Responsive changes of rumen microbiome and metabolome in dairy cows with different susceptibility to subacute ruminal acidosis

Subacute ruminal acidosis(SARA)represents one of the most important digestive disorders in intensive dairy farms,and dairy cows are individually different in the severity of SARA risk.The objectives of the current study were to investigate differences in the ruminal bacterial community and metabolome in dairy cattle with different susceptibility to SARA.In the present study,12 cows were initially enrolled in the experiment.Based on average ruminal pH,4 cows with the lowest ruminal pH were assigned to the susceptible group(SUS,pH=5.76,n=4)and 4 cows with the highest ruminal pH assigned to the tolerant group(TOL,pH=6.10,n=4).Rumen contents from susceptible(SUS,n=4)and tolerant(TOL,n=4)dairy cows were collected through rumen fistula to systematically reveal the rumen microbial and metabolic alterations of dairy cows with different susceptibility to SARA using multi-omics approaches(16S and 18S rRNA gene sequencing and metabolome).The results showed that despite being fed the same diet,SUS cows had lower ruminal pH and higher concentrations of total volatile fatty acids(VFA)and propionate than TOL cows(P<0.05).No significant differences were observed in dry matter intake,milk yield,and other milk compositions between the SUS and TOL groups(P>0.05).The principal coordinates analysis based on the analysis of molecular variance indicated a significant difference in bacterial composition between the two groups(P=0.01).More specifically,the relative abundance of starch-degrading bacteria(Prevotella spp.)was greater(P<0.05),while the proportion of fiber-degrading bacteria(unclassified Ruminococcaceae spp.,Ruminococcus spp.,Papillibacter,and unclassified Family_-XIII)was lower in the rumen of SUS cows compared with TOL cows(P<0.05).Community analysis of protozoa showed that there were no significant differences in the diversity,richness,and community structure(P>0.05).Metabolomics analysis revealed that the concentrations of organic acids(such as lactic acid),biogenic amines(such as histamine),and bacterial degradation products(such as hypoxanthine)were significantly higher in the SUS group compared to the TOL group(P<0.05).These findings revealed that the higher proportion of starch-degrading bacteria/lower fiber-degrading bacteria in the rumen of SUS cows resulted in higher VFA-producing capacity,in particular propionate.This caused a disruption in metabolic homeostasis in the rumen which might be the reason for the higher susceptibility to SARA.Overall,these findings enhanced our understanding of the ruminal microbiome and metabolic changes in cows susceptible to SARA.     

亚急性瘤胃酸中毒耐受性不同的奶牛瘤胃上皮形态及功能差异研究

本试验旨在探究SARA（亚急性瘤胃酸中毒）耐受性不同奶牛的瘤胃上皮形态及其功能差异。选取12头装有永久性瘤胃瘘管的泌乳中期荷斯坦奶牛,饲喂精粗比为4∶6的日粮,并根据瘤胃pH值的高低,分为SARA易感组（SUS,n=4）和SARA耐受组（TOL,n=4）。瘤胃上皮形态及功能分析结果显示,与TOL组比较,SUS组奶牛瘤胃上皮的棘突层和基底层厚度明显增厚（P<0.05）,SUS组奶牛瘤胃上皮组织中参与挥发性脂肪酸（VFA）吸收的PAT1、MCT4和DRA基因表达量较TOL组显著下调（P<0.05）,而H+转运载体NHE1、NHE2、NHE3和调节胞内pH的vH+ATPase和Na+/K+ATPase的表达量显著升高（P<0.05）;对参与调控瘤胃VFA代谢的基因定量结果表明,SUS组PDHA1和SREBP2的表达量显著高于TOL组（P<0.05）,而HMGCL-2的表达量显著降低（P<0.05）;此外,SUS组CDK2、CDK6和Cyclin D1、Bad及Caspase-9等参与瘤胃上皮细胞增殖与凋亡的基因表达量显著高于TOL组（P<0.05）。结果说...     

