Epidemiological study of non-contagious intramammary infections in nine commercial dairy herds following a Staphylococcus aureus control programme

Changes in prevalence in intramammary infection, by pathogen type, in herds applying a stringent contagious mastitis control programme was studied. Enrollment of 1651 lactating cows and collection of milk samples was made in this ancillary study to a cohort study of the dynamics of mastitis prevalence after adoption of a strict contagious mastitis control programme that targeted the elimination of mastitis caused by Staphylococcus aureus. Nine commercial dairies in Italy were used. Aseptic collection of milk samples from all lactating cows was performed at the time of enrollment, from all cows within 7-14 days of entering the lactating herd after the date of enrollment, and from all lactating cows at 2, 4, 7, 10 and 14 months after the date of enrollment. Prevalence of intramammary infection by pathogen type was determined from culture of milk samples. Application of the strict contagious mastitis programme did not lead to an increased risk of non-contagious mastitis. The risk of coliform, environmental streptococcal and coagulase-negative staphylococcal intramammary infections decreased after adoption of the programme. The data reported herein indicate that the overall risk for any intramammary infections decreases with adoption of a strict contagious mastitis programme, and that such a programme therefore does not necessarily lead to an increase in environmental mastitis.     

The dynamics of Staphylococcus aureus intramammary infection in nine Danish dairy herds

The aim of the present study was to examine the diversity of Staphylococcus aureus isolates from bovine intramammary infections (IMI) in nine dairy herds, and compare these with isolates from other sites on the cows by phage- and ribotyping. Whether colonisation of milkers with S. aureus could be a source of infection for bovine IMI was investigated. In addition, 100 epidemiologically unrelated S. aureus isolates from asymptomatic human carriers were also phage- and ribotyped to compare the human and bovine reservoir of S. aureus in Denmark. A total of 625 S. aureus isolates from bovine IMI, bovine skin lesions, milking personnel, and non-farm-related human carriers were included in the study. Certain types predominated in one or several herds during the study period of one-and-a-half to two years, whereas the presence of other types was of a more sporadic nature. Within the individual herds, there was a close correspondence between ribo- and phage types of S. aureus isolated from bovine intramammary infections and skin lesions. Isolates from milking personnel, however, were not identical to any of the predominant intramammary strains. Furthermore, several of the isolates from milking personnel showed ribo- and phage patterns identical to S. aureus isolates from human carriers. The findings of the present study underline the importance of strict milking hygiene and improvement of current mastitis therapy. The results support the hypothesis that some S. aureus mastitis strains are more contagious, virulent or persistent than others. The human reservoir of S. aureus does not play a major role as a source of bovine intramammary infections.     

Duration of bovine intramammary infections in commercial dairy herds

Data on the infection status of cows on seven commercial dairy farms were collected over 492 full lactations. Foremilk samples were taken at an average interval of five weeks. A total of 249 streptococcal and 433 staphylococcal infections were diagnosed. Spontaneous elimination occurred in 49 per cent of all streptococcal infections and in 54 per cent of Staphylococcus aureus infections. The average duration of spontaneously eliminated infections was 10.8 weeks for Streptococcus agalactiae, 9.9 weeks for Strep dysgalactiae, 10.4 weeks for Strep uberis and 12.8 weeks for Staph aureus. The average duration of infections persisting until drying off was 19.3 weeks for Strep agalactiae, 18.7 weeks for Strep dysgalactiae, 18.5 weeks for Strep uberis and 25.2 weeks for Staph aureus. The method and rate of elimination of infection as found in this analysis are of value for estimating new infection rates and selecting quarters for dry cow therapy.     

