Bovine serum albumin and cell counts in the diagnosis of subclinical udder infection

Summary

Puncture of the milk cisterns was performed in 120 bacteriologically positive quarters of forty‐seven lactating dairy cows on three farms. This method was used to determine whether the existing infection was an infection of the teat canal or one of the udder. The results were related to the concentration of bovine serum albumin (BSA) and the cell count in the milk. Of the bacteriologically negative quarters, both BSA levels (in 91 per cent of the quarters the BSA concentration was 0.20 mg. per ml. of milk or less) and cell counts (92 per cent contained less than 500,000 cells per ml. of milk) were low. In cases of udder infection with primary pathogenic bacteria there was a marked increase in cell count (90 per cent more than 500,000 cells per ml. of milk), whereas the increase in BSA was rather small (51 per cent still contained 0.20 mg. BSA per ml. of milk or less). While the difference in cell counts of milk from quarters with udder infections and teat canal infections with primary pathogenic bacteria was significant, the difference between the BSA levels of these two groups was not. Therefore, the cell count supplies more reliable information than does the BSA level of the milk. Of all infections, 23 per cent were found to be infections of the teat canal.     

Antibody response to toxic shock syndrome toxin-1 of Staphylococcus aureus in dairy cows

Antibody response to toxic shock syndrome toxin-1 (TSST-1) of Staphylococcus aureus in dairy cows was examined by enzyme linked immunosorbent assay (ELISA). Serum antibody to TSST-1 was not detected in 39 (76.5%) of 51 calves, which were 1-6 months of age. In contrast, TSST-1 antibody was demonstrated in 1728 (72.6%) of 2380 lactating cows housed on 36 dairy farms. The ELISA values of antibody ranged from 0.2 to 3.0 OD and presented a distribution with the peak at 1.6 OD. The mean ELISA value differed between farms, and it increased slightly along with parturient history. Somatic cell counts of milk from 174 lactating cows was compared with TSST-1 antibody and tst1,000,000 cells per ml. The mean ELISA values in milk were lower than those of sera, but they rose as somatic cells increased. The tst gene of S. aureus detected in 76.0-86.2% of the milk samples containing somatic cells > 500,000 cells per ml, a level which indicates mastitis. The data suggests that many lactating cows may be infected by TSST-1- producing S. aureus.     

A clinical trial to evaluate the effectiveness of antibiotic treatment of lactating cows with high somatic cell counts in their milk

OBJECTIVE: To determine the effectiveness of treatment of lactating cows with high somatic cell counts in milk. DESIGN: Randomised clinical trial. METHODS: Single pooled quarter samples of milk were obtained from cows with somatic cell counts above 500,000 cells/mL on fifty farms. Milk samples were cultured for known mastitis bacterial pathogens. Cows were randomly allocated to treated and untreated groups. Treated cows received both intramammary cloxacillin and parenteral erythromycin. Single pooled quarter milk samples were obtained at 6 weeks after treatment and were cultured for the presence of pathogenic bacteria. The percentage of samples with no growth at the post-treatment culture was used as an estimate of the bacteriological cures for each pathogen type and for each treatment group. Somatic cell counts of cows were compared between treatment groups and within pathogen group. The number of cows that completed a full lactation were compared between each treatment group and within each pathogen group. RESULTS: Treatment had no effect upon bacteriological cures, irrespective of pathogen present or the presence of bacteria during the previous lactation. There was no effect of treatment upon somatic cell count except for cows infected with Streptococcus dysgalactiae in which treatment caused a significant lowering of cell counts. This effect was not present in the subsequent lactation. Treatment of chronically infected cows did not alter the probability of a cow completing a full lactation but did improve the probability of newly infected cows being retained for the next lactation. Twenty-eight of 214 treated cows developed clinical mastitis in more than one quarter after treatment, thus indicating a poor technique by farmers for the insertion of intramammary antibiotics. CONCLUSIONS: Treatment during lactation of cows with high somatic cell counts in milk is ineffective in reducing bacterial infections and in reducing somatic cell counts to acceptable numbers.     

