Efficacy of immunization with a recombinant S. aureus vaccine formulated with liposomes and ODN-CpG against natural S. aureus intramammary infections in heifers and cows.

The aims of the present study were to evaluate the ability of a subunit vaccine composed of recombinant molecules of α-toxin, ß-toxin, FnBPA and ClfA, formulated with cationic liposomes and CpG-ODN, to confer protection against natural S. aureus intramammary infection (IMI) and to assess the antibody response against the vaccine components. A stringent criterion based on molecular identification of the isolates was used to define IMI. The proportion of animals that developed new S. aureus IMI was higher in the Control group compared with the Vaccine group (reduction of 60.7%), and time to new S. aureus IMI was higher for animals in the Vaccine group compared with animals in the Control group, although not statistically significant. Molecular identification of the isolates allowed the detection of S. aureus pulsotypes that appeared transiently in milk and others that were able to establish IMI, providing a new perspective to define parameters related to the definition of new IMI and cures. Specific IgG, IgG1 and IgG2 levels against the four recombinant proteins included in the vaccine were significantly increased in the vaccinated group and the recombinant α-toxin included in the vaccine generated antibodies that reduced significantly the haemolytic activity of native α-toxin. Data reported in the present study indicate a possible effect on both the proportion of animals developing new IMI and the time to new S. aureus IMI, but the incidence of disease within the study was too low to provide statistical confirmation.

Evaluation of the humoral immune response to a multicomponent recombinant vaccine against S. aureus in healthy pregnant heifers.

Staphylococcus aureus is a worldwide pathogen that causes mastitis in dairy herds. Shortcomings in control programs have encouraged the development of vaccines against this pathogen. This study evaluated the vaccine candidate VacR, which included recombinant S. aureus protein clumping factor A (rClf), fibronectin binding protein A (rFnBP) and hemolysin beta (rBt), formulated with a novel immune-stimulating complex. Comparisons were made between healthy pregnant heifers that received either VacR (n=8; VacR group) or phosphate buffered saline (PBS) plus adjuvant (control group) SC in the supramammary lymph node area on days 45 and 15 before the expected calving date. Blood and foremilk samples were collected from 7 to 60days post-calving. After calving, heifers in the VacR group produced higher total IgG (IgGtotal) titers against each component, in both serum (rBt, 3.4×105; rClf, 3.1×105; rFnBP, 2.3×105) and milk (rBt, 2.6×104; rClf, 1.3×104; rFnBP, 1.1×104), than control heifers (P<0.0001). There were increased concentrations of IgG1 and IgG2 in VacR group (P<0.05), in both serum and milk. Humoral responses remained high throughout the period most susceptible to intramammary infections (P<0.01). Antibodies produced against S. aureus rClf and rFnBP reduced bacterial adherence to fibronectin and fibrinogen by 73% and 67%, respectively (P<0.001). Milk antibodies against these adhesins inhibited S. aureus invasion of a mammary epithelial cell line (MAC-T), resulting in 15.7% of bacteria internalized (P<0.0001). There was an approximately 6-fold reduction in the hemolysis titer for the native hemolysin in the VacR group compared to the control group (P<0.0001) and a significantly increase in the proportion of positive neutrophils (VacR, 29.7%; PBS, 13.1%) and the mean fluorescent index (VacR, 217.4; PBS, 152.6; P<0.01) in the VacR group. The results suggest that VacR is a valuable vaccine candidate against S. aureus infections, and merits further field trials and experimental challenges.

Aislamiento y caracterización de Corynebacterium pseudotuberculosis biotipo ovis en Lama glama en Córdoba, Argentina / Isolation and characterization of Corynebacterium pseudotuberculosis biotype ovis in Lama glama in Córdoba, Argentina

Corynebacterium pseudotuberculosis es el agente causal de la linfadenitis caseosa en distintas especies animales. No existen reportes en Argentina de casos clínicos en camélidos. Esta comunicación describe el aislamiento y tipificación de Corynebacterium pseudotuberculosis biotipo ovis en una llama (Lama glama) en Córdoba, Argentina. (AU)

Corynebacterium pseudotuberculosis is the causal agent of caseous lymphadenitis in different animal species. There are no reports in Argentina of clinical cases in camelids. This communication describes the isolation and typing of Corynebacterium pseudotuberculosis biotype ovis in llama (Lama glama) in Córdoba, Argentina. (AU)

Intramammary inoculation of Panax ginseng plays an immunoprotective role in Staphylococcus aureus infection in a murine model.

