Udder infection of goats by coagulase-negative staphylococci

Infection of udder halves by coagulase-negative staphylococci in seven commercial goat herds was studied in conjunction with the California Mastitis Test (CMT). Nine different species were identified and only 10% of strains belonged to groups which could not be identified with any of the known Staphylococcus species. The most prevalent species were Staphylococcus epidermidis (47.7%) and Staphylococcus caprae (19.7%). About half od the coagulase-negative staphylococcal infections gave negative CMT scores. The score was independent of the species of staphylococci involved and the stage of lactation. About 60% of the coagulase-negative staphylococcal species isolated were reisolated in the identical half udder during the following lactation.     

Antimicrobial resistance and genotypic characterization of coagulase-negative staphylococci over the dry period

Coagulase-negative staphylococci (CNS) have become the most frequently isolated organisms from bovine intramammary infections in recent years. While antimicrobial resistance (AR) is not considered a major problem among mastitis pathogens, concerns over emerging AR in general are increasing worldwide. Little information exists about the association between AR and one of the most common mastitis control measures, antibiotic dry cow therapy. The primary objective of the current study was to determine the prevalence of AR in CNS isolated before and after antimicrobial dry cow therapy. An additional objective was to genotypically characterize selected CNS isolates using pulsed-field gel electrophoresis (PFGE), to assess diversity and persistence of organisms over the dry period. Resistance against 10 antimicrobials was determined using a broth microdilution method and compared between CNS isolates collected at dry-off and at calving and from cows treated or not treated with intramammary antimicrobial products at dry-off (752 cows in total). Results suggested that increasing age of a cow and dry cow treatment when combined with high milk somatic cell count at dry-off and positive clinical mastitis history, were associated with increased AR to most beta-lactam antimicrobials and sulfadimethoxine. PFGE results suggested considerable diversity among the tested isolates as well as some clusters within cows and herds. PFGE would be useful in distinguishing between potentially persisting infections and cures and reinfections with different CNS strains.     

Evaluation of two commercial systems and a new identification scheme based on solid substrates for identifying coagulase-negative staphylococci from bovine mastitis

Coagulase-negative staphylococci (CNS), mainly isolated from bovine mastitis (n = 89, representing 11 different species), were used to evaluate two commercial identification systems: ID 32 Staph and Staph-Zym. The level of agreement between the ID 32 Staph and Staph-Zym systems and conventional methods was 77 and 94%, respectively. An alternative method, based on solid biochemical substrates, is also presented. This can be used for identifying novobiocin-sensitive CNS strains from bovine mastitis.     

Infection of the bovine udder with coagulase-negative staphylococci

A commercial dairy herd with a history of chronic staphylococcal mastitis was sampled ten times during a one-year period to determine the pattern of infection with coagulase-negative staphylococci as compared to infection with Staphylococcus aureus.The attack rate (AR) for new infections with S. aureus peaked in March, dropped to a low point in June, and rose again in the autumn. There was no discernible seasonal variation in the AR with the coagulase-negative staphylococci.The prevalence and AR for S. aureus infection increased with advancing age. The AR for new infection with coagulase-negative staphylococci decreased with advancing age.There was no obvious pattern of infection related to stage of lactation with any of the staphylococci isolated.Average duration of infection was longer for S. aureus than for the coagulase-negative staphylococci. Dry period therapy appeared to be more effective in eliminating existing infections with coagulase-negative staphylococci than with S. aureus, but the udder seemed to be equally susceptible to new dry period infections with either category of organism.     

Prevalence and herd-level risk factors for intramammary infection with coagulase-negative staphylococci in Dutch dairy herds

