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Abstract
BackgroundThe pathogenesis of Haemophilus parasuis depends on the bacterium’s ability to interact with endothelial cells and invade adjacent tissues. In this study, we investigated the abilities of eight H. parasuis reference strains belonging to serovars 1, 2, 4, 5, 7, 9, 10 and 13 to adhere to and invade porcine aortic endothelial cells (AOC-45 cell line).ResultsThe strains belonging to serovars 1, 2 and 5 were able to attach at high rates between 60 and 240 min of incubation, and serovars 4, 7 and 13 had moderate attachment rates; however, the strains belonging to serovars 9 and 10 had low adherence at all time points. Strong adherence was observed by scanning electron microscopy for the strains of serovars 5 and 4, which had high and moderate numbers, respectively, of H. parasuis cells attached to AOC-45 cells after 240 min of incubation. The highest invasiveness was reached at 180 min by the serovar 4 strain, followed by the serovar 5 strain at 240 min. The invasion results differed substantially depending on the strain.ConclusionThe reference strains of H. parasuis serovars 1, 2, 4 and 5 exhibited high adhesion and invasion levels to AOC-45 porcine aorta endothelial cells, and these findings could aid to better explain the pathogenesis of the disease caused by these serovars.
Highlights
The pathogenesis of Haemophilus parasuis depends on the bacterium’s ability to interact with endothelial cells and invade adjacent tissues
To better understand the pathogenesis of Glässer’s disease, this study investigated the adherence to and invasion of Aorta endothelial cells (AOC)-45 porcine aortic endothelial cells by H. parasuis strains belonging to serovars 1, 2, 4 5, 7, 9, 10 and 13, which are considered to have different levels of virulence [2]
After the growth curves had been obtained, the infection dose was established in the following manner: a mix of colonies from a fresh H. parasuis culture harvested from a chocolate plate were used to inoculate PPLO broth supplemented as described above, and this culture allowed to grow to an OD600 of 0.7
Summary
The pathogenesis of Haemophilus parasuis depends on the bacterium’s ability to interact with endothelial cells and invade adjacent tissues. We investigated the abilities of eight H. parasuis reference strains belonging to serovars 1, 2, 4, 5, 7, 9, 10 and 13 to adhere to and invade porcine aortic endothelial cells (AOC-45 cell line). H. parasuis usually colonizes the upper respiratory tract of pigs, and it can be detected in the tonsillar area and at other respiratory sites, such as the tracheal mucosa [7]. At these sites, virulent strains are able to breach the mucosal barrier and pass into the bloodstream, leading to the severe systemic disease described above [8]. Cytolethal distending toxin, outer membrane protein 2 and heptose I and II residues in the inner core oligosaccharide have been implicated in the adherence of this bacterium to porcine alveolar macrophages, epithelial kidney cells and umbilicus vein endothelial cells [13,14,15,16]
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