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Molecular biology tools to study microbial diversity and characterization of Staphylococcus aureus in organic and conventional dairy in São Paulo

Abstract

Cheese is an important food of animal origin and can be vehicle to pathogen transmission. Among of the most prevalent microorganisms in cheese is the S. aureus that has the capacity to produce enterotoxin, to form biofilm, and some strains are resistant to antibiotics and sanitizers. The objectives of the study are to observe the microbial diversity, to perform the bacterial isolation,to identify and to characterize molecularly and phenotypically Staphylococcus aureus and to evaluate the genetic transfer of resistance genes in biofilm. For microbial diversity study, we will be performed using metagenomic sequencing of 16S rRNA gene from total DNA samples from six cheese processing plants, three organic and three conventional. From the samples will be carryied out bacterial isolation and identification of S. aureus by phenotypic analysis and the specific nuc gene. In the characterization of antibiotic resistant strains, disc infusion method will be completed testing several antimicrobial agents (oxacillin, cefoxitin, chloramphenicol, clindamycin, erythromycin, penicillin, tobramycin, tetracycline and gentamicin). Using Polymerase Chain Reaction (PCR), genes encoding biofilm formation (icaA, icaD, bap) and enterotoxins (sea-see, seg-sej) production will be identified. For molecular comparison of strains will be performed agr-type, spatyping , typing of SCCmec, and PFGE. Strains with biofilm-forming ability and with the presence of the mecA gene will be selected to the study of genetic transfer of resistance genes into biofilms, which will be perform at Department of Clinical Microbiology at Education and Research Centre Royal College of Surgeons in Ireland under supervision of Dr. Deirdre Fitzgerald Hughes. (AU)

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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ABREU, ANDERSON CLAYTON DA SILVA; CRIPPA, BRUNA LOURENCO; SOUZA, VICTORIA VILACA MARTINS ALENCAR DE; NUNEZ, KAREN VANESSA MUNIVE; ALMEIDA, JAQUELINE MILAGRES DE; RODRIGUES, MARJORY XAVIER; CIRONE SILVA, NATHALIA CRISTINA. Assessment of sanitiser efficacy against Staphylococcus spp. isolated from Minas Frescal cheese producers in Sao Paulo, Brazil. INTERNATIONAL DAIRY JOURNAL, v. 123, DEC 2021. Web of Science Citations: 0.
DA SILVA ABREU, ANDERSON CLAYTON; CARAZZOLLE, MARCELO FALSARELLA; CRIPPA, BRUNA LOURENCO; BARBOZA, GIOVANA RUEDA; RALL, VERA LUCIA MORES; ROCHA, LILIANA DE OLIVEIRA; CIRONE SILVA, NATHALIA CRISTINA. Bacterial diversity in organic and conventional Minas Frescal cheese production using targeted 16S rRNA sequencing. INTERNATIONAL DAIRY JOURNAL, v. 122, NOV 2021. Web of Science Citations: 0.
DA SILVA ABREU, ANDERSON CLAYTON; MATOS, LUIZ GUSTAVO; DA SILVA CANDIDO, TALITA JUNIA; BARBOZA, GIOVANA RUEDA; MARTINS ALENCAR DE SOUZA, VICTORIA VILACA; MUNIVE NUNEZ, KAREN VANESSA; CIRONE SILVA, NATHALIA CRISTINA. Antimicrobial resistance of Staphylococcus spp. isolated from organic and conventional Minas Frescal cheese producers in Sao Paulo, Brazil. JOURNAL OF DAIRY SCIENCE, v. 104, n. 4, p. 4012-4022, APR 2021. Web of Science Citations: 0.
DA SILVA CANDIDO, TALITA JUNIA; DA SILVA, ANDERSON CLAYTON; DE MATOS, LUIZ GUSTAVO; DO NASCIMENTO, MARISTELA DA SILVA; CAMARGO, CARLOSRI HENQUE; ZANELLA, ROSEMEIRE COBO; MORES RALL, VERA LUCIA; CIRONE SILVA, NATHALIA CRISTINA. Enterotoxigenic potential and molecular typing of Staphylococcus sp. isolated from organic and conventional fresh minas cheese in the state of Sao Paulo, Brazil. INTERNATIONAL DAIRY JOURNAL, v. 102, MAR 2020. Web of Science Citations: 0.

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