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Int. J. Mol. Sci. 2010, 11, 1048-1056; doi:10.


International Journal of

Molecular Sciences
ISSN 1422-0067 Article

Effect of Soybean Variety on Anti-Nutritional Factors Content, and Growth Performance and Nutrients Metabolism in Rat
Chunmei Gu 1, Hongbin Pan 2, Zewei Sun 3 and Guixin Qin 3,*



Institute of Food Science and Engineering, Jilin Agricultural University, Changchun, Jilin 130118, China Institute of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan Province 650201, China Institute of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin 130118, China

* Author to whom correspondence should be addressed; E-Mail:; Tel.: +86-431-8453-3506, Fax: +86-431-8453-3506. Received: 25 December 2009; in revised form: 17 January 2010 / Accepted: 20 February 2010 / Published: 9 March 2010

Abstract: The present study was undertaken to evaluate the effects of soybean varieties on content of anti-nutritional factors, growth, and nutrient digestibility in rat. For this purpose, the content of trypsin inhibitor and lectin was firstly measured in five soybean varieties. Then sixty male Wistar rats were randomly divided into six groups and fed on different diets as follows: groups 1 to 5 were fed on treatment diets containing five different varieties of soybean flour; group 6 was fed on a control diet containing casein. All animals were fed for four weeks. During this period, faeces and urine were collected to determine the nutritional efficiency of diets and body weight were measured weekly on ten rats from each group. The results showed that trypsin inhibitor and lectin content of Jilin45 was the highest, and those of Jinong7 were the lowest of the soybean varieties. In comparison, all measured parameters, that is including gain in body weight, feed utilization efficiency, nutrient digestibility, nitrogen balance and nitrogen retention, were markedly different among the five groups of animals, but were significantly lower than the control group. These findings show that soybean varieties could significantly affect trypsin inhibitor and lectin content in soybean. Keywords: soybean variety; anti-nutritional factors; rat; growth performance; nutrient digestibility

group 6. After this adaptation period their weight was 100 ± 8 g. and a fixed 12-h light: dark cycle. Consequently. relative humidity. a control group. in order to destroy or reduce this anti-nutritional content. J. 2010. and reasonable processing methods. 23 ± 1 °C. were given a balanced control diet (200 g casein/kg) for 1 week. Jilin45. Despite the heat treatment of soybeans. and also to investigate the effect of anti-nutritional factors of different soybean varieties on animal metabolism. light 0700 to 1. and it raises the cost and energy demands of soybean production. lectin. Materials and Methods 2. Sci. soybean antigen and so on [2.900 h). (Jilin parvule 6. They were then divided randomly into six equal groups of ten animals each and fed on different diets as follows: groups 1 to 5 were fed on a treatment diet containing five different varieties of soybean flour. goitrin. but the thermal treatment process could also destroy other essential nutrients. Animals and Diets Male Wistar rats. soybean must be subjected to thermal treatment before consumption by human beings or animals. it contains many kinds of anti-nutritional factors.9 MJ/kg) and were given in powdered form. causing digestive and metabolic diseases of animals [6. weighing 80 ± 5 g at the beginning of the experiment. Mol.5]. Food consumption was recorded daily. and Fengjiao7607) for four weeks. Jinong7. utilization and digestibility of soybean protein [4. Therefore. urine and faeces were collected and pooled for nine days. This collection period was arbitrarily divided into two parts of four and five days. 11 1. 2. Commercially available soybean flour was purchased from Jilin Province Agriculture Academy of Science (Jilin. utilization and breeding of soybean varieties with low-levels of anti-nutritional factors. was fed on the original reference diet. However. Rats were kept in wire-bottomed cages under a controlled atmosphere (temperature. They had ad libitum access to food and tap water. the present study was designed to analyze and compare the trypsin inhibitor and lectin content in five important cultivated soybean varieties that are generally cultivated in Jilin province. such as phytoagglutinin [9].7] as well as reduced performance traits [8].3]. soybean plays an important role in the world of animal feed and food production. These results would provide a reference for soybean breeding. To determine the nutritional efficiency of diets. The composition of the diets is shown in Table 1. α-amylase inhibiting factor. 55 ± 5%.1. such as trypsin inhibitor. respectively. . findings have shown that it still contains low-levels of anti-nutritional factors. Body weight was measured weekly on ten rats from each group before feeding in the morning. China). Jilin30. Diets were isoenergetic (16. Introduction 1049 As the main source of vegetable protein. The care and use of laboratory animals followed the institutional guidelines of Jilin Agricultural University. It is a major source of both vegetable protein for humans and animals and of edible oil [1]. The existence of these anti-nutritional factors affects the nutritional value.Int. Analyses were carried out separately for the samples collected in these two periods and the results were combined and recorded only if there were no disproportionally large differences between them.

