Pathogenicity of Aspergillus westerdijkiae to females and oothecae of Periplaneta Americana / Patogenicidade de isolado de Aspergillus westerdijkiae a fêmeas e ootecas de Periplaneta americana

ABSTRACT: Cockroach control is performed by the application of chemical insecticides which exert high selective pressure on populations and introduces synthetic substances in the environment, motivating the search for other methods of control such as entomopathogenic fungi. The objectives of this study were to investigate the pathogenicity of the JAB 42 Aspergillus westerdijkiae to females and oothecae of Periplaneta americana and to demonstrate its mechanism of action on oothecae. Suspensions containing 106 to 108 conidia/ml were used to infect females and oothecae. Mortality and other variables such as scanning electron microscopy were used to demonstrate the mechanism of action of the fungus. The isolated JAB 42 A. westerdijkiae is pathogenic to oothecae of P. americana, with low capacity to kill females. Adhesion, germination, penetration and extrusion of the fungus on the cockroach oothecae were observed.

RESUMO: O controle de baratas realizado através da aplicação de inseticidas químicos exerce alta pressão seletiva sobre as populações e introduz substâncias sintéticas no ambiente, motivando a procura por outros métodos de controle, como os fungos entomopatogênicos. Os objetivos deste trabalho foram investigar a patogenicidade do isolado JAB 42 de Aspergillus westerdijkiae a fêmeas e ootecas de Periplaneta americana e demonstrar o mecanismo de ação sobre ootecas. Suspensões contendo 106 a 108 conídios/mL do isolado foram usadas para infectar fêmeas e ootecas. A ação do fungo foi analisada pela mortalidade e outras variáveis, e por microscopia eletrônica de varredura. O isolado JAB 42 de A. westerdijkiae é patogênico a ootecas de P. americana, tendo baixa capacidade de matar fêmeas. Foi observada a adesão, germinação, penetração e extrusão do fungo sobre ootecas da barata.

Composition of Extracellular Polymeric Substances (EPS) produced by Flavobacterium columnare isolated from tropical fish in Brazil

Thirty nine isolates of Flavobacterium columnare from Brazilian fish farms had their carbohydrate composition of EPS evaluated by high efficiency liquid chromatography, using the phenol-sulfuric acid method of EPS. The occurrence of capsules on F. columnare cells was not directly related to biofilm formation, and the predominant monosaccharide is glucose.

Xylella fastidiosa: an in vivo system to study possible survival strategies within citrus xylem vessels based on global gene expression analysis

Xylella fastidiosa inhabits the plant xylem, a nutrient-poor environment, so that mechanisms to sense and respond to adverse environmental conditions are extremely important for bacterial survival in the plant host. Although the complete genome sequences of different Xylella strains have been determined, little is known about stress responses and gene regulation in these organisms. In this work, a DNA microarray was constructed containing 2,600 ORFs identified in the genome sequencing project of Xylella fastidiosa 9a5c strain, and used to check global gene expression differences in the bacteria when it is infecting a symptomatic and a tolerant citrus tree. Different patterns of expression were found in each variety, suggesting that bacteria are responding differentially according to each plant xylem environment. The global gene expression profile was determined and several genes related to bacterial survival in stressed conditions were found to be differentially expressed between varieties, suggesting the involvement of different strategies for adaptation to the environment. The expression pattern of some genes related to the heat shock response, toxin and detoxification processes, adaptation to atypical conditions, repair systems as well as some regulatory genes are discussed in this paper. DNA microarray proved to be a powerful technique for global transcriptome analyses. This is one of the first studies of Xylella fastidiosa gene expression in vivo which helped to increase insight into stress responses and possible bacterial survival mechanisms in the nutrient-poor environment of xylem vessels.

Requirement for cobalamin by Salmonella enterica serovars Typhimurium, Pullorum, Gallinarum and Enteritidis during infection in chickens

Salmonella enterica serovar Typhimurium synthesizes cobalamin (vitamin B12) only during anaerobiosis. Two percent of the S. Typhimurium genome is devoted to the synthesis and uptake of vitamin B12 and to B12-dependent reactions. To understand the requirement for cobalamin synthesis better, we constructed mutants of Salmonella serovars Enteritidis and Pullorum that are double-defective in cobalamin biosynthesis (ÃcobSÃcbiA). We compared the virulence of these mutants to that of their respective wild type strains and found no impairment in their ability to cause disease in chickens. We then assessed B12 production in these mutants and their respective wild type strains, as well as in S. Typhimurium ÃcobSÃcbiA, Salmonella Gallinarum ÃcobSÃcbiA, and their respective wild type strains. None of the mutants was able to produce detectable B12. B12 was detectable in S. Enteritidis, S. Pullorum and S. Typhimurium wild type strains but not in S. Gallinarum. In conclusion, the production of vitamin B12 in vitro differed across the tested Salmonella serotypes and the deletion of the cbiA and cobS genes resulted in different levels of alteration in the host parasite interaction according to Salmonella serotype tested.

