奶牛乳房炎无乳链球菌的分离及PCR鉴定

利用THB(Todd-Hewitt Broth)固体培养基和色素试验培养基选择培养无乳链球菌,参照GenBank中无乳链球菌参考菌株(Accession:AF015927.1、JQ289582.1)16SrRNA和种属特异性基因cfb(CAMP因子)序列设计引物,对奶样中分离的12株疑似无乳链球菌进行鉴定。结果显示,经PCR扩增后,被检测的12株细菌均可扩增出预期大小的16S rRNA基因序列,而12株中有8株可以扩增出预期大小的cfb基因序列,条带单一,特异性好。序列BLAST显示,12株菌的16S rRNA基因序列与NCBI上报道的无乳链球菌相应序列高度同源(>99.0%),各分离菌株间的16S rRNA基因序列也高度同源(99.0%～100.0%);cfb基因序列与NCBI上已报道的无乳链球菌相应序列具有高度同源性(>99.0%),各菌株间cfb基因序列也高度同源(100.0%)。经选择培养与PCR鉴定结果可以确定12株疑似菌株中有8株为无乳链球菌。     

Lactoferrin concentrations in milk from normal and subclinical mastitic cows

The concentrations of lactoferrin (Lf) in quarter milk from normal lactating cows and subclinical mastitic cows were measured to determine whether the Lf concentration in milk is influenced by the age of the cow, the stage of lactation, number of milk somatic cells and the presence of pathogens. Lf concentrations in 111 quarter milk samples from 28 normal lactating cows and 270 quarter milk samples from 198 subclinical mastitic cows were measured by means of a single radial immunodiffusion test. Lf concentrations (means +/- standard deviations; logarithmic form) in normal cows and subclinical mastitic cows were 2.23 +/- 0.39 and 2.70 +/- 0.39, respectively. The mean milk Lf concentration (log) in subclinical mastitic cows was significantly (p<0.01) higher than that in normal cows. The mean milk Lf concentration (log) in normal lactating cows aged 5 years was lower than those in normal lactating cows aged 2 years (p<0.01) and 3 years (p<0.05). The results showed that the milk Lf concentration (log) is associated with age of the dairy cow (one-way analysis of variance test, p<0.01). The mean milk Lf concentration (log) in the latter lactational period tended to be higher than those in the peak and middle periods. Milk Lf concentrations (log) tended to be proportional to the level of the somatic cell count (SCC) score. Mean milk Lf concentrations (log) in subclinical mastitic cows infected with Staphylococcus aureus and with other streptococci species were significantly (p<0.01) higher than those in cows infected with coagulase-negative staphylococci and with Corynebacterium bovis.     

Zinc concentrations in skimmed milk and whole milk samples from healthy and mastitic cows

Zinc concentrations were determined in skimmed milk and whole milk samples from healthy and mastitic cows. During spontaneous and induced mastitis, a marked decrease of Zn concentration in skimmed milk from the mastitic quarters was observed. Seemingly, there are 2 mechanisms involved in the lowering of Zn concentration in skimmed milk from mastitic quarters: a decline in Zn probably as a consequence of decreases in plasma Zn concentrations and an abrupt redistribution of Zn in the inflamed quarters.     

Comparison of fat and cream content in normal and mastitic milk of cows

Two methods for estimating fat and cream content in cow's milk were compared in normal and mastitic milk. Fat content in milk from periparturient cows was estimated simultaneously by the method of Gerber and by simply spinning small samples of untreated milk in a haematocrit centrifuge. Mastitis was experimentally induced by inoculating mammary glands with Escherichia coli. On the average, cream content in milk from normal, and uninfected mammary glands measured by the haematocrit method was 20 to 23% higher than milk fat measured by the method of Gerber. In mastitic milk from infected mammary glands haematocrit readings were 40% higher than those of Gerber. Milk fat values as measured by both methods were highly correlated. It is concluded that the haematocrit method can be used for rapid estimation of milk fat content in normal milk of cows. The greater difference between cream and fat content in mastitic milk compared to normal milk can probably, at least partly, be ascribed to the presence of large clots of cells and fat in the milk during mastitis. During centrifugation of the viscous mastitic milk samples, milk cells and fat probably are not completely separated. This results in overestimated readings of milk cream content with the haematocrit method. It therefore seems advisable to use the Gerber method for accurate measurements of milk fat in mastitic milk.     