Errors associated with milk cell counting

The accuracy of somatic cell counts in milk samples was investigated in four studies. First, the counts recorded by one milk buyer in one supply over six months ranged from 105,000 to 401,000 cells/ml with no apparent changes in the volume of milk consigned or the level of mastitis in the herd that would explain this wide range. Secondly, the counts in daily samples from one bulk milk supply for 28 days ranged from 84,000 to 282,000 cells/ml, again with no apparent changes in the performance of the herd to explain the wide range. Thirdly, the replicated counts recorded for one sample by three separate laboratories agreed closely; however, when a sample with a high cell count was interspersed then two of the three laboratories reported high cell counts suggestive of 'carry-over' in excess of the 2 per cent 'allowable' Finally, cell count data from three separate laboratories on samples from 21 cows for 33 days revealed problems with the misidentification of samples on the farm in 1 per cent of the samples, and misidentification and mishandling of 1 to 2.6 per cent of the samples in the laboratories. All three laboratories differentiated samples from cows with subclinical and clinical mastitis, but the mean cell count of the uninfected cows varied between the laboratories with one of them recording statistically significantly higher counts over the period.     

Survey of the incidence and aetiology of mastitis on dairy farms in England and Wales

A survey of clinical and subclinical mastitis was carried out on 97 dairy farms in England and Wales, selected at random from members of a national milk recording scheme. The farmers were asked to collect aseptic milk samples from five consecutive cases of clinical mastitis and from five quarters with high somatic cell counts using a defined protocol, and they completed a questionnaire that included information on the cows sampled, the herd and the history of mastitis in the herd. The samples were collected throughout the year. The mean incidence of clinical mastitis was 47 cases per 100 cows per year (estimated from historic farm records) and 71 cases per 100 cows per year (estimated from the samples collected). Streptococcus uberis and Escherichia coli were isolated in pure culture from 23.5 per cent and 19.8 per cent, respectively, of the clinical samples; 26.5 per cent of the clinical samples produced no growth. The most common isolates from the samples with high cell counts were coagulase-negative staphylococci (15 per cent), S uberis (14 per cent) and Corynebacterium species (10 per cent). Staphylococcus aureus and coagulase-positive staphylococci together accounted for 10 per cent of the samples with high somatic cell counts; 39 per cent produced no bacterial growth.     

Somatic cells count in cow's bulk tank milk

The objective of this study was therefore to present factors affecting somatic cell counts in bovine bulk milk as a result of intramammary infections as well as non-infectious factors. The paper presents also the impact of on-farm management practices on the level of bulk milk somatic cell counts and presents quality indicators in bulk tank milk. At the farm level bulk milk bacterial infection takes place through three main sources: bacterial contamination from the external surface of the udder and teats, from the surface of the milking equipment, and from mastitis microorganisms within the udder. The threshold of 200,000 cells/ml identifies bacteriological negative quarters of the udder. The counts of mammary pathogens in bulk tank milk are relatively low, on average not exceeding 1,000 cfu/ml. Environmental pathogens predominate in bulk tank milk samples with somatic cells count <300 × 10(3) ml.     

Somatic cell counts of ewes' milk.

The somatic cell counts of ewes' milk were determined by an electronic particle counter (Coulter Counter). Of 1408 apparently normal milk samples, 98.2% had a somatic cell count lower than 1.0 x 10(6) cells/ml and 85.8% of 254 bacteriologically positive samples had a count higher than 1.0 x 10(6) cells/ml. Values exceeding 1.0 x 10(6) cells/ml are indicative of subclinical mastitis, if samples were collected from clinically healthy mammary glands.     

Use of domestic detergents in the California mastitis test for high somatic cell counts in milk

The California mastitis test (CMT) is used on farms to identify subclinical mastitis by an indirect estimation of the somatic cell count (SCC) in milk. Four commercially available detergents were compared with a bespoke cmt fluid for their ability to detect milk samples with a scc above 200,000 cells/ml; differences between the interpretation of the results of the tests by eight operators were also investigated. The sensitivity and specificity of the test were affected by the type of detergent, and by the operators' interpretations. When used by the most sensitive operator, suitably diluted Fairy Liquid performed almost identically to cmt fluid in identifying milk samples with more than 200,000 cells/ml. The average sensitivities achieved by the eight operators for detecting this threshold were 82 per cent for Fairy Liquid and 84 per cent for cmt fluid, and the specificities were 93 and 91 per cent respectively. The other detergents contained less anionic surfactants and were less sensitive but similarly specific.     

Aetiology of clinical mastitis in six Somerset dairy herds.