The immunoprotective effect of Panax ginseng (Pg) extract was investigated in a mouse mastitis model. Lactating female mice were intramammarily inoculated with Pg or placebo, and then were challenged with S. aureus, while other group was inoculated with S. aureus alone. The number of bacteria recovered from mammary glands was significantly lower in Pg-treated S. aureus-infected mice (group I) compared with placebo-treated S. aureus-infected mice (group II) and S. aureus-infected mice (group III). The mRNA expression of TLR2, TLR4, IL-1α and TNF-α was influenced by treatment; being the transcript levels for all genes higher in group I compared with group II and III. Activation of NF-κB and the number of monocytes-macrophages in mammary gland tissue was significantly increased in group I compared with group II and III. Pg extract was able to trigger an adequate immune response to confront an infection demonstrating its protective effect and potential for preventing bovine intramammary infections.

Which are important targets in development of S. aureus mastitis vaccine?

Staphylococcus aureus represents one of the leading causes of mastitis in dairy cows worldwide. S. aureus IMI have variable outcomes due to virulence of the strain involved, immune defenses of the host, and by antibiotic resistance. The difficulty in eradication and the increasing concerns on antibiotics usages underscore the interest in developing new tools to control S. aureus mastitis. Vaccination represents one of the most studied of these tools but, so far, no vaccine seems to provide reliable protection. This review summarizes current knowledge on the major vaccine targets, including surface proteins, capsular polysaccharides, biofilm, and toxins. Finally, the present status of vaccination against S. aureus and the future of vaccine design were discussed, including how differences among in vivo models may influence vaccines development.

Immune response and functional role of antibodies raised in heifers against a Staphylococcus aureus CP5 lysate and recombinant antigens vaccine formulated with Iscom Matrix adjuvant.

Staphylococcus aureus is the most frequently isolated pathogen from bovine intramammary infections worldwide. Commercially available vaccines for mastitis control are composed either of S. aureus lysates or inactivated whole-cells formulated with traditional adjuvants. We recently showed the ability of a S. aureus CP5 lysate vaccine adjuvanted with Iscom Matrix to generate a longer lasting specific antibody response in blood and milk, with improved opsonic capacity, compared with a S. aureus CP5 whole-cell formulation. The aim of the present study was to obtain an experimental immunogen composed of lysed cells of a CP5 S. aureus strain supplemented with recombinant clumping factor A, fibronectin binding protein A and ß-toxin formulated with Iscom Matrix, characterize the immune response generated when immunizing pregnant heifers and assess the functional role of antibodies raised against this immunogen in experimental models. Both a lysate vaccine and a lysate+recombinant antigens vaccine elicited antibodies that promoted neutrophil phagocytosis and inhibited internalization into mammary epithelial cells, in vitro. Incorporation of defined antigenic molecules to the lysate formulation elicited a strong specific humoral immune response against both lysate and recombinant antigens and was associated with higher expression of regulatory and pro-inflammatory cytokines. In addition, antibodies were efficient for blocking S. aureus binding to bovine fibrinogen and fibronectin, and neutralizing ß-toxin effect in vitro, placing these antigens as candidates to be included in a formulation directed to prevent staphylococcal bovine mastitis.

Genotypic and phenotypic detection of capsular polysaccharides in Staphylococcus aureus isolated from bovine intramammary infections in Argentina

Staphylococcus aureus (n=157) isolated from intramammary infections in Argentine dairy areas were evaluated for presence of cap5 and cap8 loci. Isolates carrying cap5 and cap8 were serotyped using specific antisera. Sixty four percent of the isolates were genotyped as cap5 or cap8 and 50% of them expressed CP5 or 8.

Genotypic and phenotypic detection of capsular polysaccharides in Staphylococcus aureus isolated from bovine intramammary infections in Argentina.

Staphylococcus aureus (n=157) isolated from intramammary infections in Argentine dairy areas were evaluated for presence of cap5 and cap8 loci. Isolates carrying cap5 and cap8 were serotyped using specific antisera. Sixty four percent of the isolates were genotyped as cap5 or cap8 and 50% of them expressed CP5 or 8.

Proinflammatory cytokines and CD14 expression in mammary tissue of cows following intramammary inoculation of Panax ginseng at drying off.