The epidemiology of Staphylococcus aureus infections from subclinical mastitis in dairy cows during a control programme

The objective was to investigate the spread of S. aureus in seven dairy herds. Milk samples were taken to determine mastitis pathogens and somatic cell count. An approved hygiene programme was established to control the spread of S. aureus from quarter to quarter. S. aureus isolates were differentiated by geno- and phenotyping to trace their spread within a herd.In two herds S. aureus showed a high prevalence, but were eliminated from the herds by the control programme. In these herds and a third herd typing results identified a particular type that was found much more frequently than all other types. The frequent types were repeatedly detectable during the study. In three other herds with a lower S. aureus prevalence and also in one herd with a long history of S. aureus and high prevalence subclinical mastitis was caused by several distinguishable S. aureus types. These types occurred simultaneously in the particular herd and showed little or no tendency to spread from quarter to quarter. They seemed to circumvent the control procedures resulting in a relatively high rate of new infected animals and therefore were hardly eliminated. The typing results and the clinical observations indicated that strains differed in their tendency to spread and their ability to infect udder quarters. In three herds typing methods identified a predominant type with the common epidemiological features of a contagious mastitis pathogen, while in the other herds the S. aureus type patterns were similar to that of environmental pathogens.     

Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds.

Sixty-eight methicillin-resistant Staphylococcus aureus strains were isolated from mastitis milk samples originating from 20 Belgian dairyherds. All these strains appeared to be representatives of one single strain which was probably of human origin. Evidence is presented indicating a rapid in vivo evolutionary change in this strain. The following characteristics were found to be variable: the production of beta haemolysin inversely connected with fibrinolysin (staphylokinase) activity; the production of lipase, enterotoxin B and delta haemolysin; the resistance to neomycin, chloramphenicol, tetracyclines, methicillin and spectinomycin associated with constitutive or inducible macrolide resistance.     

Efficacy of vaccination and antimicrobial treatment to eliminate chronic intramammary Staphylococcus aureus infections in dairy cattle

OBJECTIVE: To determine whether a combination of vaccination and extended intramammary antimicrobial treatment would eliminate chronic intramammary Staphylococcus aureus infections in lactating dairy cows. DESIGN: Randomized controlled clinical trial. ANIMALS: 50 dairy cows with chronic mastitis caused by S aureus. PROCEDURE: Cows were identified and paired within herd on the basis of days in milk, lactation number, milk production, and numbers of quarters infected. Treated cows (n=20) received 3 doses of a polyvalent S aureus bacterin on days 1, 15, and 21 of the study along with intramammary administration of pirlimycin in all 4 quarters once daily for 5 treatments (days 16 to 20). Control cows (n=23) received no treatment. Follow-up samples for bacteriologic culture were collected for at least 3 months after treatment to determine treatment success rates. RESULTS: Significantly more S aureus infections were eliminated from treated cows (8/20 [40%]), compared with control cows (2/23 [9%]). The proportion of infected quarters that yielded negative results throughout the follow-up period was also significantly higher in treated cows (13/28 [46%]) than in control cows (2/41 [5%]). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that a combination of vaccination and antimicrobial treatment can be successful in eliminating some cases of chronic intramammary S aureus infections in dairy cattle. However, it is important to consider extended treatment protocols carefully because many cows are likely to remain infected with S aureus despite treatment and vaccination.     

Systematic sanitation of dairy herds with mastitis caused by Staphylococcus aureus

Several strategies are known for sanitizing dairy herd problems caused by Staphylococcus (S.) aureus. They mostly consist of general management measures but specific decision-making at an individual animal level has not been described. A sanitation program in the form of a process chart developed by the Bern Clinic for Ruminants was undertaken in 10 dairy herds with this problem. In an affected herd the cows were divided into 3 groups: healthy, suspect, infected. Three milk samples (MS), taken at two-week intervals were cultivated. The cows were grouped according to the culture results. To measure the success of the sanitation program, the key figures ?theoretical tank somatic cell count? (target < 150,000 SCC/ml) and ?percentage of cows over limit? (limit: 150'00 SCC/ml, target < 20 %) were used. These were compared with the corresponding key figures from dairy herds, which were followed-up by the Bern Clinic for Ruminants (control herds). The problem herd sanitation program lasted between 2 and 21 months. A total of 1598 MS were analyzed, of which 241 (15 %) were S. aureus positive (15 %). At the end of the sanitation the key figures between problem herds and control herds were similar. The sanitation program has proved to be practical. The detection of S. aureus positive cows proved to be reliable and the udder health of the herd could be significantly improved.     

