title: Microbicidal action of indole-3-acetic acid combined with horseradish peroxidase on Prototheca zopfii from bovine mastitis.
abstract: We investigated the toxic effect of indole-3-acetic acid (IAA) combined with horseradish peroxidase (HRP) on Prototheca zopfii from bovine mastitis. P. zopfii isolates were identified and characterized by morpho-physiological parameters; presences of P. zopfii genotype 2 were also investigated. Subsequently, P. zopfii was incubated in the absence (control) or presence of IAA/HRP and examined for: (i) cell viability; (ii) colonies number formation; (iii) antioxidant enzyme activity; and (iv) DNA integrity. Significance of differences was calculated using ANOVA and Tukey's test (P < or = 0.05). As evidenced by Trypan blue exclusion and colony formation in Sabouraud dextrose agar, IAA/HRP addition to the culture reduced respective P. zopfii viability and P. zopfii colony formation in a concentration- and time-dependent manner. IAA/HRP specifically reduced cell viability in 10, 15, 20, 25, and 32% after 4, 6, 8, 10, and 12 h of incubation, respectively, compared with the control at the same time. The number of colony formation was inhibited (45, 82, and 88%) by IAA/HRP after 4, 6, and 9 h of incubation, respectively, compared with the control at the same time. In addition, P. zopfii antioxidant activity increased measurably in the presence of IAA/HRP (6 h); superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase increased by 90, 120, 150% and 3.4 times, compared with the controls. IAA/HRP did not appear to effect P. zopfii DNA integrity when examined by electrophoresis. In conclusion, IAA/HRP appears to function as a microbicidal mechanism on P. zopfii genotype 2 from bovine mastitis.
authors.affiliation: Department of Basic Science, Faculty of Animal Science and Food Engineering (FZEA), University of São Paulo, Av. Duque de Caxias Norte, 225, Pirassununga, SP, 13635-000, Brazil.
authors.forename: Luciane T
authors.initials: LT
authors.lastname: Cunha
authors.forename: Silvana M P
authors.initials: SM
authors.lastname: Pugine
authors.forename: Marcia R M
authors.initials: MR
authors.lastname: Silva
authors.forename: Ernane J X
authors.initials: EJ
authors.lastname: Costa
authors.forename: Mariza Pires
authors.initials: MP
authors.lastname: De Melo
mesh_terms: D020418:Algal Proteins / Q000378:metabolism; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000378:metabolism; D002417:Cattle; D015169:Colony Count, Microbial; D048028:DNA, Algal / Q000235:genetics; D005260:Female; D006735:Horseradish Peroxidase / Q000494:pharmacology*; D007210:Indoleacetic Acids / Q000494:pharmacology*; D008414:Mastitis, Bovine / Q000382:microbiology*; D050296:Microbial Viability / Q000187:drug effects; D010088:Oxidoreductases / Q000378:metabolism; D011525:Prototheca / Q000187:drug effects* / Q000302:isolation & purification; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D014343:Trypan Blue / Q000378:metabolism
chemical_list: D020418:Algal Proteins; D000890:Anti-Infective Agents; D000975:Antioxidants; D048028:DNA, Algal; D007210:Indoleacetic Acids; C030737:indoleacetic acid; D010088:Oxidoreductases; D006735:Horseradish Peroxidase; D014343:Trypan Blue
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