IN VITRO EFFECT OF OZONE ON PROTOTHECA SPECIES DIAGNOSED BY MASS SPECTROMETRY AND CYTB GENE PCR, IN COWS WITH CLINICAL MASTITIS

Abstract

Microalgae from the genus Prototheca are eucaryotic microorganisms, saprophytic and achlorophylled organisms, with opportunistic nature, which losses the photosynthetic ability, a fact that probably enabled the human and animal infections. Bovine mastitis is the most frequent clinical picture of protothecosis in domestic animals. Nonetheless, to date, no effective treatment is known, resulting in severe losses to farmers due to segregation, discharge, and culling of infected animals. This fact has motived different in vitro and in vivo studies using a set of antifungals, algaecidal, antimicrobials, and sanitizing products against Prototheca isolates. In this regards, ozone (O3) has been used in therapy approaches of nonresponsive clinical bovine mastitis cases treated with conventional drugs. The microbicide effect of O3 has been attributed to action in the cell wall of microorganisms, enabling the safe and effective use in therapy of cutaneous lesions and mucous membranes, in addition to disinfection of utensils and the environment. Also, the routine diagnosis of algae has been based on microbiological culture, phenotypic aspects of colonies and staining methods, and micromorphology of algae, although these procedures are not unequivocal for identification of species-level of algae. Currently, molecular methods, e.g., mass spectrometry and PCR of cytB gene have allowed the taxonomic reclassification and a valuable diagnosis of the pathogen. Considering this scenario, the current study intends to investigate the algaecide effect of ozonized oil and water in 44 Prototheca sp. isolates obtained of clinical mammary infections in cows (assessing the severity clinical scores of cases), from ten dairy farms of Sao Paulo and Minas Gerais States of Brazil, where diagnosis of algae will be based on mass spectrometry (MALDI-TOF MS) and PCR of cytB gene.