添加CO2对原料乳物理性质及稳定性的影响

在实验室条件下研究了添加CO2的原料乳在脱气前后的物理性质和稳定性,以确定适宜的CO2添加量。试验设计分为CO2处理组和脱气处理组2组,4个水平,以未处理的原料乳做对照组,检测每组样品CO2浓度、感官性质、pH值、酸度和稳定性。结果表明,二氧化碳浓度为39.86 mmol/L时,原料乳开始产生酸味,脱气后,感官性质正常,pH值为6.61,滴定酸度18.6 T°,酒精稳定性68%,热稳定时间8.2 min,符合正常原料乳标准;二氧化碳浓度为49.43 mmol/L时,脱气后的原料乳pH值为6.58,滴定酸度20.7 T°,酒精稳定性66%,热稳定时间6.9 min,已不能用于生产。因此,原料乳中二氧化碳添加量以不高于39.86 mmol/L为宜。     

唾液乳杆菌利用低聚果糖的代谢过程研究

对唾液乳杆菌代谢低聚果糖的代谢过程进行探讨,采用阴离子交换色谱检测唾液乳杆菌代谢低聚果糖时各组分的变化规律,采用高效液相色谱方法检测其代谢产物。结果表明:唾液乳杆菌能够代谢低聚果糖,发酵液OD600值达到0.303时,总糖质量浓度从最初的10.00g/L降至8.82g/L,pH值从7.00降至6.12。在与低聚果糖组分同时存在的情况下,唾液乳杆菌优先利用葡萄糖、果糖和蔗糖;低聚果糖组分中能快速利用蔗果三糖,蔗果四糖代谢效率不高,不能代谢蔗果五糖。代谢产物HPLC分析结果显示,唾液乳杆菌代谢低聚果糖的代谢产物主要有乳酸和乙酸。     

双缩脲比色法分析三氯乙酸沉淀乳蛋白效果探讨

通过对双缩脲比色法分析加热条件、不同浓度的三氯乙酸(TCA)沉淀牛乳蛋白的效果,以及pH影响双缩脲比色法分析TCA沉淀牛乳蛋白的效果进行分析。结果表明,采用双缩脲比色法分析TCA沉淀乳蛋白的效果:当使用标准双缩脲试剂时,受TCA浓度的影响较大,需TCA浓度≤15%;当TCA浓度>15%时,需调整pH至11以上;pH≥11时,碱性强度微弱的变化对OD值结果无明显影响;加热煮沸与未加热煮沸对TCA沉淀牛乳蛋白的效果差别无明显规律。     

基于磁性免疫传感器的牛乳中四环素残留检测方法的构建

建立一种高灵敏磁性光学免疫传感检测方法,可用于生鲜乳中四环素残留的快速检测。方法的检出限为0.03 ng/mL,定量限为0.20 ng/mL;四环素质量浓度在0.25～32 ng/mL时,荧光信号值与质量浓度之间具有良好的线性关系,其回归方程决定系数（R2）为0.995 9;本方法准确度高,四环素不同添加水平条件下的回收率在82.6%～97.0%之间;该方法具有良好的重复性,测定结果的日内精密度和日间精密度均在5.0%以内;本方法与高效液相色谱-串联紫外检测结果高度吻合。     

采用HPLC测定乳铁蛋白质量浓度的方法研究

建立一种灵敏度高、操作简便的定量分析牛初乳或乳粉中乳铁蛋白的高效液相色谱方法.样品经脱脂、酸法去除酪蛋白和硫酸铵沉淀富集、纤维素滤膜过滤净化等方法进行预处理,采用Shodex IEC CM-825弱阳离子色谱柱分离,以农度为0.02 mol/L,pH值为7.0的磷酸缓冲盐溶液和浓度为0～1.5 mol/L氯化钠的混合溶液作为流动相,进行梯度洗脱,流速为0.5 mL/min,检测波长为280 nm.结果表明:LF的标准曲线方程为y=494439x+25044 R2=-0.9991.LF在质量浓度为0.05～4.0 g/Lg围内呈线性关系,且LF的平均加标回收率为96.883%,以上结果说明此方法方便、快捷、准确、可靠、重现性好.可用于牛初乳质量浓度或乳粉中乳铁蛋白质量分数的检测.     