益生菌对奶牛亚急性瘤胃酸中毒的保护作用

亚急性瘤胃酸中毒对奶牛的生产性能有很大影响,使用益生菌来稳定奶牛分娩后-产奶过渡期瘤胃pH可以减轻这种代谢紊乱的症状。因此,本试验选择了体重(741±55)kg,产奶期在(212±19.5)d的奶牛4头,试验采用4×4拉丁方设计,共4种日粮:对照组饲喂基础日粮,处理1组为日粮添加0.5 g/d米曲菌,处理2组为日粮添加2.5 g/d米曲菌,处理3组为日粮添加2 g/d粪肠球菌和酿酒酵母菌的混合物。每组经过3 w的适应期、4 d酸中毒期以及3 d恢复期。结果显示:瘤胃最大pH范围在5.6～6.0,占比最高,与适应期和中毒期的结果相似。中毒期间,其他pH范围表现为显著差异(P <0.05)。中毒期,处理3组较对照组显著提高了瘤胃pH(P <0.05),同时处理3组瘤胃pH在5.6～6.0的占比也显著高于对照组(P <0.05)。日粮添加2.5 g/d米曲菌显著降低了产奶量(P <0.05)。奶牛遭受瘤胃酸中毒后,随着时间的推移,丙酸、丁酸和戊酸含量显著升高(P <0.05),而乙酸、异丁酸和异戊酸含量显著降低(P <0.05)。除了处理3组外,其他组瘤胃乳酸含量随时间的增加而升高(P <0.05)。饲喂12 d后,处理3组较对照组显著提高了乳酸含量(P <0.05)。瘤胃白球菌和大肠杆菌含量在饲喂2 h后显著升高(P <0.05),之后在6 h达到稳定。结论 :粪肠球菌和酿酒酵母菌复合物可以缓解奶牛亚急性瘤胃酸中毒的症状,米曲菌可以调控瘤胃pH,但高剂量米曲菌添加水平对瘤胃pH无显著影响。     

奶牛亚急性瘤胃酸中毒的发生与危害

亚急性瘤胃酸中毒（SARA）是一个大多数牛场普遍存在的奶牛代谢性疾病。围产期、干物质采食量（DMI）过高、日粮配制差及采食模式变化过大等奶牛均易发生亚急性瘤胃酸中毒,继而造成干物质采食量下降、纤维素消化率降低、乳成分改变及引发腹泻等疾病。本文对奶牛亚急性瘤胃酸中毒的病因及危害等方面进行了简述。     

奶牛瘤胃酸中毒的发病机理与营养调控

瘤胃酸中毒主要是由于饲喂大量的极易发酵的碳水化合物,引起瘤胃内酸度升高,从而导致机体功能紊乱的一种营养代谢病。发病快、病程短、病症重、死亡率高,给奶牛养殖造成严重的损失。本文主要就奶牛瘤胃酸中毒的发病机制及预防的研究进展作了简要综述。合理调配饲料,日粮中添加缓冲物质,调控瘤胃内微生物区系,加强饲养管理等手段的综合运用可有效的防止瘤胃酸中毒的发生。     

异麦芽糖对泌乳早期奶牛瘤胃发酵及生产性能的影响

为了研究异麦芽糖对泌乳早期奶牛瘤胃发酵及生产性能的影响,选择4头泌乳天数为20 d、日产奶量30 kg左右、体重550 kg左右的经产(二胎)中国荷斯坦泌乳奶牛,随机分为2组,对照组饲喂基础日粮,试验组在基础日粮中添加60 g异麦芽糖,采用2×2交叉试验设计,每期21d。结果表明:添加异麦芽糖可以显著提高奶牛瘤胃中挥发性脂肪酸(VFA)的含量(P0.05),显著降低氨态氮(NH_3-N)的含量(P0.05),其中瘤胃液中丁酸含量有极显著差异(P0.01),乙酸和丙酸的含量呈上升趋势,但无显著差异(P0.05)。对奶样进行分析,添加异麦芽糖可以显著提高牛奶的乳脂率(P0.05),显著降低乳体细胞数(P0.05),产奶量、乳蛋白和尿素氮浓度均无显著差异(P0.05)。综上所述,异麦芽糖有助于改善奶牛的瘤胃发酵功能,提高牛奶的乳脂率。     

