RESUMO
Enterohemorrhagic E. coli (EHEC) is associated with severe gastrointestinal disease. Upon entering the gastrointestinal tract, EHEC is exposed to a fluctuating environment and a myriad of other bacterial species. To establish an infection, EHEC strains have to modulate their gene expression according to the GI tract environment. In order to explore the interspecies interactions between EHEC and an human intestinal commensal, the global gene expression profile was determined of EHEC O103:H25 (EHEC NIPH-11060424) co-cultured with B. thetaiotaomicron (CCUG 10774) or grown in the presence of spent medium from B. thetaiotaomicron. Microarray analysis revealed that approximately 1% of the EHEC NIPH-11060424 genes were significantly up-regulated both in co-culture (30 genes) and in the presence of spent medium (44 genes), and that the affected genes differed between the two conditions. In co-culture, genes encoding structural components of the type three secretion system were among the most affected genes with an almost 4-fold up-regulation, while the most affected genes in spent medium were involved in chemotaxis and were more than 3-fold up-regulated. The operons for type three secretion system (TTSS) are located on the Locus of enterocyte effacement (LEE) pathogenicity island, and qPCR showed that genes of all five operons (LEE1-LEE5) were up-regulated. Moreover, an increased adherence to HeLa cells was observed in EHEC NIPH-11060424 exposed to B. thetaiotaomicron. Expression of stx2 genes, encoding the main virulence factor of EHEC, was down-regulated in both conditions (co-culture/spent medium). These results show that expression of EHEC genes involved in colonization and virulence is modulated in response to direct interspecies contact between cells, or to diffusible factors released from B. thetaiotaomicron. Such interspecies interactions could allow the pathogen to recognize its predilection site and modulate its behaviour accordingly, thus increasing the efficiency of colonization of the colon mucosa, facilitating its persistence and increasing its virulence potential.
Assuntos
Bacteroides/metabolismo , Escherichia coli Enteropatogênica/patogenicidade , Regulação Bacteriana da Expressão Gênica , Consórcios Microbianos , Escherichia coli Enteropatogênica/genética , Escherichia coli Enteropatogênica/metabolismo , Células HeLa , Humanos , Óperon , Toxina Shiga II/genética , Toxina Shiga II/metabolismo , Transcriptoma , Virulência/genéticaRESUMO
The closely related bacterial species Bacillus cereus and Bacillus weihenstephanensis are adapted to the mesophilic and the psychrotrophic temperature range, respectively. While B. cereus strains are associated with foodborne diseases, B. weihenstephanensis strains are so far not, although similar virulence genes are found in both species. Our investigations show that both species were virulent in the insect model, Galleria mellonella, following infection via oral and haemocoel routes. However, virulence of B. weihenstephanensis was much higher at 15°C than at 37°C. Furthermore, a temperature-dependent difference between the species was seen in a cell culture cytotoxicity assay. In summary, our results demonstrate for the first time virulence of B. weihenstephanensis strains in an in vivo model. In addition, we found that G. mellonella is a useful model for studies of the psychrotolerant species of the B. cereus group, suggesting that insects might be an ecological growth niche for several members of this bacterial group.
Assuntos
Bacillus cereus/patogenicidade , Bacillus/patogenicidade , Insetos/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Chlorocebus aethiops , Temperatura , Células Vero , Virulência/genética , Virulência/fisiologiaRESUMO
Bacillus cereus is widespread in nature and frequently isolated from soil and growing plants, but it is also well adapted for growth in the intestinal tract of insects and mammals. From these habitats it is easily spread to foods, where it may cause an emetic or a diarrhoeal type of food-associated illness that is becoming increasingly important in the industrialized world. The emetic disease is a food intoxication caused by cereulide, a small ring-formed dodecadepsipeptide. Similar to the virulence determinants that distinguish Bacillus thuringiensis and Bacillus anthracis from B. cereus, the genetic determinants of cereulide are plasmid-borne. The diarrhoeal syndrome of B. cereus is an infection caused by vegetative cells, ingested as viable cells or spores, thought to produce protein enterotoxins in the small intestine. Three pore-forming cytotoxins have been associated with diarrhoeal disease: haemolysin BL (Hbl), nonhaemolytic enterotoxin (Nhe) and cytotoxin K. Hbl and Nhe are homologous three-component toxins, which appear to be related to the monooligomeric toxin cytolysin A found in Escherichia coli. This review will focus on the toxins associated with foodborne diseases frequently caused by B. cereus. The disease characteristics are described, and recent findings regarding the associated toxins are discussed, as well as the present knowledge on virulence regulation.
Assuntos
Bacillus cereus/metabolismo , Enterotoxinas/metabolismo , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Trato Gastrointestinal/microbiologia , Microbiologia do Solo , Animais , Bacillus cereus/classificação , Bacillus cereus/isolamento & purificação , Bacillus cereus/patogenicidade , Diarreia/microbiologia , Enterotoxinas/química , Enterotoxinas/genética , Regulação Bacteriana da Expressão Gênica , Humanos , Proteínas Citotóxicas Formadoras de Poros/química , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Vômito/microbiologiaRESUMO
Characteristics concerning diarrhoeal potential were investigated in B. cereus dairy strains. The thirty-nine strains, isolated from whipping cream, were tested for cytotoxicity after culturing at human body temperature as well as 25 degrees C and 32 degrees C. At 37 degrees C, none of the strains were highly cytotoxic. This observation suggests that those strains should be considered to pose a minor risk with regard to diarrhoeal food poisoning. However, some strains were moderately or highly cytotoxic when grown at 25 degrees C and 32 degrees C. While the majority of the strains were able to grow at refrigeration temperatures, only four B. weihenstephanensis strains were identified among them when subjected to discriminative PCR assays and growth temperatures which delimit this species.