Phenotypic antimicrobial susceptibility of environmental bacteria from mastitic milk of pastured dairy cows of S. Miguel (Azores)

Tropical Animal Health and Production - The aim of this study was to evaluate the phenotypic (in vitro) antimicrobial susceptibility of milk pathogens isolated from subclinical and clinical...     

Fenbendazole-related drug residues in milk from treated dairy cows

Oral administration of [¹⁴C] fenbendazole (FBZ) at a dose of 5.Omg/kg leads to the presence of radiolabel in the milk of lactating dairy cows. However, the maximum mean concentration of total FBZ equivalents quantitated to one-third of the recommended safe concentration in milk (1.67 μg/mL). The label is equally distributed to the fat and aqueous portions of the milk. The maximum level, in general, is attained approximately 24-36 h after drug administration, with the highest levels ranging from 24 to 48 h after administration. The residues rapidly deplete, attaining levels of 10-20ng/mL by day 5, and are essentially undetectable by radiolabel monitoring by day 6. Extraction of the milk by matrix solid phase dispersion indicated that the label was distributed between traces of the parent drug, FBZ, and predominantly, the FBZ sulphoxide (SO) and sulphone (SO₂) metabolites. No other radiolabelled peaks were observed. Based on these data the metabolites of FBZ, FBZ-sulphone and FBZ-sulphoxide, could be used as marker residues for monitoring the administration of FBZ to lactating dairy cows.     

复合微生态制剂对奶牛泌乳性能和乳品质的影响

本研究旨在探讨复合微生态制剂对奶牛泌乳性能和乳品质的影响,为微生态制剂在奶牛养殖中的推广应用提供参考依据。将90头荷斯坦奶牛随机分成3组(对照组、试验Ⅰ组、试验Ⅱ组),对照组饲喂基础日粮,试验Ⅰ组、Ⅱ组分别在基础日粮中添加0.5 g·kg~(-1)、1.0 g·kg~(-1)的复合微生态制剂。通过测定各处理组的产奶量、采食量、料奶比等泌乳性能指标和乳脂率、乳蛋白率、乳糖率、乳干物质率、体细胞数等乳品质指标,比较分析复合微生态制剂不同添加水平对奶牛泌乳性能和乳品质的影响。结果显示,试验Ⅱ组与对照组相比,日产奶量显著提高(P0.05),料奶比显著降低(P0.05),乳脂率、乳蛋白率均显著提高(P0.05),体细胞数显著降低(P0.05)。该结果表明奶牛基础日粮中添加1.0 g·kg~(-1)的复合微生态制剂能显著改善奶牛的泌乳性能和乳品质。     

检测奶牛乳汁孕酮的免疫生物传感器优化

为了完善已建立的孕酮免疫生物传感器的检测性能,本试验应用碳化二亚胺法将11α-羟基孕酮半琥珀酸和OVA制备成孕酮完全抗原,其浓度达0.6 mg/m L。在此基础上建立了以乳汁为基质的孕酮生物传感器的标准曲线(y=-4.9461x+8.7982,R2=0.993 2),进一步确立了检测乳汁孕酮的线性范围为0.31 ng/m L~50ng/m L,检测限为0.31 ng/m L,灵敏度为0.15 ng/m L,而且孕酮与皮质酮、皮质醇、雄烯二酮的免疫交叉反应较低,表明特异性较好。批内、批间变异系数分别为7.7%和7.5%,回收率为95.8%~115.7%,生物传感器稳定性在4℃保存4 d。该优化的奶牛乳汁孕酮免疫生物传感器的性能参数符合规定的标准,为检测妊娠、乏情和繁殖障碍疾病的奶牛乳汁孕酮含量提供了工具。     

Relationship between oxidant stress and milk productivity in dairy cows

An imbalance between formation and detoxification of oxygen radicals leads to oxidant stress that may increase in more intense oxidative metabolism caused by a high intake of metabolizable energy to provide metabolic intermediates for the milk synthesis and secretion. This hypothesis was tested using dairy cows and the concentration of hydroperoxides in lipids (LHP) extracted from circulative lipoprotein particles of low and very low density (LDL and VLDL/chylomicrons) as oxidant stress indicator. The particles were prepared by ultracentrifugation of serum obtained by coccygeal bleeding (13 cows, 1. parity, n=8 and 2. parity, n=5, lactation stage, 53 +/- 1.4 days post partum) and purified by precipitation. Concentrations of LHP-LDL/mg Lipoprotein correlated significantly with daily milk yield (r = 0.73, P = 0.004) or daily milk energy output (r = 0.77, P = 0.003) in contrast to LHP of VLDL/chylomicron particles. Thus, some evidence was obtained for an almost linear, positive relationship between milk productivity and oxidant stress occurring in LDL.     