复合酶制剂对瘤胃发酵及泌乳早期奶牛生产性能的影响

本试验旨在研究添加不同水平复合酶制剂对瘤胃发酵及奶牛生产性能的影响。试验一以奶牛全混合日粮作为底物进行体外瘤胃发酵试验,分为4组,即对照组不添加酶制剂,试验1、2和3组的酶制剂添加量分别为日粮浓度的0.10%、0.15%和0.20%,每组设9个重复。每个重复准确称取0.500 g底物,在体外发酵产气自动记录装置上发酵48 h,测定其发酵参数和营养物质降解率。结果表明:复合酶制剂显著提高发酵液中总挥发性脂肪酸和乙酸浓度(P<0.05);试验组中粗蛋白(P<0.05)和中性洗涤纤维(P<0.01)降解率显著高于对照组。试验二选择体重、胎次、泌乳天数和产奶量相近的泌乳早期荷斯坦奶牛36头,采用随机区组设计分为4组,即对照组和试验1、2和3组,对照组不添加酶制剂,试验1、2和3组分别添加0.10%、0.15%和0.20%的酶制剂,每组9个重复,试验期8周,测定产奶量和乳成分含量,计算3.5%乳脂校正乳。结果表明:复合酶制剂显著提高3.5%乳脂校正乳产量(P<0.05),0.10%、0.15%和0.20%组比对照组分别提高3.88、4.27和2.26 kg·d^-1。0.15%组的乳脂率显著高于对照组(P<0.05),比对照组高12.7%。结论添加复合酶制剂有利于瘤胃发酵和提高生产性能,且添加量为0.15%时效果较好。     

硫酸钠对奶牛瘤胃发酵的影响

本文旨在研究不同添加水平的硫酸钠对泌乳中期奶牛瘤胃发酵的影响。试验选用4头体重、胎次、产奶量均相近的荷斯坦奶牛,采用4×4拉丁方设计,试验分4个处理组,添加不同水平的硫酸钠,即0%(对照组)、0.4%(试验1组)、0.8%(试验2组)与1.2%(试验3组),试验分为4期,每期18 d(预试期15 d,正试期3 d)。结果表明:(1)饲粮中添加硫酸钠对瘤胃液pH、NH3-N平均浓度无显著影响(P>0.05);(2)饲粮中添加硫酸钠后,瘤胃内原虫数均有所提高,其中试验2组和试验3组与对照组相比均差异显著(P<0.05);(3)添加硫酸钠的试验2组瘤胃液菌体蛋白浓度与对照组相比显著提高(P<0.05)。结果说明,日粮中添加适量的硫酸钠可以改变瘤胃发酵环境,提高瘤胃液菌体蛋白浓度。     

不同瘤胃细菌培养物添加水平对奶牛瘤胃发酵及BCP含量的影响

本文旨在研究不同剂量的瘤胃细菌培养物对荷斯坦奶牛瘤胃发酵及菌体蛋白(BCP)含量的影响。试验采用单因素试验设计,选用4头装有永久性瘤胃瘘管,体况相近的荷斯坦奶牛,取其瘤胃液混匀,添加不同剂量的瘤胃细菌培养物进行体外培养,分别为:对照组(0 CUF/g),试验Ⅰ组(添加量为3%,4.4×108CUF/g),试验Ⅱ组(添加量为5%,7.8×108CUF/g),试验Ⅲ组(添加量为10%,1.2×109CUF/g),每组分别设2、4、8、12、24、36、48 h 7个指标测定时间点,每个时间点3个重复。结果表明:(1)添加瘤胃细菌培养物使瘤胃液pH有所降低,但始终在6.5～7.0,对瘤胃内环境无不良影响;(2)添加3%的瘤胃细菌培养物显著降低氨态氮(NH3-N)的含量(P<0.05),显著增加了培养液BCP的浓度(P<0.05);(3)添加3%的瘤胃细菌培养物可一定程度增加总挥发性脂肪酸(TVFA)和乙酸浓度,但对其他各挥发性脂肪酸及乙酸/丙酸比值无显著影响。由此可见,添加3%剂量的瘤胃细菌培养物可改变瘤胃的发酵环境,提高BCP浓度。     

