果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及风味影响的研究

试验旨在研究果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及培养液风味的影响。试验选用自制的奶牛乳腺上皮细胞,设5个处理组,果香味剂的添加剂量分别为0、5、10、15和20μg/mL。结果表明:①当培养基中果香味剂添加量为15μg/mL时,乳腺上皮细胞培养液中总蛋白水平、甘油三酯及乳糖含量均达到最高,其中对脂肪含量影响显著(P<0.05),而对蛋白和乳糖含量影响不显著(P>0.05),之后呈下降趋势,说明果香味剂可能会诱导奶牛乳腺上皮细胞乳蛋白、乳脂和乳糖分泌水平的升高;②运用PT/GC-MS分析乳腺培养液中的风味成分,与对照组相比,各试验组中除具有原有乳腺培养液的风味外,还有乙酸异丁酯、乙酸异戊酯、丙酸异戊酯、丙酸戊酯、丁酸异丁酯、丁酸戊酯、丁酸乙酯、邻苯二甲酸二异丁酯等物质成分,而这些物质成分正好是果香味剂中进入到乳腺培养液中的风味物质,且当添加剂量为15μg/mL时,各物质成分含量为最高(P<0.05),之后呈下降趋势。果香味剂能显著影响奶牛乳腺上皮培养液中乳成分含量和风味。     

果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及风味影响的研究

试验旨在研究果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及培养液风味的影响.试验选用自制的奶牛乳腺上皮细胞,设5个处理组,果香味剂的添加剂量分别为0、5、10、15和20 μg/mL.结果表明:①当培养基中果香味剂添加量为15 μg/mL时,乳腺上皮细胞培养液中总蛋白水平、甘油三酯及乳糖含量均达到最高,其中对脂肪含量影响显著(P＜0.05),而对蛋白和乳糖含量影响不显著(P＜0.05),之后呈下降趋势,说明果香味剂可能会诱导奶牛乳腺上皮细胞乳蛋白、乳脂和乳糖分泌水平的升高;②运用PT/GC-MS分析乳腺培养液中的风味成分,与对照组相比,各试验组中除具有原有乳腺培养液的风味外,还有乙酸异丁酯、乙酸异戊酯、丙酸异戊酯、丙酸戊酯、丁酸异丁酯、丁酸戊酯、丁酸乙酯、邻苯二甲酸二异丁酯等物质成分,而这些物质成分正好是果香味剂中进入到乳腺培养液中的风味物质,且当添加剂量为15μg/mL时,各物质成分含量为最高(P＜0.05),之后呈下降趋势.果香味剂能显著影响奶牛乳腺上皮培养液中乳成分含量和风味.     

果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及风味影响的研究

试验旨在研究果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及培养液风味的影响。试验选用自制的奶牛乳腺上皮细胞,试验设5个处理组,果香味剂的添加剂量分别为0、5、10、15和20μg/ml。结果表明:①当培养基中果香味剂添加量为15μg/ml时,乳腺上皮细胞培养液中总蛋白水平、甘油三酯及乳糖含量均达到最高,其中对脂肪含量影响显著(P<0.05),而对蛋白和乳糖含量影响不显著(P>0.05),之后呈下降趋势,说明果香味剂可能会诱导奶牛乳腺上皮细胞乳蛋白、乳脂和乳糖分泌水平的升高;②运用PT/GC-MS分析乳腺培养液中的风味成分,与对照组相比,各试验组中除具有原有乳腺培养液的风味外,还有乙酸异丁酯、乙酸异戊酯、丙酸异戊酯、丙酸戊酯、丁酸异丁酯、丁酸戊酯、丁酸乙酯、邻苯二甲酸二异丁酯等物质成分,而这些物质成分正好是果香味剂中进入到乳腺培养液中的风味物质,且当添加剂量为15μg/ml时,各物质成分含量为最高(P<0.05),之后呈下降趋势。果香味剂能显著影响奶牛乳腺上皮培养液中乳成分含量和风味。     