The lack of efficacy of conventional strategies for the maintenance of healthy udders in domestic cattle has prompted studies on the use of immunomodulators or biological response modifiers (BRM) for this purpose. These compounds are agents that modify the host's response to pathogens leading to beneficial effects on disease outcome. The objective of this study was to evaluate the effects of a single intramammary infusion of Panax ginseng (GS) extract on the amount of pro-inflammatory cytokines and the number of monocytes/macrophages present in bovine mammary tissues at drying off. Eight mammary quarters from six nonpregnant cows in late lactation were infused with 10 mL of GS (3mg/mL), six quarters were treated with 10 mL of placebo (vehicle alone) and six quarters were maintained as uninoculated controls. The analyses of tumor necrosis factor-alpha (TNF-α) by immunohistochemistry revealed that the production of this proinflammatory cytokine significantly increased (P<0.05) in the inoculated mammary glands of cows following BRM inoculation, whereas the interleukin-1 alpha (IL-1α) and IL-6 staining area was not affected by BRM treatment. The number of monocytes/macrophages detected with CD14 antibody was significantly higher (P<0.05) in BRM-treated quarters than in placebo and uninoculated control quarters. These results indicated an immunomodulator potential of the BRM used. The beneficial effect of the extract could be used as alternative therapy in the control of mastitis at drying off, either alone or in conjunction with dry cow antibiotic therapy.

Effect of Panax ginseng on cytokine expression in bovine mammary glands at drying off.

Biological response modifiers (BRM) are agents that modify the host's response to pathogens with resultant beneficial prophylactic or therapeutic effects. The objectives of this study were to describe the immunomodulatory effects of Panax ginseng (GS) on bovine mammary glands at the end of lactation. Eight mammary quarters from six nonpregnant cows in late lactation were infused with 10mL of BRM (3mg/mL), six quarters were treated with placebo (vehicle alone) and six quarters were maintained as uninoculated controls. Milk samples were collected at different time points for detection of specific cytokines mRNA by RT-PCR and Western blotting assay. A significant increase of IL-1α, IL-1ß and TNF-α mRNA expression was observed in BRM-treated compared with placebo-treated quarters at 48h post-treatment (pt) (P<0.05). A 17kDa TNF-α band expressed a sharp elevation at 24h and reduction in its level at 48h pt in BRM-treated quarters. Differences in this cytokine level between 24 and 48h pt times were significant (P<0.05). GS extract inoculation at drying off was associated with somatic cell counts increase, cytokines mRNA transcription and the presence of TNF-α in milk and can therefore exert immunomodulating effects in bovine mammary gland at drying off.

The effect of a single intramammary infusion of a biological response modifier in cows at drying off.

Biological response modifiers (BRM) are compounds that interact with the immune system to regulate specific aspects of host response. The objective of this study was to describe clinical and morphological changes during involution of bovine mammary gland following a single-dose infusion of a BRM containing lipopolysaccharide and cellular fractions of Escherichia coli incorporated into liposomes. A massive leukocyte response and increased subepithelial stroma infiltration of mononuclear cells, eosinophils and mast cells was observed in BRM-treated quarters compared with untreated controls; however, morphologic parameters assessed at 11 days post infusion were indicative of only slightly accelerated involution compared with untreated controls. In addition, BRM infusion at the end of lactation did not interfere with mammary epithelial cell proliferation and caused only mild systemic effects.

Effect of a biological response modifier on expression of CD14 receptor and tumor necrosis factor-alpha in Staphylococcus aureus-infected mammary glands at drying off.

Agents that increase natural protective mechanisms have been proposed for prevention and treatment of intramammary infections. The objectives of this study were to get an insight of innate immune mechanisms that occur during bovine mammary involution in both uninfected and chronically Staphylococcus aureus-infected glands and to describe the effects on those mechanisms of a single intramammary infusion of a LPS-based biological response modifier (BRM) at the end of lactation. Three groups of 12 cows, each one including 6 S. aureus-infected and 6 uninfected, were infused in two mammary quarters with BRM or placebo and sacrificed at 7, 14 and 21 d of involution. In uninfected and S. aureus-infected quarters treated with a BRM, the number of monocytes/macrophages detected with CD14 antibody was significantly higher (P<0.05) than in placebo-treated quarters at every sampling evaluation period. In uninfected quarters, the TNF-alpha staining area was not affected by BRM treatment. However, in infected quarters, the immunostained area for TNF-alpha was significantly higher than in uninfected quarters and BRM treatment was associated with increased staining at 21 d of involution.

