Prevalence of contagious mastitis pathogens in bulk tank milk in Prince Edward Island

The purpose of this study was to 1) estimate the herd prevalence of contagious mastitis pathogens in bulk milk from Prince Edward Island (PEI) dairy farms, 2) determine the association between bulk milk culture results and mean bulk milk somatic cell count (BMSCC), and 3) investigate the agreement of repeated bulk milk cultures. Three consecutive bulk milk samples were obtained at weekly intervals from all 258 PEI dairy herds and were cultured using routine laboratory methods. Cumulative prevalence of Staphylococcus aureus, Streptococcus agalactiae, and Mycoplasma spp. (M. bovis and M. alkalescens) was 74%, 1.6%, and 1.9%, respectively. Bulk milk somatic cell count of Staph. aureus-positive herds was higher than that of negative herds. Agreement for Staph. aureus isolation between 3 consecutive tests was moderate (kappa = 0.46). Mycoplasma bovis and M. alkalescens in bulk milk are being reported for the 1st time in PEI ever and in Canada since 1972.     

Culture of bulk tank milk as a mastitis screening test: A brief review

The culture of a sample of bulk tank milk may be a useful technique by which to screen herds for major mastitis pathogens. Staphylococcus aureus and Streptococcus agalactiae, if identified on a culture of a sample of bulk milk, reliably indicate infection of the udder. Environmental bacteria, such as the other streptococci and coliforms, are unlikely to be indicative of the proportion of cows infected with these organisms.Samples of bulk milk are readily obtainable and can be rapidly and inexpensively cultured to screen large numbers of herds for mastitis-causing bacteria, yet the performance of the test has only recently been formally assessed for its ability to correctly classify herds according to infection status.A single culture of bulk tank milk has been found to be a test with low sensitivity and high specificity for determining the presence of S. agalactiae or S. aureus in the herd. This means that many infected herds will be called negative, but few uninfected herds will be classified as positive.The literature assessing the performance of bulk tank milk culture in comparison with other mastitis screening tests, the use of bulk milk culture for prevalence surveys, and factors affecting these results is discussed.     

Management factors associated with the incidence of clinical mastitis over the non-lactation period and bulk tank somatic cell count during the subsequent lactation

Aim: To evaluate associations between management decisions related to the control of mastitis, including the infusion of antibiotics at the end of lactation (dry-cow therapy; DCT), on the incidence of clinical mastitis over the non-lactating period and the bulk tank somatic cell count (BTSCC) in the subsequent lactation. Methods: Dairy herd owners (n=158) provided information via a retrospective survey about (a) the proportion of their herds treated with DCT; (b) DCT management, including: number of occasions on which cows were dried off; manipulation of feed and water intake around drying off; infusion technique (partial vs full depth insertion of cannula); and hygiene before and after DCT infusion; (c) occurrence of mastitis and frequency of occurrence following drying off and in the subsequent lactation; (d) number of cows culled for mastitis-related conditions; (e) reasons for culling; (f) incidence of clinical mastitis; and (g) stock purchase policy with regard to mastitis. The BTSCC for each vat of milk supplied for the 1999/2000 and 2000/2001 seasons, and records of antibiotic purchases were collated for each herd. The probability that >2% of cows within a herd were diagnosed with clinical mastitis over the dry period was initially examined using univariate analysis (i.e. chi2 or logistic regression) and associated factors (p<0.2) were offered to a reverse stepwise logistic regression model. Factors hypothesised as being associated with the average lactation log10 BTSCC for the 2000/2001 season were initially examined using univariate analysis (i.e. ANOVA or linear regression analysis) and associated factors (p<0.2) were then tested using a forward manual model-building approach. Results: Increasing the percentage of the herd treated with DCT at the end of lactation was associated with reduced probability that >2% of a herd would be diagnosed with clinical mastitis over the non-lactating period and with a lower BTSCC in the subsequent lactation (p<0.01). A lower BTSCC was associated with small herds (<150 cows; p<0.05), not reducing feed intake around drying off (p<0.05), checking for clinical mastitis over the dry period in the milking parlour rather than at pasture (p<0.05), partial insertion of the DCT cannula (p<0.01), and use of 'change in udder shape' during lactation as a diagnostic criterion for mastitis (p<0.05). The incidence of clinical mastitis over the dry period was positively associated with reduced feeding around drying off (p=0.05) and the estimated volume of milk being produced at the time of drying off (p=0.014). Conclusions: Use of dry cow therapy was associated with fewer cases of clinical mastitis over the non-lactating period and reduced BTSCC over the subsequent lactation. Reduced BTSCC was also associated with smaller herds, use of partial (compared with full depth) insertion of the DCT cannula, not reducing feed intake at the time of drying off, checking for clinical mastitis over the dry (non-lactation) period in the milking parlour, and use of udder shape for diagnosis during lactation. Control of clinical mastitis and BTSCC involves a range of management practices that need to be used in conjunction with DCT. Keywords: Dairy cows, mastitis, dry-cow therapy, somatic cell count, management practices.     