2720.4. inositol. 2010. cyanocobalamine. Sci.4. isoenergetic (16. The nutrient’s digestibility was measured by the method of Noreen and Salim [11].005. J. 0. Trypsin Inhibitor and Lectin Determination The content of soybean trypsin inhibitor in soybean was measured by the procedure of Liu and Markakis [10]. 12. folic acid. 4. 1995). nicotinic acid. K. 300.018.25) by micro Kjeldahl apparatus.12. 0. Mg. 2.5 40 20 Casein Soybean proteinc Maize starchd Soybean carbohydratesc Soybean oild Soybean fatc Cellulose Soybean fibrec Salt mixe Vitamin mixf b f Diets were semipurified. I. 2. ash.00004. 0. pantothenic acid. Mol. Shanghai. The dry matter was determined by oven-drying at 105 °C for 16 h. 0. then 30% acetic acid was added to terminate the reaction. Composition of the diets (g/kg) a.9 MJ/kg) and were given in powdered form. 0. 2. Nutrients Digestibility Determination The samples of experimental diets and faeces were homogenized using a motor and pestle and analyzed by standard methods (AOAC. The salt mixture provides the following amounts (g/kg diet-1): Ca. (dL) α-tocopherol.1. crude fat. 0.6. Fe. China. pyridoxine. crude fat by petroleum ether extraction method. Analytical Methods 2. riboflavin. 0. crude fiber and subtracting this from 100. 200.2. Changchun. 100. 30. China. Co.2. crude protein (N  6. 0. 140.6. Ingredients Protein Carbohydrates Fat Fibres Others a b c d e 1050 Control diet 200 560 120 60 40 20 Soybean flour 200 435 125 15 105 42. 80. ascorbic acid.2. Lectin content in soybean was assayed according to the method of Pusztai and Grant [9] using an enzyme linked immunosorbent assay.2. cholecalciferol.00002. Nitrogen Free Extract (NFE) was calculated by taking the sum of values for crude protein. 2. 11 Table 1. Zn. Growth Performance Assay Body weight and feed consumption were weighted in rats before feeding in the morning once a week. 40. 0. Na. The vitamin mixture provides the following amounts (mg/kg diet-1): retinol. choline. .5 17. thiamin.3.1.125. menadione. and then inhibited trypsin was monitored spectrophotometrically at 410 nm. In this procedure. 340. Ingredients originating from the soybean flour.2. 1600.032. biotin. 1.Int. 20. 0. trace elements: Mn. Cu. and body weight gain and feed conversion ratio (F:G) were calculated. trypsin inhibitor in samples reacts with trypsin for 2–3 minutes. 10. paraaminobenzoic acid.

while that of Jinong7 group was the highest when compared with other treatment groups.2. Differences between mean values were determined by ANOVA followed by comparisons using the Newman-Keuls multiple range test.05). Statistical Analysis Data Analysis: Data are reported as means ± SD. while Jilin parvule 6 (3.1.22 ± 0.01A 2.01A Values are means ± SD.52 mg/g) was significantly higher than other varieties (P < 0.fecal nitrogen)/intake nitrogen 2. Soybean trypsin inhibitor and lectin content(mg/g).05 were considered significant. The Effects of Soybean Varieties on Growth Performance in Rats As a result of the presence of anti-nutritional factors the body-weight and feed conversion ratios were strikingly decreased (P < 0. values without a common superscript are significantly different. . Differences with P < 0. J. n = 10. feedstuff/gain in weight in Jilin45 group was the highest and that of Jinong7 group was the lowest.05).89 ± 0. Within an array.03B 2.4.46 ± 1. 2010. they were significantly different from other treatment groups (P < 0. Nitrogen Balance and Nitrogen Retention Assay 1051 Intake nitrogen.84 ± 0.67 ± 0.51 ± 0.51C 28.urinary nitrogen . n = 10.01A 6.fecal nitrogen Nitrogen retention = (intake nitrogen .36B 59.2. fecal nitrogen and urinary nitrogen were determined by the method of Noreen and Salim [11].46 mg/g) was the highest compared with that of other varieties. that of Jilin 45(6. 3.05).05). That of Jinong 7 was the lowest and markedly different from the other three species (P < 0. For the content of lectin. P < 0.51 mg/g) was rated next to Jilin 45 and both were markedly different from the other varieties (P < 0. as is evident from Table 3. Mol. Content of Trypsin Inhibitor and Lectin in Five Varieties of Soybean Table 2 shows significant differences in the content of anti-nutritional factors among the five tested soybean varieties.30B Lectin 3.Int.52 ± 0. 11 2.05.25B 38. Sci. Results 3. The content of trypsin inhibitor in Jilin 45 (59. Table 2. Soybean variety Jilin parvule6 Jilin30 Jilin45 Jinong7 Fengjiao7607 Trypsin inhibitor 37.81 ± 0.3. Body-weight gain in Jilin45 group was the lowest. and nitrogen balance and nitrogen retention were calculated according to the following formulas: Nitrogen balance = intake nitrogen .47 ± 0.02C 2.43A 38.urinary nitrogen . 3.61 ± 0.05).