体外法研究不同硒源及硒水平对瘤胃挥发性脂肪酸的影响

试验旨在利用体外厌氧发酵培养技术研究不同硒源及不同硒水平的添加对奶牛瘤胃挥发性脂肪酸(VFA)含量的影响。试验按照3×4完全随机试验设计,第一因素为硒源,即亚硒酸钠、硒代蛋氨酸和酵母硒3种,第二因素为硒添加量,即每毫升培养液中设0μg、0.1μg、0.2μg和0.3μg共4个添加水平,其中0μg/mL添加水平为对照组,共计10个处理组,每个处理组均进行3h、6h、9h、12h、24h和48h的体外批次培养,每个处理组设3个重复。试验结果表明,体外培养液中添加3种不同来源的硒对乙酸、丁酸、TVFA的影响组间差异显著(P<0.05),对丙酸、乙酸丙酸比影响较小(P>0.05),其中以添加酵母硒的乙酸、丙酸、丁酸、TVFA的效果最好,添加硒代蛋氨酸次之;体外培养液中添加0.3μg/mL硒,可显著提高乙酸、丁酸、TVFA的含量(P<0.05),对丙酸及乙酸丙酸比影响组间差异不显著,但均以0.3μg/mL硒组的效果较好。     

有机硒与无机硒对奶牛体外瘤胃发酵特性的影响

本试验主要研究有机硒和无机硒对奶牛体外瘤胃发酵特性的影响。试验按照3×4二因子完全随机试验设计,其中无机硒源为亚硒酸钠(SS),有机硒源为硒代蛋氨酸(SM)和酵母硒(SY),硒浓度分别为0μg/mL、0.1μg/mL、0.2μg/mL和0.3μg/mL 4个剂量,共12个处理组,0剂量为对照组。各处理组均进行3h、6h、9h、12h和24h的体外批次培养,每个处理组设3个重复。试验结果表明,与无机硒源SS相比,有机硒源SM和SY可显著增加发酵24h体外培养液中的BCP浓度(P=0.000 6),尤以SY最好;体外培养液中添加0.3μg/mL硒,可显著提高氨氮(NH_3-N)、菌体蛋白(BCP)、硒、乙酸、丙酸、丁酸和总挥发性脂肪酸(TVFA)的浓度(P 0.05),并显著降低乙酸丙酸比(P 0.05),且添加0.1μg/mL和0.2μg/mL的硒也具有一定的促进作用;不同的硒源对体外培养液中的pH、NH_3-N、硒、乙酸、丙酸、丁酸、TVFA浓度以及乙酸丙酸比均无显著影响(P 0.05),但SY组的pH有下降趋势,NH_3-N、BCP、乙酸、丙酸、丁酸、TVFA浓度有升高趋势。说明适宜浓度的硒可促进奶牛的体外瘤胃发酵,且当硒源为SY、添加剂量为0.3μg/mL时,促进效果较好。     

微生物青贮添加剂与尿素对青贮玉米体外消化VFA的影响

微生物青贮添加剂与尿素处理的全株青贮玉米经体外消化处理,测定挥发性脂肪酸总量及乙酸、丙酸和丁酸含量。结果显示,未经任何处理组其挥发性脂肪酸总含量最低,为4.60mmol/mL;乙酸、丙酸、丁酸单项含量均低于其他各样;添加微生物青贮添加剂与尿素组的挥发性脂肪酸总含量最高,为7.32mmol/mL,高于未处理组37.16%,且丁酸含量为1.19mmol/mL介于只添加尿素组和只添加微生物青贮添加剂组之间;只添加微生物青贮添加剂组丁酸含量为1.13mmol/mL;只添加尿素组丁酸含量为1.22mmol/mL,为4个处理组中最高。结果表明,添加微生物青贮添加剂与尿素能提高青贮饲料消化后VFA的总含量,降低丁酸的含量。     

漏芦乙醇提取物对奶山羊乳腺上皮细胞乳成分合成的影响

以体外培养的奶山羊乳腺上皮细胞为模型,利用β-酪蛋白、甘油三酯及乳糖/半乳糖检测试剂盒测定漏芦乙醇提取物处理的乳腺上皮细胞培养液中β-酪蛋白、甘油三酯和乳糖含量;采用荧光定量RT-PCR检测β-酪蛋白、甘油三酯和乳糖合成代谢相关酶和蛋白因子mRNA表达的变化。结果显示,25、50、100μg/ml的漏芦乙醇提取物以浓度依赖的方式显著提高乳腺上皮细胞合成分泌β-酪蛋白、甘油三酯及乳糖含量(P<0.05)及乳腺上皮细胞中乳成分合成代谢相关酶和蛋白因子的mRNA表达水平(P<0.05)。实验结果表明漏芦乙醇提取物能促进奶山羊乳腺上皮细胞合成分泌β-酪蛋白、甘油三酯和乳糖,并能提高乳腺上皮细胞乳成分合成代谢相关酶和蛋白因子的基因表达,进而影响奶山羊乳腺上皮细胞乳蛋白、乳脂和乳糖的合成。     

