UPLC-MS/MS测定生乳与豆制饮品中咪唑烟酸残留

该研究考察生乳与豆制饮品中咪唑烟酸的前处理条件,旨在建立一种超高效液相色谱-串联质谱（UPLC-MS/MS）检测生乳与豆制饮品中咪唑烟酸农药残留的方法。结果表明,该方法的前处理条件为采用甲酸∶乙腈=5∶995(V/V)提取,HLB固相萃取柱净化,20%乙腈水溶液定容。选择电喷雾离子源正离子（ESI+）多反应监测（MRM）模式,采用UPLC-MS/MS测定咪唑烟酸,外标法定量。在此条件下,咪唑烟酸在2.0～30.0μg/L质量浓度范围内线性关系良好,相关系数R2>0.99,生乳和豆制饮品中咪唑烟酸的检出限（LOD）和定量限（LOQ）均分别为0.002 mg/kg、0.005 mg/kg,加标回收率为80.77%～90.35%,精密度试验结果的相对标准偏差（RSD）为1.43%～5.27%,说明该方法简便、快速、准确,具有一定的推广价值,可用于生乳与豆制饮品中咪唑烟酸残留量检测。     

Effects of goat milk enriched with oligosaccharides on microbiota structures,and correlation between microbiota and short-chain fatty acids in the large intestine of the mouse

In this study, we explored the effects of combining goat milk and oligosaccharides on the large intestine environment of mice. A combination of goat milk with each of 3 oligosaccharides—stachyose, fructo-oligosaccharide (FOS), and a prebiotics mix—were independently fed to mice. We investigated composition changes in the microbiota of the large intestine using 16S rRNA gene sequencing; measured short-chain fatty acid content using gas chromatography–mass spectrometry; and performed a Spearman correlation analysis between microorganisms and short-chain fatty acids. Our results showed that microbial diversity in the large intestine decreased significantly in the FOS group. In terms of α diversity, microbial richness significantly declined in all 3 treatment groups; in terms of β diversity, the intestinal microbial structures clearly changed in the FOS group. The abundance of Bifidobacterium and Lactobacillus increased markedly in the FOS group compared with the other groups. Functional predictions showed that FOS reduced intestinal bacterial infections and improved the endocrine and immune systems. Spearman correlation analysis showed that propionic, isobutyric, and valeric acids were all positively correlated with certain microbiota. Our findings suggest that FOS-enriched goat milk is beneficial for improving the large intestine environment in the host.     

Performance of Holstein and Swedish-Red × Jersey/Holstein crossbred dairy cows within low- and medium-concentrate grassland-based systems

This 2 × 2 factorial design experiment was conducted to compare the performance of spring-calving Holstein dairy cows (HOL, n = 34) with Swedish Red × Jersey/Holstein crossbred (SR × J/HOL, n = 34) dairy cows within low and medium concentrate input grassland-based dairy systems. The experiment commenced when cows calved and encompassed 1 full lactation. Cows were offered diets containing grass silage and concentrates [70:30 dry matter (DM) ratio, and 40:60 DM ratio, for low and medium, respectively] until turnout, grazed grass plus either 1.0 or 4.0 kg of concentrate/d during the grazing period (low and medium, respectively), and grass silage and concentrates (85:15 DM ratio, and 70:30 DM ratio, for low and medium, respectively) from rehousing and until drying off. No significant genotype × system interactions were present for any of the feed intake or full-lactation milk production data examined. Full-lactation concentrate DM intakes were 769 and 1,902 kg/cow for the low and medium systems, respectively, whereas HOL cows had a higher total DM intake than SR × J/HOL cows in early lactation, but not in late lactation. Although HOL cows had a higher lactation milk yield than SR × J/HOL cows, the latter produced milk with a higher fat and protein content, and thus fat plus protein yield was unaffected by genotype. Milk produced by the SR × J/HOL cows had a higher degree of saturation of fatty acids than milk produced by the HOL cows, and the somatic cell score of milk produced by the former was also higher. Throughout the lactation, HOL cows were on average 30 kg heavier than SR × J/HOL cows, whereas the SR × J/HOL cows had a higher body condition score than the HOL cows. Holstein cows had a higher incidence of mastitis and ovarian dysfunction that SR × J/HOL cows.     

