Modifications in plasma cholesterol and apolipoproteins of hypercholesterolaemic rats induced by ethanol-soluble factors of Vicia faba

High-fat-high-cholesterol diets containing casein or a Vicia faba bean (faba bean) protein concentrate as the protein source were given to rats for 5 weeks. When the faba bean protein concentrate or its ethanol extract was present in the diet, a marked decrease was found in the level of circulating cholesterol associated with the lower-density lipoproteins (very-low-, intermediate- and low-density lipoproteins) compared with the level found on the diets containing casein or the faba bean protein concentrate deprived of ethanol-soluble factors. Alterations in apoprotein pattern were detected after the different dietary treatments. In particular, apoA-I appeared in an unusual form with electrophoretic mobility faster than normal in all lipoprotein fractions after feeding the diets that did not lower plasma cholesterol. When the diets contained the faba bean protein concentrate or its ethanol extract, the apoA-I disappeared from the lower-density lipoproteins but its normal form and the unusual one were apparent in the high-density lipoproteins. A moderate increase in faecal excretion of acidic steroids was found after feeding the diets containing the ethanol-soluble factors, irrespective of the protein source. The results are discussed in relation to the presence of saponin and polyunsaturated lecithin in the ethanol extract of the faba bean protein concentrate.     

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.     

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.     

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.     

Nutritional effects of field bean (Vicia faba L.) proteinase inhibitors fed to rats.

1. Weanling rats were fed on purified-casein diets containing active and inactive (autoclaved) field bean (Vicia faba L.) proteinase inhibitors (PI) at levels of 1.25, 2.5, 5.0 and 10 g/kg. Diets containing raw- and autoclaved-field-bean meal (FBM) were also given. The body-weight gain, protein efficiency ratio (PER); i.e. the ratio of the live weight gain in g/g of protein consumed and apparent nitrogen digestibility were determined 7, 14 and 21 d after the start of the trial. 2. Significant growth depression (P less than 0.001) was observed in rats given diets containing 2.5 g/kg PI (27--35%). In rats fed on raw FBM growth depression was severe after 7 d (57%). 3. Food intake was depressed at PI inclusion levels of 5 g/kg and higher between 14 and 21 d. 4. PER was depressed significanlty (33-44%) in rats given diets containing the active PI at all levels after 7 d. The protein utilization of all casein diets was higher than that of the FBM diets. 5. Apparent N digestibility was depressed in rats fed on diets containing 10 g/kg PI by 9 and 7% at 14 and 21 d respectively. The N digestibility of both raw and heated FBM was similar at approximately 84%. 6. It was concluded that the PI are not the main factors responsible for the growth depression observed when raw FBM is fed to rats.     

Effect of dietary methionine and polychlorinated biphenyls on cholesterol metabolism in rats fed a diet containing soy protein isolate

The effects of supplementation of methionine to a 20% soy protein isolate diet on serum level of cholesterol. 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity, cholesterol 7 alpha-hydroxylase activity, and biliary and fecal steroids in rats with or without receiving polychlorinated biphenyls (PCB) were investigated. Supplementation of methionine and PCB did not affect the growth. Serum level of cholesterol was higher in rats fed PCB than in controls. In rats fed PCB, addition of methionine elevated serum level of cholesterol synergistically. The activity of HMG-CoA reductase was higher in rats fed the methionine-supplemented diet than in those fed the unsupplemented diet when PCB was included in the diets. Cholesterol 7 alpha-hydroxylase activity (nmol/h.100 g body weight) was higher in rats fed PCB than in controls. Biliary secretion of bile acids was higher in rats fed PCB than in controls. On the other hand, fecal excretion of bile acids decreased in PCB-treated rats, but total steroids were not affected by PCB. In rats fed PCB, the addition of methionine did not alter cholesterol 7 alpha-hydroxylase activity and biliary and fecal steroid output. The data suggest that the increase in serum level of cholesterol due to dietary addition of methionine together with PCB would be mediated through the stimulation of hepatic synthesis of cholesterol.     

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 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.     

Influence of whole or skim milk on cholesterol metabolism in rats.

Male Wistar rats were fed commercial ration and given whole milk, skim milk, or water to drink. After 3 weeks the control group showed the greatest weight gain. Rats given whole milk had the smallest livers. Serum cholesterol levels were significantly lower in rats fed either whole or skim milk, but other serum lipids were unaffected. Liver triglyceride levels were lowest in the rats on skim milk. Activities of hepatic fatty acid synthetase, hydroxymethyl-glutaryl coenzyme A reductase and cholesterol 7alpha-hydroxylase were similar in the two milk-fed groups and considerably lower than in the controls.     

Effects of Japanese torreya (Torreya nucifera) seed oil on lipid metabolism in rats

OBJECTIVE: We investigated effects of Japanese torreya (Torreya nucifera) seed oil containing non-methylene-interrupted polyunsaturated fatty acid of all-cis-5,11,14-eicosatrienoic acid (sciadonic acid) on rat lipid metabolism. METHODS: Male Sprague-Dawley rats were fed the experimental diets based on AIN-93 containing 10% corn, soybean, or torreya oil for 4 wk. Blood and tissues were recovered from each rat, and concentrations of triacylglycerol, cholesterol, and phospholipid in plasma and liver were determined by enzymatic assays. Moreover, fatty acid composition was analyzed for triacylglycerol, cholesterol ester, and phospholipid isolated from plasma and liver lipids by gas liquid chromatography. RESULTS: Plasma triacylglycerol level in rats fed torreya oil was lower than that in rats fed corn or soybean oil, although there were no significant differences in plasma cholesterol and phospholipid levels in all rats. Liver triacylglycerol level was also lower in rats fed torreya oil, whereas liver cholesterol and phospholipid levers were same for all rats. omega-3 Polyunsaturated fatty acids such as 22:6 (omega-3) were lower in plasma and liver lipids of torreya and corn oil groups, whereas omega-6 polyunsaturated fatty acids such as 22:4 (omega-6) and 22:5 (omega-6) were higher. Considerable amounts of sciadonic acid were detected in cholesterol ester, triacylglycerol, and phospholipid in plasma and liver of rats fed torreya oil. CONCLUSION: These observations suggest that torreya seed oil can modify lipid metabolism, resulting in lower triacylglycerol levels in plasma and liver of rats.     