Dynamics of Staphylococcus aureus infections during vaccination with an autogenous bacterin in dairy cattle

The effect of an autogenous vaccine against Staphylococcus aureus on S. aureus prevalence and mastitis, as well as on somatic cell count (SCC), was studied in a dairy herd with a high prevalence of S. aureus. The vaccination group (n = 35; 22 cows and 13 heifers) and the control group (n = 36; 23 cows and 13 heifers) received the vaccine or a placebo, respectively, according to the following protocol: all animals: basic immunization (twice, 3 weeks apart); cows: booster dose at the time of drying off, 5 and 2 weeks before calculated calving date; heifers: booster dose 2 and 5 weeks before calculated calving date. The vaccine or the placebo was administered subcutaneously in the area of the supramammary lymph nodes. Quarter milk samples were collected monthly and subjected to SCC and bacteriological evaluation. At this time, the animals were also checked for signs of clinical mastitis. Non-clinical S. aureus mastitis diagnoses were based on udder quarter SCC and a positive S. aureus culture. In order to compare the SCC in individual whole milk samples, records from the monthly milk quality testing were evaluated. Cow and udder quarter prevalence of S. aureus intramammary infections calculated for the experimental animals and quarters, respectively, did not differ between groups. However, during the lactation period following the boostcr dose, the prevalence of S. aureus increased in both groups (P < 0.05). The cumulative incidence of various mastitis diagnoses (clinical, subclinical, latent infection) due to S. aureus on an animal basis did not differ between groups. On an udder quarter basis, the cumulative incidence of subclinical mastitis was higher in vaccinated animals than in control animals (33.8 versus 26.0%; P < 0.05). This was mainly due to a higher cumulative incidence of subclinical mastitis in vaccinated than control heifers. The SCC in composite milk samples did not differ between groups, but increased as lactation progressed. The herd prevalence of S. aureus differed considerably throughout the study period, but declined consistently to below 10% at the end of the study period. Recent herd checks revealed a prevalence of S aureus infections of < 5%. It is concluded that the autogenous bacterin tested in this study did not have the desired effect on the prevalence of S. aureus infections and mastitis or SCC. The decline in S. aureus prevalence was very probably due to other factors than specific immunization against S. aureus.     

Use of partial budgeting to determine the economic outcome of Staphylococcus aureus intramammary infection reduction strategies in three Ohio dairy herds

Efforts to reduce the prevalence of Staphylococcus aureus intramammary infection were monitored in 3 Ohio dairy herds. Bacteriologic culturing of milk from all lactating cows in each herd was completed multiple times to identify infected cows and monitor reduction. Partial budgeting techniques were used to determine the economic outcome of the reduction program. Of particular emphasis was the economic impact of culling to maintain or achieve milk quality premium payments on the basis of bulk tank somatic cell counts. The prevalence of S aureusinfected cows was reduced in each herd. Culturing of milk from all lactating cows appeared to be an effective method to identify infected cows. Although numbers were limited, it also appeared that culturing of composite quarter samples was effective as a herd screening test to identify S aureus-infected cows. Bacteriologic culturing had a negative financial impact in all 3 herds. Using partial budgeting to assess the economic impact of the programs, it was determined that 2 herds experienced negative financial impacts as a result of an excess culling rate when compared with a 12-month baseline period prior to the initiation of the project. All herds had increased milk production per cow during the study as measured by the mature-equivalent method. However, when actual production was considered, increased milk production in each herd was not as great as that of other Ohio herds enrolled on Dairy Herd Improvement Association testing programs.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genetic diversity of Staphylococcus aureus isolated from ovine intramammary infections in Norway

Three hundred and eighty-four Staphylococcus aureus isolates obtained from mammary secretions from 332 ewes kept for meat production were typed by pulsed-field gel electrophoresis (PFGE). The ewes were from 242 flocks located in 13 counties distributed in four regions of Norway. In total, 64 different pulsotypes were identified, 31 of these were represented by a single isolate. Fifty-nine percent of the isolates belonged to one of five closely related pulsotypes. This group of pulsotypes occurred in all the counties. Although widely disseminated, the proportions of the prevalent and closely related pulsotypes differed between the regions. Nine pulsotypes were unique to single regions but the number of isolates belonging to each of these pulsotypes was low. Resistance to penicillin was found in only 3 of the 384 S. aureus isolates. These represented three different single banding patterns, not related to any of the prevalent pulsotypes found.     

