Insights from in vitro exploration of factors influencing iron,zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereals

Developments in genetics, agronomics and processing has positioned staple cereals as important sources of iron, zinc and provitamin A (pVA) carotenoids for nutritionally vulnerable populations. Significant effort has been placed on understanding the bioavailability of these micronutrients from cereal foods, including the exploration of underlying mechanisms by which their bioavailability can be modified. While micronutrient bioavailability is preferably assessed in clinical trials, relevant in vitro digestion and intestinal cell culture models have been applied to study effects of genetic, agronomic, post-harvest and food processing on micronutrient bioavailability. This review (1) critically assesses the application of in vitro models in the exploration of mechanisms associated with iron, zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereal foods, and (2) identifies remaining gaps in order to frame future strategies to improve the nutritional impact of cereal foods.     

不同碘添加水平对牦牛瘤胃体外发酵的影响

本试验旨在研究饲粮中添加碘酸钙对牦牛瘤胃体外发酵的影响。以碘酸钙作为添加形式,以牦牛饲粮为底物进行体外瘤胃发酵,底物碘含量分别为0.1、0.3、0.5、0.7、0.9 mg/kg,发酵48h,测定发酵后的产气量、瘤胃发酵指标及消化酶活力。结果表明:当底物碘含量为0.3mg/kg,NH_3-N浓度、微生物蛋白质(MCP)含量、脂肪酶(LPS)活力、胰蛋白酶(TYS)活力、纤维素酶(CLS)活力达到最大值,分别为14.267 mg/100mL、4.619g/L、0.462U/mL、65.463U/mL、69.527U/mL,碘含量为0.3mg/kg时,乙酸(aceticacid)、丙酸(propionicacid)、总酸(TVFA)浓度达到最高值,分别为:56.142mmol/L、30.877mmol/L、87.460mmol/L,各组之间差异均不显著(P>0.05),乙酸与丙酸的比例达到最小值1.819,显著低于其它所有组乙酸与丙酸的比例(P<0.05);在底物碘含量为0.5mg/kg时,淀粉酶(AMS)活力与纤维素酶(CLS)活力达到最大值分别为1.832U/mL、69.527U/mL;在底物碘含量为0.7mg/kg时,总产气量与甲烷产量达到最大值分别为80.8mL、11.88mL。综合以上结果,在体外条件下,在底物碘含量为0.3～0.7mg/kg时,有利于瘤胃发酵和饲草料降解。     

Variation in seed protein digestion of different pea (Pisum sativum L.) genotypes by cecectomized broiler chickens: 1. Endogenous amino acid losses, true digestibility and in vitro hydrolysis of proteins

The aim of this study was to investigate the cause of variation in the digestibility of pea protein in poultry and to find a tool to select genotypes with high digestibility potential by using an in vitro hydrolysis assay. Eight pea genotypes were selected for their difference in seed protein content and composition. To reduce the variation due to tannins and particle size, seeds from these 8 genotypes were dehulled and micro-ground. They were incorporated as the only protein source in 8 different experimental isoproteinaceous diets with similar metabolisable energy content. The amino acid digestibility was studied in cecectomized chickens. A balance method was used to obtain apparent digestibility, and the isotope dilution technique was used to determine endogenous losses and true digestibility, after feeding a double labelled test meal containing chromic oxide and ¹⁵N-labelled peas. The 8 diets showed differences in apparent amino acid digestibility. The average apparent digestibility for all amino acids varied between 79.5 and 86.3%, with the highest values for arginine (85.2 to 90.8%) and glutamic acid (85.2 to 90.5%), and the lowest values for cystine (63.3 to 69.7%) and tryptophan (69.1 to 80.3%). This variability of apparent amino acid digestibility was due to variations in endogenous losses and true digestibility among the 8 pea genotypes. The average endogenous losses as determined for 9 amino acids ranged from 3.6 to 5.4% of ingested amino acids, with the highest value for threonine (8.0 to 11.0%). The average true digestibility varied between 84.4 and 90.2%, with the highest values for lysine (89.0 to 95.0%), and the lowest for isoleucine (81.0 to 88.7%) and valine (82.4 to 88.7%). In vitro hydrolysis of protein from micro-ground seeds was performed for the 8 pea genotypes using three proteases (pepsin, trypsin and chymotrypsin). The quantity of small peptides (< 3 kDa) that appeared after the combined hydrolysis with pepsin (3 h) followed by trypsin and chymotrypsin (15 min) was significantly correlated with the average true digestibility of the 8 genotypes (R = 0.74; P < 0.05).     