values without a common superscript are significantly differ.57 ± 0. P < 0.12B Feedstuff/gain in weight 4.Int. The striking difference in these parameters in the five treatment groups was attributed to the different soybean varieties.15 ± 0. 11 Table 3. Nitrogen balance and nitrogen retention among treatment groups were the highest in the Jinong7 group and the lowest in the Jilin45 group. The Effect of Soybean Varieties on Nitrogenous Metabolism Table 5 presents the effects of the anti-nutritional factor on nitrogen balance and nitrogen retention.02B Fat 83. Groups Control diet Jilin parvule 6 Jilin 30 Jilin45 Jinong7 Fengjiao7607 Body-weight gain(g) 93. Mol.15B Values are means ± SD. n = 10.61D 79.81 ± 0.05).43D 88.19C Nitrogen free extract 91. and there were significant differences between the values (P < 0.01B 6.23 ± 0.07C 85.32B 70. The digestibility of dry matter.49 ± 0.65 ± 0.28C 65.97 ± 0.07B 87.33A 82. 2010.21B 77. Within an array. .37 ± 0. Sci.33 ± 0.19C 88. Within an array.3.29 ± 0. Main nutrient digestibility of diet (%).12C 87. and these were significantly different from other three treatment groups (P < 0.25 ± 0.02C 1052 Values are means ± SD. J.23B 88.61 ± 0. The presence of trypsin inhibitor and lectin resulted in a significant increase in fat digestibility.20E 66. protein and nitrogen free extract were the lowest in Jilin45 group and the highest in Jinong7 group.02A 6.75 ± 0.00 ± 0.61D 65.39 ± 0.05.01D 5. which was highest in the Jinong7 group and the lowest in the Jilin45 group.4.70 ± 0.81 ± 0. 3. and significantly different between values (P < 0.21 ± 0.90 ± 0.05. n = 10.22A 89.11 ± 0.69C 78.29 ± 0.25D 87.94 ± 0.42 ± 0.31 ± 0.95B 77.05).01C 6. values without a common superscript are significantly differ. The nitrogen balance and retention were significantly lower in treatment groups compared with the control group. Table 4.35 ± 0.28A 72.82 ± 0.00 ± 0.03D 77.87 ± 0.13B 89.45 ± 0.73C 78. The Effect of Soybean Varieties on Nutrient Digestibility The anti-nutritional factors lowered the nutrient digestibility of the experimental diets compared to control group. except for fat digestibility (Table 4).36 ± 0.05).17 ± 0.15A 87. Groups Control diet Jilin parvule6 Jilin 30 Jilin45 Jinong7 Fengjiao7607 Dry matter 89.88 ± 0. Body-weight gain of rat and feed conversion ratio.66A 87.01C 8.03 ± 0.06B 69.27 ± 0.29 ± 0. 3. P < 0.22 ± 0.26B 62.43B Protein 80.