Decision analysis model for paratuberculosis control in commercial dairy herds

A previous economic test-and-cull decision analysis model has been strengthened and updated with current epidemiologic information. Created using Excel and PrecisionTree software, the model incorporates costs and benefits of herd management changes, diagnostic testing, and different management actions based on test results to control paratuberculosis in commercial dairy herds. This novel "JD-Tree" model includes a herd management decision node (four options), a test/no test decision node (two options), a diagnostic test choice decision node (five options), test result chance nodes (four levels of possible results), and test action decision nodes (three options; cull, manage, no action). The model culminates in a chance node for true infection status. Outcomes are measured as a net cost-benefit value to the producer. The model demonstrates that improving herd management practices to control infection spread (hygiene) is often more cost-effective than testing; not all herds should test as part of a paratuberculosis control program. For many herds, low-cost tests are more useful than more sensitive, higher cost tests. The model also indicates that test-positive cows in early stages of infection may be retained in the herd to generate farm income, provided they are managed properly to limit infection transmission. JD-Tree is a useful instructional tool, helping veterinarians understand the complex interactions affecting the economics of paratuberculosis control and to define the accuracy and cost specifications of better diagnostic tests.     

Somatic cell count in milk of selenium-supplemented dairy cows after an intramammary challenge with Staphylococcus aureus

The objective of this study was to evaluate the effect of selenium (Se) supplementation on milk somatic cell count (SCC) in dairy cows. Twelve multiparous Holstein-Friesian cows were fed a diet containing a suboptimal Se concentration (<0.05 ppm, dry basis) starting 2 months before calving. Supplemented cows (n=6) received a single s.c. injection of barium selenate (1 ml/50 kg BW) 45 days prior to calving, whereas control group was kept unsupplemented. Twenty weeks after calving, two mammary quarters (right side) of each cow were challenged with 205,000 cfu/ml of Staphylococcus aureus (strain Newbould 305). Blood was collected bi-weekly until day 150 of lactation for the analysis of blood glutathione peroxidase (GPx1; EC 1.11.1.9) activity. To re-isolate the challenging pathogen and to evaluate SCC, aseptic milk samples were collected daily starting on the day of challenge, and finishing 7 days after inoculation. Unsupplemented cows had a lower activity of GPx1 through the experiment (P<0.001). Natural log SCC (lnSCC) was higher in unsupplemented than Se-supplemented cows (P=0.04), showing evidence of significance after 5 days. Selenium supplementation of dairy cows fed a diet containing a suboptimal Se concentration, resulted in higher blood activity of GPx1, and lower mean lnSCC after an intramammary challenge with Staph. aureus.     

Intramammary infections in a dairy herd with a low incidence of Streptococcus agalactiae and Staphylococcus aureus infections.

In a dairy herd with a low incidence of intrammary infections due to Streptococcus agalactiae and Staphylococcus aureus, clinical mastitis remained a serious problem despite good control of nonclinical mastitis through postmilking teat disinfection and antibiotic therapy of known infected quarters at the end of lactation. During the 2-year study, the incidence of clinical mastitis was 0.88 cases/cow-year; 32.2% were caused by streptococcal species other than Str agalactiae and 33.5% by gram-negative organisms. Among all new infections detected, 54.1% were caused by streptococcal species other than Str agalactiae and 25.7% by gram-negative bacteria. Among new infections, 41.6% occurred during the nonlactating period or within a few days of calving. Incidence of clinical mastitis was highest in the 1st month of lactation. Among 84 gram-negative infections, 42.8% were caused by Klebsiella pneumoniae, 20.2% by Escherichia coli, and 23.8% by Enterobacter spp. Among the many serotypes of K pneumoniae and E coli, none was predominant.     

