奶牛隐性乳腺炎病原菌的分离鉴定和体细胞数及酶的相关性研究

对330个乳样分别用加州乳腺炎检验(California Mastitis Test,CMT),体细胞计数(Somatic Cell Count,SCC)和细菌的分离培养(Isolation of bacteria)法进行隐性乳腺炎的检测,结果表明3种方法检测的结果差异不显著(P>0.05);对检测患有隐性乳腺炎的241乳区进行了细菌的分离培养,结果表明引起隐性乳腺炎的最主要致病菌为金黄色葡萄球菌、凝固酶阴性葡萄球菌、链球菌、酵母菌、假单胞菌和棒状杆菌、其它细菌的检出率差异根显著(P<0.05)。对分离出优势细菌的部分乳样进行了SCC与LDH,NAGase,ALP和ACP几种酶的测定和相关性分析,结果表明:LDH与SCC有显著的相关性(r=0.7936),并且LDH与NA-Gase,ALP,ACP都有明显的相关性;NAGase与ACP,LDH有明显的相关性,但与ALP没有相关性;ALP只与ACP和LDH有显著的相关性外,与SCC,NAGase没有相关性;ACP与SCC,LDH,NAGase,ALP都有极显著的相关性。     

Experimental intramammary inoculation with Mycoplasma bovis in vaccinated and unvaccinated cows: effect on milk production and milk quality.

The effect of vaccination on milk production was evaluated in vaccinated and control cows experimentally challenged in two of four quarters with live Mycoplasma bovis. During the first three weeks after experimental challenge, six of eight unchallenged quarters on vaccinated cows and seven of eight unchallenged quarters on control cows became infected. Most of these quarters secreted normal milk, with negative California Mastitis Test scores and maintained normal milk production throughout most of the study (although some quarters on control cows remained infected). All challenged quarters became infected, had strong California Mastitis Test reactions, and had a drastic (greater than 85%) loss in milk production. Thereafter, four of eight challenged quarters on control cows remained infected, had mostly positive California Mastitis Test scores, produced mostly normal-appearing milk, and recovered some productive capabilities. By the end of the study no M. bovis could be recovered from challenged quarters on vaccinated cows and the milk appeared mostly normal. The California Mastitis Test scores on these quarters, however, remained elevated and milk production remained very low.     

Pathogenicity of different species of staphylococci in caprine udder

Aseptic foremilk samples were collected from Finnish landrace goats. Ten different species of staphylococci, causing subclinical infections were detected. Twelve goats with persistent subclinical staphylococcal infection were followed on a monthly basis and compared with foremilk samples of nine goats suffering from clinical mastitis. Parameters of inflammation based on the activity of the California Mastitis Test (CMT). N-acetyl-beta-D-glucosaminidase (NAGase) and antitrypsin were determined from the milk. Staphylococci were further classified using the API STAPH system. On the basis of elevations of activities of CMT, NAGase and antitrypsin, Staphylococcus aureus was the most pathogenic in clinical and subclinical mastitis, S. hyicus showed only marginal pathogenicity. Subclinical infections were persistent and the infective organism was not always detected from milk by culture. The biochemical reactions of subclinical staphylococci seemed to vary within the same gland by time. Antitrypsin was most effective in differentiating between subclinical and clinical infection. A teat cistern puncture technique was found to be suitable for the goat.     

Cell free mitochondrial DNA in serum and milk associated with bovine mastitis: a pilot study

Mastitis is inflammation of mammary gland affecting all the species of domestic animals. Fragments of the mitochondrial genome released from dying cells are considered surrogate markers of mitochondrial injury. We hypothesized that bovine mastitis would be associated with increased cell free mitochondrial DNA (mtDNA) content in serum and milk. Milk and serum samples were collected from sub-clinical mastitic and normal animals. Mastitis was confirmed by California mastitis test and bacterial isolation. Oxidative stress, nitric oxide and inflammatory cytokines were also estimated. Real time polymerase chain reaction was conducted in serum and milk from sub-clinical mastitic animals and compared with healthy animals targeting the mtDNA genes cytochrome b. Mastitis animals showed higher oxidative stress markers and nitric oxide along with higher level of inflammatory cytokines. Cell free mtDNA was significantly higher in serum and milk of mastitic animals comparing to that of healthy control. The higher cell free relative mtDNA content in mastitis animals indicates injury to the mammary epithelial cells and thereby releasing the mtDNA in the milk and blood. This mtDNA may be a bio-marker of oxidative stress and tissue injury in bovine mastitis.     

