In the present study, polymerase chain reaction (PCR) using random primer (E-20) was used to characterize and identify strains included in this study. Strains included 4 vaccinal reference strains of Pasteurella multocida, CU strain and 4 field isolates of Pasteurella multocida isolated from diseased turkeys which were identified biochemically and serologically as A:1, A:3, A3x4 and D:11. The obtained results revealed that all strains were reacted positively and in different manner with the E20 primer except the 2 field isolates. The results of these reactions demonstrated in terms of bands of different molecular weight specific to each strain. This can be used as a base for characterization and differentiation of strains involved in the present study as the 2 field strains A:1 and A:3 react with primer. Mouse protection test was performed by vaccination of mice with local fowl cholera oil adjuvant vaccine then challenge with virulent field strains A:1, A:3, D:12 and untypable isolates. Results revealed that the local fowl cholera adjuvant vaccine could protect mice against virulent challenge with A:1, A:3 and D:12 field strains but it could not be protect mice against untypable isolates

Objective: The objective of this study was to evaluate some hematological parameters in commercial layers inoculated with two virulent Pasteurella multocida serotypes.Materials and Methods: A total of 84 twenty-week-old black Harco layers were randomly assigned to seven groups (A, B, C, D, E, F and G) with 12 birds per group. 1mLof live attenuated fowl cholera (FC) vaccine was administered subcutaneously at 24 weeks of age to groups A and B, emulsified inactivated (killed) FC vaccine was administered dosed at 0.5 mL per bird subcutaneously at 24 weeks of age to groups C and D, groups E and F were not vaccinated, while group G served as control. Groups A, C and E were inoculated with P. multocida serotype A:1 and groups B, D and F were inoculated with P. multocida serotype A:3. Using McFarland Standard, each bird received a dose of 0.5 mL (0.1 mL intranasally and 0.4 mL intramuscularly) containing 4.5 x 108 cfu/bird. Results: For PCV (P≤0.2692 and P≤0.7643) and HB (P≤0.2806 and P≤0.7266) on day 2 and 10 post inoculation, there was no significant difference between the vaccinated, non-vaccinated groups and control group G. However, there was a highly significant difference P≤0.05 in the mean concentrations of ALP between the control group G (67.67±1.453 u/l) vaccinated groups A (80.33±4.98 u/l), B (81.33±2.60 u/l), C (75±6.35 u/l), and D (84±5.132 u/l) and unvaccinated groups E (104±1.528 u/l ), and F (78 ±3.512 u/l) post inoculation.Conclusion The PCV significantly decrease P≤0.05 in layers vaccinated and inoculated with P. multocida but increase in unvaccinated layers inoculated P. multocida. The mean serum ALP concentration significantly increase P≤0.05 in unvaccinated layers inoculated with P. multocida when compared to layers vaccinated and inoculated with P. multocida. [J Adv Vet Anim Res 2017; 4(3.000): 234-240]

Pasteurella multocida type A is the etiologic agent of fowl cholera, a highly contagious and fatal disease of chickens. The present research work was performed for the isolation, identification and molecular detection of P. multocida Type A from chickens. Liver, heart and spleen of suspected dead chicken (n=35) were collected from Gazipur and Pabna districts in Bangladesh. The targeted bacteria from the samples were isolated, identified and characterized based on their morphology, staining, cultural, biochemical characters, pathogenicity test, histopathological study and Polymerase Chain Reaction (PCR). The P. multocida organism was isolated from 11.42% (n=4/35) samples. The organisms were gram negative, non-spore forming rod, non-motile, occurring singly or pairs in Gram staining, whereas in Leishman's stain, bipolar shaped organisms were observed. All the isolates were found positive for oxidase and catalase tests, produced indole, and fermented glucose, mannitol and sucrose. Necrotic foci in liver and congestion with hemorrhages in heart were found on necropsy. After pathogenicity test, the pathological changes were reconfirmed by histopathology depicting congestion, hemorrhage and lymphocyte infiltration in heart, liver and spleen tissues. In type specific PCR reaction, the organisms were confirmed as P. multocida Type A. In conclusion, P. multocida type A is prevalent among poultry in the studied regions; thus, care must be taken to control of the disease. [J Adv Vet Anim Res 2015; 2(3.000): 338-345]