Clinical mastitis was monitored in six Somerset dairy herds for one year. The herds all had three-month geometric mean bulk milk somatic cell counts of less than 250,000 cells/ml. Escherichia coli was the predominant pathogen isolated on all the farms and in all months of the year. Environmental pathogens accounted for 61.4 per cent of all cases of clinical mastitis and for 79.3 per cent of the mastitis cases in which an aetiological agent was identified. The mean annual incidence was 41.6 cases per 100 cows (range 14 to 75). Affected cows suffered a mean of 1.5 cases and 16.4 per cent of quarters suffered at least one repeat case. Mastitis due to E. coli was more severe than mastitis due to other causes and it tended to be more severe in early lactation and during the housing period. Mastitis was significantly more severe (grades 2 and 3) in the herd with the lowest bulk milk somatic cell count and in the herd which was kept indoors throughout the year than in the other four herds. Mastitis was fatal in 2.2 per cent of cases and resulted in the death of 0.6 per cent of the lactating cows.     

Costs associated with selected preventive practices and with episodes of clinical mastitis in nine herds with low somatic cell counts

Nine dairy herds (mean size, 149 cows) with bulk-tank milk somatic cell counts of less than 300,000 cells/ml and greater than 80% of cows with Dairy Herd Improvement Association linear somatic cell counts less than or equal to 4 were selected for study. Each herd was monitored for 12 consecutive months. Duplicate quarter-milk specimens were collected from each cow for bacteriologic culturing at beginning of lactation, cessation of lactation, and at the time of each clinical episode of mastitis. Streptococcus agalactiae was never isolated and Staphylococcus aureus was isolated from less than 1% of all quarters. There were 554 episodes of clinical mastitis. During the year of study, the incidence rate of clinical mastitis varied from 15.6 to 63.7% of cows among the 9 herds. Mean costs per cow per year in herd for mastitis prevention were: $10 for paper towels, $3 for nonlactating cow treatment, and $10 for teat disinfectants. Mean cost associated with clinical mastitis was $107/episode. Approximately 84% ($90) of the costs attributed to a clinical episode were associated with decreased milk production and nonsalable milk. Costs of medication and professional veterinary fees per clinical episode varied significantly among the 9 herds. Three of the herds did not have a veterinarian treat a clinical episode of mastitis during the year of study even though 2 of these herds had the first and third highest incidence rates of clinical mastitis. When calculated on a per cow in herd basis, mean costs of $40/cow/year were attributed to clinical mastitis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seasonal variation of bulk milk somatic cell counts in UK dairy herds: investigations of the summer rise

Individual cow somatic cell count (SCC) patterns were explored over a one year period in 33 dairy herds to investigate the reason for a summer rise in bulk milk somatic cell counts (BMSCC). Cow test day somatic cell counts were categorised according to the magnitude of change since the previous test day reading, to examine which categories were responsible for the summer increase. Multilevel models using Markov chain Monte Carlo methods were specified to estimate the number of somatic cells/ml produced by different cell count categories. Stage of lactation and parity were accounted for in the models. There was an increase in the proportion of cows that remained above 200,000 cells/ml for two consecutive recordings in summer and this group of cows were responsible for 70.8% of the increase in somatic cells/ml produced from May to September compared with October to March. There was no evidence that a greater new infection rate (somatic cell counts moving from below 100,000 cells/ml to over 200,000 cells/ml) contributed to the increased summer bulk milk somatic cell counts. There was no indication that a general small increase in all somatic cell counts played an important role in the increased summer somatic cell counts. Markov chain Monte Carlo methods provided a valuable and flexible platform for parameter estimation in reasonably complex multilevel models.     

Effects of a two-year dairy herd health management programme on udder health, use of antibiotics and longevity

Since 2003, the Research Institute of Organic Agriculture (FiBL) is realizing a herd health management programme ("pro-Q" project) focussing on udder health. The objectives of the project are: (1) to reduce antibiotic mastitis treatments, (2) to optimise udder health and (3) to improve longevity, measured as averaged number of herd lactations. The farms get expert advice in prevention and treatment on herd- and animal-level. After 2 years, treatment recordings of the 65 investigated farms showed that antibiotic mastitis therapies were reduced from 38 to 26 treatments per 100 cows and year (equals a reduction of 32%). Lactation numbers of the herds increased significantly by 0.2 lactations from 3.3 to 3.5 lactations per cow. Udder health remained constant over all farms during 2 years: theoretical bulk milk cell counts averaged constantly at around 180000 cells/ml. Improvement of udder health on farm level was significantly influenced by higher somatic cell count when the project started and enhanced by farmer's motivation and farm-veterinarians' commitment to the project.     