高效液相色谱法检测牛奶及其制品中的溶菌酶

目的建立一种测定牛奶及奶酪制品中低含量溶菌酶的高效液相色谱-荧光检测新方法。方法样品经p H 6.0的氯化钠溶液活化,再在低p H值条件下除蛋白后,采用反相色谱柱(PLRP-S 250 mm×4.6 mm,300?,5μm)用A水(0.1%的三氟乙酸(trifluoroacetic acid,TFA)),B乙腈(0.1%TFA)体系作为流动相进行梯度洗脱,使用荧光检测器在激发波长(λ_(ex))276 nm,发射波长(λ_(em))345 nm处检测。结果最优实验条件下,溶菌酶在2.0~30.0 mg/L浓度范围内线性良好,加标实验结果显示回收率和相对标准偏差(relative standard deviation,RSD)分别在92.3%~104.3%和0.81%~3.26%之间,对牛奶和奶酪制品的检出限分别为20和40 mg/kg。结论该方法操作简便,结果准确、可靠,可用于乳制品中溶菌酶含量测定。     

Alterations in ruminal bacterial populations at induction and recovery from diet-induced milk fat depression in dairy cows

Ten ruminally cannulated Holstein cows were used in a crossover design that investigated changes in ruminal bacterial populations in response to induction and recovery from diet-induced milk fat depression (MFD). Further, the effect on the ruminal microbiota of the cows with diet-induced milk fat depression inoculated with rumen contents from non-milk fat-depressed donor cows was evaluated. Milk fat depression was induced during the first 10 d of each period by feeding a low-fiber, high-starch, and high-polyunsaturated fatty acid diet (26.1% neutral detergent fiber, 28.1% starch, 5.8% total fatty acids, and 1.9% C18:2), resulting in a 30% decrease in milk fat yield. Induction was followed by a recovery phase, where all cows were switched to a high-fiber, low-starch, and low-polyunsaturated fatty acid diet (31.8% neutral detergent fiber, 23% starch, 4.2% total fatty acids, and 1.2% C18:2) and were allocated to (1) control (no inoculation) or (2) ruminal inoculation with donor cow digesta (8 kg/d for 6 d). Ruminal samples were collected at the end of induction (d 10) and during recovery (d 13, 16, and 28), separated to solid and liquid fractions, extracted for DNA, PCR- amplified for the V1-V2 region of the 16S rRNA gene, and analyzed for bacterial diversity. Results indicated that bacterial communities were different between fractions. In each fraction, differences were significant between the induction (d 10) and recovery (d 13, 16, and 28) periods; however, differences were less apparent with time during the recovery period. The MFD (d 10) was typified by a reduction in the relative sequence abundance of Bacteroidetes and an increase in the relative sequence abundance of Firmicutes and Actinobacteria across both fractions. At the genus level, relative sequence abundance of unclassified Lachnospiraceae, Butyrivibrio, Bulleidia, and Coriobacteriaceae were higher on d 10 and were positively correlated with trans-10,cis-12 CLA and the trans-10 isomer, suggesting their potential role in altered biohydrogenation reactions. A switch to the recovery diet resulted in a sharp increase in the Bacteroidetes lineages and a decrease in Firmicutes members on d 13; however, this shift appears to stabilize by d 28, indicating the restoration process for ruminal bacteria from an altered state is gradual and complex. Inoculation of 10% of rumen contents from non-MFD donor cows to MFD cows revealed this procedure had transient effects on only a few bacterial populations, and such effects disappeared after d 16 following cessation of inoculation. It can be concluded that alterations in milk FA profiles at induction are preceded by microbial alterations in the rumen driven by dietary changes.     

Management practices associated with Johne's bulk tank milk ELISA positivity

Johne's disease (JD) control is often based on the culling of positive animals and the adoption of management practices that minimize exposure of young stock to the pathogen (Mycobacterium avium ssp. paratuberculosis). Throughout 2010 to 2013, the province of Ontario, Canada, instituted a voluntary Johne's control program consisting of whole-herd testing and risk assessment. The JD risk assessment evaluated 5 management areas to characterize herd JD risk. Using a modified milk ELISA technique with an optical density cut-off of 0.089, province-wide bulk tank milk (BTM) testing was used to assess the prevalence of JD high-risk herds at the end of the control program and again 4 yr after its completion. Approximately 71% of Ontario bulk tanks were classified as positive in 2017 compared with roughly 46% in 2013. In 2019, the same JD risk assessment used in the original program was readministered on 180 Ontario dairy farms. Using this cross-sectional approach, logistic regression models were built using data from the original program risk assessment and follow-up risk assessment as well as the BTM ELISA results to determine management factors associated with the control of JD. We demonstrated that management of the maternity area is an important factor in the control of Johne's disease. Although it is believed that the highest risk group for JD infection is calves under 6 mo, the cleanliness scores of older heifers and their exposure to mature cow manure was significantly associated with JD control; farms with highly contaminated weaned and bred heifers and those that had exposure to mature cow manure were more likely to be unsuccessful in their JD control efforts. Careful management of young calves appears to be important for JD control, and this management should continue even after calves have left the maternity area.     

