ABSTRACT
Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.
Subject(s)
Humans , Cell Movement/physiology , rho GTP-Binding Proteins/physiology , Virulence Factors/genetics , Epithelial Cells/microbiology , Enteropathogenic Escherichia coli/pathogenicity , Type III Secretion Systems/physiology , Blotting, Western , Apoptosis , Virulence Factors/physiology , Real-Time Polymerase Chain Reaction , Flow CytometryABSTRACT
This study reports a co-infection of Escherichia coli and Salmonella in a free-living ruddy ground dove (Columbina talpacoti) received at the Laboratory of Ornithological Studies of the State University of Ceará, Brazil. The bird presented diarrhea, leg paralysis and anorexia, and died shortly after. Necropsy was then performed and samples from lung, kidney, liver and intestine were collected for microbiological and histopathological analyses. Escherichia coli was isolated from cloacal swab, lung and kidney samples. Salmonella ser. Saintpaul was identified in liver and spleen samples. Escherichia coli isolates were tested for the presence of eight diagnostic genes for diarrheagenic pathotypes (STEC, ETEC, EPEC, EIEC, EAEC) with conventional polymerase chain reaction (PCR). EAEC was detected in the lung and kidney, and STEC in the intestine. In conclusion, Columbina talpacoti is susceptible to enteroaggregative Escherichia coli and Salmonella ser. Saintpaul infection, which may have public health implications.(AU)
Este estudo relata um caso de coinfecção por Escherichia coli e Salmonella ser. Saintpaul em uma rolinha-roxa (Columbina talpacoti) recebida pelo Laboratório de Estudos Ornitológicos da Universidade Estadual do Ceará, Brasil. A ave apresentava diarreia, paralisia nas pernas e anorexia, indo a óbito rapidamente. A necropsia foi realizada e amostras de pulmão, rim, fígado e intestino foram coletados para isolamento microbiológico e análise histopatológica. Escherichia coli foi identificada em amostras de suabe cloacal, pulmão e rim. Salmonella ser. Saintpaul foi identificada no fígado e baço. Isolados de E. coli foram testados para a presença de oito genes de diagnóstico para patotipos diarreiogênicos (STEC, ETEC, EPEC, EIEC, EAEC) através de reação em cadeia de polimerase (PCR) convencional. EAEC foi detectada no pulmão e rim, e STEC foi identificada no intestino. Em conclusão, Columbina talpacoti é suscetível a infecção por Escherichia coli enteroagregativa e Salmonella ser. Saintpaul, o que pode implicar em risco para a saúde pública.(AU)
Subject(s)
Animals , Coinfection/veterinary , Columbidae , Drug Resistance, Bacterial , Escherichia coli Infections/veterinary , Salmonella Infections, Animal/diagnosisABSTRACT
Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.