Abstract

The aim of this investigation was the assessment of toxicity of two new isolates of Bacillus thuringiensis, and the aqueous extract of Melia azedarach through in vivo assays in CF1 mice. Bt 1958-2, Bt 2014-2 and the BTh Thuricide 63 standard isolates were grown in liquid usual glicosed medium, and Cry proteins were purified by centrifugation on a sucrose gradient. The supernatant was autoclaved at 121º C, 15min. to maintain the exotoxins. Dehydrated leaves of M. azedarach were used to prepare a 10% aqueous extract. Mice were treated either orally or intraperitoneally with a whole bacterial suspension (1.10(10) UFC/mL), a culture supernatant or purified crystal protein (50 µg/mL), and with the plant extract (50 µg/mL). The stomachs of the mice were collected and observed in stereomicroscopy, and the stomach contents were analyzed in 10% SDS-PAGE. Results showed that none of the oral treatments were toxic to mice, but intraperitoneal bacterial suspensions were lethal to the animals 6 - 24 hours after injection. In conclusion, the Cry proteins of the new B. thuringiensis isolates must be evaluated for their use as tools in the biotechnology field, since they do not show toxicity against mammals, intragastrically or peritoneally, just like the M. azedarach aqueous extract (10%), with those being indicated for the biological control of pest insects.

Highlights

  • The entomopathogen Bacillus thuringiensis (Berliner, 1911), a Gram-positive bacterium, is naturally found in the soil (Höfte, Whiteley, 1989)

  • The supernatant was autoclaved at 121o C for 15 min, as described by Perani et al (1998), since that is a procedure which preserves the b-exotoxins if present in the culture medium

  • Follow-up analyses of symptoms from chemical substances in animals showed no change in the stomach mucosa of CF1 mice when treated with B. thuringiensis and M. azedarach, when compared with the control

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Summary

Introduction

The entomopathogen Bacillus thuringiensis (Berliner, 1911), a Gram-positive bacterium, is naturally found in the soil (Höfte, Whiteley, 1989). It is characterized by crystal production during sporulation, containing Cry proteins, encoded by the cry genes, with a wide division into classes and. Besides the Cry proteins, known as d-endotoxins, B. thuringiensis isolates can synthesize other toxins, such as b-exotoxin, phospholipases, proteases, chitinases, (Schenpf et al, 1998; Rabinovitch et al, 1998; Vilas-Bôas et al, 2012), and enterotoxins (Zahner et al, 2005). That toxin is analogous with ATP, being identified as an inhibitor of the rRNA synthesis (Mackedonski; Hadjiolov, 1972), resulting in dispersion and decreased number of chromosomes as well, double and micronuclei, tetraploid cells, among other effects of in vitro assays with Alium cepa (Sharma; Sahu, 1977)

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