不同方法分离奶牛乳腺上皮细胞体外培养的研究

从荷斯坦奶牛乳房无菌采取乳腺组织,通过不同的方法分离、培养、纯化牛乳腺上皮细胞,研究乳腺上皮细胞的体外培养效果.结果表明:组织块接种、0.25%的胰酶和100 u/mL透明质酸酶的混合液37℃消化组织块3 h.可以得到大量细胞用于原代培养.在以DMEM/F12为基础培养液,在培养液中添加10%的胎牛血清、100μg/mL的双抗、表皮生长因子、胰岛素、转铁蛋白、硒酸钠等,组成的完全培养液中奶牛乳腺上皮细胞生长良好.上皮细胞显微结构显示:奶牛乳腺上皮细胞在体外培养过程中舍有不同的细胞类型,大多数上皮细胞呈多角形,细胞之间连接成片,细胞界限明显,部分细胞界限不明显.     

气相色谱-质谱联用法测定瘤胃液中的甲酸和其他6种挥发性脂肪酸

本试验旨在建立一种测定瘤胃液中甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸的气相色谱-质谱联用方法。样品选用湘东黑山羊和荷斯坦奶牛的瘤胃液,18000 r/min离心10 min,取上层清液。将上层清液与25%的偏磷酸按照9∶1(v∶v)的比例混合均匀,静置0.5 h后,4℃条件下18000 r/min离心10 min,取上清液过0.22μm滤膜,进行气相色谱-质谱分析。以DB-FFAP毛细管柱进行分离,甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸在9 min内达到基线分离。结果表明:这7种组分色谱峰保留时间的相对标准偏差均小于0.5%,峰面积的相对标准偏差均小于5.0%。甲酸、乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸分别在0.8~121.0μg/mL、0.3~3081.0μg/mL、0.4~1165.2μg/mL、0.4~275.1μg/mL、0.2~543.0μg/mL、0.5~129.9μg/mL和0.4~271.5μg/mL内线性关系良好,线性相关系数均达到0.998以上,检出限(3倍信噪比)在0.069~0.252μg/mL。山羊和奶牛瘤胃液样品的加标回收率分别为91.8%~103.8%和78.7%~95.2%。本试验建立的测定瘤胃液中甲酸以及其他6种挥发性脂肪酸的气相色谱-质谱联用方法选择性强、样品用量少、操作简单,可为反刍动物瘤胃碳水化合物代谢及甲烷生成的相关研究提供技术支撑。     

芦丁对体外培养奶牛乳腺上皮细胞生长的影响

本实验旨在研究芦丁对奶牛乳腺上皮细胞生长的影响,实验将奶牛乳腺上皮细胞分成5组,每组细胞培养基中分别含有0、25、50、100、150μg/mL芦丁,细胞在37℃,5%CO2条件下培养48 h和96 h后测定相关指标。结果表明:细胞培养48 h,与0μg/mL芦丁组相比,100μg/mL芦丁组的细胞活性、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)活性均显著提高,而丙二醛(MDA)含量和乳酸脱氢酶(LDH)活性均显著降低;细胞培养96 h,100μg/mL和150μg/mL芦丁组的细胞活性、GSH-Px和SOD活性均显著提高,而MDA含量和CAT、LDH活性均显著降低。与0μg/mL芦丁组相比,100μg/mL芦丁组细胞的Bcl-2表达显著升高,而Caspase3和P53表达均显著降低。综上,芦丁能够提高奶牛乳腺上皮细胞的抗氧化能力和抑制细胞凋亡,进而促进细胞的增殖,其中添加100μg/mL芦丁的效果最好。     

果味剂对奶风味及奶牛生产性能的影响

试验选择了24头泌乳前中期的同一胎次、健康无病、具有相近体重、产奶量与泌乳期的荷斯坦奶牛,将其分为4组,每组6头牛,在奶牛日粮中添加苹果味果味剂、香蕉味果味剂、混合香型果味剂,并设置对照组(不添加果味剂)进行奶牛饲养试验。对奶牛的生产性能以及奶样中风味物质进行评定。试验结果表明:①在奶牛生产性能方面,与对照组相比,3种果味剂都有减缓产奶量下降的趋势,其中香蕉味的作用效果最好(产奶量下降:香蕉味0.5%,苹果味11.9%,混合味8.86%,对照13.5%)。②对奶样进行感官评价,在单独的色泽、组织状态、奶香味、果香味、甜味、酸味、苦味、涩味、饲草味和牛体味中,4种样品基本没有差异,4个样品的综合风味存在显著差异(P<0.05),在风味的总得分上,混合风味>苹果风味>香蕉风味=对照组。     