Effects of feed intake on in situ rumen microbial contamination and degradation of feeds

An in situ study was conducted using four rumen cannulated wethers and five feeds (corn grain (CG), soybean meal (SBM), sunflower meal (SFM), vetch–oat hay (VH), and lentil straw (LS)) to determine (using ¹⁵N infusion techniques) the effects of two intake levels (40 and 80 g DM/kg W^0.75) on the microbial contamination and rumen degradation of DM or CP. Degradation studies were also performed for neutral and acid detergent fibres (NDF and ADF, respectively) of VH. In most cases, the evolution with time of the microbial DM adhered to particles fitted well to exponential curves, which diverged between both levels of intake due to the lower values attained at high intake. However, as a consequence of the large and progressive increase of the variability of microbial contamination, statistical evidence is only partial. This behaviour was less evident for CP contamination, especially in forages. The lack of these corrections underestimated (P < 0.001) the effective degradability (ED) of all tested fractions, with important differences between feeds. The intake level had not relevant effects on the distribution of soluble, insoluble available and unavailable fractions of DM or CP (or fibres in VH), but determined some changes in their degradation rates (k_d). Direct relations between these last values and rumen outflow rates (k_p) were shown in some feeds for the degradation of CP and also of DM in concentrates, whereas negative relations were shown for the degradation rates of DM in forages (or NDF and ADF of VH). All ED estimates decreased with the intake rise, but for CP this effect (P < 0.05) was only detected in SBM and VH. These decreases represent 40.8 (CG), 80.8 (SBM), 62.1 (SFM), 66.8 (VH), and 19.5% (LS) than those estimated considering only k_p changes, which shows the interest to consider conjointly the variations of both rates to study the effect of diet changes on ED.     

Mango explant browning: Effect of ontogenic age,mycorrhization and pre-treatments

The in vitro performance of mango shoot culture is delimited by several factors, of which explant necrosis and media browning are considered as major constraints for successful exploitation of such an important commercial crop. Our results showed that source of explants had a significant effect on the performance of in vitro cultures of mango. The variations in survival of explants collected from glasshouse grown seedlings and field-grown stock plants indicated towards differences between their physiological/ontogenic age. Though, chronologically, both are young, the glasshouse grown shoots are ontogenetically younger and; therefore, responded better over field-grown shoots. Improved performance of explants harvested from mycorrhizal plants suggests integration of mycorrhizal biotechnology with tissue culture biotechnology in order to achieve better results as mycorrhiza being a well-known stress alleviator may help explants mitigate in vitro stresses. Furthermore, shoot tip explants of field-grown ‘Amrapali’ mango was assayed for their browning propensity at pre- and post-culture stages. Status of different bio-chemicals such as in vivo phenol, in vitro phenol exudation, phenylalanine ammonia lyase, peroxidase and polyphenol oxidase as affected by various pre-treatments were estimated to establish the relationship of these bio-chemicals with necrosis of mango shoot cultures. The different pre-treatments comprised etiolation of newly emerged shoots (5–7 days old) of stock plants using black polythene for 7 days, etiolation treatment + agitation of explants in antioxidant solution (antioxidants: ascorbic acid at 100 mg l⁻¹ + citric acid at 50 mg l⁻¹), etiolation treatment + agitation of explants in polyvinylpyrrolidone solution (PVP) at 0.2% and non-etiolated control. Of these, explants treated with PVP proved to be superior to other treatments with regards to explant necrosis percentage as such explants exhibited least activities of phenols and oxidative enzymes. Correlation studies indicated existence of high positive correlation between explant necrosis and these bio-chemicals.     

