牛乳过敏原β-乳球蛋白检测方法的研究进展

目的食物过敏已成为全球性的食品安全问题,欧美等发达国家均要求对食品中的过敏原成分进行标识,其中就包括作为八大过敏性食物之一的牛乳及其乳制品。β-乳球蛋白是牛乳中的主要过敏原,约占乳清蛋白的50%,牛乳总蛋白的10%,并且约有82%的牛乳过敏患者对β-乳球蛋白过敏,因此其可以作为检测食品中是否含牛乳蛋白的一个有效的标志物。建立灵敏、可靠的β-乳球蛋白检测方法,对牛乳过敏原标识及保障牛乳过敏人群的安全消费具有重要意义。本文主要综述了牛乳β-乳球蛋白的高效液相色谱法、超高效液相色谱法、液相色谱-质谱联用法、酶联免疫吸附法、免疫层析法、电化学免疫法、蛋白微阵列法、等离子体共振法等色谱学和免疫学检测方法的研究进展,并对未来发展方向作出展望,以期促进牛乳β-乳球蛋白等过敏原检测方法的研究与开发。     

牛乳中脂肪测定不确定度评定及质控图 建立的研究

目的本研究旨在借助测量不确定度评定用以表征合理地赋予被测量之间的分散性,了解测量结果所处的范围,利用质控图法分析影响测量结果的主要因素,从而减少测量的误差。方法根据GB/T5413.3-2010《食品安全国家标准婴幼儿食品和乳品中脂肪的测定》第1法。结果利用纯牛奶CS样品(核查样品Check the sample,即核查样品指质量控制测量中附有参考量值的物料,指定实验室检测样品的检测值)。在期间精密度测量条件下,对牛乳检测中的脂肪进行不确定度评定。结论该不确定度评价方法在实验室质控分析工作中有较强的实用价值。     

Foods for Special Dietary Needs: Non-dairy Plant-based Milk Substitutes and Fermented Dairy-type Products

A growing number of consumers opt for plant-based milk substitutes for medical reasons or as a lifestyle choice. Medical reasons include lactose intolerance, with a worldwide prevalence of 75%, and cow's milk allergy. Also, in countries where mammal milk is scarce and expensive, plant milk substitutes serve as a more affordable option. However, many of these products have sensory characteristics objectionable to the mainstream western palate. Technologically, plant milk substitutes are suspensions of dissolved and disintegrated plant material in water, resembling cow's milk in appearance. They are manufactured by extracting the plant material in water, separating the liquid, and formulating the final product. Homogenization and thermal treatments are necessary to improve the suspension and microbial stabilities of commercial products that can be consumed as such or be further processed into fermented dairy-type products. The nutritional properties depend on the plant source, processing, and fortification. As some products have extremely low protein and calcium contents, consumer awareness is important when plant milk substitutes are used to replace cow's milk in the diet, e.g. in the case of dairy intolerances. If formulated into palatable and nutritionally adequate products, plant-based substitutes can offer a sustainable alternative to dairy products.     

Milk fatty acids in dairy cows fed whole crude linseed, extruded linseed, or linseed oil, and their relationship with methane output

This experiment studied the effect of 3 different physical forms of linseed fatty acids (FA) on cow dairy performance, milk FA secretion and composition, and their relationship with methane output. Eight multiparous, lactating Holstein cows were assigned to 1 of 4 dietary treatments in a replicated 4 × 4 Latin square design: a control diet (C) based on corn silage (59%) and concentrate (35%), and the same diet supplemented with whole crude linseed (CLS), extruded linseed (ELS), or linseed oil (LSO) at the same FA level (5% of dietary dry matter). Each experimental period lasted 4 wk. Dry matter intake was not modified with CLS but was lowered with both ELS and LSO (−3.1 and −5.1 kg/d, respectively) compared with C. Milk yield and milk fat content were similar for LSO and ELS but lower than for C and CLS (19.9 vs. 22.3 kg/d and 33.8 vs. 43.2 g/kg, on average, respectively). Compared with diet C, CLS changed the concentrations of a small number of FA; the main effects were decreases in 8:0 to 16:0 and increases in 18:0 and cis-9 18:1. Compared with diet C (and CLS in most cases), LSO appreciably changed the concentrations of almost all the FA measured; the main effects were decreases in FA from 4:0 to 16:0 and increases in 18:0, trans-11 16:1, all cis and trans 18:1 (except trans-11 18:1), and nonconjugated trans 18:2 isomers. The effect of ELS was either intermediate between those of CLS and LSO or similar to LSO with a few significant exceptions: increases in 17:0 iso; 18:3n-3; trans-11 18:1; cis-9, trans-11 conjugated linoleic acid; and trans-11, trans-13 conjugated linoleic acid and a smaller increase in cis-9 18:1. The most positive correlations (r = 0.87 to 0.91) between milk FA concentrations and methane output were observed for saturated FA from 6:0 to 16:0 and for 10:1, and the most negative correlations (r = −0.86 to −0.90) were observed for trans-16+cis-14 18:1; cis-9, trans-13 18:2; trans-11 16:1; and trans-12 18:1. Thus, milk FA profile can be considered a potential indicator of in vivo methane output in ruminants.     

