Abstract
BackgroundPasteurella multocida B:2 causes haemorrhagic septicaemia in cattle and buffaloes. However, buffaloes are found to be more susceptible to the infection than cattle. Upon infection, the pathogen rapidly spread from the respiratory tract to the blood circulation within 16-72 h, causing septicaemia. So far, limited study has been conducted to evaluate the response of endothelial cells of buffalo towards P. multocida B:2 and its lipopolysaccharide (LPS). This study aimed to evaluate the ultrastructural changes in the aortic endothelium of buffaloes (BAEC) following exposure to P. multocida B:2 and its endotoxin. The endothelial cells were harvested from the aorta of healthy buffaloes and were prepared as monolayer cell cultures. The cultures were divided into 3 groups before Group 1 was inoculated with 107 cfu/ml of whole cell P. multocida B:2, Group 2 with LPS, which was extracted earlier from 107 cfu/ml of P. multocida B:2 and Group 3 with sterile cell culture medium. The cells were harvested at 0, 6, 12, 18, 24, 36, and 48 h post-inoculation for assessment of cellular changes using transmission electron microscopy.ResultsThe BAEC of Groups 1 and 2 demonstrated moderate to severe endothelial lysis, suggestive of acute cellular injury. In general, severity of the ultrastructural changes increased with the time of incubation but no significant difference (p > 0.05) in the severity of the cellular changes between Groups 1 and 2 was observed in the first 18 h. The severity of lesions became significant (p < 0.05) thereafter. Both treated Groups 1 and 2 showed significantly (p < 0.05) more severe cellular changes compared to the control Group 3 from 6 h post-inoculation. The severity reached peak at the end of the study period with score 3 for Group 1 and score 2.8 for Group 2.ConclusionsThis study revealed that both whole cells P. multocida B:2 and LPS endotoxin showed similar moderate to severe cellular damage, but whole-cell P. multocida B:2 appeared to be more potent in causing much severe damage than LPS alone.
Highlights
Pasteurella multocida B:2 causes haemorrhagic septicaemia in cattle and buffaloes
The present study describes and evaluate for the first time the severity of ultrastructural lesions shown by the buffalo aortic endothelial cells (BAEC) following exposure to P. multocida B:2 and its LPS endotoxin to better understand the pathogenesis of haemorrhagic septicaemia (HS)
The early lysis of BAEC observed in this study following exposure to P. multocida B:2 and its endotoxin provide an elaboration of the rapid development of circulatory failure and death of buffalo with HS
Summary
Pasteurella multocida B:2 causes haemorrhagic septicaemia in cattle and buffaloes. buffaloes are found to be more susceptible to the infection than cattle. This study aimed to evaluate the ultrastructural changes in the aortic endothelium of buffaloes (BAEC) following exposure to P. multocida B:2 and its endotoxin. There is no study, on the response by endothelial cells of buffaloes to in-vitro infection with live P. multocida and the endotoxin LPS. Since buffalo is more susceptible to HS, a study using buffalo would be beneficial in evaluating the ultrastructural changes, especially pertaining to the severity of endothelial cell damage. Different strains of P. multocida isolated from different hosts with different diseases are known to have different degree of invasiveness [10] Considering this paucity of information and high susceptibility of buffaloes to P. multocida B:2, this experiment was conducted to evaluate the in-vitro damages on the endothelial cells of buffalo following exposure to whole cell and endotoxin of P. multocida B:2
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