In this study, the prevalence of intramammary infection (IMI) with coagulase-negative staphylococci (CNS) in The Netherlands was estimated on 49 randomly selected herds with at least 40 lactating cows. In total, 4220 quarter milk samples were collected. The prevalence of CNS IMI in The Netherlands was estimated at 10.8% at quarter level and 34.4% at cow level, making it the most frequently isolated group of pathogens. Fourteen species of CNS were identified; the most frequently isolated species was Staphylococcus chromogenes (30.3%) followed by Staphylococcus epidermidis (12.9%) and Staphylococcus capitis (11.0%). Prevalence of CNS IMI was higher in heifers compared to older cows. Geometric mean quarter SCC of CNS-positive quarters was 109,000 cells/ml, which was approximately twice as high as culture-negative quarters. Quarters infected with S. chromogenes, S. capitis and Staphylococcus xylosus had a higher SCC (P<0.05) than culture-negative quarters, while quarters that were culture-positive for S. epidermidis and Staphylococcus hyicus tended to have a higher SCC than culture-negative quarters. An increased prevalence of CNS IMI was associated with the herd-level variables source of drinking water not being tap water, housing of dry cows in one group instead of multiple groups, measurement of cow SCC every month, udder health monitoring by the veterinarian, pasturing during outdoor season, percentage of stalls contaminated with milk, and BMSCC>250,000 cells/ml. Although a causal relation between these factors and prevalence of CNS is not proven and for some factors not even likely, knowledge of the associations found may be helpful when approaching CNS problems on dairy farms.     

Species identification and some characteristics of coagulase-negative staphylococci isolated from bovine udders.

The species of 203 strains of coagulase-negative staphylococci (CNS), isolated from bovine udder quarters was determined; all were tested for hydrophobicity and encapsulation, attributes that may relate to virulence. Twelve species were identified, of which Staphylococcus simulans, (34.5%), S. chromogenes (16.7%), S. epidermidis (13.8%) and S. xylosus (8.9%) were the most frequent. The majority of strains possessed a hydrophilic cell surface. However, strains from two species (S. chromogenes and S. epidermidis) were more hydrophobic than the others. Only five strains were encapsulated (S. xylosus, 3; S. saprophyticus, 1; and S. sciuri, 1). Judging from the low frequencies of hydrophobic and encapsulated strains, and comparing with strains isolated from clinical cases, it is suggested that these properties are not major virulence determinants of CNS.     

Haemolytic and proteolytic activity of coagulase-negative staphylococci isolated from mastitis cows

The aim of the present study was to assess the haemolytic and proteolytic activity of coagulase-negative staphylococci (CNS) isolated from cows with mastitis. The study was conducted on 100 CNS strains: S. xylosus (n=28), S. chromogenes (n=26), S. haemolyticus (n=25), S. sciuri (n=14), S. warneri (n=4), S. hominis (n=2), S. saprophyticus (n=1); 22 CNS were isolated from cows with clinical mastitis and 78 from those with subclinical mastitis. The CNS studied showed the ability to produce only alpha-haemolysin and belonged to one strain - S. haemolyticus (21.0% of isolated CNS strains). Haemolysin-positive CNS were responsible for both clinical and subclinical mastitis (22.7% and 20.5%, respectively). The ability to produce protease was found in 31.0% of CNS belonging to two strains: S. chromogenes and S. sciuri. Protease-positive CNS were the etiological factor of both clinical and subclinical mastitis (31.8% and 30.8%, respectively). All S. xylosus, S. warneri, S. hominis, and S. saprophyticus strains were found protease-negative and haemolysin-negative, irrespective of whether they caused clinical or subclinical mastitis in cows.     

Comparison of the API Staph-Ident and DMS Staph Trac micromethods for the identification of coagulase-negative staphylococci.

Two rapid identification micromethods, the API Staph-Ident and the DMS Staph Trac, were compared for the identification of coagulase-negative staphylococci. Seventy-five isolates, mainly of bovine origin, were used as the test organisms. Species identification and profile numbers assigned to isolates by each system were compared. However, no clear correlation patterns emerged, indicating the two methods were not comparable.     

Classification of coagulase-negative staphylococci isolated from bovine clinical and subclinical mastitis

One hundred and forty eight strains of coagulase-negative staphylococci (CNS) from bovine clinical and subclinical mastitis were classified by conventional biochemical testing. Staphylococcus simulans was the most frequently isolated species. The distribution of CNS species in the two groups was similar regardless of whether the cow suffered from clinical or subclinical mastitis. There was no significant difference between the grades of severity of the clinical mastitis associated with the various CNS. The small difference in pathogenicity observed between species in this study, and the similarity of distribution in the various species of clinical and subclinical mastitis could indicate that the species per se is of minor importance in the development of mastitis.     

