近红外反射光谱(NIRS)技术分析奶粉品质的研究

奶粉中蛋白质和脂肪是影响奶粉营养品质的主要因素,利用近红外反射光谱分析技术对来自国内不同地区的奶粉共900份样品进行蛋白质和脂肪成分测定分析。研究了不同的样品数日、光谱预处理和散射校正技术对发展奶粉近红外测定定标模型的影响。结果表明．在样品数目为200—400范围内建立的定标分析模型较理想;数学预处理中以一阶导数较好,且以“1,4,4,1”的处理组合最为理想;光谱散射校正中采用“标准正态变量转换(SNV) 趋势变换法(De—trending)”的组合建立回归方程效果较好。利用改进最小二乘法回归技术(Modified PLS)建立多种定标模型,并进行交叉验证(cross—Validation)来分析各种因素对定标模型的影响。同时筛选出较理想的蛋白质和脂肪定标分析回归方程,其中蛋白质和脂肪含量的相关系数高速0．973和0．850。探讨了NIRS技术在建模应用中的一些影响因素,以及由NIRS技术建立奶粉分析模型用于快速分析和在线检测的可行性。     

家蚕核型多角体病毒酪氨酸蛋白磷酸酯酶基因核苷酸序列分析

从家蚕核型多角体病毒中国镇江株 (BmNPV ZJ)基因组DNA中克隆出酪氨酸蛋白磷酸酯酶基因 (ptp) ,该基因的编码部分由 5 0 7个核苷酸组成 ,其中A为 16 3、C为 99、G为 113、T为 132 ,G +C含量约为 4 2 %。根据其核苷酸序列推演的蛋白质由 16 8个氨基酸残基组成 ,其中含有酪氨酸蛋白磷酸酯酶催化活性区的 11个氨基酸“HC”基序。该基因与苜蓿银纹夜蛾核型多角体病毒 (AcMNPV)ptp和BmNPV T3株 (日本 )的ptp核苷酸序列的同源性分别为 96 8%和 98 2 %。BmNPV ZJ酪氨酸蛋白磷酸酯酶 (BmNPV ZJPTPase)的氨基酸全序列与AcMNPV、BmNPV T3、芹菜夜蛾核型多角体病毒 (AfMNPV)PTPase和黄杉毒蛾核型多角体病毒 (OpMNPV)PTPase 1的氨基酸全序列的同源性分别为 97%、97 6 %、96 %和 6 0 % ,而与OpMNPVPTPase 2的同源性仅为 2 0 %。在NCBI数据库中查找BmNPV ZJPTPase的同源性序列 ,查找到的 5 99381个序列中发现至少有 14种mRNA加帽酶其N端部分存在PTPase催化活性区的“HC”基序 ,但其氨基酸全序列的同源性只有 31%～ 32 %。该基因序列已被GenBank数据库收录 ,登录号为AF316 871     

黄皮种子脱水敏感性与核酸、蛋白质代谢的关系

黄皮种子发育晚期,胚内核酸和蛋白质合成能力增强,而花生种子发育晚期则呈下降趋势。脱水处理使生理成熟期黄皮胚核酸和蛋白质合成能力急剧下降,核酸水解酶活性增强。生物大分子代谢能力的变化是黄皮种子脱水敏感性的分子基础。     

Influence of dietary lysine level on whole-body protein turnover, plasma IGF-I, GH and insulin concentration in growing pigs

Four growing pigs (initial liveweight 25.9 ± 0.54 kg, final liveweight 43.0 ± 1.06 kg) were used to study the effect of dietary lysine level on nutrient digestibility, whole-body protein turnover, plasma insulin-like growth factor-I (IGF-I), growth hormone (GH), insulin, glucose, and urea nitrogen (PUN). Four diets, containing 7.0 g (L1), 9.5 g (L2), 12.0 g (L3) and 14.5 g (L4) lysine per kg diet respectively, were formulated as experimental treatments. The animals and diets were allocated in a 4 × 4 Latin square design. Nitrogen (N) metabolism and whole-body protein turnover were measured by classical method and single-dose ¹⁵N end-product method, respectively. The blood samples were taken at the end of each experimental period. Results showed that N retention (NR) and N biological value (NBV) were significantly increased from L1 to L4 (P < 0.05). However, differences in NR and NBV between L2, L3 and L4 were not significant (P > 0.05). There was no significant difference on dry matter (DM) digestibility, organic matter (OM) digestibility and N digestibility between different treatments (P > 0.05). Whole-body protein synthesis, protein degradation and protein accretion increased markedly from L1 to L2 (P < 0.05), but did not increase further from L2 to L4. Whole-body protein accretion (y, g/kg W^0.75/d) increased with dietary lysine (x, g/kg) in a quadratic manner: y = − 0.09x² + 2.12x − 5.14 (r² = 0.96, n = 4, P < 0.05).The results also showed that differences in plasma IGF-I, GH, glucose and PUN concentration between different treatments were not significant (P > 0.05). Plasma insulin concentration (y, μIU/ml) was increased with dietary lysine (x, g/kg) in a quadratic manner: y = 0.23x² − 4.10x + 32.25 (r² = 0.99, n = 4, P < 0.05), but it was not found that plasma insulin concentration was related to NR. A significant correlation was found between NR (y, g/d) and plasma IGF-I (x, ng/ml): y = − 3.1 × 10^− 3x² + 1.31x − 122.28 (r² = 0.99, n = 4, P < 0.05).It was concluded that dietary lysine level had a significant influence on NR and whole-body protein turnover but not on plasma IGF-I and GH concentration. Plasma IGF-I may be an important factor controlling N metabolism of growing pigs. Further research was needed to study the mechanism.     

