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Ultrastructural changes in endothelial cells of buffaloes following in-vitro exposure to Pasteurella multocida B:2
Yulianna Puspitasari1,2, Annas Salleh3, Mohd Zamri-Saad1
1Research Centre for Ruminant Diseases, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400, Serdang, Malaysia.
Background:
Pasteurella 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.
Results:
The 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.
Conclusions:
This 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.
Insights
Pasteurella multocida B:2 infection causes severe damage to buffalo aortic endothelial cells (BAEC). Both whole bacteria and their lipopolysaccharide (LPS) induced significant cellular injury, with whole bacteria being more potent.
Area of Science:
- Veterinary Pathology
- Microbiology
- Cellular Biology
Background:
- Pasteurella multocida B:2 is a pathogen causing hemorrhagic septicaemia in livestock, with buffaloes exhibiting higher susceptibility.
- Limited research exists on the response of buffalo aortic endothelial cells (BAEC) to P. multocida B:2 and its lipopolysaccharide (LPS).
Purpose of the Study:
- To investigate the ultrastructural changes in buffalo aortic endothelial cells (BAEC) upon exposure to whole-cell P. multocida B:2 and its LPS.
- To compare the pathogenicity of whole-cell P. multocida B:2 versus its LPS in causing endothelial cell damage.
Main Methods:
- Cultured BAEC were exposed to live P. multocida B:2 (Group 1) or extracted LPS (Group 2), with a control group receiving sterile medium (Group 3).
- Transmission electron microscopy was used to assess ultrastructural changes at various time points (0-48 hours post-inoculation).
Main Results:
- Both P. multocida B:2 and LPS induced moderate to severe endothelial lysis and acute cellular injury in BAEC.
- Cellular damage severity increased with incubation time, with significant differences observed after 18 hours.
- Whole-cell P. multocida B:2 exposure resulted in more severe cellular damage compared to LPS alone.
Conclusions:
- Both whole-cell P. multocida B:2 and its LPS cause significant ultrastructural damage to buffalo aortic endothelial cells.
- Whole-cell P. multocida B:2 demonstrates greater potency in inducing endothelial cell injury than LPS alone.
