Study of the interaction of the algae Prototheca zopfii with neutrophils recovered from bovine milk and action of the IAA/HRP on this pathogen / Estudo da interação da alga Prototheca zopfii com neutrófilos recuperados de leite bovino e ação do sistema AIA/HRP sobre este patógeno

AUTOR(ES)
DATA DE PUBLICAÇÃO

2010

RESUMO

Studies have shown the incidence of bovine mastitis associated with the algae Prototheca zopfii. The objective of this work was to study the interaction of P. zopfii with neutrophils recovered from bovine milk and to evaluate the effect of system indole-3-acetic acid/horseradish peroxidase (IAA/HRP) on the viability of this microorganism in vitro experiment. P. zopfii was recovered from cows with clinical mastitis and both the molecular and morphological characterization were performed besides the evaluation of exponential growth of the microorganism in the laboratory. Next, neutrophils recovered from bovine milk were incubated in the absence and presence of opsonized P. zopfii and were evaluated the production of hydrogen peroxide, antioxidant enzymes on neutrophils and microorganism, and phagocytic capacity. In another study, P. zopfii was incubated with the system IAA/HRP and the viability assessed by colony forming units (CFU), antioxidant enzymes activity, membrane integrity by exclusion with Trypan blue and DNA integrity. The results were analyzed by analysis of variance with a 5% significance using the Tukey test. Results from P. zopfii characterization showed various cellular sizes, presence of autofluorescence, exponential microorganism growth throughout the incubation time and was not possible to determine the beginning of the death. Moreover it was found genotypes 1, 2 and 3 in the isolates in study. The production of hydrogen peroxide by neutrophils in the presence of algae was stimulated 5 times compared to the control, increase the activity of catalase (CAT) in 21% and glutathione reductase (GR) in 27% was seen in neutrophils; and there was no significant difference in CAT, GR and superoxide dismutase (SOD) activity produced by P. zopfii. P. zopfii was not engulfment by neutrophils. The system IAA/HRP inhibited the growth of the microorganism in 45, 82 and 88% in the times of 4, 6 and 9 hours of incubation, the activity of SOD, CAT, glutathione peroxidase (GPx) and GR increased respectively by 90, 120, 150%, and 3.4 times, decreased the viability of P. zopfii 10, 15, 20, 25 and 32% after the times of 4, 6, 8, 10 and 12 hours of incubation, and did not affect the integrity of DNA after 6 hours of incubation. As a conclusion, P. zopfii is highly resistant to the neutrophils and demonstrated to be susceptible to the effect microbicidal of system IAA/HRP.

ASSUNTO(S)

mastitis microbicida Ácido indol-3-acético horseradish peroxidase mastite peroxidase de raiz forte p. zopfii microbicide indole-3-acetic acid p. zopfii

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