原料奶中尿的掺假定性检测方法

研究了原料乳中尿的检测方法,尿中含有肌酐,在pH值为12的条件下,肌酐与苦味酸反应生成红色或橙红色的复合苦味酸肌酐,通过颜色的变化判断鲜乳中是否有尿的存在.结果表明,该方法用于检测原料乳中尿含量在4％以上含量具有明显的颜色变化,同时方法操作简单,结果判定较直观.适用于乳品企业进行尿定性的检测.     

酒花中异α酸对短乳杆菌49胞外pH、胞内ATP及细胞膜脂肪酸组成的影响

以实验室从腐败啤酒中分离得到的酒花抗性菌株短乳杆菌49为出发菌株,考察酒花胁迫对其生理应激反应的影响,探究相关酒花胁迫机制。研究结果表明:酒花对短乳杆菌49有显著的抑制作用,60 mg/L为短乳杆菌49最高耐受酒花异α酸浓度;酒花会使短乳杆菌49胞外pH值呈下降趋势,且高浓度的酒花导致胞外pH值升高;酒花胁迫下的短乳杆菌49比无酒花胁迫胞内ATP低1个数量级。随着酒花质量浓度的升高,短乳杆菌49细胞膜脂肪酸碳链长度增加,尤其是C_(20)增长较为明显,平均链长也随酒花质量浓度的提高而提高;环状脂肪酸C_(19)相较于对照组显著增加;在高酒花质量浓度下细胞膜脂肪酸不饱和脂肪酸以及支链脂肪酸所占比例随之增加,饱和直链脂肪酸减少;同时细胞膜脂肪酸流动指数(U/S)值也随酒花质量浓度的升高而升高。本文为啤酒污染菌的抗酒花胁迫机制以及对啤酒腐败菌的控制提供理论依据。     

崂山奶山羊乳的热稳定性影响因素研究

本文以崂山奶山羊乳为研究对象,对山羊乳的一般理化性质进行分析,并重点探讨了山羊乳的热稳定性。采用热凝固时间法(HCT)测定不同泌乳期、温度、pH值及相关助剂对山羊乳热稳定性的影响。结果表明,山羊乳的pH接近中性、酸度为11~23°T,且与泌乳期呈正相关关系。羊乳的稳定性随泌乳期的延长而增强,随加热温度的升高而急剧减弱。pH对羊乳的热稳定性影响较大,pH越大,HCT值越大,当待测乳样的pH值超过6.5时,乳样不再发生沉淀。在pH=6.0、140℃加热条件下,加入适量浓度为0.01~0.20mol/L的柠檬酸盐、磷酸盐、EDTA均可改善羊乳的稳定性,且添加物浓度越大,改善效果越好。添加钙离子亦可改变羊乳的热稳定性,其稳定性随钙离子浓度的升高而降低。羊乳热稳定性随蔗糖浓度的增大呈现出先升高后降低的趋势,添加浓度为35%的蔗糖时,待测羊乳的HCT值最高,羊乳的热稳定性最好。     

去除胆固醇细菌的分离、鉴定及其性能的初步研究

从金华火腿、光明酸奶、光明奶酪、味全乳酸菌饮品及泡菜等样品中分离得到100余株乳酸菌菌株,利用高胆固醇MRS-CHOL培养基测定其体外胆固醇去除能力,从中筛选得到一株高效去除胆固醇的菌株JH-1,去除率达到48.68%。通过形态学特征、生理生化特征和16S rDNA序列同源性分析,鉴定菌株JH-1为短乳杆菌(Lactobacillus brevis)。研究了培养时间、接种量、培养温度、培养基初始pH以及基质中胆固醇胶束质量浓度对胆固醇去除率的影响,发现短乳杆菌JH-1分别在时间为48 h,接种量体积分数为3%,培养温度为37℃,初始pH值为6,胆固醇胶束质量浓度为1 mg/mL时,胆固醇去除率达到最大。     