Effect of frequent milkings on milk NAGase, plasmin, trypsin inhibitory capacity and the quality of whey as the growth medium for mastitis pathogens

Unequal milking intervals affected milk somatic cell count and N-acetyl-beta-D-glucosaminidase (NAGase) activity. The total daily output of milk NAGase and plasmin decreased, if quarters were emptied frequently during the day. Mastitis pathogens showed stimulated growth in whey prepared from filled quarters as compared with growth in whey from quarters emptied frequently during the day. The quality of whey as growth medium for mastitis pathogens paralleled plasmin activity in respective milk samples. Adaptation of mastitis pathogens to grow in whey had an enhancing effect on bacterial growth during subsequent inoculations in whey. Bacteria probably "learn" to overcome the effect from endogenous antibacterial factors and to use nutrients present in whey.     

N-acetyl-beta-D-glucosaminidase (NAGase) in porcine milk

Sow milk contains a high basal level of the lysosomal enzyme N-acetyl-β-D-glucosaminidase (NAGase). The level is extremely high in colostrum. High basal levels seem to hide inflammatory changes. Therefore NAGase did not prove to be as good an indicator for porcine agalactia syndrome as reported for mastitic bovine or ovine milk.     

Inhibition of lacteal leukocyte phagocytosis by colostrum, nonlactating secretion, and mastitic milk

The Fc receptors on leukocytes obtained from normal milk, colostrum, nonlactating gland secretion, and mastitic milk were detected by rosette formation, using sensitized erythrocytes. The percentage of leukocytes bearing Fc receptors from normal milk was significantly (P less than 0.01) greater than that from other secretions. Fc receptors were found primarily on polymorphonuclear leukocytes from normal milk, mastitic milk, and colostrum. However, in nonlactating gland secretion obtained 6 weeks after milking was completed, Fc receptors were predominantly on macrophages. The low percentage of leukocytes bearing Fc receptors obtained from colostrum, nonlactating gland secretion obtained 7 days after milking was completed, and mastitic milk was associated with the presence of a blocking factor in these secretions, which specifically attached to Fc receptors. These secretions significantly (P less than 0.01) blocked the Fc receptors on leukocytes from normal milk and on other cells bearing FC receptors. The presence of Fc receptors on leukocytes obtained from normal milk was related to a high percentage of phagocytizing leukocytes through Fc receptors and a large number of phagocytized bacteria (phagocytic activities). In contrast, the low percentage of leukocytes bearing Fc receptors from colostrum, from nonlactating gland secretion (7 days after milking), and from mastitic milk was associated with depressed phagocytic activities. Preincubation of leukocytes from normal milk with whey from colostrum, nonlactating gland secretion (7 days after milking), and mastitic milk significantly (P less than 0.01) inhibited phagocytosis. This effect was associated with the blocking of Fc receptors by these secretions. Possible mechanisms for and implications of these findings are discussed.     

N-acetyl-beta-D-glucosaminidase test for screening milk samples for subclinical mastitis.

The use of the N-acetyl-beta-D-glucosaminidase (NAGase) test for detecting subclinical mastitis was investigated in surveys of milk samples from 20 farms. A milk sample was considered to be mastitic if it had a milk cell count above 400,000 cells/ml, and the NAGase test results were graded accordingly. The test gave an average of 16.6 per cent false positives and 2.0 per cent false negatives per herd. It was concluded that the NAGase test could be used as a rapid screening method for selecting suspect samples for further analysis by standard methods.     

Treatment of Streptococcus agalactiae mastitis in dairy cows: comparative efficacies of two antibiotic preparations and factors associated with successful treatment

A commercially available, penicillin-novobiocin, intramammary infusion product and a solution of procaine penicillin G (1.2 X 10(6) IU) in 10 ml of sterile saline solution were evaluated for their comparative efficacies against Streptococcus agalactiae mastitis in 3 California dairy herds. After composite milk samples from each cow in each herd were bacteriologically cultured, cows infected with S agalactiae (n = 228) were assigned randomly to 2 treatment groups. Milk samples were reevaluated bacteriologically 21 to 25 days after treatment. Both preparations were highly effective against S agalactiae in first-lactation cows and in cows scored negative or trace by use of the California Mastitis Test. Efficacy was significantly decreased in cows with California Mastitis Test scores of 1, 2, or 3. Herd and treatment were associated significantly with treatment success or failure. Most treatment failures were in one herd in cows that were given procaine penicillin G in sterile saline solution. Milk production and lactation stage were not associated with success or failure of treatment.     

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.     