The effect of dietary crude protein concentration and inulin supplementation on nitrogen excretion and intestinal microflora from finisher pigs

A 2 × 2 factorial experiment was used to investigate the interaction between dietary crude protein (CP) concentration (200 vs 140 g/kg) and inulin supplementation (0 vs 12.5 g/kg) on nitrogen (N) excretion and intestinal microflora from 16 boars (n = 4, 74.0 kg live weight). The diets were formulated to contain similar concentrations of digestible energy and lysine. Pigs offered the high CP diets had a higher excretion of urinary N (P < 0.01), faecal N (P < 0.01) and total N (P < 0.001) than the pigs offered the low CP diets. Inulin supplementation increased faecal N excretion (P < 0.05) and decreased urine: faeces N ratio (P < 0.05) compared to the inulin free diets. There was no significant effect (P > 0.05) of dietary treatment on N retention. There was an interaction (P < 0.05) between dietary CP concentration and inulin supplementation on caecal E.coli. Pigs offered the diet containing 200 g/kg CP plus inulin decreased the population of E.coli compared to the inulin supplemented 140 g/kg protein diet. However, CP concentration had no significant effect on the population of E.coli in the unsupplemented diets. Inulin supplementation increased caecal bifidobacteria (P < 0.01) compared to the inulin free diets. In conclusion, inulin supplementation favourably altered N excretion and lowered the population of E.coli at high CP concentrations.     

Dietary fibre and indigestible protein increase ileal glycoprotein output without impacting colonic crypt goblet cells in weaned piglets

Mucus plays an important role in gut health by favouring colonisation resistance. The aim of this study was to quantify the impact of fibre and protein on mucin recovery in ileal digesta and on goblet cell histochemistry in the proximal colon. A control diet with highly digestible protein and low fibre and three complex diets containing indigestible protein associated with low, soluble or insoluble fibre sources were tested for 14 days in piglets weaned at 28 days of age. Mucin concentration was determined by ethanol precipitation. Goblet cell subtypes in colonic crypts were analysed by histochemistry. The ileal mucin output was higher with the complex diets than with the control diet (31.6 on average vs 21.7 g/kg DM intake). Increases observed with the low and soluble fibre diets were similar (34 g/kg DM intake). This suggests a limited effect of soluble fibre in presence of indigestible protein. Surprisingly, the observed increase was lower with insoluble fibre (27.2 g/kg DM intake). DM and N output and digestibility were not affected by the diets, but a linear relationship was found between DM and mucin output. In proximal colon, crypt depth, goblet cell numbers per crypt and glycosylation subtypes were not affected by the diet suggesting no change in the capacity of mucus to protect the gut. To conclude, introducing highly indigestible protein in the diet increased ileal mucin output, without effects of these factors on crypt goblet cell patterns in the proximal colon. Introducing fibre in a diet containing highly indigestible protein had a marginal effect.     

Effect of feeding fermentable fiber on synthesis of total and mucosal protein in the intestine of the growing pig

In a previous study, a reduced efficiency of ileal digestible threonine (THR) use for body protein deposition was observed in growing pigs when pectin was included in the diet. This response was not due to increased physical endogenous ileal THR loss. Our aim was to explore the contribution of diet-induced increases in protein synthesis in the colon, especially mucins, to dietary THR requirements. Twelve barrows (21 kg mean BW) were fed either a cornstarch–soybean meal-based diet (Control) or Control with 12% pectin (Pectin). Pigs were given intravenously 1.5 mmol/kg BW of L-1-¹³C valine (40 mol%) to measure fractional and absolute synthesis rates (FSR, ASR, respectively) of mucosal and whole intestinal protein in the jejunum and colon. Dietary pectin inclusion increased plasma levels of glucose, isoleucine and glutamine (P < 0.05) but had no effect on insulin or urea nitrogen (P > 0.10). There were no differences in FSR and ASR of whole intestinal protein in jejunum and colon (P > 0.10). The FSR of mucosal proteins in colon, not in jejunum, was increased with dietary pectin supplementation (P < 0.05). Assuming mucosal protein mass is constant, these results imply that the higher protein synthesis in colon mucosa contributes to the reduced THR efficiency observed in pectin-supplemented diet.     

牛传染性鼻气管炎病毒gD基因的克隆与表达

构建pET32a重组表达载体,转化到BL21(DE3),诱导表达牛传染性鼻气管炎病毒重组gD蛋白。用已发表的IBRV的gD基因序列设计一对特异性引物,通过PCR方法扩增一条766bp的目的基因gD,将gD基因克隆到原核表达载体pet32a,得到重组表达载体pet32-gD,转化到BL21(DE)中,通过IPTG诱导获得融合蛋白。重组表达蛋白经纯化后,免疫印迹分析。结论证明表达的重组蛋白具有良好的免疫原性。