Herd characteristics and management practices associated with bulk-tank somatic cell counts in herds in official Dairy Herd Improvement Association programs in Ohio

OBJECTIVE: To identify herd characteristics and management practices associated with bulk-tank somatic cell counts (BTSCC) in dairy herds in Ohio enrolled in official Dairy Herd Improvement Association (DHIA) programs. SAMPLE POPULATION: 186 dairies in Ohio. PROCEDURE: All herds in official DHIA programs in 9 counties were asked to participate. Extensive information regarding herd characteristics and management practices was obtained, using a standardized questionnaire. Bulk-tank milk samples were requested from all participating herds for bacterial culture. Official DHIA test-day records for January 1997 were obtained from all herds enrolled in official DHIA programs in the 9 counties. Potential associations were identified, using multivariable ANOVA. RESULTS: Participation was 186 of 479 (39%) herds. Streptococcus agalactiae and Mycoplasma spp were not isolated from bulk-tank milk samples. Staphylococcus aureus was isolated from 64 of 172 (37%) of the herds. The BTSCC were inversely associated with peak daily milk production, postmilking teat disinfection, percentage of eligible cows in the herd detected in estrus, and directly related to the extent to which BTSCC was perceived as a herd problem during the preceding 2 years. Type of housing for nonlactating cows and product used for treatment of nonlactating cows also were significantly associated with BTSCC. CONCLUSIONS AND CLINICAL RELEVANCE: Consideration of herd characteristics and implementation of management practices associated with BTSCC could result in increased milk yield and production of milk with lower BTSCC.     

Testing of bulk tank milk for Salmonella Dublin infection in Danish dairy herds.

The usefulness of enzyme-linked immunosorbent assay (ELISA) was investigated as a simple method to screen for Salmonella Dublin infection in dairy herds, examining bulk tank milk samples for lipopolysaccharide (O:1,9,12) antibodies. The cut-off value for the ELISA on bulk tank milk was established based on individual milk samples (n = 2887) and bulk tank milk from 52 herds. Bulk tank milk samples (n = 5108) were collected from 1464 dairy herds located in 19 different areas. About 10% of the dairy herds in Denmark participated in the study. The percentage of herds changing from test-negative to test-positive in each area was correlated with the incidence of S. Dublin outbreaks in the corresponding county (r = 0.48, n = 19; P < 0.025). The mean level of the OD values obtained in the first and third test rounds was not constant (Pr /t/ = 0.0001). The study demonstrated that the probability of being test-negative in the third test round was 0.926 for a herd with 2 previous test-negative results. It was concluded that the investigated ELISA method was in general accordance with the cases of clinical S. Dublin infection recorded, and that the method has a potential for national screening purposes.     