猪乳风味物质的SDE-GC-MS和SPME-GC-MS分析鉴定

研究采用同时蒸馏-气相色谱-质谱(SDE-GC-MS)和固相微萃取-气相色谱-质谱(SPME-GC-MS)法对猪乳风味物质进行分析鉴定。结果表明:同时蒸馏提取鉴定出分子量较小的挥发性物质21种,其中顺-1-甲基-4-(1-甲基乙基)-环己烷、萜烯醇和反-1-甲基-4-(1-甲基乙基)-环己烷为3种主要成分;而固相微萃取提取鉴定出分子量相对较大的挥发性物质22种,其中芥酸、环己基甲酸十二醇酯和十六烷基环己基甲酸酯为主要的3种;同时蒸馏提取的风味物质出峰时间均早于固相微萃取,综合两者分析显示,猪乳中主要风味成分是酯、脂肪酸、烷烃、醛、醇、酮等化合物。研究结果对模拟猪乳乳香风味,开发仔猪用饲料香味剂具有一定的参考辅助作用。     

Research Progress on Influence Factors of Mutton Flavor

The flavor of mutton affects the acceptance of consumers.Adipose tissue is the most obvious source of mutton flavor, and the compounds presenting mutton flavor mainly include branched chain fatty acids, aldehyde compounds, phenolic compounds, ketone substance and so on.Mutton flavor is affected by genetic factors such as breed, sex, age, nutrition factors such as the nutrient level of diet and additive, feeding model and disease, etc.The effect of breed on the flavor may be achieved by the genetic control on fat composition and metabolism, and the amount of fat oxidation products is the main reason for different flavor among species.There are differences in fatty content and composition because of different sex, and it also has an effect on the flavor of mutton.Along with the growth of sheep, the deposits of subcutaneous fat, the proportion of saturated fatty acid, and the concentration of the lipid oxide increase, all these led to strong flavor of the lamb.Each parts of the lamb has different flavor because of the differences kind and content of the volatile compounds.The types of diet have a certain influence on mutton flavor, such as high protein feed, high energy feed, flavor plant, seeds with high content of unsaturated fatty acid and other additives.Feeding mode and disease factor also can make a change of mutton flavor.Other factors, such as the feeding density, illumination and so on, also have effects on the flavor of mutton in varying degrees.The flavor of mutton can be improved through breed selection, nutrition regulation and scientific feeding and management during sheep production.     

影响羊肉风味的因素研究进展

羊肉的风味在一定程度上影响消费者的接受度,因此对羊肉风味的研究备受关注。脂肪组织是羊肉最明显的风味来源,而呈现羊肉风味的化合物主要有支链脂肪酸、醛类化合物、酚类化合物及酮类物质等。品种、性别和年龄等遗传因素、饲料营养水平及各种添加物质等营养因素、饲养方式及疾病等因素均可影响羊肉风味。脂肪组成和代谢的遗传控制可以形成品种特有的风味,而脂肪氧化产物碳酰化合物种类和数量的差异是种间风味差异的主要原因。性别不同在脂肪含量及组成上存在差异,对羊肉风味也有影响。随着动物年龄的增长,皮下脂肪沉积增多,饱和脂肪酸比例增大,脂质氧化物的浓度增加,羊肉的风味也会变得强烈。不同部位的羊肉因挥发性化合物的种类和含量不同而在风味上存在差异。羊肉的风味受到采食饲料类型的影响,高蛋白饲料、高能量饲料、风味植物、不饱和脂肪酸含量高的籽实及其他添加剂均在一定程度上影响羊肉的风味。饲养方式与疾病因素也会使羊肉风味发生变化。饲养密度、光照等环境因素对羊肉风味也有不同程度的影响。养羊生产中可以通过品种选择、营养调控及科学饲养管理等途径来改善羊肉气味和风味。     

牦牛乳及其风味物质研究进展

牦牛主要分布于我国青藏高原及其周边地区，其特殊的生存环境使牦牛乳与普通牛乳相比，在营养价值、风味物质等方面具有独特性，使其受到越来越多的消费者关注。本文从牦牛乳商业化的产业链角度出发，对牦牛乳年产奶量及影响因素、营养成分、风味物质以及风味物质的浓缩和鉴定方面进行总结，旨在为进一步开发利用生鲜牦牛乳作为高品质商业无菌乳提供参考。     

