Epidemiologia molecular aplicada ao monitoramento de estirpes de Staphylococcus aureus na produção de queijo Minas frescal

We studied the molecular epidemiology of Staphylococcus aureus strains potentially toxigenic, isolated from the production process of Minas frescal cheese in a small dairy plant in the state of São Paulo. For this, samples were taken during the period from June 2008 to July 2009. Samples were collected from the surface of the receiving and storage tanks of raw milk, the surface of the balance tank of pasteurized milk, the water supply system, the pipes and equipments, the hands of the handler and from the packaged cheese, totaling 140 samples. The colonies isolated on Baird-Parker Agar confirmed as Gram positive and positive for catalase, coagulase and acetoin production, were submitted to extraction of bacterial DNA using the Invitek - Uniscience® kit. Confirmation of the isolated species and enterotoxins SEA, SEB, SEC, SED and TSST-1 toxin was carried out through the amplification of specific fragments of chromosomal DNA. Among the 74 strains of isolated coagulase-positive staphylococci, only 41 (55.4%) strains were confirmed as Staphylococcus aureus, of which 25 (61.0%) were positive to the presence of staphylococcal toxins. The most frequently identified enterotoxin was SEA. The toxigenic strains of Staphylococcus aureus were more frequently isolated from hands of the handler (16.0%), raw milk receiving tank (12.0%), pasteurized milk for cheese making (12.0%) and fresh white cheese ready for consumption (12.0%).

Detecção dos genes de virulência e identificação do perfil clonal de isolados de Staphylococcus aureus colonizantes de nasofaringe otbtidos em estudo de base populacional

Pós-graduação em Doenças Tropicais - FMB

Previous infection with Staphylococcus aureus strains attenuated experimental encephalomyelitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dynamics of biofilm formation and the interaction between candida albicans and methicillin-susceptible (MSSA) and -resistant staphylococcus aureus (MRSA)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genetic determinants of methicillin resistance and virulence among Staphylococcus aureus isolates recovered from clinical and surveillance cultures in a Brazilian teaching hospital

Aims. To quantify the presence of SCCmec types and virulence genes among Staphylococcus aureus colonizing and infecting patients from a teaching hospital. Methods. We analyzed 225 and 84 S. aureus isolates recovered from surveillance and clinical cultures, respectively. Strains were studied for the presence and type of SCCmec, as well as for several virulence genes. Univariate and multivariable analysis were performed in order to identify predictors of invasiveness (defined as isolation from clinical cultures). Results. The presence of SCCmec types III (OR, 2.19, 95% CI, 1.08-4.45) and IV (OR, 5.28 95% CI, 1.35-20.63) and of genes coding for exfoliative toxin B (etb, OR, 6.38, 95% CI, 1.48-27.46) and Panton-Valentine leukocidin (pvl, OR, 2.38, 95% CI, 1.16-4.86) was independently associated with invasiveness. Conclusions. SCCmec types III and IV and virulence genes are associated with greater invasiveness of S. aureus. Patients colonized with methicillin-resistant S. aureus, as well as with strains harboring etb or pvl, may be prone to develop invasive disease. Infection-preventing strategies should be more intensively applied to this group.

Epidemiologia molecular e estudo dos fatores de virulência de Staphylococcus aureus resistentes à oxacilina isolados de feridas em pacientes atendidos em unidades básicas de saúde da cidade de Botucatu

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fluorescent amplified fragment length polymorphism genotyping of human and animal Staphylococcus aureus isolates from dairy farms with manual milking

Fluorescence amplified fragment length polymorphism (fAFLP) was used to assess the genetic relatedness of 40 Staphylococcus aureus strains isolated from human and animal skin samples in seven dairy farms with manual milking. S. aureus was isolated from 11 out of 30 (36%) human skin samples and from 29 out of 100 (29%) teat skin samples from apparently healthy cows. Genomic DNA from each isolate was double-digested with EcoRI and MseI and complementary oligonucleotide adaptors were ligated to the restriction fragments. Pre-selective and selective, amplification reactions were performed, the amplified fragments were separated by electrophoresis in an ABI377 sequencer and analysed using GeneScan 3.1 and Genotyper 2.5. Three single isolates (a-c), a predominant cluster with 35 isolates (d) and another cluster with two isolates (e) were identified. Both clusters d and e included human and animal isolates genetically related, because the profiles had 90-100% homology. Since no cluster was comprised uniquely of human or animal isolates and given the close genetic relatedness among human and animal samples in the farms, the present findings support the. hypothesis that dairy workers can spread S. aureus through manual milking. (C) 2005 Elsevier B.V. All rights reserved.