功能低聚糖对植物乳杆菌ZDY2013发酵乳发酵特性及冷藏效果的影响

为探究功能低聚糖对植物乳杆菌ZDY2013发酵乳的发酵特点和冷藏期功能的影响,选取具有益生元特性的低聚木糖、低聚异麦芽糖及其组合物为发酵乳中碳水化合物,评价发酵乳中植物乳杆菌发酵特性和pH值变化,同时对发酵乳的持水性及冷藏期抗氧化活性进行解析。结果表明：植物乳杆菌能利用不同浓度功能低聚糖进行代谢;相比葡萄糖,发酵乳中添加低聚糖更有利于植物乳杆菌的生长,尤其是其组合物浓度为3.0%时,能显著提高发酵乳中活菌数及降低发酵乳pH值;冷藏期（21 d）内,各组发酵乳活菌数呈下降趋势,但均高于108 cfu/g,pH值在7 d后比较稳定,而持水性不断增强;发酵乳DPPH自由基清除能力在第7 d最强,而ABTS+和羟自由基的清除能力呈下降趋势,且添加同浓度的组合物优于葡萄糖。该研究结果将为功能低聚糖及植物乳杆菌ZDY2013在乳品开发中的应用提供理论依据。     

Milk urea concentration as an indicator of ammonia emission from dairy cow barn under restricted grazing

Bulk milk urea concentration was evaluated to assess its potential as an indicator of ammonia emission from a dairy cow barn in a situation with restricted grazing. An experiment was carried out with a herd of, on average, 52 Holstein-Friesian dairy cows. The cows were housed in a naturally ventilated barn with cubicles and a slatted floor, were fed ensiled forages and feed supplements, and each day were allowed 8.5 h of grazing. The experiment was a balanced randomized block design, replicated 3 times. The experimental factor was the bulk milk urea level, which was adjusted to levels of 15, 35, and 55 mg of urea per 100 g of milk, respectively, by changing the level of nitrogen fertilization of the pasture, the herbage mass and grass regrowth age, and the level and type of feed supplement. Ammonia emission from the barn was measured using sulfur hexafluoride as the tracer gas. Ammonia emission generally increased upon an increase in adjusted milk urea levels. A dynamic regression model was used to predict ammonia emission from bulk milk urea concentration, temperature, and a slurry mixing index. This model accounted for 66% of the total variance in ammonia emission and showed that emission increases exponentially with increasing milk urea concentration. At levels of 20 and 30 mg of urea per 100 g of milk, ammonia emission increased by about 2.5 and 3.5%, respectively, when milk urea concentration increased by 1 mg/100 g. Furthermore, emissions from the barn increased 2.6% when temperature increased by 1°C. The study showed that bulk milk urea concentration is a useful indicator for ammonia emissions from a dairy cow barn in a situation with restricted grazing.     

Phase Feeding of Urea after Feeding Soybean Meal to Dairy Cows

Urea-free control, medium-, and high-urea concentrates containing 0, 1.6, and 2.3% urea, respectively, were applied as main treatments, each to a group of nine lactating cows during three 8-wk periods. Split plot treatments consisted of feeding each concentrate normally or the urea portion in phase 2 or 4 h after feeding the soybean meal component. Dry matter intake, milk yield, milk lipid composition of butyric through capric acids were decreased, but milk linoleic acid was increased with high-urea rather than medium-urea or control concentrate. Ammonia in blood serum was reduced with 2-h phase feeding and urea was reduced with 2- or 4-h phase feeding as compared to normal feeding. Ammonia was unaffected by normal or phase feeding of the control concentrate. Normal feeding of high-urea concentrate increased ammonia in serum, but phase feeding reverted concentration comparable to the control group (160 to 122 μ/100 ml). Ammonia was nearly the same with normal or phase feeding of medium-urea concentrate.     

Effect of rumen-degradable protein balance and forage type on bulk milk urea concentration and emission of ammonia from dairy cow houses