Bulk tank milk analysis: factors associated with appearance of Mycoplasma sp. in milk

Factors associated with the presence of Mycoplasma sp. in bulk tank milk samples were evaluated from 664 herds during 2.25 years. Milk quality components were not strongly related to the presence of Mycoplasma sp. in bulk tank milk. The presence of other contagious mastitis pathogens, Staphylococcus aureus and Streptococcus agalactiae, was also not related to the presence of mycoplasma, suggesting that the aetiology and transmission of mycoplasma mastitis were different from transmission of other contagious mastitis pathogens. The occurrence of the first isolation of mycoplasma from a bulk tank was not correlated to season of the year. Mycoplasma in bulk tank milk samples were more likely to be found in herds shipping more milk, an indirect measure of herd size. This suggests that larger herds are more likely to have mycoplasma mastitis. However, the first appearance of mycoplasma mastitis in a bulk tank sample was followed by a sample without this pathogen in more than 60% of herds. Mycoplasma sp. was not detected in any herd a year after first isolation. These findings suggest that this pathogen could be controlled and eliminated from herds.     

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.     

Streptococcus agalactiae mastitis: a review.

Streptococcus agalactiae continues to be a major cause of subclinical mastitis in dairy cattle and a source of economic loss for the industry. Veterinarians are often asked to provide information on herd level control and eradication of S. agalactiae mastitis. This review collects and collates relevant publications on the subject. The literature search was conducted in 1993 on the Agricola database. Articles related to S. agalactiae epidemiology, pathogen identification techniques, milk quality consequences, and control, prevention, and therapy were included. Streptococcus agalactiae is an oblique parasite of the bovine mammary gland and is susceptible to treatment with a variety of antibiotics. Despite this fact, where state or provincial census data are available, herd prevalence levels range from 11% (Alberta, 1991) to 47% (Vermont, 1985). Infection with S. agalactiae is associated with elevated somatic cell count and total bacteria count and a decrease in the quantity and quality of milk products produced. Bulk tank milk culture has, using traditional milk culture techniques, had a low sensitivity for identifying S. agalactiae at the herd level. New culture methods, using selective media and large inocula, have substantially improved the sensitivity of bulk tank culture. Efficacy of therapy on individual cows remains high. Protocols for therapy of all infected animals in a herd are generally successful in eradicating the pathogen from the herd, especially if they are followed up with good udder hygiene techniques.     

The selenium status of dairy herds in Prince Edward Island

Bulk tank milk selenium (Se) concentration was compared with mean serum Se concentration in 15 herds and was found to be an accurate reflection of the herd Se status. The Se status of 109 Prince Edward Island (PEI) dairy herds was monitored for 1 year using bulk tank milk Se concentration. Fifty-nine percent of the herds surveyed were, at some point, found to be marginal or deficient in Se, putting them at risk of disease and suboptimal production. The periods of greatest risk of deficiency were fall and winter, at which time 5% and 4%, respectively, of herds sampled fell in the range considered truly deficient in Se. Herds in which Se supplementation was provided in the form of a commercial dairy concentrate were over 4 times more likely to be Se-adequate than herds not using this method, and adjusted average daily milk yield was 7.6% greater in herds determined to be Se-adequate when compared with Se-marginal herds. We conclude that many dairy producers in PEI are providing insufficient supplementary Se in the ration to meet the recommended Se intake for lactating cows.     

A longitudinal survey of anti-Ostertagia ostertagi antibody levels in individual and bulk tank milk in two dairy herds in Normandy

The Ostertagia-specific antibody levels in milk were monitored in 2 dairy herds to investigate seasonal variations and the relationship between individual and bulk tank milk antibody levels. Bulk tank and individual milk samples from all lactating animals were collected over a 1-year period at weekly and monthly intervals, respectively. The Ostertagia-specific antibody levels were measured with an indirect ELISA and the test results were expressed as optical density ratios (ODR). A clear seasonal pattern that followed the expected intake of infectious larvae was observed in the individual and bulk tank milk antibody levels of both herds. Within each herd, there was a large variation in the individual ODRs. This variation remained large when the distribution of individual ODRs was plotted according to high and low bulk tank milk ODR categories. The results suggest that the effect of seasonal variations on cut-off levels that predict production responses after anthelmintic control, needs to be assessed.     