Effects of dry cow intramammary therapy on quarter infections in the dry period

Quarter milk samples were taken from 150 cows from three dairy farms in south-east Queensland at drying off, two, four and six weeks after drying off, at calving, and one, two and three weeks after calving. In each of the herds, the cows were randomly allocated to three groups of approximately equal size. One group had all the quarters of all the cows treated at drying off with a dry cow antibiotic infusion containing cloxacillin; the second group was given no treatment, and the third group had selected quarters treated on the basis of their high activity of N-acetyl-beta-D-glucosaminidase at drying off. Dry cow treatment resulted in a marked reduction in the number of infected quarters at two and four weeks after drying off, so that the comprehensively treated group had significantly less infected quarters at these times (P<0.02). Twelve dinical cases of mastitis were detected two weeks after drying off in the untreated groups, 10 in the untreated quarters of the selectively treated groups, and no cases in the comprehensively treated groups. These cases were due mainly to Streptococcus uberis and Streptococcus dysgalactiae. The number of infected untreated quarters increased markedly between drying off and two weeks later, but in all three groups there was a marked decrease in the number of infected quarters between six weeks after drying off and calving, suggesting that the mammary glands were more able to overcome infections at this time.     

Identification of coagulase-negative staphylococci from bovine mastitis using RFLP-PCR of the groEL gene

Coagulase-negative staphylococci (CNS) have become the predominant pathogens causing bovine mastitis in many countries. CNS infections are associated with damage to milk secretory tissue of the mammary gland by increased connective tissue stroma, moderate increases of somatic cells count in milk and significant production decreases. These consequences impose serious economic losses for the farmers and the dairy industry. Routine veterinary laboratories do not usually identify CNS at the species level. Thereby, the aims of this study were to identify the most common staphylococcal pathogens involved in bovine mastitis using PCR-restriction fragment length polymorphism (RFLP) analysis of a partial groEL gene sequence and to compare our results with the identification carried out by the conventional method. A total of 54 isolates of Staphylococcus, involved in bovine mastitis, were analyzed by this method. The size and number of the fragments obtained by either AluI or HindIII/PvuII digestions made possible to form clear patterns differentiating, among the isolates, 11 of the most common species of animal staphylococcal pathogens. Most of the isolates clustered together with the reference strain of Staphylococcus chromogenes (28) and the type strain of Staphylococcus epidermidis (8). Besides, some isolates clustered together with the type strain of Staphylococcus aureus (5). All patterns were confirmed by the conventional biochemical method, showing concordant results. Thus, the PCR-RFLP of the groEL gene constitutes a reliable and reproducible molecular method for identification of CNS species responsible for bovine mastitis.     

Antibiotic susceptibility of methicillin-resistant and methicillin-susceptible coagulase-negative staphylococci isolated from bovine mastitis

The aim of the present study was to evaluate the antibiotic susceptibility of methicillin-susceptible (MS) and methicillin-resistant (MR) coagulase-negative Staphylococcus (CNS) strains isolated from milk of cows with mastitis. The study was conducted on 100 CNS strains (20 MRCNS and 80 MSCNS) isolated from milk samples of 86 cows from the Lublin (Poland) region farms. Antibiotic susceptibility of microorganisms was evaluated using the disc-diffusion method on the Mueller-Hinton agar according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The highest efficacy against MSCNS was demonstrated for cephalosporin antibiotics, i.e. cefacetril (91.3%), ceftiofur (67.5%), cefoperazone (66.3%) and cephalexin (60.0% of susceptible MSCNS strains). Moreover, a high percentage of vancomycin-susceptible strains was demonstrated (83.8%). The activity of combination of amoxicillin with clavulanic acid and gentamicin was found weaker (63.8% and 61.3% of susceptible strains, respectively). About 50.0% of MSCNS were susceptible to erythromycin, enrofloxacine and amoxicillin. A large proportion of CNS was resistant to neomycin, penicillin, tetracycline, streptomycin, lincomycin and ampicillin (28.8%, 30.0%, 31.3%, 31.3%, 33.8% and 33.8% of susceptible strains, respectively). The highest percentage of MRCNS was susceptible to vancomycin (75.0%), erythromycin (65.0%) and streptomycin (50.0%). Their susceptibility to enrofloxacine (35.0%) as well as gentamicin and tetracycline (30.0%) was markedly lower. The lowest activity was found for lincomycin and neomycin (20.0% of susceptible MRCNS strains, each).     

Management of the dry cow in control of peripartum disease and mastitis.