J.Int.88 ± 0. [19] also reported that the effect of lectin on growth performance of animals changed with its dosage. environment.01C 0.99 ± 0. Fu et al. Many studies have confirmed that daily gain in body weight and feed utilization efficiency are lower in animals fed with diets containing trypsin inhibitor and lectin [14–16].0 mg/g. the growth of animals was decreased by 13% and with the addition of 7.82 ± 0. values without a common superscript are significantly differ. the main anti-nutritional factors in soybean that inhibit growth in animals are trypsin inhibitor and lectin.2 mg/g.01B 0. the lower the body weight gain and feed utilization efficiency are.83 ± 0. Schulze et al. Mol. [12] reported that the trypsin inhibitor content in different soybean sources was different. Li [20] observed that when the concentration of lectin in diet was between 0–1. the results showed that soybean variety had a marked effect on the content of anti-nutritional factors like trypsin inhibitor and lectin.05). . body weight gain and feed utilization efficiency with a diet of Jilin45 containing high levels of trypsin inhibitor and lectin were the lowest.01B 1053 Values are means ± SD.62 ± 0.5 units/g.72 ± 0. and lead to significant differences in body weight gain and feed conversion ratio in the five treatment groups. and decreased by 23% compared with control rats. Discussions Qin et al.05.72 ± 0. n = 10.78 ± 0. the other is that lectin could combine with small intestine epithelium and induce constitutional and functional changes in the small intestine [17].02D 0. [8] discovered that by adding 2. Sci. 2010. that was decreased by 32%. This result shows that the higher the anti-nutritional factor content is.90 ± 0. Therefore.6 units/g. while those animals in the Jilin45 group had the lowest values. In the present study.4 g/kg Kunitz Trypsin Inhibitor (KTI) to a control diet. which could affect the content and activity of the soybean anti-nutritional factors. Groups Control diet Jilin parvule 6 Jilin 30 Jilin45 Jinong7 Fengjiao7607 Nitrogen balance 0. and 10% to others. In the present study. the body weight gain and feed utilization efficiency of rats fed with raw soybean diet were significantly lower.02A 0.01B Nitrogen retention 0.81 ± 0. climate. The content of anti-nutritional factors in Jilin 45 was markedly higher than in the other four tested soybean varieties (P < 0. Nitrogen balance and nitrogen retention. but when above 2.70 ± 0.71 ± 0. P < 0.01A 0. and was lowest in Jinong7. 40% to trypsin inhibitor. while those with a diet of Jinong7 containing low levels of anti-nutritional factors were the highest. 11 Table 5. growth of rats was obviously lower.01D 0. Within an array. 4. but that in L81-45907 it was 17. In the present study. no obvious change of growth performance in rats was found in 20 days. geographical factors.01B 0.01C 0. This indicates that the effect of soybean trypsin inhibitor on animal growth is related to its levels in diet. and this further demonstrates that soybean varieties have an obvious effect on the content of trypsin inhibitor and lectin.02B 0. Liener [18] reported that 50% of growth inhibition in rat fed with raw soybean diet was attributed to lectin. [13] found that trypsin inhibitor content in L81-4387 was 16. These results may be attributed to genetic differences in soybean varieties. Grant et al. One of the possible reasons for this result is that trypsin inhibitor leads to the loss of endogenous nitrogen.2 g/kg KTI.01B 0.

Trypsin inhibitor and lectin led to a significant decrease of nitrogen balance and nitrogen retention in treatment groups compared to those of control group. In addition. the level of trypsin inhibitor also affects these two parameters. . Many studies have demonstrated that the effect of lectin on nitrogen metabolism was obvious. chicken. similar results were obtained. Li [20] discovered that with increase of lectin content in diet. Hagemeister and Barth [29] reported that with an increase of trypsin inhibitor content in the diet. On the basis of above results. In the present study. Nutrient digestibility was decreased in animals fed with trypsin inhibitor. rat and mouse and so on. 11 1054 Digestion and absorption of nutrients could be measured by two parameters. and the decrease of protein digestibility was pronounced the biggest. the results showed that protein digestibility was lowered in rats fed with lectin. endogenous nitrogen in chymus was obviously increased. Qin [25] reported that lectin could combine with a specific receptor (polyose) of the epithelial cell surface in the small intestine wall. destroying the brush border mucosa structure of the small intestine. nitrogen balance and nitrogen retention are.Int. similar results were reported [8. raw soybean led to a significant decrease in dry matter. In the present study. the present study discovered that fat digestibility was significantly higher in treatment groups compared with control group. for example pig. J. the extent of the decrease in the Jilin45 group was the highest. the nitrogen loss showed a linear increase. These data show that the higher the trypsin inhibitor and lectin content in soybean is. it could be concluded that different soybean varieties have different anti-nutritional factors contents. interfering with the function of many enzymes in the brush border mucosa. raw soybean was used as the source of trypsin inhibitor.21]. the study of the differences in anti-nutritional factor content in different soybean varieties and their effects on animals have important significance. thereby affecting digestion and absorption of the nutrient. These results indicate that trypsin inhibitor and lectin could lower nitrogen deposit. No. Sci. and nitrogen balance and nitrogen retention were obviously decreased. so as to decrease of protein utilization efficiency. In the present study. including nutrient digestibility and deposit. Many studies have found that protein digestibility was decreased by 20–40% in animals fed with diets containing raw soybean or high levels of trypsin inhibitor compared with those fed with diets containing heated soybean or soybean meal [22–24]. which was possibly attributed to a longer retention time of chymus in digestive tract and thorough digestion of fat. In previous studies. Acknowledgments The authors want to thank National Natural Science Foundation of China (NSFC. while that of the Jinong7 group was the smallest. Therefore. Mol.56% in the Jilin45 treatment group containing the highest level of trypsin inhibitor compared with the control group. In addition. and the results showed that protein digestibility was significantly decreased by 22. protein and nitrogen free extract digestibility. but the nitrogen balance and nitrogen retention showed a linear decrease. the lower the nutrient digestibility. and it mainly increases the effluence of endogenous nitrogen so as to decrease nitrogen balance and nitrogen retention [26–28]. 30471251) for the funding. 2010. In the present study.

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