高效液相色谱法测定乳粉及液态奶中牛磺酸含量

建立高效液相色谱法测定乳粉和液态奶中牛磺酸含量。样品用水溶解,超声提取10 min,提取液经亚铁氰化钾和乙酸锌沉淀蛋白后,离心,上清液用异硫氰酸苯酯和三乙胺衍生化,室温衍生1 h,加入正己烷为衍生化终止剂,静置分层,下层溶液用水定容后过0.22μm微孔滤膜,过滤液经Diamonsil AAA氨基酸分析柱(4.6×250 mm,5μm)分离,以0.05 mol/L乙酸钠水溶液(pH值=6.50)和甲醇/乙腈(1/1,V:V)为流动相进行梯度洗脱。采用外标法定量,在5.0～40μg/mL线性范围内相关系数(r)大于0.999,线性关系良好,定量限为5 mg/100 g(S/N≥10),添加水平为5、10、25 mg/100 g时,回收率范围在90%～108%之间,相对标准偏差范围为1.16%～2.24%。结果显示,本方法准确、可靠,满足乳粉和液态奶中牛磺酸含量的有效确定,特别对于特膳奶粉比GB方法更适合样品中牛磺酸含量的有效检测。     

Bedding and bedding management practices are associated with mesophilic and thermophilic spore levels in bulk tank raw milk

Mesophilic and thermophilic spore-forming bacteria represent a challenge to the dairy industry, as these bacteria are capable of surviving adverse conditions associated with processing and sanitation and eventually spoil dairy products. The dairy farm environment, including soil, manure, silage, and bedding, has been implicated as a source for spores in raw milk. High levels of spores have previously been isolated from bedding, and different bedding materials have been associated with spore levels in bulk tank (BT) raw milk; however, the effect of different bedding types, bedding management practices, and bedding spore levels on the variance of spore levels in BT raw milk has not been investigated. To this end, farm and bedding management surveys were administered and unused bedding, used bedding, and BT raw milk samples were collected from dairy farms (1 or 2 times per farm) across the United States over 1 yr; the final data set included 182 dairy farms in 18 states. Bedding suspensions and BT raw milk were spore pasteurized (80°C for 12 min), and mesophilic and thermophilic spores were enumerated. Piecewise structural equation modeling analysis was used to determine direct and indirect pathways of association among farm and bedding practices, levels of spores in unused and used bedding, and levels of spores in BT raw milk. Separate models were constructed for mesophilic and thermophilic spore levels. The analyses showed that bedding material had a direct influence on levels of spores in unused and used bedding as well as an indirect association with spore levels in BT raw milk through used bedding spore levels. Specific bedding and farm management practices as well as cow hygiene in the housing area were associated with mesophilic and thermophilic spore levels in unused bedding, used bedding, and BT raw milk. Notably, levels of spores in used bedding were positively related to those in unused bedding, and used bedding spore levels were positively related to those in BT raw milk. The results of this study increase the understanding of the levels and ecology of mesophilic and thermophilic spores in raw milk, emphasize the possible role of bedding as a source of spores on-farm, and present opportunities for dairy producers to reduce spore levels in BT raw milk.     

高蛋白饮用型纯益生菌发酵乳的研制

以鲜牛乳和浓缩牛奶蛋白为主要原料,以酸度和感官评价为评价指标,通过单因素试验和正交试验确定高蛋白饮用型纯益生菌发酵乳的最佳发酵工艺条件。结果表明,高蛋白饮用型纯益生菌发酵乳的最佳发酵工艺条件为浓缩牛奶蛋白添加量6.0%,发酵剂接种量2.0%、发酵温度43 ℃、二次无菌均质压力4 MPa。在此优化条件下,制得的高蛋白饮用型纯益生菌发酵乳感官评分达到89分,酸度129.1 °T,口感清爽顺滑,组织状态良好,理化及微生物指标均符合国家相关标准。