PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Staphylococcus spp. plays a significant role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most important species due to the high prevalence and the difficulty of in vivo treatment that is related to the expression of virulence factors and biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolated from bovine mastitis and to evaluate the expression and regulation of genes involved in its production. MALDI-TOF and phenogenotypic identification assays were performed to characterize the isolates. The phenotypic biofilm production and the presence of icaA and icaD and bap genes were evaluated. The Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Furtherly, Real-time PCR (qPCR) was performed at chosen times to quantify the expression of icaA, icaD and hld genes in three selected isolates. All 20 strains were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II showed a prevalence of 70%. Transcriptional analysis revealed increased expression of ica genes at eight hours of growth. These results confirm that polysaccharides production mediated by the icaADBC operon genes is an essential mechanism to the biofilm formation and contributes to the early stages of bacterial growth.

• Gene biofilm Staphylococcus aureus bovine subclinical mastitis bovine icaA icaD hld Agr types

Abstract in Portuguese:

Staphylococcus spp. desempenham um papel significativo na etiologia da mastite bovina. Staphylococcus aureus é considerada a espécie mais importante devido a alta prevalência e a dificuldade de tratamento in vivo que está relacionado à expressão dos fatores de virulência e formação de biofilme. Este estudo teve como objetivo detectar a expressão fenotípica da formação de biofilme em 20 cepas de S. aureus isoladas de mastite bovina e avaliar a expressão e regulação de genes envolvidos em sua produção. MALDI-TOF e ensaios de identificação fenogenotípica foram realizados para caracterizar os isolados. A produção fenotípica de biofilme e a presença dos genes icaA, icaD e bap foram avaliadas. O sistema Agr foi tipificado (agr I, agr II, agr III e agr IV) e seu regulador (agr RNAIII) foi detectado. Além disso, a PCR em tempo real (qPCR) foi realizada nos tempos determinados para quantificar a expressão dos genes icaA, icaD e hld em três isolados selecionados. Todas as 20 linhagens foram produtoras de biofilme e a maioria apresentava os genes icaA e icaD. Apenas um isolado apresentou o gene bap. O gene agr do tipo II mostrou uma prevalência de 70%. A análise transcricional revelou aumento da expressão de genes ica às oito horas de crescimento. Estes resultados confirmam que a produção de polissacarídeos mediada pelos genes do operon icaADBC é um mecanismo essencial para a formação do biofilme e contribui para os estágios iniciais do crescimento bacteriano.

• Gene biofilme Staphylococcus aureus mastite subclínica bovina bovinos icaA icaD hld tipos de Agr

Abstract in English:

Zafalon L.F., Nader Filho A., Oliveira J.V. & Resende F.D. 2005. [Electrical conductivity and chloride concentration of milk as auxiliary diagnostic methods in bovine subclinical mastitis.] Comportamento da condutividade elétrica e do conteúdo de cloretos do leite como métodos auxiliares de diagnóstico na mastite subclínica bovina. Pesquisa Veterinária Brasileira 25(3):159-163. Depto Medicina Veterinária Preventiva e Reprodução Animal, FACVJ, Unesp - Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/no, Jaboticabal, SP 14884-900, Brazil. E-mail: zafalon@iz.sp.gov.br Electrical conductivity measured by a hand-held meter and chloride concentration of milk were studied as auxiliary methods for diagnosis of bovine subclinical mastitis in the identification of affected mammary quarters where Staphylococcus aureus and Corynebacterium sp were later isolated. Tests were made during 2 years in Holstein cows of a dairy farm producing type C milk, where milking was performed once a day. Sensitivities of electrical conductivity and chloride concentration tests from mammary quarters, where Corynebacterium sp was isolated (65.3% and 78.3%, respectively), were superior to the found in mammary quarters where S. aureus was identified (55.4% and 68.2%, respectively). The efficacies of the two diagnostic tests were similar. Statistical significance was demonstrated with regression analysis of both tests of healthy mammary quarters and subclinical mastitis quarters infected with Staphylococcus aureus.

• Bovine subclinical mastitis electric conductivity chloride level Staphylococcus aureus Corynebacterium sp.

Abstract in Portuguese:

Zafalon L.F., Nader Filho A., Oliveira J.V. & Resende F.D. 2005. [Electrical conductivity and chloride concentration of milk as auxiliary diagnostic methods in bovine subclinical mastitis.] Comportamento da condutividade elétrica e do conteúdo de cloretos do leite como métodos auxiliares de diagnóstico na mastite subclínica bovina. Pesquisa Veterinária Brasileira 25(3):159-163. Depto Medicina Veterinária Preventiva e Reprodução Animal, FACVJ, Unesp - Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/no, Jaboticabal, SP 14884-900, Brazil. E-mail: zafalon@iz.sp.gov.br Electrical conductivity measured by a hand-held meter and chloride concentration of milk were studied as auxiliary methods for diagnosis of bovine subclinical mastitis in the identification of affected mammary quarters where Staphylococcus aureus and Corynebacterium sp were later isolated. Tests were made during 2 years in Holstein cows of a dairy farm producing type C milk, where milking was performed once a day. Sensitivities of electrical conductivity and chloride concentration tests from mammary quarters, where Corynebacterium sp was isolated (65.3% and 78.3%, respectively), were superior to the found in mammary quarters where S. aureus was identified (55.4% and 68.2%, respectively). The efficacies of the two diagnostic tests were similar. Statistical significance was demonstrated with regression analysis of both tests of healthy mammary quarters and subclinical mastitis quarters infected with Staphylococcus aureus.