As the Dutch government and dairy farming sector have given priority to reducing ammonia emission, the effect of diet on the ammonia emission from dairy cow barns was studied. In addition, the usefulness of milk urea content as an indicator of emission reduction was evaluated. An experiment was carried out with a herd of 55 to 57 Holstein-Friesian dairy cows housed in a naturally ventilated barn with cubicles and a slatted floor. The experiment was designed as a 3 x 3 factorial trial and repeated 3 times. During the experiment, cows were confined to the barn (no grazing) and were fed ensiled forages and additional concentrates. The default forage was grass silage. The nutritional experimental factors were: (1) rumen-degradable protein balance of the ration for lactating cows with 3 levels (0, 500, and 1000 g/cow per d), and (2) proportion of corn silage in the forage ration for lactating cows with 3 levels (0, 50, and 100%) of forage dry matter intake. Several series of dynamic regression models were fitted. One of these models explained emission of ammonia by the nutritional factors and the temperature; another model explained ammonia emission by the bulk milk urea content and the temperature. The ammonia emission from the barn increased when levels of rumen-degradable protein balance increased. Furthermore, at a given level of rumen-degradable protein balance, the emission of ammonia correlated positively with the corn silage content in the forage ration. However, this correlation was not causal, but was the result of interaction between corn silage proportion and intake of ileal digestible protein. The bulk milk urea content and the temperature correlated strongly with the ammonia emission from the barn; the selected model accounted for 76% of the variance in emission. It was concluded that the emission of ammonia from naturally ventilated dairy cow barns was strongly influenced by diet. The emission can be reduced approximately 50% by reducing the rumen-degradable protein balance of the ration from 1000 to 0 g/cow per d. The milk urea content is a good indicator of emission reduction.     

Effects of breed,farm intensiveness,and cow productivity on infrared predicted milk urea

Milk urea content is receiving growing interest from science and industry as a tool to infer the protein adequacy of dairy cows' diets, nitrogen excretion and its environmental impact, and efficiency of animals' protein metabolism. Fourier-transform infrared (FTIR) prediction is a high-throughput method for rapidly and cheaply evaluating milk urea content at the population level. Existing knowledge of the major sources of variation (e.g., year, season, farming system, individual herd, and the cow's breed, parity, stage of lactation, and productive potential) is fragmentary. The objective of this work was to study at the population level the simultaneous effects of all the major sources of variation and their most important interactions. Milk urea content in 1,759,706 test day milk samples collected from 291,129 lactations of 115,819 cows from 6,430 herds over 8 yr was predicted by FTIR. The milk urea content data (and also milk protein percentage, for comparison) were analyzed using a linear model that included the effects of parity, days in milk (DIM) class, year, month, herd intensiveness level, cow productivity level, breed, and herd intensiveness and cow productivity levels within breed. All sources of variation of milk urea content proved highly significant, the most important in terms of F-value being breed > year > herd intensiveness level > parity. The ranking for milk protein was very different (DIM class > herd intensiveness level > parity > breed). The patterns of the least squares means for urea and protein contents of milk were also very different and sometimes contrasting. The seasonal variation in urea was sinusoidal. Urea content increased during the first 4 mo of lactation and then remained almost stable before decreasing after 11 mo. Specialized dairy breeds had lower average milk urea content than dual-purpose breeds; in the former case it was lower in Holsteins than in Brown Swiss, and in the latter it was lower in Simmentals than in Alpine Greys. The effect of herd intensiveness level was much stronger than the effect of cow productivity level; the increase in milk urea with increasing herd average daily milk yield was almost linear in the case of dairy breeds but curvilinear in dual-purpose breeds. The large differences in breed and the modest relationships with the cow's productive potential require further analysis at the genetic level to obtain information of potential use in genetic improvement of the dairy cow populations.     

Natural variations of citrate and calcium in milk and their effects on milk processing properties

Natural variations among milk constituents, and their relations to each other as well as to processing parameters, represent possibilities for differentiation of milk to produce high-quality natural products. In this study, we focused on natural variations in milk citrate and its interplay with calcium distribution in milk, in relation to processing properties. Milk samples from individual cows from farms varying in feeding and management practices were collected from April to June 2017 to maximize natural variations in citrate and calcium. Chemical composition, rennet coagulation properties, and ethanol stability were analyzed for all milk samples. We focused particularly on calcium distribution and citrate content and the correlation of these to other milk parameters. No significant change in citrate content was observed during the sampling period, which suggests that mechanisms other than feeding affect citrate levels in milk. Several significant correlations were found, including a positive correlation between complexed serum calcium and citrate, and a negative correlation between urea and ionic calcium. These are both of interest in relation to further processing, as with regard to the stability of UHT milk and in cheese making. Although the correlation between complexed serum calcium and citrate may be explained by their affinity, the underlying driver for the negative relationship between natural milk urea and ionic calcium needs to be clarified by further studies. Furthermore, milk from the different farms varied not only with regard to organic versus conventional farming systems; feeding practices between farms also play an important role in milk composition and functionality. However, none of the differences in milk composition between farms were found to decrease milk functionality and thus would probably not cause any processing problems.     

