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Articles by Simon Menanteau-Ledouble in JoVE
JoVE Immunology and Infection
Использование флуоресцентных Иммуно-химия для обнаружения Edwardsiella ictaluri В канале сома ( I. punctatus) Образцов
Department of Basic Sciences, Mississippi State University
Здесь мы опишем процедуру, позволяющую маркировки
Other articles by Simon Menanteau-Ledouble on PubMed
Development of Bioluminescent Edwardsiella Ictaluri for Noninvasive Disease Monitoring
Edwardsiella ictaluri is a facultative intracellular bacterium that causes enteric septicemia of catfish (ESC). In this study, we aimed to develop bioluminescent E. ictaluri that can be monitored by noninvasive bioluminescence imaging (BLI). To accomplish this, the luxCDABE operon of Photorhabdus luminescens was cloned downstream of the lacZ promoter in the broad host range plasmid pBBR1MCS4. Edwardsiella ictaluri strain 93-146 transformed with the new plasmid, pAKlux1, was highly bioluminescent. pAKlux1 was stably maintained in E. ictaluri without any apparent effect on growth or native plasmid stability. To assess the usefulness of the bioluminescent strain in disease studies, catfish were infected with 93-146 pAKlux1 by intraperitoneal injection and by bath immersion, and in vivo bacterial dissemination was observed using BLI. This study demonstrated that bioluminescent E. ictaluri can be used for real-time monitoring of ESC in live fish, which should enable observation of pathogen attachment sites and tissue predilections.
Importance of Skin Abrasion As a Primary Site of Adhesion for Edwardsiella Ictaluri and Impact on Invasion and Systematic Infection in Channel Catfish Ictalurus Punctatus
The route of entry of Edwardsiella ictaluri into its catfish host has been a subject of investigation since the pathogen was first discovered. There is evidence to support entry through the intestinal tract, the nares, and the gills. Here, we evaluated the role of skin abrasion through a series of experimental challenges using bioluminescent E. ictaluri carrying the plasmid pAKLux1. Our results show that E. ictaluri is able to colonize abrasion sites on catfish skin and that catfish with abrasions developed systematic infection faster. We also found that abrasions are associated with significantly increased mortalities following experimental immersion exposure. Finally, a protocol was developed during this study that allowed for immunohistochemical examination of the tissue layers underneath the abrasion sites, confirming the presence of E. ictaluri in subdermal tissues from abrasion sites. This study constitutes the first report on the role of channel catfish skin as a portal of entry for E. ictaluri and further illustrates how versatile this pathogen can be in its mechanisms of entry.