亚急性瘤胃酸中毒对泌乳奶牛血浆生化指标及代谢组的影响研究

本试验旨在探究亚急性瘤胃酸中毒（subacute rumen acidosis, SARA）对泌乳奶牛血液生化指标和血浆代谢组的影响。试验选用8头装有永久性瘤胃瘘管的泌乳中期奶牛,随机分成对照组（CON）（n=4）和处理组（SARA）（n=4）,CON组与SARA组分别饲喂精粗比为4∶6和6∶4的全混合日粮,试验周期为3周。于试验期每周最后一天晨饲后0、2、4、6、8和12 h进行瘤胃pH的测定,同时在pH测定当天晨饲后6 h采集颈静脉血样,用于血液生化指标和代谢组的分析。结果表明,与CON组相比,SARA组的瘤胃pH显著降低（P=0.002）,两组干物质采食量没有显著差异（P=0.524）。血浆生化指标结果显示,与CON组比较,SARA组奶牛的β-羟丁酸浓度显著降低（P=0.007）,甘油三酯浓度显著升高（P=0.014）。采用液相-质谱（LC-MS）联用技术对血浆进行代谢组学分析,主成分分析（PCA）和偏最小二乘法分析（PLS-DA）结果显示,SARA组奶牛血浆代谢物组成较CON组发生明显变化,两组间共检测到26种差异代谢物（VIP>1 & FDR<0.05）,与CON组相比,SARA组奶牛血浆中的7-酮脱氧胆酸、脱氧胆酸、胆酸、12-酮脱氧胆酸、12（13）Ep-9-KODE、12,13-DHOME和L-天冬酰胺代谢物含量显著升高（FDR<0.05）,而十一烷二酸、十六烷二酸、9-HODE、血氧烷B3、PGE2、L-精氨酸、L-色氨酸、L-苏氨酸、异丁酰甘氨酸、异戊酰甘氨酸、马尿酸、4-羟基马尿酸和6-磷酸-2-脱氢-D-葡萄糖酸酯等代谢物含量显著降低（FDR<0.05）。综上,与CON组比较,SARA组奶牛的脂代谢、氨基酸代谢和糖代谢发生显著变化,这些物质可作为SARA潜在的生物标志物。     

饲喂复合益生菌对泌乳中期奶牛产奶量及乳成分的影响

本文旨在研究不同剂量的复合益生菌制剂对泌乳中期奶牛产奶量及乳成分的影响。试验选用18头体重、胎次、产奶量均相近的中国荷斯坦奶牛,采用完全随机设计,分为3个处理组,即(对照组)、0.6×1011 cfu/d(试验1组)与1.2×1011 cfu/d(试验2组),试验期70 d(预试期10 d,正试期60 d)。结果表明:试验1组、2组与对照组相比产奶量分别提高3.11%、4.80%,4%乳脂校正乳提高5.14%、8.07%,能量校正乳提高4.74%、8.76%,差异均显著(P<0.05);试验2组较对照组乳脂率和乳蛋白率分别提高5.56%、5.86%,并显著提高乳脂、乳蛋白、非脂乳固体物的产量(P<0.05)。两试验组牛奶中的乳糖率及乳糖产量与对照组无显著差异(P>0.05)。这表明,日粮中饲喂复合益生菌可以显著提高奶牛生产性能,从而提高饲料利用效率。     

Signals for identifying cows at risk of subacute ruminal acidosis in dairy veterinary practice

Controlling rumen disorders is critical to ensure successful dairy herd health management. Lactation diets of dairy cows are commonly rich in concentrates and low in physically effective fibre. Feeding of these diets increases the risk of rumen disorders with far‐reaching consequences for cattle health, welfare and sustainability of dairy production. The term subacute ruminal acidosis or SARA is often used as a synonym for poor rumen health. Being subclinical, SARA lacks of clear symptoms and is therefore difficult to diagnose and to control in the practice. This review article summarises common and identifies new direct and indirect cow signals related to SARA. We have performed a scientific evaluation and interpretation of each of these cow signals by highlighting their advantages and disadvantages from the practitioner's point of view. The gold standard of SARA cow signals still remains direct measurement of ruminal pH. However, continuous pH monitoring is cost‐intensive and often biased by sensor drift. Single‐point ruminal pH measurements by oral stomach tubing or rumenocentesis have strong limitations. Therefore, there is a need for reliable and robust markers of SARA that are easily accessible and inexpensive. Such indirect parameters are the observation of chewing and feeding activities, as well as the monitoring of milk, faecal, urine and blood variables. Also, novel technologies that allow rapid and non‐invasive measurement of the rumen mucosa thickness and ruminal motility patterns might provide advantages in SARA diagnosis. Due to several constraints of these indirect diagnostic tools, such as limited specificity and sensitivity, we strongly recommend using a combination of the signals to reliably identify cows at risk of SARA in a dairy herd.     

