Abstract
Staphylococcus spp. isolated from samples of Minas cheese traditionally manufactured following artisan procedures were identified using molecular techniques and further analyzed using PCR and specific primers for the detection of classic enterotoxins (SEA, SEB, SEC, SED, and SEE) and toxic shock syndrome toxin-1 (TSST-1). Specific sea, sec, sed, and see genes were identified using multiplex PCR, whereas seb and tst genes were detected by uniplex PCR. In vitro antagonism with Lactobacillus spp. was evaluated to assess antimicrobial susceptibility. Classic enterotoxins and TSST-1 genes were not detected. The antimicrobials sulfonamide, penicillin, ceftzadime, and oxacillin showed higher resistance rates in the antibiogram (100%, 80%, 60%, and 40%, respectively), whereas other antimicrobials were effective in percentages above 70%. Lactobacillus spp. were able to inhibit Staphylococcus spp. in vitro. Thus, our results indicated that the isolated Staphylococcus spp. were sensitive to the most common antimicrobials tested and were inhibited by Lactobacillus spp.
Highlights
Staphylococcus spp. isolated from samples of Minas cheese traditionally manufactured following artisan procedures were identified using molecular techniques and further analyzed using PCR and specific primers for the detection of classic enterotoxins (SEA, SEB, SEC, SED, and SEE) and toxic shock syndrome toxin-1 (TSST-1)
A ausência de bacteremia nessas pacientes sugeriu que a doença seria resultado de uma intoxicação por produtos elaborados pelo micro-organismo
Alto percentual de resistência também foi observado para Penicilina (80%), Ceftazidima (60%) e Oxacilina (40%)
Summary
Foram avaliadas quinze cepas de Staphylococcus spp. previamente isoladas de queijo produzido a partir de leite cru provenientes da região da Serra da Canastra (MG). A identificação dos micro-organismos foi realizada por meio de técnicas moleculares baseadas na amplificação de um fragmento do rDNA 16S14 e os produtos da PCR purificados foram sequenciados em aparelho ABI3130, utilizando-se polímero POP7 e BigDye v.3.1, realizado pela Valid Biotecnologia do Laboratório de Genética Animal do Departamento de Zootecnia da Escola de Veterinária - UFMG. Para a pesquisa dos genes seb e tst, foram realizadas duas reações de PCR-Uniplex. A reação do fragmento para o gene tst foi programada para 94oC por 5 minutos, seguido de 30 ciclos térmicos, cada um consistindo de 95oC por 1 minuto, 55oC por 1 minuto e 72oC por 2 minutos, e, finalmente, 72oC por 7 minutos. Iniciadores usados para a detecção por PCR dos genes sea-see e tst em Staphylococcus spp. isolados de amostras de queijos artesanais da região da Serra da Canastra (Minas Gerais, Brasil)
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