雪莲果离体培养及植株再生体系的建立

对雪莲果幼嫩叶片和腋芽离体培养与植株再生过程进行了研究,建立了雪莲果的再生体系.该体系以雪莲果叶片和腋芽为材料进行离体培养,结果表明:MS可作为雪莲果再生体系的基本培养基;叶片、腋芽诱导愈伤组织和不定芽培养基为MS 6-BA 2/0 mg/L NAA 0.2mg/L 蔗糖3%;继代培养基为MS 6-BA 1.0 mg/L NAA 0.1 mg/L 蔗糖3%;生根培养基为1/2 MS IAA 0.3 mg/L 蔗糖3%.     

寒富苹果叶片离体再生及四倍体诱导

为建立寒富苹果高效的离体再生体系和多倍体诱导体系,以试管苗叶片为外植体,研究了培养基中激素对寒富叶片再生芽的影响及适宜的四倍体诱导方法。结果表明,当培养基中BA质量浓度为0.5mg/L时,再生频率最高达35.7%;当培养基中BA质量浓度为2.5mg/L时,再生频率超过90%,平均再生芽数在4以上。在培养基中附加1.0mg/LTDZ,再生频率达100%,平均再生芽数达19.47。以附加15、30、60、120mg/L秋水仙素的液体再生培养基处理叶片5d,各个质量浓度处理均诱导出四倍体植株,诱变率在5.3%～22.2%之间;寒富苹果叶片在附加50mg/L秋水仙素固体再生培养基上处理5d亦获得了四倍体植株。研究结果表明寒富苹果叶片具有极强的不定芽再生能力,叶片再生过程中进行秋水仙素处理是获得其四倍体的一个有效途径。     

利用胚性细胞悬浮系研究香蕉枯萎病抗性离体筛选技术

以香蕉(Musa spp. AAA group)胚性细胞悬浮系(Embryogenic Cell Suspension,ECS)为试材,进行抗枯萎病离体筛选技术研究。首先测试不同的γ射线(60Co)辐射剂量对ECS植株再生能力的影响;在此基础上,以不同浓度的枯萎病病原菌毒素粗滤液(FOC)为选择压力,研究FOC对ECS植株再生能力的影响,并进行离体抗性筛选。结果表明,ECS的植株再生能力与辐射剂量之间呈负相关,半致死剂量为70～80Gy。同样地,未经辐射处理的ECS其植株再生能力也随着FOC体积分数的增加而急剧下降,但经辐射处理过的ECS其植株再生能力与CK相比成倍地增加;当FOC体积分数达到12%后,只有从经80Gy辐射后的ECS获得了再生植株。这表明ECS耐FOC的能力随着辐射剂量的增加而增强。     

小鼠胚胎体外培养的研究——从囊胚期培养发育到体节期

在小鼠胚胎体外培养研究中,为筛选出适合囊胚期胚胎发育的最佳培养液,本研究分三个试验组进行。第一试验组的培养液主要由CMRL-1066和人脐带血清（HCS）组成;第二试验组的培养液是CMRL-1066加上胎牛血清（FCS）;第三试验组用TCM-199添加少量碳酸氢钠做培养液。在适宜的体外培养条件下,不同培养液获得了不同的培养成绩。试验结果表明：具有HCS的CMRL-1066培养液是把小鼠从囊胚期胚胎在体外条件下培养至体节期的最佳培养液;添加FCS的CMRL-1066培养液不够理想;而加进少量碳酸氢钠的TCM-199培养液则完全不适合培养囊胚期小鼠胚胎。     

美铁芋的离体培养与快速繁殖研究

试验结果表明:美铁芋离体培养的最佳外植体是成熟的叶片,0.5%丙酸钠可有效控制初代培养中内源细菌污染,1/2MS比MS和2/3MS更适合愈伤组织诱导与芽增殖;在增殖培养中,VB1浓度以0.4mg/L为佳,KT不适合芽的增殖与生长,6-BA以5.0mg/L为佳;诱导愈伤组织的最佳培养基为1/2MS+6-BA3.0mg/L+NAA0.2mg/L,芽增殖的最佳培养基为1/2MS+6-BA5mg/L+NAA0.2mg/L,最佳生根培养基为12MS+NAA0.5mg/L。