A simple methodology to collect culturable bacteria from feces of Anopheles darlingi (Diptera: Culicidae).

A simple methodology based in a modified mosquito cage with a Petri dish containing culture medium was successfully used as an alternative method to the traditional digestive tract dissection protocol to collect bacteria from the feces of the mosquito Anopheles darlingi.

Adherence and virulence genes of Escherichia coli from children diarrhoea in the Brazilian Amazon.

The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli . Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli . Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene . EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg , aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea ( P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.

Adherence and virulence genes of Escherichia coli from children diarrhoea in the Brazilian Amazon

The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli. Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli. Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene. EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg, aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea (P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.

The Hem and Has haem uptake systems in Serratia marcescens.

Serratia marcescens, like several other Gram-negative bacteria, possesses two functional haem uptake systems. The first, referred to as the Hem system, can transport haem present at a concentration equal to or above 10(-6) M. It requires an active outer-membrane receptor which uses proton-motive force energy transmitted by the inner-membrane TonB protein. The other system, Has, takes up haem at lower concentrations and utilizes a small secreted haem-binding protein (haemophore) and its cognate TonB-dependent outer-membrane receptor HasR. Various combinations of mutations were used to examine haem uptake activity by the two systems in S. marcescens. The Hem uptake system enables S. marcescens to take up haem at a concentration of 10(-6) M in the presence of various levels of iron depletion. The Has system, which enables such uptake even in the presence of lower haem concentrations, requires higher iron depletion conditions for function. Has haem uptake requires the presence of HasB, a TonB paralogue encoded by the has operon. These two systems enable S. marcescens to take up haem under various conditions from different sources, reflecting its capacity to confront conditions encountered in natural biotopes.

HasB, the Serratia marcescens TonB paralog, is specific to HasR.

Serratia marcescens possesses two functional TonB paralogs, TonB(Sm) and HasB, for energizing TonB-dependent transport receptors (TBDT). Previous work had shown that HasB is specific to heme uptake in the natural host and in Escherichia coli expressing the S. marcescens TBDT receptor HasR, whereas the S. marcescens TonB and E. coli TonB proteins function equally well with various TBDT receptors for heme and siderophores. This has raised the question of the target of this specificity. HasB could be specific either to heme TBDT receptors or only to HasR. To resolve this question, we have cloned in E. coli another S. marcescens heme receptor, HemR, and we show here that this receptor is TonB dependent and does not work with HasB. This demonstrates that HasB is not dedicated to heme TBDT receptors but rather forms a specific pair with HasR. This is the first reported case of a specific TonB protein working with only one TBDT receptor in one given species. We discuss the occurrence, possible molecular mechanisms, and selective advantages of such dedicated TonB paralogs.