Persistent shedding of Salmonella enteritidis from the udder of a cow

Salmonella enteritidis phagetype 8 was isolated from ill humans, milk-line filters, milk from a bulk tank, and milk from the right hind quarter of a five-year-old Holstein cow on a dairy farm in southern Alberta. The affected animal was removed from the herd and continued to shed S. enteritidis from this quarter during a seven-month interval in an isolation facility. Milk from the affected quarter was visually normal, and no other pathogen was isolated from the udder during the investigation. After removal of the infected cow from the herd, milk from the bulk tank was culturally negative for Salmonella sp. during the succeeding 15 months.     

Impact of intramammary infections in dairy heifers on future udder health, milk production, and culling

Dairy heifers represent the future of a dairy herd, and are expected to freshen with a healthy and well-developed udder, capable of producing an optimal amount of high quality milk. A high proportion of heifers have infected mammary quarters at calving, with coagulase-negative staphylococci (CNS) being the most common cause. Staphylococcus aureus and environmental pathogens are also found. The aim of this paper is to summarize how intramammary infections during (late) gestation and early lactation impair the development of the mammary gland and negatively affect future udder health and milk production. Heifers calving with either subclinical or clinical mastitis are also at a higher risk to be culled in first lactation. The magnitude of the effect is most likely related to the virulence of the causative pathogen, the persistence of the infection when milk production has started, and the time of onset of infection. Histological changes in udder tissue from quarters infected with S. aureus are more pronounced than those in udder tissue from CNS-infected quarters. The longer the infections exist and the longer they persist into lactation, the larger the impact on heifers' future udder health and milk production will be. In general, CNS infections are cleared early in lactation and some studies show that CNS do not have a large impact on future milk production and udder health. Future research should elucidate to what extent pathogen-specific as well as host-related factors affect the persistence of IMI in early lactating heifers.     

Prevalence study of Staphylococcus aureus in quarter milk samples of dairy cows in the Canton of Bern, Switzerland

In the present study, the prevalence of S. aureus in mammary gland quarters of dairy cows in Switzerland was estimated and a risk factor analysis was carried out. Dairy cows were selected by one-step-cluster sampling with stratification by herd size. Forty-seven of 50 randomly chosen farms participated in the study, resulting in 603 cows and 2388 quarter samples. Milk samples were collected in all herds on two occasions two weeks apart. In 6% of cows (95% CI: 2.7-9.3%) at least one milk sample was positive for S. aureus and from 2% (0.8-3.2%) of all quarters, S. aureus was cultured at least once. In four quarters a latent S. aureus infection (agent detected and somatic cell count (SCC) <100,000cell/ml) was diagnosed. Multivariable hierarchic logistical regression analysis yielded five significant risk factors for observing S. aureus in a milk sample: high SCC, a S. aureus-positive neighbouring quarter, a palpable induration in the quarter, and a wound, scar tissue or crush injury affecting the teat. The type of housing (P=0.1596) was also a factor that remained in the model. The mentioned risk factors must be considered during the evaluation of herds with S. aureus problems. The occurrence of latent S. aureus infections emphasises that not only quarters with a high SCC but all quarters of all cows must be cultured for control measures to be effective.     

