Lucia Esther Alcaraz

Species identification, slime production and oxacillin susceptibility in coagulase-negative staphylococci isolated from nosocomial specimens

Ninety-two coagulase negative staphylococci (CNS) (forty-five of clinical origin and forty-seven of environmental origin), collected in a hospital in San Luis, Argentina, from March to June, 1999, were identified to species level by the ID 32 Staph and API Staph System (bioMerieux). Slime production was investigated by the quantitative and qualitative methods. Oxacillin susceptibility was determined by the disk diffusion test (1 µg), the agar dilution method (0.125 to 4 mg/ml) and agar screen (6 µg/ml). The presence of mecA gene was investigated by PCR. The clinical CNS species most commonly isolated were S. epidermidis, S. haemolyticus, S. hominis and S. saprophyticus. The frequency of slime production by clinical and environmental isolates was similar (25/45 and 27/47, respectively) and the results obtained by the quantitative and the qualitative methods correlated well. The mecA gene was detected in all S. epidermidis, S. haemolyticus and S. hominis isolates, which were resistant to oxacillin by the phenotypic methods. However, this gene was not present in S. klossii, S. equorum, S. xylosus and S. capitis strains. The gene was neither found in two out of the six S. saprophyticus isolates, in two out of three S. cohnii subsp. urealyticum isolates and in two out of five S. cohnii subsp. cohnii isolates, all of which resulted oxacillin resistant according to MIC. The gene was not found in oxacillin-susceptible strains either. Most of the CNS isolates (enviromental and clinical) that were slime producers were found to be oxacillin resistant, which makes the early detection of these microorganisms necessary to prevent their dissemination in hospitals, particularly among immunocompromised patients.

Antibacterial activity of extracts of Acacia aroma against methicillin-resistant and methicillin-sensitive Staphylococcus

Antibacterial activity of organic and aqueous extracts of Acacia aroma was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), methicillin sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus epidermidis. Inhibition of bacterial growth was determined using agar diffusion and bioautographic methods. Among all assayed organic extracts only ethanolic and ethyl acetate extracts presented highest activities against all tested Staphylococcus strains with minimal inhibitory concentration (MIC) values ranging from 2.5 to 10 mg/ml and from 2.5 to 5 mg/ml respectively. The aqueous extracts show little antibacterial activity against Staphylococcus strains. The bioautography assay demonstrated well-defined growth inhibition zones against S. aureus in correspondence with flavonoids and saponins. A. aroma would be an interesting topic for further study and possibly for an alternative treatment for skin infections.

IFN-γ plays a detrimental role in murine defense against nasal colonization of Staphylococcus aureus

The anterior nares are the major reservoir in humans of Staphylococcus aureus with the risk of developing endogenous infections or transmitting infections to susceptible persons. The mechanisms that mediate attachment of staphylococci to the nasal mucosa are little known. The purpose of the present work was to study some factors that could influence the nasal colonization in an animal model of mice. We investigated the possible role of IFN-gamma. We used S. aureus ATCC 35556 (SA113) slime-producing and ATCC 25923 non-slime-producing strains. Male 6-week-old BALB/c, C57BL/6 (wild-type, WT), and gene-deficient IL-12p40 (IL-12p40-/-) or IL-4 (IL-4-/-) mice on C57BL/6 background were infected with a dose of S. aureus of 10(6) CFU in 10mul of saline. The total number of S. aureus CFU per nose and lung, specific IgA response and IFN-gamma levels were evaluated. Significant higher CFU were recovery from the narines of C57BL/6 compared with BALB/c mice either after ATCC 35556 (p<0.0001) and ATCC 25923 (p<0.02) strain infection. Low IgA response correlated with high bacterial counting in the C57BL/6 nasal region. Moreover, C57BL/6 mice showed major colonization of slime-producing S. aureus ATCC 35556 than non-slime-producing ATCC 25923 S. aureus strain (p<0.02). IL-12p40-/-mice clarified the bacteria from their nose more efficiently that WT mice after slime-producing S. aureus (p<0.0001). Accordingly, significant lower level of IFN-gamma were detected in IL-12p40-/- compared with WT mice after infection with this strain (p<0.03). The results suggested the influence of the slime production in nasal colonization of S. aureus, and indicate at first time that IFN-gamma may play a detrimental role in this mucosal infection. These results could contribute to elucidate mucosal immune mechanisms involved in S. aureus colonization and then control infections in susceptible persons.

Presence of enterotoxigenic Staphylococcus aureus in artisan fruit salads in the city of San Luis, Argentina

An increase in the consumption of fruit juices and minimally processed fruits salads has been observed in recent years all over the world. In this work, the microbiological quality of artisan fruit salads was analysed. Faecal coliforms, Salmonella spp, Shigella spp, Yersinia enterocolitica and Escherichia coli O157:H7 were not detected; nevertheless, eleven strains of Staphylococcus aureus were isolated. By multiplex PCR, all isolates showed positive results for S. aureus 16S rRNA gene and 63.6% of them were positive for sea gene. Furthermore, PCR sea positive strains were able to produce the corresponding enterotoxin. Finally, the inactivation of these strains in fruit salads by nisin, lysozyme and EDTA, was studied. EDTA produced a total S. aureus growth inhibition after 60 h of incubation at a concentration of 250 mg/L. The presence of S. aureus might indicate inadequate hygiene conditions during salad elaboration; however, the enterotoxigenicity of the strains isolated in this study, highlights the risk of consumers’ intoxication. EDTA could be used to inhibit the growth of S. aureus in artisan fruit salads and extend the shelf life of these products.