体细胞数对牛奶品质及产量的影响

根据宁夏地区某奶牛场的DHI检测获得的体细胞数据(SCS),利用SAS软件实现单因素方差分析,探讨不同乳中SCS对产奶量和乳中主要成分的影响,并对它们之间的相关性进行分析。结果表明:乳中SCS与日产奶量呈显著负相关(P0.01);与乳糖和胎次呈正相关(P0.01);乳中SCS水平与乳脂率、乳蛋白率和尿素氮没有明显的相关性。本试验通过对SCS的研究为乳品质的测定提供一定依据和参考。     

Pharmacokinetics of phenylbutazone in plasma and milk of lactating dairy cows

Phenylbutazone was administered intravenously (i.v.) to a group of four lactating cows at a dosage of 6 mg/kg body weight. Whole plasma, protein-free plasma and milk were analysed for phenylbutazone residues. Pharmacokinetic parameters of total and free phenylbutazone in plasma were calculated using a non compartmental method. In regards to whole plasma data, the mean volume of distribution at steady state ( V _ss), was 147 mL/kg body weight, with a mean (± SEM) terminal elimination half-life ( t _1/2) of 40 ± 6 h. The mean clearance ( Cl ) was 3 mL/h/kg body weight. The V _ss as determined from the protein-free plasma fraction was 50 021 mL/kg body weight. This larger V _ss of free phenylbutazone compared to total plasma phenylbutazone was attributed to a high degree of plasma protein binding, as well as the greater penetration of free phenylbutazone into tissues. The mean t _1/2 of free phenylbutazone was 39 ± 5 h. This similarity to the t _1/2 estimated from total plasma phenylbutazone data is attributed to an equilibrium between free and plasma phenylbutazone during the terminal elimination phase. Mean t _1/2 as determined from milk, applying a urinary excretion rate model, was 47 ± 4 h. Milk clearance of phenylbutazone was 0.009 mL/h/kg body weight, or about 0.34% of total body clearance. Furthermore, evidence suggests that phenylbutazone either binds to milk proteins, or is actively transported into milk, as its concentration in milk was greater than that predicted due to a simple partitioning from plasma into milk.     

Loss of milk yield due to Akabane disease in dairy cows

Akabane disease is an infection with clinical signs of congenital malformation and abortion in ruminants. Abnormal parturitions caused by Akabane disease result in great economic loss. The purpose of this study is to estimate the reduction in the milk yield from abnormal parturition due to Akabane disease. The data were collected from 33 Holstein cows on 11 farms. The animals had abnormal parturitions during the period from September 1998 to March 1999, and were diagnosed as having Akabane disease. The mean and standard deviation of the rate of reduction in the milk yield of 33 cows after abnormal parturition caused by Akabane disease was -11.4 +/- 14.9%. The means and standard deviations of the rate of reduction of four cows calving abnormally at 220-239 days of gestation, nine cows calving abnormally at 240-270 days of gestation, and 20 cows calving abnormally at 271-300 days of gestation were -26.6 +/- 24.7%, -14.7 +/- 11.0%, and -6.9 +/- 12.3%, respectively. In this study, we demonstrated that the rate of reduction in the milk yield in cows affected by Akabane disease was -11.4 +/- 14.9%, but values as high as -26.6 +/- 24.7% were reached in the comparison with the milk yield obtained after normal parturition.     

Impact of iodine supplementation of dairy cows on milk production and iodine concentrations in milk

