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      Biofilm formation by Prototheca zopfii isolated from clinical and subclinical bovine mastitis in distinct growth conditions under different dyes Translated title: Formação de biofilme por Prototheca zopfii isolados de mastite bovina clínica e subclínica em distintas condições de crescimento sob diferentes corantes

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          Abstract

          ABSTRACT: Prototheca spp. have been reported as an emergent environmental mastitis pathogen in several countries. Biofilm formation is a significant factor associated with different degrees of virulence developed by many microorganisms, including Prototheca spp. The present study aimed to compare two growth conditions and two staining dyes to determine which combination was more appropriate to evaluate qualitatively and quantitatively the production of biofilm by P. zopfii. Biofilm formation was evaluated in polystyrene microplates under static and dynamic growth conditions and staining with crystal violet or cotton blue dye. All P. zopfii isolates from cows with mastitis were classified as biofilm-producers in all growth conditions and staining. The cotton blue dye proved to be more appropriate method to classify the intensity of P. zopfii biofilm production.

          Translated abstract

          RESUMO: Prototheca spp. tem sido relatado como um patógeno ambiental causador de mastite bovina em vários países. A formação de biofilme é um fator associado a diferentes graus de virulência desenvolvidos por muitos microrganismos, incluindo Prototheca zopfii. O presente estudo teve como objetivo comparar duas condições de crescimento e dois corantes para determinar a combinação mais adequada para avaliar qualitativa e quantitativamente a produção de biofilme por P. zopfii. A formação de biofilme foi avaliada em microplacas de poliestireno sob condições estáticas e dinâmicas de crescimento e coloração com cristal violeta ou azul de algodão. Todos os isolados de P. zopfii de vacas com mastite foram caracterizados como produtores de biofilme, independentemente das condições de crescimento e coloração. O corante azul de algodão demonstrou ser o método mais adequado para classificar a intensidade de produção de biofilme de P. zopfii.

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          Formation of biofilms by Staphylococcus aureus on stainless steel and glass surfaces and its resistance to some selected chemical sanitizers

          The objectives of this work were to verify the capability of Staphylococcus aureus of forming bio-film on stainless steel and glass surfaces; to evaluate the efficiency of sodium dichloroisocyanurate, hydrogen peroxide and peracetic acid in inactivating Staphylococcus aureus cells adhered onto these surfaces; and to visualize biofilm development by scanning electron microscopy before and after sanitizer treatment. The surfaces studied consisted of 10x20mm chips immersed in Petri dishes containing BHI broth inoculated with S. aureus ATCC 25923. Biofilm formation was observed after 15-day incubation, when the cells were removed using the swab technique, followed by Baird Parker agar plating. Also, the efficiency of the chemical sanitizers on the chip surfaces was tested and the non-removed cells were counted on the Baird-Parker agar. After biofilm formation and use of sanitizers, the chips were respectively observed by scanning electronic microscopy following a pre-existing protocol. The obtained results showed biofilm formation on both surfaces, with bacterial count in the order of 10(7) CFU/cm² on and 10(8) CFU/cm² on stainless steel and glass surfaces, respectively. Peracetic acid was the most efficient in removing adhered cells, presenting 5.26 and 4.5 decimal reduction for adhered cells on stainless steel and glass surfaces, respectively.
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            Diversity within the current algal species Prototheca zopfii: a proposal for two Prototheca zopfii genotypes and description of a novel species, Prototheca blaschkeae sp. nov.

            In order to clarify the intraspecies taxonomic position of the non-photosynthetic algal species Prototheca zopfii, as well as the aetiology of bovine mammary protothecosis, a selection of P. zopfii strains isolated from clinical cases of protothecal mastitis and from various environmental habitats was characterized using a polyphasic molecular approach. Based on sequence analysis of the 18S rRNA gene, which showed distinct differences between the three currently known biotypes or 'variants', specific oligonucleotides were designed and used in biotype-specific PCRs. Furthermore, the pattern of cellular fatty acids was evaluated. Typing by means of these techniques revealed that the previously defined biotypes of P. zopfii were clearly different. Based on sequence analysis, the pattern of fatty acids and physiological characteristics, it is proposed that biotype 3 should be reclassified as representing a novel species, Prototheca blaschkeae sp. nov. (type strain, RZIII-3(T) = SAG 2064(T)). Furthermore, it is proposed that P. zopfii merits reclassification as a species comprising at least two genotypes that in future could probably be considered to represent two subspecies.
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              Influence of the incubation temperature, atmosphere and dynamic conditions on biofilm formation by Salmonella spp.

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                cr
                Ciência Rural
                Cienc. Rural
                Universidade Federal de Santa Maria (Santa Maria, RS, Brazil )
                0103-8478
                1678-4596
                February 2019
                : 49
                : 2
                : e20180574
                Affiliations
                [1] Santa Maria Rio Grande do Sul orgnameUniversidade Federal de Santa Maria orgdiv1Departamento de Medicina Veterinária Preventiva (DMVP) orgdiv2Programa de Pós-graduação em Medicina Veterinária (PPGMV), Programa de Pós-Graduação em Ciências Farmacêuticas (PPGCF), Centro de Ciências Rurais (CCR) Brazil
                [3] Santa Maria Rio Grande do Sul orgnameUniversidade Federal de Santa Maria orgdiv1Departamento de Fisiologia e Farmacologia Brazil
                [4] Botucatu orgnameUniversidade Estadual Paulista orgdiv1Departamento de Higiene Veterinária e Saúde Pública orgdiv2Faculdade de Medicina Veterinária e Zootecnia Brazil
                [2] Frederico Westphalen RS orgnameInstituto Federal de Educação, Ciência e Tecnologia Farroupilha (IFFar) orgdiv1Laboratório de Microbiologia e Doenças Infecciosas Brazil
                [5] Petrolina orgnameUniversidade Federal do Vale do São Francisco Brazil
                Article
                S0103-84782019000200451
                10.1590/0103-8478cr20180574
                645c3f77-91d4-4f6b-8beb-357da689d96e

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 20 July 2018
                : 02 January 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 25, Pages: 0
                Product

                SciELO Brazil

                Categories
                Microbiology

                Microbiology & Virology
                Prototheca zopfii,bovine mastitis, intramammary infections, microplate biofilm assay, virulence factor,Prototheca zopfii, dairy cattle,fator de virulência,ensaio de biofilme em microplaca,infecções intramamárias,mastite bovina,bovino de leite

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