Ultrastructural changes in endothelial cells of buffaloes following in-vitro exposure to Pasteurella multocida B:2

Abstract

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 10 7 cfu/ml of whole cell P. multocida B:2, Group 2 with LPS, which was extracted earlier from 10 7 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 postinoculation 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.

Effect of Exposure to Pasteurella multocida B:2 and its Immunogens on Clinical, Hematological and Serum Biochemical Parameters in Neeli Ravi Buffaloes

“…During natural infection, the pathogen enters through the oral or intranasal route. It causes acute infection in the lungs and then crosses blood capillaries through endothelial cells to produce septicemia (Puspitasari et al, 2020). This disease causes very acute respiratory and systemic signs (De Alwis, 1999).…”

“…Another study on oral and subcutaneous inoculation of OMP extract of P. multocida type B: 2 has resulted in significant differences in hematology and biochemistry profiles of experimental buffaloes (Chung et al, 2016). In a recent study, in-vitro exposure of P. multocida type B:2 and its LPS endotoxins has resulted in ultrastructural changes in buffaloes' endothelial cells (Puspitasari et al, 2020).…”

Immune Protective Efficacy of China’s Traditional Inactivated and Attenuated Vaccines against the Prevalent Strains of Pasteurella multocida in Mice

“…In China, as in other parts of the world, P. multocida is a major causative agent of respiratory infection in farm and wild animals [23], causing the different symptoms of pasteurellosis, which range from minor local infection to fatal septicemia. It is a serious and highly contagious disease, affecting domesticated animals, especially buffalos [24], poultry [25], rabbits [26], and pigs. Vaccination is a successful method of controlling P. multocida infection [27], and inactivated vaccines, live attenuated vaccines, recombinant vaccines, subunit vaccines, and DNA vaccines against P. multocida have been extensively studied as preventive strategies [28][29][30].…”

Transboundary Animal Diseases, an Overview of 17 Diseases with Potential for Global Spread and Serious Consequences

“…Mathematical models to evaluate outbreak data and potential intervention strategies have been described [347]. In vitro assays have been performed using macrophages and aortic endothelial cells [348][349][350][351]. Mouse models can play an important role in investigation of pathogenesis and vaccine development [317,[351][352][353][354][355][356][357][358][359][360][361][362][363].…”

“…Mouse models can play an important role in investigation of pathogenesis and vaccine development [317,[351][352][353][354][355][356][357][358][359][360][361][362][363]. Rats with or without immunosuppression have also been used to explore pathogenesis and novel vaccines [349][350][351][364][365][366]. Rabbits are occasionally used for vaccine evaluation and, along with mice, were recently used to evaluate a novel phage lysate marker vaccine along with a DIVA ELISA [358].…”