The compound paraquat dichloride hydrate significantly affects the in vitro growth rate of a Chromobacterium violaceum wild type strain / O composto hidrato-dicloreto de paraquat afeta a taxa de crescimento in vitro de uma cepa do tipo selvagem de Chromobaterium violaceum

Introduction: the toxicity of pesticides on bacterial cell growth is still limited. Objectives: The current study aimed to assess the in vitro growth rate of the C. violaceum wild type strain ATCC12472 exposed to the herbicide paraquat dichloride hydrate at different incubation times and final concentrations. Methodology: bacterial inocula were incubated in a nutrient broth medium containing the compound paraquat at final concentrations 100 and 1.000 µg mL-1 under aeration conditions. Spectrophotometric readings at different incubation times were carried out to estimate the in vitro bacterial growth rate. Moreover, the number of viable bacteria cells in the samples was also estimated in the presence of the paraquat at two concentrations tested based on colony-forming units grown on the nutrient broth agar. Results: significant decreases in the C. violaceum growth rate were detected, after one hour of paraquat exposure at a final concentration of 1,000 µg mL-1 (p<0.05) compared to all treatments tested. After two hours of paraquat exposure, significant decreases were progressively found at all final concentrations of 100 (p<0.01) and of 1,000 µg mL-1 (p<0.001). These data were also corroborated by counting the total number of colony-forming units at final concentrations tested. Conclusion: the findings described in current study suggest that the compound paraquat dichloride hydrate exerts significant effects on the in vitro growth rate of a C. violaceum wild type strain.

Introdução: a toxicidade de pesticidas sobre o crescimento de células bacterianas ainda é limitada. Objetivos: o presente estudo objetivou avaliar a taxa de crescimento in vitro de uma cepa selvagem de C. vilaceum ATCC12472 exposta ao herbicida hidratodicloreto de paraquat em diferentes tempos de incubação e concentrações finais. Metodologia: inóculos bacterianos foram incubados em um caldo nutritivo contendo o composto paraquat nas concentrações 100 e 1.000 µg mL-1 sob condições de aeração. Leituras espectrofotométricas em diferentes tempos de incubação foram realizadas para estimar a taxa de crescimento in vitro bacteriano. Além disso, o número de células bacterianas viáveis nas amostras foi também estimado na presença do paraquat nas duas concentrações testadas, baseado no número de unidades formadoras de colônias crescidas em meio nutritivo ágar. Resultados: diminuições significativas na taxa de crescimento da C. violaceum foram detectadas, após uma hora de exposição ao paraquat na concentração final de 1.000 µg mL-1 (p<0,05) em comparação aos demais tratamentos testados. Com duas horas de exposição ao paraquat, diminuições significativas foram progressivamente encontradas em todas as concentrações finais de 100 (p<0,01) e de 1.000 µg mL-1 (p<0,001). Tais dados foram corroborados pela contagem do número total de unidades formadoras de colônias nas concentrações analisadas. Conclusão: as descobertas descritas aqui sugerem que o composto hidrato-dicloreto de paraquat exerce efeitos significativos sobre a taxa de crescimento in vitro de uma cepa selvagem da bactéria C. violaceum

Temporal expression pattern of the insulin-like growth factor II and fibroblast growth factor transcripts in avian embryogenesis

In this study, the abundance of IGF-II and bFGF transcripts was estimated in the chicken embryos using the competitive RT-PCR analysis. Significant enhancements in the abundance of IGF-II mRNA were observed at stages HH1 and 5, and a new accumulation in these levels was observed at stage HH18 in comparison to the basal levels. The abundance of bFGF mRNA increased significantly at stages HH18 and 20, followed by an upregulation in the expression of these transcripts at stage HH26. These findings provided important information about the temporal expression pattern of IGF-II and bFGF transcripts in the whole chicken embryos during in ovo development.

Fatores de crescimento coordenam múltiplas vias de sinalização durante o desenvolvimento embrionário. Neste estudo, a abundância de mRNA dos genes IGF-II e bFGF foi estimada em embriões de galinha por análises de RT-PCR competitiva. Aumentos na abundância de mRNA de IGF-II foram observados nos estádios HH1, 5. Os níveis de mRNA de bFGF exibiram aumentos a partir dos estádios HH18 e 20, seguido por uma acentuada redução a níveis basais no estádio HH24 e por um segundo pico na expressão destes transcritos no estádio HH26. Tais descobertas proporcionam importantes informações sobre o padrão de expressão destes fatores de crescimento durante a embriogênese de aves

Regulation of gene expression in Mycoplasmas: contribution from Mycoplasma hyopneumoniae and Mycoplasma synoviae genome sequences

This report describes the transcription apparatus of Mycoplasma hyopneumoniae (strains J and 7448) and Mycoplasma synoviae, using a comparative genomics approach to summarize the main features related to transcription and control of gene expression in mycoplasmas. Most of the transcription-related genes present in the three strains are well conserved among mycoplasmas. Some unique aspects of transcription in mycoplasmas and the scarcity of regulatory proteins in mycoplasma genomes are discussed.

Internalin-expressing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells.

The use of the food-grade bacterium Lactococcus lactis as antigen delivery vehicle at the mucosal level is an attractive vaccination strategy intensively explored during the last decade. In this study, we developed L. lactis strains which could be used as a DNA delivery vector to combine both advantages of mucosal delivery and of DNA vaccination. To render lactococci capable of invading epithelial cells, the Listeria monocytogenes inlA gene was cloned and expressed in L. lactis under transcriptional control of the native promoter. Western blot and immunofluorescence assays revealed that recombinant lactococci efficiently displayed the cell wall anchored form of InlA. We demonstrated that this expression promotes internalization of L. lactis inlA+ into the human epithelial cell line Caco-2. Gentamicin assay showed that invasiveness of L. lactis in these cells is approximately 100-fold higher for L. lactis inlA+ than for wild type (wt) L. lactis strains. Moreover, we showed that L. lactis inlA+ is able to enter intestinal cells in vivo, after oral inoculation of guinea pigs. After internalization, L. lactis inlA+ was able to deliver a functional eukaryotic gfp gene into epithelial Caco-2 cells; GFP was detected in 1% of internalized cells. The L. lactis inlA+ strain will be a useful bacterial vector for the development of new live oral DNA vaccines. It also constitutes an interesting new model to study the role of internalin in bacterial localization in the animal host.

Hepatic mRNA expression and plasma levels of insulin-like growth factor-I (IGF-I) in broiler chickens selected for different growth rates

The hepatic expression and plasma concentrations of IGF-I were investigated in three broiler chicken strains selected for different growth rates (HP-Hubbard-Pettersen, a fast growing strain; NN-Naked-neck, a strain with an intermediate growth rate and a heterozygous genotype, and C-Caipira, a slow growing crossbred strain). The chickens were studied at 1, 21 and 42 days of age and had free access to food throughout the study. Hepatic IGF-I mRNA expression was assessed by dot blot analysis using a randomly labeled chicken IGF-I cDNA as the probe and plasma IGF-I concentrations were assayed by radioimmunoassay. The hepatic levels of IGF-I mRNA increased from 1 to 21 days of age in all strains, with NN chickens showing a higher (p < 0.05) IGF-I expression than the other strains. Plasma IGF-I concentrations increased (p < 0.05) with broiler chicken age, but there were no significant differences among the strains. These results indicate that despite differences in the growth rates among the strains, the changes in the expression of IGF-I mRNA in liver and in the plasma levels of IGF-I were independent of broiler chicken strain, but varied with chicken age.