Molecular differentiation between Salmonella enterica subsp enterica serovar Pullorum and Salmonella enterica subsp enterica serovar Gallinarum / Diferenciação molecular entre Salmonella enterica subsp enterica serovar Pullorum e Salmonella enterica subsp enterica serovar Gallinarum

S. Pullorum (SP) and S. Gallinarum (SG) are very similar. They are the agents of pullorum disease and fowl typhoid, respectively, and the two diseases are responsible for economic losses in poultry production. Although SP and SG are difficult to be differentiated in routine laboratory procedures, the ability to metabolize ornithine is a biochemical test that may be used to achieve this aim. While SP is able to decarboxylate this amino acid, SG is not. However, the isolation of strains showing atypical biochemical behavior has made this differentiation difficult. One of the genes associated with the metabolization of the amino acid ornithine is called speC, and is found in both serovars. The analysis of 21 SP and 15 SG strains by means of PCR did not enable the differentiation of the two serovars, because fragments produced were identical. However, after enzymatic treatment with restriction enzyme Eco RI, the band pattern of each serovar showed to be different, even in samples of atypical biochemical behavior. This fact enabled the standardization of the technique for a quick and safe differentiation of serovars Pullorum and Gallinarum.

A S. Pullorum (SP) é muito semelhante à S. Gallinarum (SG), agentes da Pulorose e Tifo aviário, respectivamente, sendo que as duas enfermidades são responsáveis por perdas econômicas no setor avícola. SP e SG são de difícil diferenciação em procedimento laboratorial rotineiro, mas uma prova bioquímica muito utilizada na distinção das duas refere-se à capacidade de assimilar o aminoácido ornitina: SP descarboxila este aminoácido enquanto SG não. No entanto, o isolamento de cepas com comportamento bioquímico atípico, tem dificultado tal diferenciação. Um dos genes relacionados à assimilação do aminoácido ornitina, denomina-se gene speC, o qual está presente nos dois sorovares. Analisando 21 amostras de SP e 15 de SG com a utilização da PCR não foi possível realizar a diferenciação dos dois sorovares pois os fragmentos gerados eram idênticos. Posteriormente, com o uso da técnica de tratamento enzimático com a enzima de restrição Eco RI, foi possível observar que o padrão de bandas gerado em cada sorovar era diferente, mesmo quando amostras que apresentavam comportamento bioquímico atípico eram analisadas. Tal fato permitiu a padronização da técnica para ser utilizada na diferenciação entre os sorovares Pullorum e Gallinarum de maneira rápida e segura.

Nitrogen assimilation in Citrus based on CitEST data mining

Assimilation of nitrate and ammonium are vital procedures for plant development and growth. From these primary paths of inorganic nitrogen assimilation, this metabolism integrates diverse paths for biosynthesis of macromolecules, such as amino acids and nucleotides, and the central intermediate metabolism, like carbon metabolism and photorespiration. This paper reports research performed in the CitEST (Citrus Expressed Sequence Tag) database for the main genes involved in nitrogen metabolism and those previously described in other organisms. The results show that a complete cluster of genes involved in the assimilation of nitrogen and the metabolisms of glutamine, glutamate, aspartate and asparagine can be found in the CitEST data. The main enzymes found were nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthetase (GOGAT), glutamate dehydrogenase (GDH), aspartate aminotransferase (AspAT) and asparagine synthetase (AS). The different enzymes involved in this metabolism have been shown to be highly conserved among the Citrus and Poncirus species. This work serves as a guide for future functional analysis of these enzymes in citrus.

Fluorescent amplified fragment length polymorphism (fAFLP) analyses and genetic diversity in Litopenaeus vannamei (Penaeidae)

The Pacific white shrimp, Litopenaeus vannamei (Penaeidae), represents about 95 percent of all Brazilian shrimp production. The Brazilian L. vannamei foundation broodstock was made up of specimens collected from different American Pacific sites, but little information was collected on the genetic structure of the broodstock. We used the fluorescence amplified fragment length polymorphism (fAFLP) method to study the genetic diversity of L. vannamei broodstock lines 03CMF1 and 03CBF1 originally produced by breeder-shrimps imported mainly from Panama and Ecuador, although wild individuals from other localities may also have been used in producing these two lines. Our results showed a total of 93 polymorphic bands ranging from 50 to 500 bp, the mean Nei's genetic diversity calculated for the total sample was 13.4 percent and identity and genetic distance analyses indicated high genetic homogeneity within and between both the broodstock lineages studied which suggests that they had similar genetic structure. These results may represent an important tool for the appropriate management of L. vannamei broodstocks.

Elimination of a Staphylococcus aureus plasmid by 8-methoxypsoralen plus near UV light

Em uma linhagem de Staphylococcus aureus portadora de um plasmídio para resistência à tetraciclina, mostrou-se que a 8-metoxipsoraleina associada à luz UV longa (-365 nm) pode eliminar plasmídio