Antimicrobial susceptibility of Staphylococcus aureus causing bovine mastitis in Argentine dairy herds.

We assessed the in vitro activity of selected antimicrobial agents against 95 Staphylococcus aureus strains causing both clinical and subclinical bovine mastitis belonging to 61 dairy farms from the Central dairy area of Argentina. Minimal inhibitory concentrations (MICs) of penicillin, oxacillin, gentamicin, erythromycin, enrofloxacin and florfenicol were estimated. In addition, the agar diffusion test was performed. MIC50 and MIC90 were as follows: penicillin, 0.05 and 4 microg/ml; oxacillin, 0.25 and 0.25 microg/ml; gentamicin, 0.25 and 0.5 microg/ml; erythromycin 0.125 and 0.25 microg/ml; enrofloxacin 0.25 and 0.5 microg/ml, and florfenicol 4 and 8 microg/ml. Beta-lactamase activity was detected in 89% of 46 penicillin-resistant strains. Apart from penicillin, antimicrobial resistance in S. aureus causing bovine mastitis remains rare in Argentine dairy farms.

Antimicrobial susceptibility of Staphylococcus aureus causing bovine mastitis in Argentine dairy herds / Sensibilidad a los antimicrobianos de Staphylococcus aureus causantes de mastitis bovina en rodeos lecheros de Argentina

We assessed the in vitro activity of selected antimicrobial agents against 95 Staphylococcus aureus strains causing both clinical and subclinical bovine mastitis belonging to 61 dairy farms from the Central dairy area of Argentina. Minimal inhibitory concentrations (MICs) of penicillin, oxacillin, gentamicin, erythromycin, enrofloxacin and florfenicol were estimated. In addition, the agar diffusion test was performed. MIC50 and MIC90 were as follows: penicillin, 0.05 and 4 µg/ml; oxacillin, 0.25 and 0.25 µg/ml; gentamicin, 0.25 and 0.5 µg/ml; erythromycin 0.125 and 0.25 µg/ml; enrofloxacin 0.25 and 0.5 µg/ml, and florfenicol 4 and 8 µg/ml. b-lactamase activity was detected in 89% of 46 penicillin- resistant strains. Apart from penicillin, antimicrobial resistance in S. aureus causing bovine mastitis remains rare in Argentine dairy farms.

Se evaluó la actividad in vitro de un grupo seleccionado de antimicrobianos contra 95 aislamientos de Staphylococcus aureus obtenidos de casos de mastitis bovina clínica y subclínica, en 61 rodeos lecheros de la cuenca central de Argentina. Fueron estimadas las concentraciones inhibitorias mínimas (CIM) de penicilina, oxacilina, gentamicina, eritromicina, enrofloxacina y florfenicol. Además se realizó la prueba de difusión en agar. Las CIM50 y CIM90 obtenidas fueron: penicilina 0,05 y 4 µg/ml; oxacilina 0,25 y 0,25 µg/ml; gentamicina 0,25 y 0,5 µg/ml; eritromicina 0,125 y 0,25 µg/ml; enrofloxacina 0,25 y 0,5 µg/ml y florfenicol 4 y 8 µg/ml. Se detectó actividad de b-lactamasa en el 89% de las cepas resistentes a la penicilina. A excepción de lo observado para penicilina, la resistencia a los antimicrobianos en S. aureus causantes de mastitis bovina en Argentina parece ser un fenómeno poco frecuente.

Characterization of a new variant of IS257 that has displaced the capsule genes within bovine isolates of Staphylococcus aureus.

Many bovine Staphylococcus aureus isolates from Argentina are nontypeable (NT), i.e., they do not produce serotype 5 or 8 capsular polysaccharides (CPs). Some of these NT strains have a deletion of the cap5(8) gene cluster mediated by a variant of IS257, now designated IScap. IScap showed 93% amino acid identity to S. aureus ORF49 but only 85% identity to IS431 from S. aureus N315 and 88% identity to an IS257-like element from bovine strain RF122. Thirty-six (53%) of 68 bovine isolates, drawn from a previously described S. aureus strain collection, carried some variant of IS257, including IScap. Of these 36 IS+ isolates, 6 were CP5+, 1 was CP8+, and 29 were NT. Forty-four of the 68 isolates were NT, and 24 of these 44 NT isolates (55%) exhibited IScap-mediated deletion of the cap5(8) gene cluster. IScap was not found among 20 human NT S. aureus isolates bearing the cap5HIJK genes, which suggests that IScap-mediated deletion of the capsule locus is restricted to bovine strains of S. aureus. We were unable to identify a precursor strain in which IScap flanked the cap5(8) capsule locus, nor were we able to select for deletion of the cap5(8) locus in vitro. Our results support the hypothesis that deletion of the cap5 locus occurred in the distant past and that the relative abundance of these NT strains may be a result of their ability to persist in subclinical mastitis infection in cows.