Subacute rumen acidosis in lactating cows: an investigation in intensive Italian dairy herds

Subacute rumen acidosis (SARA) represents one of the most important metabolic disorders in intensive dairy farms that affects rumen fermentations, animal welfare, productivity and farm profitability. The aim of the present study was to study the occurrence of SARA in intensive Italian dairy herds and to determine the relationship between diet composition, ruminal pH and short chain fatty acids (SCFA) concentration. Ten commercial dairy herds were investigated; twelve cows in each herd were selected randomly among animal without clinical signs of disease, with good body condition and between 5 and 60 day-in-milk (DIM), to perform rumenocentesis and obtain rumen fluid. Ruminal pH was determined immediately after sampling and concentration of SCFA in ruminal fluid was determined on samples after storage. An other objective of this research was to study in detail the effects of rumenocentesis on animal health: this study could confirm the extreme validity of this technique as ruminal sampling. Results were subject to anova and correlation analysis using SIGMA STAT 2.03. The results indicated the presence of SARA in three herds (more than 33% cows with rumen pH < 5.5), a critical situation (more than 33% cows with rumen pH < 5.8) in five farms and a normal rumen pH condition in two herds. In particular, dairy herds show on average SCFA concentration of 150, 145, 123 mmol/l for low pH, critical pH and normal pH herds respectively. There were not significant differences among diet composition even if herds with SARA showed a light discordance between initially chemistry composition and residual feed. In the affected herds it was not possible to understand the exact causes of SARA. Animal management seems to be one of the most important factors in developing SARA including total mixed ration preparation.     

甘露寡糖对泌乳早期奶牛瘤胃发酵及生产性能影响的研究

试验旨在研究甘露寡糖(mannan-oligosaccharides MOS)对泌乳早期奶牛瘤胃发酵及生产性能的影响。选择4头泌乳天数为20 d,日产奶量30 kg左右,体重550 kg左右的经产(二胎)中国荷斯坦泌乳奶牛,随机分为两组,对照组饲喂基础日粮,试验组饲喂在基础日粮中添加60 g的试验日粮,采用2×2交叉试验设计,每期21 d。结果表明:添加甘露寡糖可以显著提高奶牛瘤胃中总的挥发酸和氨态氮的含量(P<0.05),其中瘤胃液中乙酸含量有极显著差异(P<0.01),丙酸、丁酸的含量无显著差异(P>0.05)。对奶样进行分析,添加甘露寡糖可以显著提高乳脂率(P<0.05),并且明显降低乳中体细胞数(P<0.05),产奶量、乳蛋白和乳中尿素氮浓度均无显著差异(P>0.05)。试验表明,在日粮中添加甘露寡糖有助于提高奶牛瘤胃乙酸发酵比例,提高奶牛乳脂率。     

一例牛瘤胃酸中毒的诊治

瘤胃酸中毒多发生于牛,特别是奶牛.主要是精饲料喂量过多,精粗饲料比例不当所造成.以1～3胎的奶牛发病最多,7胎后的发病较少.一年四季均可发生,但以冬春季较多.临产牛和产后3 d内的发病较多.产奶量愈多,发病率愈高.笔者于2005年夏季带学生在临夏市某兽医站实习时遇到一病例,现将诊治情况总结如下.     

Relationships of metabolic profiles to milk production and feeding in dairy cows

The aim of this study was to assess the practicality of the metabolic profile test (MPT) for feeding evaluation in dairy cattle. Stepwise regression analysis was used to evaluate the relationships of MPT to feeding and milk production of 4,679 cows in 343 commercial dairy herds. Significant explanatory variables were determined by forward set-up selection, among the deviated values from the reference values of 10 blood metabolites and body condition score, to predict dependent variables, i.e., milk production and the rate of feeding to nutrient requirements, in each or all lactation stages and the dry period. The milk production model of the all-lactation stage showed the greatest goodness-of-fit (adjusted R(2)=0.214, p<0.0001) with high positive regression coefficients for serum cholesterol, magnesium, urea nitrogen and albumin, and negative for glucose and calcium. In the feeding models, goodness-of-fit of crude protein was relatively high (R(2)=0.072, p<0.0001) with a positive relationship to blood urea nitrogen. Although the other feeding models were low in goodness-of-fit, several significant explanatory variables to feeding were found. All feeding models in the late lactation stage and the dry period, in which the feeding was stable, had greater goodness-of-fit than those in the early lactation stage in which milk production varied. It was concluded that the values which deviated from the reference values for the MPT components could assess milk production and feeding, and the MPT is a practical tool for auxiliary feeding evaluation.     