Phenotypic characteristics of Staphylococcus aureus isolated from bovine mastitis in Israeli dairy herds

A study of the characterization of the phenotypic patterns of Staphylococcus aureus strains isolated from bovine subclinical mastitis in Israeli dairy herds and their correlation with the severity of the disease was undertaken. A total of 400 chronically S. aureus-infected Israeli-Holstein cows, from 15 dairy herds were included in this study. Based on the results of the biochemical reactions, of the anti-biogram and phage typing, one major type of S. aureus was determined in each herd, its prevalence being between 54 and 100% of the total isolates from that same herd. The majority of the isolates were found to be non-haemolytic (62.7%). The most common phage type was 3/A,3/C,55,71, which was predominant in five herds. In two herds none of the isolates (24) were typable by this set of phages. All isolates were susceptible to methicillin, erythromycin, cephalotin, norfloxacin, trimethoprin-sulphamethoxazole and novobiocin. Most isolates were resistant to penicillin (96.6%) and 52% to oxytetracyclin. Differences in protein patterns between 50 and 36 kDa were found by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. No correlation between any combination of the phenotypic characteristics was found when correlation was done with milk yield and somatic cell count, corresponding to the 6 months before sampling. Otherwise, a positive correlation was found between type of haemolysis and the N-acetyl-beta-glucosaminidase (NAGase) values. In milk from quarters infected with the-non-haemolytic strains, the level of NAGase was significantly lower (P < 0.05) than that from quarters infected with the haemolytic strains (69.7 and 105.9, respectively). However, the level of NAGase activity in the milk of the quarters infected with the non-haemolytic strains was significantly higher (P < 0.05) when compared to the milk of quarters infected with coagulase-negative staphylococci (43.5).     

Population diversity of Staphylococcus aureus isolated from bovine mastitis in Brazilian dairy herds

Population diversity was evaluated in strains of Staphylococcus aureus isolated from bovine mastitis in Brazilian dairy herds by PCR-RFLP and sequencing of the 3'-terminal portion of the coagulase gene, and the susceptibility of strains to antimicrobials. The results showed great diversity in S. aureus population studied and the existence of predominant clones that account for most infections. No associations between the predominant types observed in the PCR-RFLP and the forms of presentation of the mastitis or to any of the different patterns of antimicrobial resistance were observed.     

In vitro antimicrobial susceptibility of Staphylococcus aureus strains from dairy herds in KwaZulu-Natal

Staphylococcus aureus is 1 of the most important causes of bovine mastitis and is responsible for significant economic losses to the dairy industry worldwide. One of the principal approaches used in treating intramammary infections is the administration of antimicrobials. Due to the propensity of S. aureus to develop resistance, antimicrobial susceptibility monitoring is necessary to ensure that treatment regimens are effective. As part of this investigation, 90 S. aureus strains isolated from mastitis cases submitted to Allerton Provincial Veterinary Laboratory during 2008 and 2009 were evaluated for their susceptibility to a panel of 10 antimicrobials. Only 8 of the 90 S. aureus isolates tested (8.9%) were found to be susceptible to all of the antimicrobials evaluated. A very high level of resistance to the beta-lactam antibiotics was noted: 47.8% of the isolates were resistant to penicillin and 65.6% were resistant to ampicillin. Minimal resistance to oxacillin, cephalothin and trimethoprim-sulfamethoxazole (1.1%) was found. Seventeen (18.9%) of the isolates tested were found to be resistant to 3 or more antimicrobials. The need for vigilant monitoring of bacterial resistance trends in the dairy industry is warranted as the potential public health implications are significant.     

