in the rats' blood at the beginning of the experiment averaged 13.79 +/- 0.33 g Hb/dl; at the end of the depletion period it ranged from 5.62 +/- 0.38 to 6.30 +/- 0.39 g Hb/dl blood, difference which was not statistically significant (p < or = 0.05). The bean iron relative biological value (RBV, relative to FeSO4 = 100) evaluated by the slope ratio method, was 74% for the WBF and 77% for the DBF. The hemoglobin regeneration efficiency was 60.4 +/- 6.5, 50.8 +/- 3.2, and 52.7 +/- 5.7%, respectively, for the iron sulfate (reference source), the WBF, and the DBF.     

Effects of the whole seed and a protein isolate of faba bean (Vicia faba) on the cholesterol metabolism of hypercholesterolaemic rats

The aim of the present work was to analyse the hypocholesterolaemic efficiency of a Vicia faba-protein isolate in relation to the intact legume. In addition, the mechanisms underlying the effects of this isolate were investigated. Hypercholesterolaemic rats were divided into three groups and fed high-fat diets rich in cholesterol-containing casein, whole seeds of Vicia faba or the protein isolate of faba beans as protein source, for 2 weeks ad libitum. The protein isolate was prepared by isoelectric precipitation and spray dried. Analyses of serum, liver and faeces, as well as of the activity of hepatic 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase, were assessed by enzymatic methods. The rats fed on Vicia faba diets showed significantly lower body weights and energy intakes than rats fed on casein diets. The whole-seed diet induced a significant reduction in plasma triacylglycerol. Feeding rats on diets containing faba bean seeds, or the protein isolate, induced a significant decrease in plasma (LDL+VLDL)-cholesterol but not in HDL-cholesterol. Hepatic cholesterol and triacylglycerol were also reduced. The hypocholesterolaemic effects of Vicia faba were not the result of a reduction in cholesterol synthesis as assessed from HMG-CoA reductase activity, but the result of an increase in steroid faecal excretion. The faba bean-protein isolate obtained under our experimental conditions was useful in improving the metabolic alterations induced by feeding with a hypercholesterolaemic diet compared with casein. The effectiveness of the whole seeds was higher than that of the protein isolate.     

Mushroom tyrosinase inhibitors from mung bean (Vigna radiatae L.) extracts

A seventy percent ethanol from mung bean (Vigna radiatae L.) was extracted further with CH(2)Cl(2), EtOAc and n-BuOH to afford four fractions: CH(2)Cl(2)-soluble, EtOAc-soluble, n-BuOH-soluble and residual extract fractions. When using l-3,4-dihydroxyphenylalanine as the substrate for mushroom tyrosinase, the EtOAc-soluble fractions showed the highest inhibitory activity. Two pure flavonoid compounds, vitexin and isovitexin, were isolated (using the enzyme assay-guided fractionation method) from the EtOAc-soluble fractions. Vitexin and isovitexin showed high inhibitory activities, with IC(50) values of 6.3 and 5.6 mg/ml, respectively. This is the first study on the active compositions of azuki beans against mushroom tyrosinase.     

Mushroom tyrosinase inhibitors from mung bean (Vigna radiatae L.) extracts

A seventy percent ethanol from mung bean (Vigna radiatae L.) was extracted further with CH₂Cl₂, EtOAc and n-BuOH to afford four fractions: CH₂Cl₂-soluble, EtOAc-soluble, n-BuOH-soluble and residual extract fractions. When using l-3,4-dihydroxyphenylalanine as the substrate for mushroom tyrosinase, the EtOAc-soluble fractions showed the highest inhibitory activity. Two pure flavonoid compounds, vitexin and isovitexin, were isolated (using the enzyme assay-guided fractionation method) from the EtOAc-soluble fractions. Vitexin and isovitexin showed high inhibitory activities, with IC₅₀ values of 6.3 and 5.6 mg/ml, respectively. This is the first study on the active compositions of azuki beans against mushroom tyrosinase.     

Studies with chicks on the growth depressing factor(s) in faba beans (Vicia faba L. var. minor).

The growth response of broiler chicks was improved when they were fed diet containing high levels (greater than 85%) of heat treated as compared to raw faba beans. Pelleting and microwave treatment of faba beans containing 10% moisture resulted in maximal improvements in chick growth rates and feed efficiency of only 6% and 8%, respectively. Steam autoclaving (121 degrees for 20 minutes), extrusion (130 degrees or 152 degrees), or microwave treatment of faba beans containing 18% added water resulted in maximal improvements of 16% in growth rates and 19% in feed: gain ratios. the above study, a time course study of the relationship between faba beans autoclaving time anc chick growth response, and studies with individual faba bean fractions (starch isolate and protein isolate) suggests that processing improves the utilization of faba bean mainly through its effects on components associated with the protein and possibly the hull portions of faba beans. The utilization of faba bean starch is not improved with processing. Although the nature of the growth depressing factor(s) has not been established, the results from this study indicated that the major component does not appear to be a tryspin inhibitor, hemagglutinin, some other heat labile protein, or vicine.