Prevalence of subclinical udder infections and individual somatic cell counts in three dairy goat herds during a full lactation

For dairy goats, both the determination of the somatic cell counts (SCC) and the interpretation of these values may be a problem. Several investigations have shown that SCC for goat's milk, even from not infected mammary halves, are often higher than for cows milk. In the three herds examined about 40% of mammary halves and 30% of the goats were infected. However large differences between the three herds could be observed. In most cases, infections were caused by coagulase negative staphylococci (CNS) or corynebacteria. The SCC of individual milk samples from goats without any udder infection hardly differed from those of goats with at least one udder half infected with CNS. In 20% and 30% of the cases the SCC was higher than 750'000 cells/ml, respectively. The relation between California Mastitis Test (CMT) reactions and udder infections was not very close. Over 20% of mammary halves infected with CNS showed negative CMT reactions. On the other hand, 25% of samples from mammary halves without a proven infection reacted positively. The large differences in individual cell counts on herd and animal level indicate that production and breeding systems might be important reasons for the higher SCC. As a consequence, the most common methods for or the control of udder health and udder infections (SCC, California Mastitis Test) are of limited value for goats. Since there was only a weak relation between milk quality properties and SCC, any arguments for the introduction of legal limits below 1 million cells per ml can hardly be found.     

Antitrypsin and N-acetyl-beta-D-glucosaminidase as markers of mastitis in a herd of Ayshire cows

Microplate analyzers for N-acetyl-beta-D-glucosaminidase (NAGase) and antitrypsin in milk were high-capacity systems for determination of subclinical mastitis on a mammary-quarter basis in a herd of 80 Ayshire cows. Infected quarters could be better differentiated from non-infected quarters by interquarter evaluation of milk NAGase and antitrypsin, than by adaptation of threshold values based on measured NAGase and antitrypsin values. This differentiation between healthy quarters and mastitic quarters was further improved by using absolute values and interquarter evaluations in combination. Therefore, quarter-based sample collection in combination with microplate technology and computerization is proposed as the method of choice for monitoring udder health in herds.     

Reduced Milk Production in Udder Quarters with Subclinical Mastitis and Associated Economic Losses in Crossbred Dairy Cows in Ethiopia

The objective of the study was to estimate the losses associated with subclinical mastitis (SCM) in crossbred dairy cows in the Central Highlands of Ethiopia. A split udder investigation was performed with 30 cows to determine production losses associated with SCM. Each quarter of the study cows was examined using the California Mastitis Test (CMT) and quarter milk production was measured over a period of 8 days. Production losses were determined for different CMT scores by comparing production of quarters with CMT score 0 to quarters with CMT scores trace, 1, 2 and 3, respectively. Using data from a recently published study, economic losses were determined for different farm sizes and production subsystems by multiplying the prevalence of the respective CMT scores with the production losses associated with these CMT scores. Mean quarter milk production was 0.82 + 0.40 kg per milking in the split udder trial. Milk production was reduced by 1.2%, 6.3%, and 33% in quarters with CMT scores 1+, 2+, and 3+, respectively. Using data from the published study, a quarter with SCM lost an average of 17.2% of its milk production. Production losses associated with SCM were estimated at 5.6% for the Addis Ababa Milk Shed. Stratified losses were highest (9.3%) in urban dairy farms (UDF) and small-scale farms (6.3%). The estimates of the financial losses ranged from US$29.1 in dairy herds in secondary towns (DHIST) to US$66.6 in UDF. A total loss of US$38 was estimated for each cow per lactation. Reducing mastitis in UDF (highest prevalence) to the level of DHIST (lowest prevalence) could reduce the loss by US$35. As this does not include costs associated with treatment or culling of diseased cows, this figure probably underestimates the possible benefits of control measures.     

Risks factors associated with subclinical mastitis in water buffaloes in Pakistan

The present study was carried to ascertain the association of various risk factors of mastitis in water buffaloes. The milk samples from buffaloes were collected and screened through California Mastitis Test for the presence of mastitis. In the present study, 15.2 % prevalence of subclinical mastitis was recorded both at the government (13.4 %) and private farms (15.5 %). The chi-square analysis showed significantly higher involvement of the right rear and front quarters. The analysis of variance technique showed significant difference in live body weight, milk yield, teat end to floor distance (P?<?0.001), udder depth, teat length, and teat diameter in mastitic and healthy buffaloes. The frequency analysis also revealed significant difference between various groups including lactation stage, teat and/or udder pathology, teat shape, and udder shape (P?<?0.001). The logistic regression analysis revealed significant positive association of mastitis with milk leakage, live body weight, milk yield, parity, calf suckling, pendulous udder, number of attendants at the farm, dirty hind legs, and udder depth.     

