Fumigant antifungal activity of Corymbia citriodora and Cymbopogon nardus essential oils and citronellal against three fungal species.

Corymbia citriodora and Cymbopogon nardus essential oils samples were analyzed by GC and GC-MS and their qualitative and quantitative compositions established. The main component of essential oils of C. citriodora and C. nardus was citronellal, at 61.78% and 36.6%, respectively. The essential oils and citronellal were tested for their fumigant antifungal activity against Pyricularia (Magnaporthe) grisea, Aspergillus spp., and Colletotrichum musae. The minimum inhibitory concentration (MIC) ranged from 100 to 200 ppm for the essential oils and 25 to 50 mg · mL(-1) for citronellal. The contact assay using the essential oils and citronellal showed growth inhibition of the three fungal species. However, a concentration of 1.47 mg · mL(-1) only reduced the inhibition of Aspergillus growth to 90% at 14 days of exposure. For the fumigant assay, 0.05, 0.11, and 0.23 mg · mL(-1) of essential oils and citronellal drastically affected growth of P. grisea, Aspergillus spp., and C. musae. Harmful effects on the sporulation and germination of the three fungi were seen, and there was complete inhibition at 0.15 mg · mL(-1) with both oils and citronellal. This showed that the crude component of essential oils of C. citriodora and C. nardus markedly suppressed spore production, germination, and growth inhibition of P. grisea, Aspergillus spp., and Colletotrichum musae.

Cloning and expression of Aujeszky's disease virus glycoprotein E (gE) in a baculovirus system

Aujeszky' s disease (AD) is an infectious disease causing important economic losses to the swine industry worldwide. The disease is caused by an alpha-herpesvirus, Aujeszky' s disease virus (ADV), an enveloped virus with a double stranded linear DNA genome. The ADV genome encodes 11 glycoproteins, which are major targets for the immune system of the host in response to the infection. The glycoprotein E (gE) is a non-essential protein and deletion of the gE gene has been used for the production of marker vaccines. Aiming to develop molecular reagents for the production of a gE specific ELISA test, the gE gene was amplified by PCR, cloned and expressed into a baculovirus expression system. The recombinant DNA vector pFastBac-gE-ADV was used for site-specific transposition into the recombinant baculovirus (bacmid). Colonies with recombinant bacmid-pFastBac-gE-ADV were selected by antibiotic and color selection and the presence of the gE gene was confirmed by PCR. The recombinant bacmid-pFastBac-gE-ADV was cotransfected in insect Trichoplusia ni and the presence of the recombinant DNA and gE protein were detected by PCR, SDS-PAGE and Western blotting, respectively.

A doença de Aujeszky (DA) é uma enfermidade infecto-contagiosa que causa grandes perdas econômicas ao produtor e à agroindústria suinícola em todo o mundo. É causada pelo vírus da doença de Aujeszky (VDA), um alfaherpesvírus envelopado com genoma DNA de fita dupla e linear. O genoma do VDA codifica 11 glicoproteínas, as quais são os maiores alvos do sistema imune do hospedeiro em resposta a infecção. A glicoproteína E (gE) é uma proteína não essencial e a deleção do gene da gE é muito utilizada para a produção de vacinas com marcadores. Com o objetivo de desenvolver insumos moleculares para a produção de um teste de ELISA específico para gE do VDA, a seqüência do gene da gE foi amplificada, clonada e expressa no sistema de expressão em baculovírus. O produto da amplificação foi clonado no vetor pGem®-T Easy e subclonado no plasmídeo de expressão pFastBac™1. O DNA recombinante pFastBac-gE-VDA foi usado para a transposição sítio-específica no baculovírus recombinante (bacmid). Após seleção por antibióticos e cor, as colônias com o recombinante bacmid-pFastBac-gE-VDA foram selecionadas e a presença do gene da gE foi confirmada por PCR. O DNA recombinante viral, bacmid-pFastBac-gE-VDA, foi usado para cotransfecção de células de inseto Trichoplusia ni e a presença do recombinante e a proteína gE foi determinada por PCR, por SDS-PAGE e Western blotting, respectivamente.