Abstract Benzoic acids preservatives may be converted to benzene in soft drinks. The use of alternative antimicrobial compounds, specifically nisin and natamycin, would reduce benzene formation. Initially, doses of nisin and natamycin were tested against Lactobacillus plantarum lactic bacteria and Zygosaccharomyces bailii yeast. Using 167 UI/mL of nisin and 0.017% w/v of natamycin no microbial growth was reached on lemon-flavored soft drinks. The benzene formation was analyzed in a standard formulation (380 mg/L of sodium benzoate and 180 mg/L of potassium sorbate) and nisin and natamycn formulation (167 UI/mL of nisin and 0.017% w/v of natamycin). The soft drinks were stored without UV exposure at 20 °C and with UV sunlight exposure at 30 °C, for 120 days at PET bottles. The initial benzene content was 1.29 μg/kg for standard and 0.09 μg/kg for nisin and natamycin formulation; these values increased to 11.00 μg/kg and 0.18 μg/kg, respectively. The sensory attributes did not differ from each formulation, either initially or after 120 days of storage. The lower benzene concentrations obtained using nisin and natamycin suggest that these compounds are attractive alternatives to benzoic acids, keeping the sensory quality and microbial stability during shelf life.

Visando a seleção de seis grupos de genes cry de Bacillus thuringiensis (Bt), que codificam proteínas ativas para coleópteros e lepidópteros pragas do arroz, 46 isolados de Bt provenientes de amostras de solos das regiões orizícolas do Rio Grande do Sul (RS), foram testados por PCR. Os isolados de Bt foram crescidos em Ágar Nutriente durante 12 h e submetidos a extração de DNA total. Os fragmentos amplificados foram analisados em géis de agarose (1-1,5%). Os resultados referentes ao total de isolados selecionados mostraram que 56,51% foram potencialmente específicos a lepidópteros (cry1, cry2 e cry9) e 21,73% a coleópteros (cry3 e cry7/8), tendo sua distribuição homogênea entre as regiões orizícolas do RS. Apenas os genes cry2 foram localizados somente na região Litoral. Nos bioensaios com lagartas de Spodoptera frugiperda o isolado Bt 2027-1 obteve a maior mortalidade corrigida (25%), o qual havia sido pré-selecionado pela presença de genes cry9. Para a mesma espécie, os testes de toxicidade através de proteínas purificadas de Bt aizawai HD68 revelaram uma DL50 de 0,95 mg/larva. Dois isolados de Bt causaram 100% de mortalidade às larvas de Oryzophagus oryzae, tendo esses sido pré-selecionados pela presença de genes cry3. Os resultados dos bioensaios confirmam a predição da atividade de Bt por PCR, a qual deve estar diretamente relacionada aos genes cry que codificam as proteínas inseticidas específicas.

Bacillus thuringiensis (Bt) isolates from soil samples of rice-fields in Rio Grande do Sul (RS) were tested through PCR, aiming at the screening of six groups of Bt cry genes, which codify active proteins for coleopterans and lepidopterans rice pests, and their bioinsecticide potential regarding their use in IPM system, as well. Forty six Bt isolates were grown in Agar Nutrient for 12 h and submitted to total DNA extraction. The amplified fragments were analyzed in agarose gel (1-1.5%). The screening isolates showed that 56.51% were potentially lepidopterans specific (cry1, cry2 and cry9) and 21.73% were coleopteran specific (cry3 and cry7/8), with a homogeneous distribution in the rice-field areas in Rio Grande do Sul. Cry2 genes were found just once and in the Litoral area. Bioassays against Spodoptera frugiperda larvae showed the highest corrected mortality (25%) with Bt 2027-1 isolate selected by the presence of cry9 genes. The toxicity bioassays carried out with S. frugiperda using purified proteins of Bt aizawai HD68 indicated a LD50 of 0.95 µg/larvae. Two Bt isolates carrying the cry3 genes (PCR detection) caused a 100% mortality to Oryzophagus oryzae larvae. Bioassay results confirmed the prediction of Bt activity by PCR, which must have a straight relationship with the cry genes that codify those specific insecticidal proteins.

A bactéria Bacillus thuringiensis (Bt) caracteriza-se pela produção de proteínas tóxicas a representantes de diversas ordens de insetos, as quais são codificadas por genes cry. Devido a esta característica, mais de 40.000 cepas de Bt foram isoladas e cerca de 190 genes cry, caracterizados. Como os dados sobre Bt são limitados no Rio Grande do Sul, essa pesquisa objetivou avaliar a distribuição de seis famílias de genes cry de Bt, desse estado, que codificam proteínas ativas contra insetos-praga. O perfil dos 46 isolados de solos do Rio Grande do Sul foi avaliado, por PCR com os primers que detectam os genes cry1, cry2, cry3, cry7 cry8 e cry9 e suas respectivas proteínas foram analisadas por SDS-PAGE a 10%. A presença de genes cry9 foi detectada em 47,82% dos isolados, seguido de cry3 (15,21%), cry1 e cry7 (ambos com 6,52%) e cry2 (2,17%). Oito perfis genéticos foram identificados, sendo o perfil cry9 (39,13%) o mais freqüente. A análise protéica de Bt identificou 14 famílias de proteínas Cry possivelmente codificadas por genes presentes nos isolados, além de proteínas desconhecidas que podem caracterizar novos genes cry. Esses isolados revelam a presença de genes que codificam proteínas específicas contra lepidópteros e coleópteros, as quais poderão ser avaliadas quanto à toxicidade in vivo contra insetos-praga das plantas cultivadas.

The Bacillus thuringiensis (Bt) bacterium is characterized by the production of toxic protein to representatives of several insect orders, which are coded by cry genes. Due to this characteristic, more than 40.000 Bt strains were isolated and around 190 cry genes characterized. As the data on Bt are limited in Rio Grande do Sul, this research aimed the evaluation of the distribution of six Bt families of cry genes, in this state, that codify active proteins against insect-pest. The 46 isolates profiles of soil samples from Rio Grande do Sul were evaluated, by PCR with primers that detect cry1, cry2, cry3, cry7, cry8 and cry9 genes, and its respective proteins were analyzed by SDS-PAGE 10%. The presence of cry9 genes were detected in 47.82% of the isolates, followed by cry3 (15.21%), cry1 and cry7 (both with 6.52%) and cry2 (2.17%). Eight genetic profiles were identified, being the profile cry9 (39.13%) the most frequent one. The Bt proteic analysis identified 14 families of Cry protein that are possibly coded by the genes in these isolates, besides the unknown proteins that can characterize new cry genes. These isolates reveal the presence of genes that codify specific proteins against lepidopterans and coleopterans, which can be evaluated for its in vivo toxicity against insect-pest of the cultivated plants.