一种牛乳酪蛋白肽粉复合物对睡眠的改善作用

该研究探讨一包含牛乳酪蛋白肽粉、酸枣仁、百合、γ-氨基丁酸的复合物改善睡眠的功效。利用秀丽隐杆线虫模型研究复合物及复合物中各个功效成分改善睡眠的效果,然后使用体内小鼠模型评价复合物改善睡眠作用,并测定小鼠下丘脑中神经递质含量。与空白组相比,复合物组显著延长线虫睡眠时间,各指标均优于复合物中单个组分的作用结果。复合物低、中、高剂量组小鼠睡眠潜伏期分别提高18.18%、20.60%、25.60%,睡眠时长分别提高34.54%、45.24%、47.29%,入睡率分别提高25.00%、33.33%、33.33%。在旷场实验中复合物组运动总路程分别降低15.06%、13.96%、9.58%,进入中央区域次数分别提高138.10%、114.29%、114.29%,中央区域停留时间分别提高142.98%、176.89%、114.25%、静止总时间分别增加28.13%、44.42%、55.40%。与空白组相比,低中高复合物剂量组小鼠下丘脑GABA含量显著提高34.74%、66.45%、76.99%,5-HT含量显著提高32.67%、50.34%、79.72%,Glu含量显著降低33.41%、34.83%、39.55%。综上所述,牛乳酪蛋白肽粉、酸枣仁、百合、γ-氨基丁酸的复合物能够改善睡眠,通过调节下丘脑神经递质水平来改善睡眠,研究结果为新食品资源开发提供理论参考。     

Prepartum dietary management of energy intake affects postpartum intake and lactation performance by primiparous and multiparous Holstein cows

An experiment was conducted to determine the effect of plane of energy intake prepartum on postpartum performance. Primiparous (n = 24) and multiparous (n = 23) Holsteins were randomly assigned by expected date of parturition to 1 of 3 prepartum energy intakes. A moderate energy diet [1.63 Mcal of net energy for lactation (NE_L)/kg; 15% crude protein (CP)] was fed for either ad libitum intake (OVR) or restricted intake (RES) to supply 150 or 80% of National Research Council (2001) energy requirement, respectively, for dry cows in late gestation. To limit energy intake to 100% of NRC requirement at ad libitum dry matter intake (DMI), chopped wheat straw was included as 31.8% of dry matter (DM) in a control diet (CON; 1.21 Mcal of NE_L/kg of DM; 14% CP). Multiparous and primiparous cows assigned to OVR gained body condition during the dry period [initial body condition score (BCS) = 3.3], but were not overconditioned by parturition (BCS = 3.5). Multiparous cows in the OVR group lost more BCS postpartum than multiparous RES or CON cows. Primiparous cows lost similar amounts of BCS among dietary treatment groups postpartum. Addition of chopped wheat straw to CON diets prevented a large decrease in DMI prepartum in both primiparous and multiparous cows. During the first 3 wk postpartum, DMI as a percentage of BW was lower for multiparous OVR cows than for multiparous RES cows. Prepartum diet effects did not carry over through the entire 8-wk lactation period. Because of greater mobilization of body stores, OVR cows had greater milk fat percentage and greater 3.5% fat-corrected milk yield during the first 3 wk postpartum. Multiparous cows assigned to OVR experienced a 55% decrease in energy balance and primiparous cows a 40% decrease in energy balance during the last 3 wk before parturition, compared with CON or RES cows that had little change. Multiparous cows fed OVR had a greater contribution of energy from body energy reserves to milk energy output than either CON or RES cows. Overfeeding energy prepartum resulted in large changes in periparturient energy balance. Even in the absence of overconditioning, a large change in DMI and energy balance prepartum influenced postpartum DMI and BCS loss, especially for multiparous cows. Chopped wheat straw was effective at controlling energy intake prepartum, although primiparous cows did not achieve predicted DMI. Even so, controlling or restricting energy intake in primiparous cows was not detrimental to lactational performance over the first 8 wk of lactation.     