Efficacy of extended pirlimycin hydrochloride therapy for treatment of environmental Streptococcus spp and Staphylococcus aureus intramammary infections in lactating dairy cows

Fifty-one chronically infected lactating dairy cows were used to evaluate the efficacy of extended pirlimycin therapy regimens for treatment of intramammary infections by environmental Streptococcus spp and Staphylococcus aureus. Cows (n = 47) with one or more infected mammary quarters were blocked by parity and randomly allocated to one of three groups for treatment with pirlimycin (50 mg/mammary quarter) as follows: one treatment per day for 2 days (n = 36 infected mammary quarters); one treatment per day for 5 days (n = 36 infected mammary quarters); and one treatment per day for 8 days (n = 20 infected mammary quarters). Four cows with nine infected mammary quarters were included as untreated controls. Milk samples from each mammary quarter were collected 7 days before treatment, immediately before treatment, and weekly for 4 weeks after the final treatment for microbiological evaluation. A bacteriologic cure was defined as a treated, infected quarter that was bacteriologically negative for the presence of previously identified bacteria at weekly intervals after treatment. Efficacy of pirlimycin therapy against intramammary infections caused by environmental Streptococcus spp and S. aureus was 44.4%, 61.1%, and 95.0% for the 2-, 5-, and 8-day treatment regimens, respectively. None of the infections in the untreated control quarters was cured. Significant differences in efficacy were detected between all pirlimycin groups and the untreated control group, between the 8- and 2-day treatment regimens, and between the 8-day and 5-day treatment regimens (P < or = .05). Results of this study indicate that extended pirlimycin therapy was effective in eliminating intramammary infections caused by environmental streptococci and S. aureus in lactating dairy cows.     

Characterization of cytokine expression in milk somatic cells during intramammary infections with Escherichia coli or Staphylococcus aureus by real-time PCR

The expression of inflammatory cytokines, including interleukin (IL)-6, IL-8, IL-12, granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma, by milk somatic cells was characterized by real-time polymerase chain reaction (PCR) in dairy cows experimentally challenged with either E. coli (n = 8) or S. aureus (n = 8). The mRNA abundance of a target gene was calibrated with that of a reference gene (beta-actin) and expressed as fold of induction over the control quarter at each time point. At no single time point did all eight quarters challenged with the same type of bacteria demonstrated increased expression of a target gene and there was large variation among animals at each given time. As a consequence, most tested comparisons were not statistically significant except the peak time points of IL-8 expression (75- and 29- fold in glands challenged with E. coli and S. aureus, respectively). However, the average fold induction of all targeted cytokines was increased in response to both bacterial challenges with the exception of IFN-gamma. The expression of IFN-gamma was only increased in milk somatic cells isolated from E. coli, but not S. aureus, challenged mammary glands. Moreover, upregulated expression of cytokine genes had higher magnitudes and/or faster responses in glands challenged with E. coli in comparison with those challenged with S. aureus. We propose that the compromised upregulation of inflammatory cytokines in S. aureus infected glands may, at least partially, contribute to the chronic course of infection caused by this pathogen. Further research on identifying factors responsible for the differentially expressed cytokine profiles may be fundamental to developing strategies that mitigate the outcome of bovine mastitis.     

Efficacy of a 5-day extended therapy program during lactation with cephapirin sodium in dairy cows chronically infected with Staphylococcus aureus

This study determined the efficacy of a 5-day extended therapy with cephapirin sodium in dairy cows chronically infected with Staphylococcus aureus. Chronically infected cows selected from 14 dairy herds in the St-Hyacinthe region, Québec were randomly allocated to a group of 31 cows treated for 5 consecutive days with 200 mg of cephapirin per quarter BID or a group of 30 untreated control cows. Bacteriological cure was determined by 3 negative bacterial cultures at 10, 24, and 31 days after treatment. The cow cure rates were 25.8% (8/31) in the treated cows and 3.3% (1/30) in the control group (P = 0.013). The quarter cure rates at first sampling post-treatment were 77.6% (38/49) and 18% (9/50) in the treated and the control groups, respectively (P < 0.0001). A 5-day extended therapy with cephapirin is effective in treating cows chronically infected with S. aureus.