Intramammary infections during the periparturient period in Argentine dairy heifers.

Prevalence of intramammary infections at prepartum and postpartum in primigravid heifers from five dairy herds located in the central dairy area of Argentina was determined. Mammary secretion samples from 140 heifers (560 mammary quarters) were obtained 14 days prior to the expected calving day and within 7 days after parturition and subjected to bacteriological analysis. No clinical mastitis cases were detected during the study. The number of infected heifers in at least one mammary quarter at pre and postpartum was 87 (62.2%) and 53 (37.8%), respectively. The most prevalent mastitis pathogens at prepartum among samples yielding a positive bacteriological culture were coagulase-negative staphylococci (69.07%), Staphylococcus aureus (12.71%) and Streptococcus uberis (4.42%). A decrease on isolation frequency of coagulase-negative staphylococci (53.41%) and S. uberis (2.27%) was observed at postpartum, while that of S. aureus showed an increase (21.59%). Presence of intramammary infections appeared to be associated with some management conditions. These results highlighted the need to improve diagnosis and control measures in replacement heifers.

Intramammary infections during the periparturient period in Argentine dairy heifers / Infecciones intramamarias durante el periparto en vaquillonas en Argentina

Prevalence of intramammary infections at prepartum and postpartum in primigravid heifers from five dairy herds located in the central dairy area of Argentina was determined. Mammary secretion samples from 140 heifers (560 mammary quarters) were obtained 14 days prior to the expected calving day and within 7 days after parturition and subjected to bacteriological analysis. No clinical mastitis cases were detected during the study. The number of infected heifers in at least one mammary quarter at pre and postpartum was 87 (62.2%) and 53 (37.8%), respectively. The most prevalent mastitis pathogens at prepartum among samples yielding a positive bacteriological culture were coagulasenegative staphylococci (69.07%), Staphylococcus aureus (12.71%) and Streptococcus uberis (4.42%). A decrease on isolation frequency of coagulase-negative staphylococci (53.41%) and S. uberis (2.27%) was observed at postpartum, while that of S. aureus showed an increase (21.59%). Presence of intramammary infections appeared to be associated with some management conditions. These results highlighted the need to improve diagnosis and control measures in replacement heifers.

Se determinó la prevalencia al preparto y posparto de infecciones intramamarias causadas por organismos patógenos de mastitis en vaquillonas primíparas de cinco establecimientos lecheros ubicados en la cuenca central santafesina. Se tomaron muestras de secreción mamaria de 140 vaquillonas (560 cuartos mamarios) aproximadamente 14 días antes de la fecha probable de parto y dentro de los 7 días posparto, y se procesaron bacteriológicamente. No se detectaron casos de mastitis clínicas durante el estudio. El número de vaquillonas infectadas en al menos un cuarto mamario al preparto y posparto fue de 87 (62,2%) y 53 (37,8%), respectivamente. Los organismos patógenos más prevalentes al preparto entre las muestras con cultivo bacteriológico positivo fueron estafilococos coagulasa negativos (69,07%), Staphylococcus aureus (12,7%) y Streptococcus uberis (4,42%). Al posparto se observó un descenso en la frecuencia de aislamiento de estafilococos coagulasa negativos (53,41%) y S. uberis (2,27%), mientras que la de S. aureus mostró un aumento (21,59%). La presencia de infecciones intramamarias pareció estar asociada con algunas prácticas de manejo. Estos resultados ponen de manifiesto la importancia de hacer extensivo el diagnóstico y control de la enfermedad a las vaquillonas de reemplazo antes de su ingreso al rodeo en ordeño.

Effect of a biological response modifier on expression of growth factors and cellular proliferation at drying off.