Effect of the intramammary device on milk infection status, yield, and somatic cell count and on the morphological features of the lactiferous sinus of the bovine udder

Twenty primiparous heifers were fitted intramammarily with polyethylene coils in both quarters of one random right or left udder half at 5 days after parturition. Foremilk samples were collected and udder-half milk yields were measured at the afternoon milking on days - 1, 3, 7, and 14 and on every 14th day for 8 months after the device was inserted. Three weeks after the heifers were fitted with the intramammary device, 6 were euthanatized for gross observation of devices and tissues and cytologic evaluation of the gland cistern epithelium. There were significantly fewer bacterial isolations (P less than 0.01) and less clinical mastitis (P less than 0.05) in treated quarters than in the control quarters. Coagulase-negative staphylococci were isolated at frequencies of 0 and 15.2% for treated and control quarters. The reduction in isolation frequency for treated, compared with control, quarters was less marked with other organisms. Intramammary devices in no way interfered with the milking process. Milk yields per milking were 4.2 kg for treated udder halves compared with 4.4 kg for control halves; however, this 0.2 kg difference was not significant. Mean milk somatic cell counts, as determined by electronic counter, were 34 X 10(3) and 81 X 10(3) cells/ml for control and treated quarters (P less than 0.05). Mean bovine serum albumin values were 0.160 and 0.175 mg/ml for control and treated udder halves (P less than 0.05), indicating an increased capillary permeability due to the device. Quantitative morphologic analysis of gland cisterns showed a significant (P less than 0.05) change toward a single layer of epithelial cells in treated quarters compared with a double layer in control quarters.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of the lactation period on the cell content of sheep milk]

Milk samples of 201 ewes were examined in 6 week intervals during a complete lactation period. Those samples were analyzed for the presence of pathogenic bacteria and the somatic cell count was determined. Besides, the California Mastitis Test (CMT) was performed and the udder was clinically examined. The cell counts were found to depend on the lactation period. During 6 weeks following parturition the cell count was 63,000 cells/ml. This number decreased towards the 24th week of lactation to 32,000 cells/ml. At the end of lactation this value increased again to 425,000 cells/ml. The median value of ewes with normal udder health was 56,000 cells/ml milk. For samples from which pathogenic bacteria were isolated this value was 159,000 cells/ml. The most frequent pathogens isolated from the milk samples were coagulase-negative cocci (59.6% of bacteriologically positive samples), the median number being 88,000 somatic cells/ml in these sheep. Coagulase-positive cocci were isolated in 25.3% of the samples, the median value of the cell count was 295,000 cells/ml. In 12.1% of the samples streptococci were found. The median value was 167,000 cells/ml. From the remaining 3.0% of bacteriologically positive samples Pasteurellae, E. coli and Actinomycetae were isolated. The median value of the somatic cell count was 184,000 cells/ml. We consider coagulase-positive cocci therefore as the most pathogenic bacteria for the ovine udder.     

Determination of the number of somatic cells in milk using the rapid diphenylamine DNA filter method

The rapid reaction of the diphenylamine agent with DNA was used for the determination of the counts of somatic cells in cow's milk, using the DNA filter method. The method is based on the filtration of a warmed (65-70 degrees C) mixture of milk with Triton X-100 through the Synpor nitrocellulose membrane filter, pore size 2 to 5 microns, and subsequent DNA determination of the collected somatic cells by the colour reaction of diphenylamine. A 2ml quantity of distilled water and 4 ml of diphenylamine reagent were added to the membrane filters with somatic cells. The mixture is warmed in water bath at 90 to 100 degrees C for 20 min., then it is cooled, centrifuged (3500 X g, 15 min.), and the optical density is measured at 595 nm. The relation 8 micrograms = 1 million cells was used for the conversion of DNA content to the counts of cells. The average variation coefficient of the determination was 5.9% and the coefficient of correlation between the diphenylamine DNA filter method and the direct microscopy of the somatic cells on membrane filters was r = 0.997. Using the diphenylamine DNA filter method, the counts of somatic cells can also be determined from milk samples stored in frozen condition or from the filters with collected cells kept at the temperature of 4 degrees C (10 days) or 25 degrees C (3 days). Milk stabilized with formaldehyde can also be used for the determination if stored at 4 degrees C.     