On-farm feeding interventions to increase milk production in lactating dairy cows

Tropical Animal Health and Production - The objective of this study was to investigate the effect of tropical legume (Phaseolus calcaratus) mixed with ruzi grass feeding on the performance of...     

Effect of feeding Bacillus subtilis natto fermentation product on milk production and composition, blood metabolites and rumen fermentation in early lactation dairy cows

This experiment was conducted to determine the effect of Bacillus subtilis natto fermentation product supplementation on blood metabolites, rumen fermentation and milk production and composition in early lactation dairy cows. Thirty-six multiparous Holstein cows (DIM = 29 ± 6 days, parity = 2.8 ± 1.1) were blocked by DIM and parity and then randomly assigned to three treatments (12 per treatment) in a 9-week trial. Cows in control, DFM1 and DFM2 were fed TMR diets supplemented with 0, 6 and 12 g of B. subtilis natto solid-state fermentation product per day per cow respectively. Plasma non-esterified fatty acids were lower (p = 0.03) in DFM1 and DFM2 compared with control cows (633 and 639 vs. 685 μm). Ruminal propionate increased (23.9 vs. 26.3 and 26.9/100 mol, control vs. DFM1 and DFM2 respectively) and acetate decreased (64.2 vs. 62.7 and 62.1/100 mol, control vs. DFM1 and DFM2 respectively) with increasing B. subtilis natto fermentation product supplementation. DMI of the cows in three treatments was not affected by B. subtilis natto fermentation product supplementation, but milk yield was 3.1 and 3.2 kg/day higher for DFM1 and DFM2 than that for control cows on average across the 9-week trial, and significant differences were observed during weeks 5-9 of the trial, which resulted in 9.5% and 11.7% increase in feed efficiency. B. subtilis natto fermentation product supplementation did not affect milk fat percentage and protein yield but increased (p < 0.05) milk fat yield and lactose percentage (p < 0.01) and tended to decrease protein percentage (p = 0.06). The findings show that B. subtilis natto fermentation product was effective in increasing lactation performance of early lactation dairy cows possibly by altering the rumen fermentation pattern without any negative effects on blood metabolites.     

过瘤胃蛋白和丙二醇对乳牛瘤胃发酵、产乳量及乳成分的影响

用 8头荷斯坦黑白花奶牛 ,在饲粮中添加过瘤胃蛋白和丙二醇 ,研究其对乳牛瘤胃发酵、产乳量及乳成分的影响。结果表明 ,过瘤胃蛋白+丙二醇可显著提高丙酸浓度 (P <0 0 5) ,并且产乳量较对照组有增加的趋势。乳脂率、乳蛋白率、乳糖率及无脂固形物率 ,并未因添加过瘤胃蛋白和丙二醇而受到显著影响 (P >0 0 5)。乳蛋白质量、乳脂量及无脂固形物量 ,过瘤胃蛋白 +丙二醇组较对照组有增加的趋势 ,但差异均不显著 (P >0 0 5)。     

Impact of a simultaneous application of anionic salts and rumen buffer on acid-base-balance and mineral metabolism in dairy cows

In this study, the influence of simultaneous application of anionic salts (AS) and rumen buffer (RB) on the metabolism of dairy cows was examined. Eleven rumen fistulated, non-pregnant and non-lactating dairy cows received equal amounts of one AS (CaCl2 or CaSO4) and one RB (NaHCO3 or KHCO3) via rumen cannula during feeding time over a period of eight days. Before the first application of AS and RB and on day eight of the treatment period, blood, urine and rumen fluid samples were taken. The following parameters were measured: whole blood: pH, base excess, bicarbonate; serum: sodium, potassium, chloride, calcium; urine: pH, net acid base excretion, sodium, potassium, chloride, calcium; rumen fluid: pH. The changes of each parameter were compared via ANOVA.The changes in acid-base balance on day eight were very small, although significant. But p-values showed that the statistical evidence was low.The most changes occurred when NaHCO3 was fed in combination with one of the AS used. In this case a small acidogenic load was seen in blood (p < 0.05), and calcium concentrations increased slightly (p < 0.05). No alkalotic reaction could be detected when any combination of AS and RB were given to the cows. Simultaneous application of AS and RB results in a loss of effectivity of AS. Neither an adequate acidification of blood nor an activation of calcium metabolism occurred. In feed ration for cows in the last weeks of pregnancy, rumen buffer must not be fed, if anionic salts are given for prevention of parturient paresis.