Within-herd prevalence of intramammary infection caused by Mycoplasma bovis and associations between cow udder health,milk yield,and composition

Subclinical mastitis is one of the major health problems in dairy herds due to decreased milk production and reduced milk quality. The aim of this study was to examine the within-herd prevalence of subclinical intramammary infection caused by Mycoplasma bovis and to evaluate associations between M. bovis and cow daily milk yield, udder health, and milk composition. Individual cow composite milk samples (n = 522) were collected from all lactating dairy cows in 1 Estonian dairy farm in November 2014. Daily milk yield, days in milk, and parity were recorded. Collected milk samples were analyzed for somatic cell count, milk protein, fat, and urea content. The presence of M. bovis, Staphylococcus aureus, Streptococcus agalactiae, and Streptococcus uberis in the milk samples was confirmed by quantitative PCR analysis. The within-herd prevalence of M. bovis was 17.2% in the study herd. No association was observed between days in milk and parity to the presence of M. bovis in milk. According to linear regression analysis, the daily milk yield from cows positive for M. bovis was on average 3.0 kg lower compared with cows negative for M. bovis. In addition, the presence of M. bovis in milk samples was significantly associated with higher somatic cell count and lower fat and urea content compared with milk samples negative for M. bovis. In conclusion, subclinical M. bovis intramammary infection is associated with decreased milk yield and lower milk quality.     

Identification of lactose ureide, a urea derivative of lactose, in milk and milk products

With the widespread consumption of milk, the complete characterization of the constituents of milk and milk products is important in terms of functionality and safety. In this study, a novel nonreducing carbohydrate was separated from powdered skim milk and was identified using electron spray ionization-mass spectrometry (m/z 385.1[M + H⁺]), ¹H, ¹³C, ¹H¹H-correlation spectroscopy, and heteronuclear single quantum-nuclear magnetic resonance spectra. The carbohydrate was identified as a lactose derivative of urea, N-carbamoyl-o-β-d-galactopyranosyl-(1–4)-d-glucopyranosylamine (lactose ureide, LU). For the HPLC analysis of LU in milk and milk products, benzoylated LU, hepta-o-benzoyl lactose ureide (melting point 137–139 °C; m/z 1,113 [M + H⁺]; wavelength of maximum absorption, λ_max, 229 nm; molar extinction coefficient, ε, 8.1037 × 10⁷), was used as a standard. The crude nonreducing carbohydrate fraction from raw milk, thermally processed milk, and milk products such as powdered milks were directly benzoylated and subjected to HPLC analysis using an octadecylsilyl column to determine the quantity of LU. The content of LU in 10% solutions of powdered skim milk and powdered infant formula (5.0 ± 1.1 and 4.9 ± 1.5 mg/L, respectively) were almost 3-fold higher than that of UHT milk (1.6 ± 0.5 mg/L) and higher than that of low-temperature, long-time-processed (pasteurized at 65 °C for 30 min) milk (1.2 ± 0.3 mg/L) and the fresh raw milk sample (0.3 ± 0.1 mg/L). A time-course of the LU content in raw milk during heating at 110 °C revealed that LU increased with time. From these results, it is likely that LU is formed by the Maillard-type reaction between the lactose and urea in milk and milk products. Because the concentration of LU in milk increased with the degree of processing heat treatment, it could serve as an indicator of the thermal deterioration of milk. Although it is known that the human intestine is unable to digest LU, the gastrointestinal bacteria in human subjects are able to digest and utilize urea nitrogen in formation of essential amino acids that are available to the host human. These findings suggest that LU in milk might have a functional role in human health.     