Evaluation by enzyme-linked immunosorbent assay of local and systemic production of milk immunoglobulins to surface exopolysaccharide antigen in cows with staphylococcal mastitis

An enzyme-linked immunosorbent assay (ELISA) was developed to evaluate milk immunoglobulin levels to surface exopolysaccharide antigen of Staphylococcus aureus in cows with staphylococcal mastitis. Quarter milk samples were obtained from 24 lactating dairy cows on two occasions, one month apart. Cows were classified as S. aureus-positive (S. aureus cultured from at least one quarter on both sample dates) or S. aureus-negative. Individual quarter samples were tested for IgA (representing local synthesis) and IgG1 (primarily of systemic origin) specific for staphylococcal surface exopolysaccharide antigen. No significant differences were found for specific IgA or IgG1 between S. aureus-positive and S. aureus-negative cows, nor between infected and non-infected quarters of S. aureus-positive cows. The data indicate that, in cows with staphylococcal mastitis, milk immunoglobulins specific for exopolysaccharide antigen are not significantly increased by either the systemic or the local immune response.     

An investigation of bulk tank milk selenium levels in the San Joaquin Valley of California

We evaluated selenium determination of bulk milk tank samples as an alternative to testing blood selenium for evaluating herd selenium status in DHIA dairy herds in the San Joaquin Valley of California. A method of determining milk selenium levels using inductively coupled plasma spectrometry is described. Mean bulk tank milk selenium levels were 0.0224 mg/L (Range 0.0126-0.0418 mg/L). No statistically significant relationships were found between bulk tank milk selenium levels of a herd and calving interval, days open or log somatic cell counts. Mean herd blood and milk levels were directly proportional to bulk tank milk selenium levels. Within a herd milk selenium levels of a cow were directly proportional to the cow's blood selenium level. Herd selenium levels were not significantly related to soil selenium levels. Determination of bulk tank milk selenium levels has the potential to be a low cost, non-invasive means of evaluating herd selenium levels in order to determine selenium deficiency. Further studies with this technique in areas which are deficient in selenium may provide estimates of the sensitivity, specificity and predictive value of bulk milk tank selenium for determining selenium deficiency in dairy herds.     

Detection of Staphylococcus aureus in bulk tank milk using modified Baird-Parker culture media.

The purpose of this project was to evaluate the use of 2 selective/differential culture media for detecting Staphylococcus aureus in bulk tank milk. One medium was Baird-Parker agar base supplemented with egg york tellurite emulsion and acriflavine. The other medium was Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine. An increased inoculum of bulk tank milk (0.3 mL) was used to enhance the detection of S. aureus in samples containing low numbers of organisms. The sensitivity and specificity for detecting S. aureus in bulk tank milk were 94.8% and 100%, respectively, using Baird-Parker agar base supplemented with egg yolk tellurite emulsion and acriflavine, and 89.7% and 100%, respectively, using Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine. Both media are practical for detecting S. aureus in bulk tank milk and monitoring its spread in lactating dairy herds in Alberta.     

Spatial distribution of antibodies to Salmonella enterica serovar Typhimurium O antigens in bulk milk from Texas dairy herds

Environmental factors that enhance either the survivability or dispersion of Salmonella enterica serovar Typhimurium (S. Typhimurium) could result in a spatial pattern of disease risk. The objectives of this study were to: (1) describe herd status based on antibody response to Salmonella Typhimurium as estimated from bulk tank milk samples and (2) to describe the resulting geographical patterns found among Texas dairy herds. Eight hundred and fifty-two bulk milk samples were collected from georeferenced dairy farms and assayed by an indirect enzyme-linked immunosorbent assay (ELISA) using S. Typhimurium lipopolysaccharide (LPS). ELISA signal-to-noise ratios for each bulk tank milk sample were calculated and used for geostatistical analyses. Best-fit parameters for the exponential theoretical variogram included a range of 438.8 km, partial sill of 0.060 and nugget of 0.106. The partial sill is the classical geostatistical term for the variance that can be explained by the herd's location and the nugget is the spatially random component of the variance. We have identified a spatial process in bulk milk tank titers for S. Typhimurium in Texas dairy herds and present a map of the expected smoothed surface. Areas with higher expected titers should be targeted in further studies on controlling Salmonella infection with environmental modifications.     