Short communication: Diurnal profiles of conjugated linoleic acids and trans fatty acids in ruminal fluid from cows fed a high concentrate diet supplemented with fish oil, linseed oil, or sunflower oil

Trans-18:1 and 18:2 isomer composition in ruminal fluid during the daily feeding cycle was examined in 3 cows fed a high concentrate diet (35:65) with 5% (DM basis) sunflower oil (SO), 5% linseed oil (LO), or 2.5% fish oil (FO) in a 3 x 3 Latin square with 3 4-wk periods. Grass hay and concentrate mixtures were fed at 0900, 1300, and 1700 h daily. Ruminal fluid was collected at 0900, 1100, 1300, 1500, 1700, 2000, and 0000 h. Feeding SO resulted in the greatest mean concentrations (% of total fatty acids) of trans10,cis12-18:2 and cis9,trans11-18:2. In particular, trans10,cis12-18:2 with SO was greater at 1500 (0.29%), 2000 (0.34%), and 0000 h (0.25%) relative to 0900 h (0.07%). Cis9,trans11-18:2 concentration increased from 0.47% at 0900 h to a peak of 2.06% at 1100 h; it remained greater than the percentage determined at 0900 h at 1300 (1.4%) through 0000 h (1.1%). Concentration of trans11,cis15-18:2 was greatest with LO, ranging from 3.3% (0900 h) to a peak of 11.4% at 2000 h. Mean trans10-18:1 concentration ranked by diet was SO > FO > LO. Peak trans10-18:1 with SO was observed at 1700 h (14.9%) compared with 0900 h (5.1%). Trans11-18:1 did not differ with diet or time. Stearic acid decreased over time with all diets reaching minimum concentrations at 1700 to 2000 h relative to 0900 h. Feeding FO, however, decreased mean 18:0 concentration 4-fold compared with LO or SO. The moderate effect on concentration of trans-18:1 coupled with accumulation of 18:2 intermediates and the decrease of 18:0 over time suggest that oils reduced the biohydrogenation of 18:2 isomers to trans-18:1.     

具有T细胞口服免疫耐受作用的β-乳球蛋白水解物的制备及鉴定

以牛乳中主要过敏原β-乳球蛋白为研究对象,制备和鉴定具有T细胞表位和口服免疫耐受性的水解物,旨在为牛乳过敏患者口服免疫治疗方法提供理论依据。采用生物信息学方法预测β-乳球蛋白的T细胞表位,通过质谱分析6种蛋白酶水解β-乳球蛋白水解物的氨基酸序列,用T细胞增殖实验鉴定水解物中多肽免疫耐受作用,体内实验测定小鼠血清特异性抗体(IgE、IgG₁、IgG_2a)、Th1细胞因子(IFN-γ、IL-17)、Th2细胞因子(IL-4、IL-5、IL-13)、组胺、几丁质酶-3样蛋白1的水平及小鼠脾脏细胞亚群的变化水平,验证β-乳球蛋白水解物的口服免疫治疗活性。研究结果表明：胰蛋白酶、复合蛋白酶和木瓜蛋白酶水解物具有口服耐受性,其中复合蛋白酶和木瓜蛋白酶水解物具有口服耐受性是创新性发现。中性蛋白酶水解物虽然含有T细胞表位,但是仍然具有致敏性。因此含有T细胞表位的水解物不一定具有口服免疫耐受性,需要体内实验验证。研究结论以期为新型抗过敏乳基料的开发和临床治疗牛乳过敏提供参考数据。