Objectives: The objectives of this study were to isolate and identify Pasteurella multocida from fowl cholera (FC) suspected chicken, and to prepare and efficacy determination of formalin killed fowl cholera vaccine using the isolated P. multocida strain. Materials and methods: A total of five suspected dead chickens were collected from Brothers Poultry Farm located at Gazipur district, Bangladesh. The samples were processed and the P. multocida was isolated through conventional bacteriological techniques, were finally confirmed by polymerase chain reaction using P. multocida specific primers targeting cap gene. The P. multocida isolate was used to develop a formalin killed fowl cholera vaccine. The efficacy of the newly prepared vaccine was determined in Starcross-579 chickens (n=30) aging 15 weeks either by injecting 1 mL (group-A; n=10) or 0.5 mL (group-B; n=10) vaccine containing approximately 3.2x108 CFU/mL P. multocida organism; 10 birds were kept as unvaccinated control. The sera from the vaccinated and control birds were collected and were subjected for antibody titre determination by enzyme-linked immunosorbent assay (ELISA). Finally the vaccinated birds were challenged using virulent strains of P. multocida to confer the protection against FC. Results: P. multocida could be isolated from both the samples. The formalin killed vaccine prepared from the isolated bacteria was subjected for the determination of antibody titre in chicken, and found that the antibody titres in the birds of group A and group B were 4.513 and 4.07 respectively after primary vaccination, and 4.893 and 4.37 respectively after booster vaccination. Most of the vaccinated birds were found to be survived after challenging with virulent strain of P. multocida. Conclusion: It is concluded that the causal agent of FC (P. multocida) was successfully isolated from FC affected dead chickens. The prepared formalin killed fowl cholera vaccine induces protective immune response and conferred protection against challenge infection caused by the virulent strain of P. multocida. [J Adv Vet Anim Res 2016; 3(1.000): 45-50]

Fowl cholera is a contagious bacterial disease of poultry caused by Pasteurella multocida and has a significant economic importance worldwide. Fowl cholera prevention depends mainly on vaccination using live and inactivated vaccines, but they have many limitations. The present study aimed to prepare fowl cholera chicken embryo derived inactivated bacterin and to evaluate it by experimental infection and molecular evaluation. Fowl cholera chicken embryo derived inactivated bacterin prepared by in-vivo growing of Pasteurella multocida serotype 3 (P-1059) on chicken embryonating eggs, then inactivated by formalin (0.3%) and adjuvant added and then quality control parameters tested. The prepared bacterin was evaluated by experimental challenge with homologues and heterologous serotypes of Pasteurella multocida serotypes in comparison with commercial inactivated multivalent vaccine. Molecular evaluation of prepared bacterin was carried out using SDS-PAGE for in-vivo and in-vitro grown Pasteurella multocida serotypes. Results revealed that the prepared fowl cholera chicken embryo derived inactivated bacterin was free from bacterial and fungal contaminations and safe for use. Fowl cholera chicken embryo derived inactivated bacterin provided high protection rates with low mortalities against experimental infections with homologues and heterologous serotypes of Pasteurella multocida. Results of SDS-PAGE revealed that in-vivo grown Pasteurella multocida serotypes showed expression of additional specific bands (35kDa, 39kDa). It was suggested that 39kDa is a dominant structural protein in Pasteurella multocida grown in-vivo and play a role in antigenic cross protection among its serotypes. In conclusion, Fowl cholera chicken embryo derived inactivated bacterin is effective against infections of homologous and heterologous Pasteurella multocida serotypes.

Pasteurella multocida is a terrible veterinary pathogen that causes widespread infections in husbandry. To induce homologous and/or heterologous immunity against the infections, outer membrane protein Hs (OmpH) in the envelope of different strains of P. multocida are thought to be attractive vaccine candidates. Previously we cloned and characterized a gene for OmpH from pathogenic P. multocida (A:3) (In Press, Korean J. Microbiol. Biotechnol. 2005, 33, December).