The dry period has great implications on overall health and productivity in the subsequent lactation. Many anatomic, physiologic, and immunolgic changes are occurring for both the cow and the mammary gland during this time. These changes need to be understood and taken into consideration when assessing and implementing health management programs that involve this crucial time period. Specifically, nutritional and immunologic requirements of the individual cow need to be considered. The occurrence of many peripartum diseases is significantly influenced through nutritional and metabolic parameters that can be strongly influenced, controlled, and monitored in the dry period. From an udder-health perspective, the goal of the dry period can be met by recommending administration of DCT to all quarters of all cows at the end of lactation. As research continues to explore and define shortcomings of this approach and as scrutiny of the prophylactic use of antibiotics increases, however, novel approaches to preventing and eliminating IMI may become more readily available. These approaches offer new methods to improve upon and redefine what should be realistic goals of the dry period and afford an opportunity for continued improvement of udder health in today's dairy herds.     

Association of cow and quarter-level factors at drying-off with new intramammary infections during the dry period

Our objective was to describe cow and quarter-level factors associated with drying-off, and to evaluate their impacts on new intramammary infections (IMI) during the dry period. Data from 300 cows in five research herds were collected starting 2 weeks prior to scheduled drying-off. Variables of interest included daily milk production, teat-end integrity, formation of the teat-canal keratin plug, and quarter-milk bacteriological culture results. Overall, 11% of quarters developed new IMI in the dry period; this varied by herd, parity and time of the study. Most new IMI were caused by environmental streptococci and coliform organisms (34 and 30%, respectively). Quarters that had a cracked teat-end had higher odds of developing new infections than those without cracks (15 and 10%, respectively). Quarters that formed a keratin plug early in the dry period had a lower odds than those that did not close (10 and 14%, respectively). After 6 dry weeks, 23% of quarters were still open. The hazard of quarters closing if milk production on the day prior to drying-off was >21 kg 1.8-times less.     

Within herd comparison of teat dipping and dry cow therapy with only selective dry cow therapy in six herds

A within herd comparison of teat dipping and dry cow therapy (full treatment) with only selective dry cow therapy (partial treatment) was carried out in six commercial dairy herds for a two year period. In four herds, the incidence of clinical mastitis was 2 to 12 per cent higher in the partial treatment group. In another herd, in which the pattern of clinical mastitis isolates was unusual in that minor pathogens were isolated from 30 per cent of mastitis cases, the incidence was 43 per cent higher in the partial treatment group. In the remaining herd the incidence was 10 percent higher in the full treatment group. Streptococcus uberis mastitis was more common in the partial treatment groups of five herds; coliform mastitis was more common in the full treatment groups of two herds and similar in both groups in the other herds. High rates of coliform mastitis were associated with poor herd environmental conditions but this was not true for Strep uberis mastitis. Rates of staphylococcus aureus and Strep dysgalactiae mastitis were low in all herds. The level of major pathogen infection in cows completing the trial in all herds increased in the partial treatment group from 5 per cent of quarters at the start to 12 per cent at the finish of the trial. In the full treatment group, however, there was only a small increase in this level. In contrast, levels of Corynebacterium bovis infection increased by 17 per cent in both treatment groups. Continued use of teat dipping and dry cow therapy was associated with a higher rate of coliform mastitis in two of the three herds where there were poor standards of hygiene and husbandry.     

Risk factors associated with bacteriological cure, new infection, and incidence of clinical mastitis after dry cow therapy with three different antibiotics

Factors affecting bacteriological cure rates (BCR) and new intramammary infections (IMI) during the dry period as well as clinical mastitis (CM) during early lactation were investigated in 414 German Holstein dairy cows receiving dry cow therapy. Cows were treated with either benethamine benzylpenicillin (300,000 IU), penethamate hydriodide (100,000 IU), and framycetin sulphate (100 mg, n = 136), or cefquinome (150 mg, n = 135), or benzathine cloxacillin (1,280 mg, n = 143). Overall BCR, IMI, and CM at parturition were 86.4%, 20.7%, and 4.3%, respectively. The three antibiotic treatments differed only in BCR, with cloxacillin yielding better results than the others. Udder quarters from cows with > 4 lactations had a higher risk of IMI and CM at calving. Chronic changes in udder tissues were linked to a lower BCR and were associated with a higher risk of CM during early lactation. The risk of CM at calving was higher in udder quarters with unspecific or subclinical mastitis before drying off. In conclusion, with antibiotic dry cow therapy, age and health status of the udder appear to be major determinants of IMI and CM during the dry period and early lactation, while BCR was associated with the antibiotic type and udder tissue status.