Concentrations of triiodothyronine (T3), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in milk from healthy and naturally infected quarters of cows

The effect of naturally acquired bacterial infection of the bovine udder on the activity of 5'-thyroxine monodeiodinase (5'-MD), and on the concentrations of the pro-inflammatory cytokines interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha in milk, from healthy (control) and inflamed quarters, was determined. The diagnostic procedure included history and clinical examination of the udder, macroscopic evaluation of secretions, the Californian Mastitis Test, determination of somatic cell counts and bacteriological examination of milk. It has been found that the milk triiodothyronine (T3) content and the 5'-MD activity from inflamed quarters were decreased when compared with controls. The decrease in the milk T3 from subclinical mastitic quarters was manifested when somatic cell counts were > 10(6) ml(-1). TNF-alpha was on average 2-fold higher in infected milk, and the concentration of IL-6 was unchanged. These results suggest that the decreased T3 content in mammary secretions during naturally occurring mastitis is associated with the severity of inflammation, increased TNF-alpha concentration and impaired enzymatic activity of 5'-MD.     

Effect of restricted suckling on milk yield,milk composition and udder health in cows and behaviour and weight gain in calves,in dual-purpose cattle in the tropics

The study was conducted to evaluate the effects of restricted suckling (RS) in dual-purpose cows and calves compared to artificial rearing (AR). Twelve Holstein-Zebu cows with calves were assigned to each treatment. Cows were milked once daily in the morning with calves present to stimulate milk let-down. RS calves suckled 30 min after milking and 30 min in the afternoon, whereas AR calves were milk-fed from nipple bottles. The daily saleable milk yield was higher in RS than in AR cows (p < 0.01; 7.44 vs 6.50 kg/day), whereas RS cows had lower milk-fat content (p < 0.001). AR cows had higher California Mastitis Test scores (p < 0.001) and lower lactose content (p < 0.001) compared to RS cows, which indicates an improved udder health in RS cows. AR calves displayed more ‘cross-suck’ during suckling/milk feeding (p < 0.001) and during observations of general behaviour (p < 0.05), and more ‘lick and sniff interior’ during milking (p < 0.05) and suckling/milk feeding (p < 0.01), compared to RS calves. During observations of general behaviour the RS calves were more often observed to ‘walk’ (p < 0.01) and ‘lick self’ (p < 0.05) and less frequently to ‘eat concentrate’ (p < 0.05) than AR calves. Results support the hypothesis that RS increases milk yield, influences milk composition and improves udder health in cows, and decreases abnormal sucking in calves.     

N-acetyl-beta-D-glucosaminidase activities, milk somatic cell counts, and blood leukocyte and erythrocyte counts in cows after heat-induced stress or after intravenous administration of adrenocorticotropic hormone

Six midlactation, nonpregnant cows were subjected to heat-induced stress or to repeated injections of adrenocorticotropic hormone (ACTH). The heat stress (experiment 1) consisted of subjecting the cows to 7 days of fluctuating temperatures (21 C during the day, 32 C during the night). The ACTH-induced stress (experiment 2) was accomplished by giving each of the cows 100 IU of ACTH twice daily for 4 consecutive days. The cows' milk somatic cell counts (MSCC), milk N-acetyl-beta-D-glucosaminidase (NAGase) activity, quarter milk production (ie, milk production per mammary quarter), blood leukocyte counts, and plasma concentrations of NAGase were monitored in both experiments. Although heat stress did not result in significant differences in NAGase, heat stress did significantly (P less than 0.05) decrease milk production. The ACTH administrations resulted in increased milk NAGase activity, MSCC, and blood leukocyte counts, and in decreased plasma NAGase activity and quarter milk production. Therefore, milk NAGase activity and MSCC were not affected by short-term heat stress, but were increased by ACTH-induced stress. Blood erythrocyte concentrations were not affected by heat stress or by ACTH-induced stress.     

Changes in mRNA of immune factors expressed by milk somatic cells of Holstein cows with hypocalcemia after calving

Changes in immune factors expressed by milk somatic cells from Holstein cows with hypocalcemia after calving were investigated in this study. Fourteen multiparous Holstein cows after their 3rd or 4th calving in one farm were used. The cows were divided into 2 groups: 7 cows needing treatment due to onset of hypocalcemia (hypocalcemia group; age = 5.53 ± 0.27 years, parity = 3.14 ± 0.14) and 7 cows without health problems (control group; age = 5.88 ± 0.31 years, parity = 3.57 ± 0.26). Milk samples were collected aseptically using a cannula and mRNA of immune factors expressed by milk somatic cells were analyzed. Milk samples (50 mL) were collected from the right rear mammary gland of cows before milking at day 1 and weeks 1, 2, 4, and 8 after calving. All milk samples showed a negative reaction to the California Mastitis Test. Levels of relative interleukin (IL)-6 and cathelicidin in the hypocalcemia group were lower than those in the control group in weeks 1 to 8. A significant difference in relative IL-6 levels was found in week 4 (P < 0.05). These results suggest that levels of IL-6 expressed by milk somatic cells may be affected by hypocalcemia in dairy cows.