Agents that increase natural protective mechanisms have been proposed for the prevention and treatment of intramammary infections. Staphylococcus aureus is a major pathogen causing primarily subclinical chronic mastitis that responds poorly to antibiotic therapy. The objectives of this study were to describe the effects of a single intramammary infusion of a lipopolysaccharide-based biological response modifier (BRM) on mammary epithelial cellular proliferation and expression of insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor (VEGF) in uninfected and Staph. aureus-infected bovine mammary glands during involution. Three groups of 12 cows, 6 Staph. aureus-infected and 6 uninfected, were infused with BRM or placebo in 2 mammary quarters and killed at 7, 14, and 21 d of involution. The proportion of infected quarters, mammary cell proliferation, and IGF-I and VEGF expression were evaluated. Biological response modifier treatment decreased the proportion of Staph. aureus-infected mammary quarters at 7 d of involution, but a similar number of isolations were observed at 14 and 21 d of involution in either treated or control quarters. The percentage of proliferating mammary epithelial cells was higher in infected than uninfected quarters at every observation period, irrespective of the treatment administered, whereas uninfected BRM-treated quarters showed increased cell proliferation at 7 d of involution. Insulin-like growth factor-I expression in uninfected quarters was not affected by treatment and showed a decrease at 21 d of involution. Expression of IGF-I was greater in infected than uninfected quarters at every observation period, irrespective of the treatment received. Expression of VEGF was greater in BRM-treated uninfected quarters at 7 d of involution compared with controls. In infected quarters, VEGF expression was lowest in BRM-treated quarters at 7 d of involution and increased throughout the observation period. Conversely, untreated infected quarters showed the highest VEGF expression at 7 d and decreased at 21 d of involution. Mammary cell proliferation and expression of IGF-I and VEGF were increased in Staph. aureus-infected quarters. Increased mammary cell proliferation and VEGF expression were observed in BRM-treated quarters during the first week of involution.

Microbiological screening test validation for detection of tylosin excretion in milk of cows with low and high somatic cell counts.

Antibiotic residues in milk above tolerance levels interfere with dairy product processing and pose potential health risks to consumers. Residue avoidance programmes include, among other components, the observance of withdrawal times indicated in label instructions. Persistence of antibiotics in milk following treatment is influenced by drug, dosage, route of administration, body weight and mammary gland health status. Compositional changes that take place during intramammary infection (IMI) can affect antibiotic excretion in milk, thus modifying milk withdrawal time. The objectives of this study were to validate sensitivity and specificity of a qualitative microbiological method (Charm AIM-96) to detect tylosin in bovine composite milk and to determine the influence of subclinical IMI in tylosin excretion following intramuscular administration. For test validation, two groups of approximately 120 cows were used; one received a single intramuscular injection of tylosin tartrate at a dose of 20 mg/kg, while the other group remained as untreated control. Test sensitivity and specificity were 100% and 94.1% respectively. To determine the influence of subclinical IMI in tylosin excretion, two groups of seven cows, one with somatic cell counts (SCC) < or =250 000 cells/ml and the other with SCC > or =900 000, were administered a single intramuscular injection of tylosin tartrate at a dose of 20 mg/kg. Milk samples were obtained every 12 h for 10 days following treatment. Milk tylosin excretion averaged between 5 and 9 days for cows with low and high SCC respectively (P < 0.0001). Compositional changes in cows with high SCC most likely affect the pharmacokinetic characteristics of tylosin, extending the presence of the antibiotic in milk, thus influencing milk withdrawal times.

[Antimicrobial sensitivity of coagulase-positive staphylococcal strains isolated from bovine mastitis in the central dairy catchment area of Argentina ]. / Susceptibilidad a antimicrobianos de cepas de estafilococos coagulasa positivos aisladas de mastitis bovina en la cuenca lechera central de la Argentina.

The activity of antimicrobial agents frequently used for treating bovine mastitis was determined against 101 coagulase-positive staphylococci isolated from bovine mammary secretion. The isolates were obtained from 39 dairy farms located in the central dairy area of Argentina. The disk diffusion method was used and the following antimicrobial agents were tested: penicillin, ampicillin, oxacillin, cephacetrile, penicillin + novobiocin, erythromycin, pirlimycin, novobiocin and neomycin. The highest levels of resistance were observed against penicillin and ampicillin (47.6%), while the lowest against erythromycin (2%), pirlimycin (4%) and neomycin (2.9%). No resistant strains against oxacillin, cephacetrile and penicillin + novobiocin were detected.