Molecular epidemiology of Staphylococcus aureus isolates at different sites in the milk producing dairy farms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Proteinograma e teores de cobre, ferro e zinco no soro sanguíneo de ovelhas da raça Santa Inês com mastite experimental por Staphylococcus aureus

Costa N.A., Simao L.C.V., Santos R.A., Afonso J.A.B., Fagliari J.J., Cardoso E.C., Soares P.C. & Mendonca C.L. 2010. [Proteinogram and serum concentrations of copper, iron and zinc in Santa Ines ewes with Staphylococcus aureus experimentally induced mastitis.] Proteinograma e teores de cobre, ferro e zinco no soro sanguineo de ovelhas da raca Santa Ines com mastite experimental por Staphylococcus aureus. Pesquisa Veterinaria Brasileira 30(5):435-442. Clinica de Bovinos, Campus Garanhuns, Universidade Federal Rural de Pernambuco (UFRPE), Avenida Bom Pastor s/n, Boa Vista, Caixa Postal 152, Garanhuns, PE 55292-901, Brazil. E-mail: carlalopes.mendonca@gmail.comThe aim of the present study was to evaluate the effect of Staphylococcus aureus experimentally induced mastitis on proteinogram and serum concentrations of cupper, iron and zinc levels of Santa Ines primiparous ewes. The right mammary gland of ten healthy ewes was inoculated with 1,0x10(4) UFC/mL of S. aureus. Clinical examination and determination of serum concentrations of proteins by electrophoresis in polyacrylamide gel (SDS-PAGE), cupper, iron and zinc, as well plasma level of fibrinogen were measured before the inoculation (control) and 12h, 24h, 36h, 48h, 60h, 72h, 84h, 96h, 108h, 120h, 132h, 168h, 180h, 288h and 336h after bacteria inoculation. All animals experimentally infected presented clinical mastitis and subsequent loss of mammary gland function. The electrophoretogram allowed the identification of 23 proteins with molecular weights (MW) ranged from 26.000 to 185.000 daltons (Da) including acute-phase proteins, IgG and IgA. A significant increase (P<0,05) in haptoglobin, ceruloplasmin, IgG and IgA concentrations was observed. Antitrypsin and acid glicoprotein concentrations did not alter. The levels of iron and zinc decreased and the cupper concentration increased. A positive correlation between plasma fibrinogen and serum ceruloplasmin (r=0.74), haptoglobin (r=0.62) and IgA (r=0.62) was also identified. Results showed the importance of ceruloplasmin and haptoglobin as acute-phase proteins in ewes with intramammary infections and confirms fibrinogen as an inflammatory marker because its high correlation with specific proteins. The alterations in the serum levels of Cu, Fe and Zn suggest the action of inflammatory mediators triggered by S. aureus.

Occurrence of Staphylococcus aureus in raw milk produced in dairy farms in São Paulo state, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de Staphylococcus aureus em queijo tipo frescal

OBJETIVO: Verificar a ocorrência de Staphylococcus aureus em uma amostra de queijo tipo Minas frescal comercializado na cidade de Poços de Caldas, MG, de modo a obter subsídios que permitam avaliar o risco potencial que este produto pode representar para a saúde da população consumidora. MÉTODOS: Foi investigada a presença e o número de cepas de Staphylococcus aureus em 80 amostras de queijo tipo Minas frescal produzido artesanalmente e comercializado na cidade de Poços de Caldas, MG, Brasil. RESULTADOS: Os resultados obtidos evidenciaram a presença de S. aureus em 40 (50,0%) amostras, cujas contagens revelaram valores médios em torno de 10(5)/g. CONCLUSÕES: Tais achados parecem ser extremamente preocupantes, pois além de se situarem acima do limite máximo de 10³/g estabelecido pelo Ministério da Saúde, estes valores mostraram-se muito próximos dos requeridos para a produção de enterotoxinas em quantidades suficientes para a ocorrência de surtos de intoxicação alimentar estafilocócica.

Perfil eletroforético e concentração de imunoglobulinas G (IgG) do soro sanguíneo de cabras Saanen com mastite experimental induzida por Staphylococcus aureus suplementadas com vitamina E

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Oxacillin Resistance of Staphylococcus aureus Isolated from the University Hospital of Botucatu Medical School in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Risk factors for acquisition of Methicillin-resistant Staphylococcus aureus among patients from a burn unit in Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) is an important agent of colonization and infection in burn units. in order to identify risk factors for MRSA acquisition in a Brazilian burn unit, we performed two retrospective studies. In the first ("cohort" study), 175 patients who were not colonized with MRSA on admission were followed to assess risk factors for MRSA acquisition. in the second ("case-case-control" study), 143 individuals from the previous study who were negative for both MRSA and Methicillin-susceptible S. aureus (MSSA) on admission were followed. Case-control studies were performed to investigate risk factors for MRSA and MSSA acquisition. MRSA and MSSA were recovered from 75 and 23 patients, respectively. In the "cohort" study, only the number of wound excisions (Odds Ratio [OR] = 1.55, 95% Confidence Interval [CI] = 1.21-1.98, P = 0.001) was associated with MRSA acquisition. in the "case-case-control" study, burns involving head (OR=3.43, 95%CI = 1.50-7.81, P = 0.003) and the number of wound excisions (OR = 1.83, 95%CI = 1.27-2.63, P = 0.001) were significant risk factors for MRSA. Burns involving perineum were negatively associated with MSSA acquisition (OR = 0.16, 95%CI = 0.03-0.75, P = 0.02). In conclusion, the acquisition of MRSA was related to the site of the burn and to the surgical manipulation of tissues, but not to the use of antimicrobials. (C) 2009 Elsevier Ltd and ISBI. All rights reserved.