利用非线性化学指纹图谱法检测奶粉中掺杂的尿素含量

采用"硫酸-丙二酸-溴酸钠-溴化钠-硫酸铈铵"为振荡体系,利用非线性化学指纹图谱法测定奶粉中掺杂的尿素。先向奶粉中掺入不同量的尿素,配成掺杂尿素的奶粉标样,每份奶粉标样的总量为1.00 g。通过非线性化学指纹图谱法对奶粉标样进行测定,运用最小二乘法,拟合出诱导时间相对于奶粉中掺杂尿素含量之间的一元线性关系,可求出奶粉中掺杂的尿素含量。实验表明该方法可以达到以下指标:当奶粉中掺杂的尿素含量在0~40.00 mg/g范围内时,尿素含量与诱导时间线性关系良好,相关系数R2为0.996 6和0.999 5,回收率为95.48%~104.06%,相对标准偏差不大于1.82%,奶粉1#检出限为3.3×10~(-4) mg/g,奶粉2#检出限为4.5×10~(-4) mg/g,奶粉3#检出限为1.1×10~(-4) mg/g。方法准确度高,操作简单,是一种切实可行的测定奶粉中掺杂尿素的方法,也可作为复杂样本中其他成分定量分析借鉴的方法。     

Changes in milk urea around insemination are negatively associated with conception success in dairy cows

Dietary protein levels are a risk factor for poor reproductive performance. Conception is particularly impaired in cases of high blood or milk urea. The objective of this study was to investigate the association between conception and low milk urea or changes in milk urea around artificial insemination (AI). Data were obtained from the French Milk Control Program for a 4-yr period (2009–2012). Milk urea values between 250 and 450 mg/kg (4.3 and 7.7 mM) were considered intermediate (I), and values ≤150 mg/kg (2.6 mM) were considered low (L). Milk urea values before and after each AI were allocated into 4 classes representing the dynamics of milk urea (before-after; I-I, I-L, L-I, and L-L). Subclinical ketosis was defined using milk fat and protein contents before AI as proxies. A logistic regression with a Poisson correction and herd as a random variable was then performed on data from Holstein or all breeds of cows. The success of conception was decreased [relative risk (95% confidence interval) = 0.96 (0.94–0.99)] in low-urea cows compared with intermediate-urea cows after AI; no significant association was found for urea levels before AI. When combining data on urea before and after AI, I-L urea cows exhibited a 5 to 9% decrease in conception compared with I-I urea cows, and L-I urea cows showed no difference in conception success compared with I-I urea cows. A decreased conception success for L-L urea cows compared with I-I urea cows was observed for the analysis with cows of all breeds. This work revealed that a decrease in urea from intermediate (before AI) to low (after AI) is a risk factor for conception failure. Surveys of variation in milk urea in dairy cows close to breeding are highly recommended.     

Cow and herd variation in milk urea nitrogen concentrations in lactating dairy cattle

Milk urea nitrogen (MUN) is correlated with N balance, N intake, and dietary N content, and thus is a good indicator of proper feeding management with respect to protein. It is commonly used to monitor feeding programs to achieve environmental goals; however, genetic diversity also exists among cows. It was hypothesized that phenotypic diversity among cows could bias feed management decisions when monitoring tools do not consider genetic diversity associated with MUN. The objective of the work was to evaluate the effect of cow and herd variation on MUN. Data from 2 previously published research trials and a field trial were subjected to multivariate regression analyses using a mixed model. Analyses of the research trial data showed that MUN concentrations could be predicted equally well from diet composition, milk yield, and milk components regardless of whether dry matter intake was included in the regression model. This indicated that cow and herd variation could be accurately estimated from field trial data when feed intake was not known. Milk urea N was correlated with dietary protein and neutral detergent fiber content, milk yield, milk protein content, and days in milk for both data sets. Cow was a highly significant determinant of MUN regardless of the data set used, and herd trended to significance for the field trial data. When all other variables were held constant, a percentage unit change in dietary protein concentration resulted in a 1.1 mg/dL change in MUN. Least squares means estimates of MUN concentrations across herds ranged from a low of 13.6 mg/dL to a high of 17.3 mg/dL. If the observed MUN for the high herd were caused solely by high crude protein feeding, then the herd would have to reduce dietary protein to a concentration of 12.8% of dry matter to achieve a MUN concentration of 12 mg/dL, likely resulting in lost milk production. If the observed phenotypic variation is due to genetic differences among cows, genetic choices could result in herds that exceed target values for MUN when adhering to best management practices, which is consistent with the trend for differences in MUN among herds.