AIM: To determine the effect of iodine supplementation on milk production and iodine concentrations in milk for pasture-fed, seasonally calving dairy cows. METHODS: The study was run over two consecutive seasons on the same dairy farm. In Trial One, 294 Friesian dairy cows were either untreated or injected intramuscularly three times with iodised oil (2,370 mg iodine/dose) at the start of lactation (Day 1) and at about 100-day intervals thereafter. Iodine concentrations in milk were determined prior to each injection on Days 1, 98 and 210, and milk, fat and protein yields measured on Days 43, 98, 176 and 238. In Trial Two, 234 Friesian dairy cows were either untreated or injected intramuscularly with iodised oil (2,370 mg iodine/dose) in early lactation (Day 1) and 99 days later. Iodine concentrations in milk were determined prior to treatment on Days 1, 55 and 161. Serum thyroxine (T4) and triiodothyronine (T3) concentrations were determined in the latter two samples and milk, protein and fat yields were determined on Days 25, 119, 182 and 221. In both trials, pasture was sampled at 1- or 3-monthly intervals and iodine concentrations in herbage measured. RESULTS: Iodine concentrations in pasture averaged 0.24 mg/kg dry matter (DM) in both seasons and varied little over the experimental period. Iodine concentrations in milk from unsupplemented cows were <20 microg/L in both trials. Three iodine injections at about 100-day intervals increased iodine concentrations in milk to 58 microg/L for at least 98 days after each treatment. Two iodine injections at about 100-day intervals raised milk iodine concentrations to 160 microg/L and 211 microg/L at least 55 days after each treatment, but had no effect on serum thyroid hormone concentrations. Iodine supplementation had no significant effect on milk, milkfat or milk protein yield. CONCLUSIONS: Pastures in New Zealand that contained 0.24 mg iodine/kg DM appeared to provide an adequate iodine intake for dairy cows. Intramuscular injections of iodine resulted in significant increases in iodine concentrations in milk. CLINICAL SIGNIFICANCE: Increasing iodine concentrations in milk via intramuscular injection of iodine could provide a method for significantly increasing iodine intakes of humans, in particular, children.     

Plasminogen activator activity in secretions from clinically normal, mastitic, lactating and dried-off udders of dairy cows

Using the composite fibrin plates method, plasminogen activator (PA) activity was found in secretions from clinically normal, mastitic, lactating and dried-off udders of dairy cows. The possible involvement of the fibrinolytic system in the physiology and pathology of the bovine mammary gland is discussed.     

复合益生菌制剂对奶牛泌乳性能及乳品质的影响

为研究复合益生菌制剂对奶牛泌乳性能及乳品质的影响,试验选取45头泌乳期相近的荷斯坦奶牛,随机分为3组,对照组、试验1组、试验2组,每组3个重复,每个重复5头。对照组饲喂基础日粮,试验1、2组分别在基础日粮中添加1%、2%的复合益生菌制剂,试验期为60d。结果表明:试验2组在第10、30、60天奶牛产奶量较对照组分别提高2.28%、0.62%、9.08%(P<0.05),试验1组在第10、60天较对照组分别提高4.6%、8.9%(P<0.05);第30天时,试验1、2组乳中总固形物含量较对照组均显著性提高(P<0.05),乳糖率、乳蛋白率、乳脂率之间差异不显著(P>0.05);第60天时,与对照组相比,试验1、2组乳中乳糖率分别提高了4.41%、7.02%(P<0.05),乳蛋白率提高了4.16%、9.97%(P<0.05),总固形物含量提高了2.44%、3.02%(P<0.05),乳脂率含量之间差异不显著(P>0.05)。综上,日粮中添加复合益生菌制剂可有效提高奶牛产奶量及乳品质,且添加2%复方益生菌制剂效果更优。     

Elimination kinetics of chlorhexidine in milk following intramammary infusion to stop lactation in mastitic mammary gland quarters of cows

OBJECTIVE: To evaluate the elimination kinetics of chlorhexidine in milk when used as an intramammary infusion to stop lactation in cows. DESIGN: Prospective study. ANIMALS: 6 cows. PROCEDURE: The study was performed in 2 phases. Three cows were studied in each phase. All cows were treated with chlorhexidine suspension by infusion into a mastitic mammary gland quarter after 2 milkings 24 hours apart. Foremilk samples (100 mL) were collected from treated and untreated (controls) mammary gland quarters of each cow. Chlorhexidine was extracted from raw milk, and residue concentrations were quantified by use of high-performance liquid chromatography. Foremilk samples from days 2, 5, and 8 were analyzed in phase I, and samples from time 0 and days 3, 7, 14, 21, 28, 35, and 42 were analyzed in phase II. RESULTS: In phases I and II, there was no quantifiable transference of chlorhexidine to milk in untreated mammary gland quarters. Measurable chlorhexidine residues were found in milk from treated mammary gland quarters of 2 cows throughout the 42-day sample period in phase II. Estimated mean elimination half-life for chlorhexidine in milk was 11.5 days. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of the long elimination half-life of chlorhexidine in milk from treated mammary gland quarters, the lack of human dietary exposure data to suggest a food tolerance for chlorhexidine in food products, and the Food and Drug Administration's published zero tolerance for chlorhexidine in uncooked edible calf tissues, we do not recommend extralabel use of chlorhexidine suspension as a treatment to stop lactation in mastitic mammary gland quarters of cows.     

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...