The contribution of mammary infections by coagulase-negative staphylococci to the herd bulk milk somatic cell count

Quarter foremilk samples were taken at 2–3 weekly intervals for several years in a experimental herd comprising about 45 cows. The samples were submitted to bacteriological analysis and somatic cell counting. The most prevalent quarter infections from 1982 to 1988 were by coagulase-negative staphylococci (15–20% of all the quarters sampled). Most of these (75.6%) persisted until drying-off Dry cow therapy eliminated 86.5% of these infections. Comparison of udder quarters within cows, involving 775 samples from pairs of non-infected quarters and quarters infected by coagulase-negative staphylococci, yielded geometric means of somatic cell counts of 210 000 and 420 000 cells/ml, respectively. The correlation (r=0.87) between the herd bulk milk somatic cell count (SCC) and its estimation from the quarter milk somatic cell count performed on the same day allowed us to evaluate the contribution of the different categories of quarters, according to their infection status, to the herd bulk milk SCC. Quarters infected by a major pathogen (8.5% of samples) gave rise to 46.6% of the total number of cells, while quarters infected by coagulase-negative staphylococci (17.8% of samples) gave rise to 18.1%. Although coagulase-negative staphylococci represented only a secondary source of somatic cells as compared to major pathogens, they were not a negligible source considering the threshold of 300 000 somatic cells advocated for herd milk of good quality.     

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.     

酵母培养物益康XP对奶牛血浆内毒素含量及其它指标影响的研究

选取泌乳初期(泌乳天数<40d)健康奶牛12头,按胎次、泌乳天数和产奶量相同或相似的原则进行配对,分为试验组和对照组。两组日粮组成相同,但试验组牛每天添加益康XP(剂量为试验第1-7d500g/头/d,从第8d后减少到120g/头/d,分两次饲喂)。试验期为60d。于试验前1d、试验第20d、40d、60d分别采血,用鲎试剂法测定血浆内毒素含量;定期测定日均产奶量、乳脂率、乳蛋白率和体细胞数;并记录情期受胎率及疾病发生情况。结果表明:试验组奶牛在试验后第20d、40d、60d血浆内毒素均极显著低于对照组(p<0.01);产奶量在第20d、60d显著高于对照组(p<0.05);两组奶牛乳脂率、乳蛋白率、体细胞数差异均不显著(p>0.05);试验组情期受胎率极显著高于对照组(p<0.01);在疾病发生率方面,试验组极显著低于对照组(p<0.01)。     

The associations between milk production, milk composition and Salmonella in the bulk milk supplies of dairy farms in Ontario.

The purpose of this study was to assess changes in dairy herd milk production and milk composition associated with changes in Salmonella contamination of bulk milk on dairy farms in southwestern Ontario. Twenty-three dairy farms that had submitted milk filters for culture from which Salmonella were isolated (cases) and 23 farms that submitted Salmonella-negative milk filters (controls) were included in the study. The rolling herd averages for milk and fat of case and control farms for the months of December 1985, December 1986 and April 1987 were compared and no significant differences were detected. Case and control farms were divided into three groups (A,B,C) on the basis of Salmonella culture results of milk filters submitted at various time periods throughout the study. Daily and monthly changes in milk production and composition parameters that reflected the time periods of milk filter culture were compared. The following unconditional associations between a changing Salmonella infection status on dairy farms and changes in milk production or composition variables were significant (p less than or equal to 0.05): group A: case farms had higher plate loop counts than control farms; group B: case farms had younger cows than control farms; group C: case farms had cows with longer average days in lactation than control farms. After analytical control of confounding variables, the disappearance of Salmonella from bulk milk supplies of dairy farms was associated with a decrease in percent fat and in somatic cell count.     

Causative organisms and somatic cell counts in subclinical intramammary infections in milking goats in the UK

The bacterial causes of subclinical mastitis were determined in samples of milk taken from one half of the udders of 159 goats in three different herds. The mean prevalence of subclinical infection was 33 percent, with prevalences of 26 percent, 39 percent and 42 percent in the three herds. Staphylococcus aureus was isolated from seven (13 percent) of the 53 infected halves, coagulase-negative staphylococci accounted for 47 percent, Corynebacterium species for 31 percent and alpha-haemolytic streptococci for 6 percent of the infected samples. The mean somatic cell count of the uninfected milk samples was 428,000 cells/ml, and 93 percent of uninfected samples had counts less than 1,000,000 cells/ml; the mean cell count of the infected samples was 2,785,000 cells/ml.