Efficacy of bacteriophage Lysed Pasteurella marker vaccine in laboratory animal models with a novel DIVA for haemorrhagic septicaemia

Abstract

ObjectiveThe present study aimed at evaluating the efficacy of an improved phage lysate marker vaccine for haemorrhagic septicaemia in mice and rabbit model and development of a DIVA ELISA based on iron restricted outer membrane protein (IROMP).MethodThe experimental vaccine was prepared by lysing P. multocida B:2 grown under iron restricted conditions with a Pasteurella bacteriophage and addition of an alum adjuvant to enhance the immunogenicity. The vaccine was administered in mice and rabbits divided into two group each. Phage lysate vaccine (PL-VacI) was administered to group I mice and rabbits whereas group II mice and rabbits received alum precipitated HS vaccine (HS-VacII). Antibody titres were monitored 0, 30, 60, 90, 210 and 240 dpv. An IROMP (130 kDa) based indirect ELISA was also developed to differentiate between infected and vaccinated animals. The Pasteurella phage isolated in present study was sequenced at Georgia Genomic Facilty, Georgia.ResultThe sequence of PMP-GAD-IND (Pasteurella bacteriophage) was deposited in GenBank under no KY203335. The group I mice and rabbits vaccinated with Phage lysate vaccine (PL-VacI) group revealed significantly higher antibody titres than group II mice and rabbits receiving alum-precipitated bacterin (HS-VacII) by MAT, IHA and ELISA (P < 0.05 and P < 0.001). The peak log 10 values (3.46) in case of group I mice by ELISA were attained at 90DPI whereas in group II mice the peak values at 90DPI were 2.82. Mean log10 titres by ELISA in group I and II rabbits were 2.43 and 2.35 respectively at 30DPI whereas at 120DPI the titres were 3.29 and 2.75, respectively. The DIVA ELISA detected presence of a novel 137 kDa IROMP/siderophore antibody in sera of group I mice and rabbits (PL-VacI) absent in sera of mice and rabbits given HS-VacII.ConclusionThe bacteriophage based marker vaccine (PL-VacI) had a more effective and longer immune response against HS in mice and rabbit in comparison to the widely used alum precipitated HS vaccine (HS-VacII). Moreover, the development of a recombinant IROMP based indirect ELISA could serve as an excellent tool to differentiate between infected and vaccinated cattle and buffaloes for effective control of HS.

Isolation and genome analysis of a lyticPasteurella multocidaBacteriophage PMP-GAD-IND

“…27 × 24 nm and tail c . 134·5 nm (Qureshi and Saxena ,b). Similar morphologies of bacteriophages of P. multocida have been reported (Ackermann and Karaivanov ).…”

“…Bioinformatics has made it easier to predict the most immune‐dominant epitopes, B cell and T cell interactions, even to design peptide vaccines based on a neutralizing epitopes with great precision. The purpose of this study was to screen high potential immunogenic epitopes for T‐cells that might have played a role in improved immunity with our bacteriophage based phage lysate vaccine (Qureshi and Saxena ,b). We focussed on potential MHC class I and MHC class II epitopes.…”

“…Haemorrhagic Septicaemia (HS) is an acute, fatal disease of bovines caused by Pasteurella multocida caused by capsular serogroup B:2 in Asia and E:2 in central Africa (Qureshi and Saxena ,b). HS is one of the most fatal of the five important epizootic diseases affecting buffaloes and cattle in India with estimated annual economic losses due to P. multocida infection alone to the tune of Rs.…”

“…We report here the isolation and genomic characterization of a new broad acting lytic Pasteurella phage PMP‐GAD‐IND isolated from the sewage of GADVASU, dairy farm, India. PMP‐GAD‐IND was used for the development of a phage lysate marker vaccine against HS in bovines (Qureshi and Saxena ,b).…”

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

“…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]. Rats with or without immunosuppression have also been used to explore pathogenesis and novel vaccines [349][350][351][364][365][366].…”