Udder shape and teat-end lesions as potential risk factors for high somatic cell counts and intra-mammary infections in dairy cows

The association of common bacterial pathogens in milk samples during calving with udder shape or the presence of 'teat-end' lesions was investigated in 240 dairy cows from two herds. Sixty-three of 120 cows (53%) in one herd (herd A) and 54/120 animals (45%) in a second herd (herd B) had normal-shaped udders. The remaining animals had udder shapes defined as follows: large pendulous (18% herd A, 26% herd B); large between hindquarter (10% herd A, 17% herd B); overall small (8% herd A, 5% herd B); or small but pendulous (11% herd A, 7% herd B). At calving teat-end lesions were present in 63% and 76% of the quarters of herd A and B animals, respectively. There was no herd effect on udder shape or teat-end lesions. Analysis of variance revealed that udder shape and teat-end lesions did not have a significant association with quarter somatic cell count. However there was some association between mammary infection and udder shape and teat-end lesions. Compared to other udder shapes, cows with large between hindquarter shape had significantly less Staphylococcus aureus and Streptococcus uberis infection (P<0.001). There was a similar albeit less significant negative association with Escherichia coli infection (P<0.01). Infection with Streptococcus agalactiae and Streptococcus dysgalactiae was more frequent in cows with large pendulous and overall small udder conformations. The results also suggest an association between intra-mammary infection at calving and the presence of hyperkeratotic teat-end lesions, given that S. aureus, coagulase-negative staphylococci, S. uberis, S. agalactiae and E. coli were cultured from significantly more quarters with such lesions than from quarters without lesions or with other types of lesion (P<0.001).     

Molecular detection of Streptococcus agalactiae in bovine raw milk samples obtained directly from bulk tanks

Mastitis is the most common infectious disease affecting dairy cattle; in addition, it remains the most economically important disease of dairy industries around the world. Streptococcus agalactiae, a contagious pathogen associated with subclinical mastitis, is highly infectious. This bacterium can cause an increase in bulk tank bacterial counts (BTBC) and bulk tank somatic cell counts (BTSCC). The microbiological identification of S. agalactiae in samples from bulk tanks is an auxiliary method to control contagious mastitis. Thus, there are some limitations for time-consuming cultures or identification methods and additional concerns about the conservation and transport of samples. Bulk tank samples from 247 dairy farms were cultured and compared through polymerase chain reaction (PCR), directed to 16S rRNA genes of S. agalactiae, followed by BTBC and S. agalactiae isolation. The mean value of BTBC was 1.08×10(6) CFU mL(-1) and the bacterium was identified through the microbiological method in 98 (39.7%; CI(95%)=33.8-45.9%) and through PCR in 110 (44.5%; CI(95%)=38.5-50.8%) samples. Results indicated sensitivity of 0.8571±0.0353 (CI(95%)=0.7719-0.9196) and specificity of 0.8255±0.0311 (CI(95%)=0.7549-0.8827). The lack of significant difference between microbiological and molecular results (κ=0.6686±0.0477 and CI(95%)=0.5752-0.7620) indicated substantial agreement between the methods. This suggests that PCR can be used for bulk tank samples to detect contagious mastitis caused by S. agalactiae.     

Occurrence of Prototheca zopfii, a mastitis pathogen, in milk

Prototheca zopfii was isolated from cow-composite milk and bulk tank milk by a new selective Prototheca enrichment method. Repeated testing of cow-composite milk from individual cows resulted in P. zopfii isolation data indicating a strong statistical correlation of P. zopfii with specific cows. Prototheca sp. were isolated from the milk of 31 of 79 cows in a single herd, and contamination was discounted as the source. Prototheca sp. were also recovered from 28 of 787 bulk tank milk samples from the Eastern U.S.A., and 22 of 69 temperature control milk samples from pooled dairy farm milk from delivery trucks.