水牛乳汁中乳腺上皮细胞的分离培养及鉴定

试验旨在从水牛（Bubalus bubalis）乳汁中分离培养水牛乳腺上皮细胞并对其进行鉴定,建立经济、便捷的水牛乳腺上皮细胞分离方法。通过无菌收集高产奶水牛泌乳后期乳汁,将其与PBS按2：1（V/V）混匀离心;去除上层乳脂,取带少量上清的沉淀,与PBS按上述比例混匀再次离心;将脂肪层去除干净,取沉淀上方1 mL上清及沉淀分别接种于细胞培养皿。通过细胞形态、免疫荧光、生长曲线测定、细胞接种活力、RT-PCR等对分离的细胞进行鉴定。结果表明,试验成功从水牛乳汁的上清和沉淀中分离获得了水牛乳腺上皮细胞,上清部分细胞和杂质较少;而沉淀部分则细胞和杂质较多。细胞呈典型鹅卵石样,无成纤维细胞,细胞活力好,多处于分裂期。当细胞增殖高度融合时,出现乳头状结构和分层生长现象。经免疫荧光鉴定,分离获得的水牛乳腺上皮细胞表达上皮细胞标志蛋白--细胞角蛋白18。水牛乳腺上皮细胞的生长曲线呈S型,符合一般生物学规律。细胞接种存活率的测定结果显示,细胞活力好、贴壁能力强。经RT-PCR检测,水牛乳腺上皮细胞表达乳蛋白及乳脂合成相关基因。从乳汁中分离水牛乳腺上皮细胞并对其进行鉴定,成功建立了水牛乳汁分离乳腺上皮细胞的方法,为研究水牛泌乳机制、改善乳品质、提高产奶量奠定基础。     

Isolation,Culture and Identification of Buffalo Mammary Epithelial Cells from Milk

This study was aimed to set up a simple, economic and pure method to culture and identify buffalo mammary epithelial cell in vitro, which were collected from the fresh milk. The milk was collected in the late lactation period of the high yield milk buffalo, then it was mixed with the PBS in the ratio of 2:1 and centrifugated. A few suspernatants and the pellets were resuspended with the PBS when the milk fat were decanted, then recntrifuged. The final pellets and 1 mL of suspernatants were seeded in petri dish without fat, respectively. The isolated cells were mammary epithelial cells which were identified by cell morphology, immunofluorescence, growth curve, cell viability and RT-PCR. The buffalo mammary epithelial cells were successfully isolated from both the pellets and supernatants. The cells and impurities were less in the supernatants than that in the pellets. The cells were cobblestone-like without fibroblasts and most in cell division. In the post-confluent culture, mammary epithelial cells formed dome structures and layer-separated growth. The cells expressed cytokeratin 18 was identified by immunofluorescence which was the marker of mammary cells. The cell growth curve and cell survival rate were measured. The results showed that the buffalo mammary epithelial cells were activity and easy to attach. The mammary epithelial cells were expressed of milk protein and milk fat related genes detected by RT-PCR. The buffalo mammary epithelial cell line were successfully isolated and identified from fresh milk, which estabilished foundation for the study of the mechanism of lactation, the amelioration of the quality of milk and the improvement of milk yield of buffalo.     

Proteomic analysis to unravel the effect of heat stress on gene expression and milk synthesis in bovine mammary epithelial cells

Heat stress can play a negative effect on milk yield and composition of dairy cattle, leading to immeasurable economic loss. The basic components of the mammary gland are the alveoli; these alveolar mammary epithelial cells reflect the milk producing ability of dairy cows. In this study, we exposed bovine mammary epithelial cells to heat stress and compared them to a control group using isobaric tags for relative and absolute quantitation combined with liquid chromatography coupled with tandem mass spectrometry. Compared with a control group, 104 differentially elevated proteins (>1.3‐fold) and 167 decreased proteins (<0.77‐fold) were identified in the heat treatment group. Gene Ontology analysis identified a majority of the differentially expressed proteins are associated in cell‐substrate junction assembly, catabolic processes and metabolic processes. Some of these significantly regulated proteins were related to the synthesis and secretion of milk, such as milk protein and fat. This finding was further supported by the results obtained from the reduced β‐casein expression through the system of plasminogen activator – plasminogen – plasmin and decreased fatty acid synthase could partly explain why milk fat synthesis ability of dairy cows decreased under heat stress. Our results highlight the effects of heat stress on synthesis of milk protein and fat, thus providing additional clues for further studies of heat stress on dairy milk production.