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.     

铅诱发蚕豆根尖细胞的遗传损伤

目的研究醋酸铅对蚕豆根尖细胞的遗传损伤效应。方法对籽粒饱满的蚕豆种子催芽24h后选根长1.0～1.5cm的蚕豆随机分组,用浓度为0.1、1.0、5.0、25.0、50.0mg/L的醋酸铅溶液处理,时间分别为12、24、36h,进行蚕豆根尖细胞微核试验。结果醋酸铅溶液可诱发蚕豆根尖细胞微核率显著升高(P<0.01),浓度超过0.1mg/L还可明显降低蚕豆根尖细胞有丝分裂指数(P<0.05),且高浓度短时间作用和低浓度长时间作用具有等效性。结论醋酸铅呈浓度依赖和时间依赖地显著降低蚕豆根尖细胞的有丝分裂指数,并诱发蚕豆根尖细胞微核率显著升高,利用蚕豆根尖细胞微核技术检测环境铅污染是可能的。     

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.     

Effects of dietary xenobiotics on the metabolism of copper, alpha-tocopherol and cholesterol in rats

The effects of dietary addition of some xenobiotics on tissue levels of copper and alpha-tocopherol in rats were studied with special reference to serum cholesterol. The chemicals added here were 0.05% polychlorinated biphenyls (PCB), 0.3% caffeine, 0.05% 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), 0.3% flavone, 0.3% chloretone, 0.3% 2-tert-butyl-4-methoxyphenol (BHA), 0.15% sodium phenobarbital, 0.15% phenytoin, 0.2% sodium pentobarbital, and 0.15% phenylbutazone. Of these chemicals, dietary PCB, caffeine, DDT, flavone, chloretone, BHA and phenytoin raised the level of copper in liver, kidney, or serum. Serum alpha-tocopherol and cholesterol also increased with the additive or synergistic effects on these metabolisms. Moreover, these combined addition of dietary 0.02% PCB with 0.2% BHA seemed to cause additive or synergistic effects on these metabolism. Moreover, these chemicals raised serum ceruloplasmin activity. In general, serum copper, ceruloplasmin activity and alpha-tocopherol were correlated with serum cholesterol in these dietary manipulations.     

The fat and protein fractions of freshwater clam ( Corbicula fluminea) extract reduce serum cholesterol and enhance bile acid biosynthesis and sterol excretion in hypercholesterolaemic rats fed a high-cholesterol diet

We investigated whether the fat and protein fractions of freshwater clam (Corbicula fluminea) extract (FCE) could ameliorate hypercholesterolaemia in rats fed a high-cholesterol diet. We also explored the mechanism and the components that exert the hypocholesterolaemic effect of FCE. The doses of the fat and protein fractions were equivalent to those in 30?% FCE. The fat and protein fractions of FCE, two major components of FCE, significantly reduced the serum and hepatic cholesterol levels. The fat fraction more strongly reduced serum cholesterol levels than the same level of total FCE. The excretion of faecal neutral sterols increased in rats fed the total the FCE and the fat fraction of FCE. On the other hand, faecal bile acid levels were greater in rats fed the total FCE and the fat and protein fractions of FCE than in control animals. The hepatic gene expression of ATP-binding cassette transporter G5 and cholesterol 7α-hydroxylase was up-regulated by the administration of the total FCE and both the fat and protein fractions of FCE. These results showed that the fat and protein fractions of FCE had hypocholesterolaemic properties, and that these effects were greater with the fat fraction than with the protein fraction. The present study indicates that FCE exerts its hypocholesterolaemic effects through at least two different mechanisms, including enhanced excretion of neutral sterols and up-regulated biosynthesis of bile acids.     

Effect of a radical scavenger "water soluble protein" from broad beans (Vicia faba) on antioxidative enzyme activity in cellular aging

We isolated a free-radical scavenger "water soluble protein (WSP)" from broad beans. Hydrocortisone (HC) is known to inhibit superoxide generation and was used as the reference scavenger. WSP was examined for its effect on antioxidation in young (PDL 20, 25% of the maximum life span) and old (PDL 50, 62.5% of the maximum life span) human fibroblasts (TIG-1). Cells were treated with WSP or HC for 4 and 6 wk in young cells, and for 3 and 6 wk in old cells. The cytosolic superoxide dismutase activity in the cells treated with WSP or HC tended to decrease as compared with that in the non-treated cells (control) with the exception of WSP-treated young cells 4 wk after culturing. Young cells were equal in glutathione peroxidase activity to the control, but the activity level in WSP- or HC-treated young cells 6 wk after culturing was 10-50% lower than that in the control. Young and old cells treated with WSP or HC were superior to the control in catalase activity with the exception of HC-treated old cells. WSP- or HC-treated cells were higher in glutathione (GSH) concentration than the control with the exception of WSP-treated young cells 4 wk after culturing and HC-treated old cells 6 wk after culturing. Such increases in catalase activity and GSH concentration by WSP treatment may be related to the delay of cellular aging-dependent degeneration.