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Pasteurella multocida is incriminated in fowl cholera in poultry and snuffles in rabbits, causing significant economic losses in the poultry industry worldwide. This study investigated the prevalence, kmt1 gene sequencing, antibiogram, pathogenicity, and PCR detection of the virulence and resistance genes of P. multocida recovered from chickens, ducks, and rabbits. Approximately 150 samples were collected from diseased chickens, ducks, and rabbits (50 for each) from commercial farms in Ismailia government, Egypt. The collected samples were bacteriologically examined, and the recovered isolates were subsequently subjected to kmt1 gene sequencing, antibiogram, pathogenicity test, and PCR-based detection of virulence and antibiotic resistance genes. The overall prevalence of P. multocida was 20% (30/150). The highest prevalence was recorded in rabbits (24%; 12/50), followed by ducks (20%; 10/50) and chickens (16%; 8/50). The lung was the most predominant infected organ. Moreover, the kmt1 phylogenetic analysis emphasized that the tested P. multocida strains presented remarkable genetic identity with other P. multocida strains from the USA, Denmark, China, India, and Iran. Chloramphenicol and ciprofloxacin exhibited promising results in treating P. multocida. Moreover, the prevalence of oma87, ptfA, luxS, tadG, and nanB virulence genes was 100%, 86.7%, 76.6%, 73.3%, and 56.7% respectively. Herein, 13.3% of the tested strains were multidrug-resistant (MDR) to 5 antibiotic classes and carried bla ROB-1 and tetH genes. Besides, 10% of the tested P. multocida strains were extensively drug resistant (XDR) to 7 antibiotic classes and harbored the bla ROB-1 , tetH, and ermX genes. Moreover, 10% of the recovered P. multocida strains were MDR to 5 antibiotic classes and had tetH and ermX genes. Pathogenicity test revealed that the mortality rate was 20% at 1 day post-infection, reaching its highest point (100%) at 2 days post-infection. Briefly, this study highlights the occurrence of MDR and XDR P. multocida in domestic birds and rabbits, revealing a public health threat. The oma87, ptfA, luxS, and tadG virulence genes and the bla ROB-1 , tetH, and ermX antibiotic resistance genes are usually associated with emerging MDR P. multocida strains., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The prevalence of Pasteurella multocida strains among 275 backyard chickens from different regions of Upper Egypt was studied. A total of 21 isolates of P. multocida were recovered in 21 out of 275 chickens tested (7.6%) and were confirmed using phenotypic characterisation. Somatic serotyping of the 21 isolates resulted in 12 isolates being classed as serotype A:1 (57.14%), 4 as serotype A:3 (19.05%) and 5 could not be typed (23.8%). Capsular typing, using multiplex polymerase chain reaction (PCR), demonstrated that 18 strains were capsular type A (85.7%), and 3 were type D (14.3%). The present findings suggest that a multiplex capsular PCR could be valuable for the rapid identification of P. multocida in cases of fowl cholera infection. A total of 5 isolates of P. multocida were selected to study their pathogenicity in embryonated chicken eggs instead of conducting a study in mature chickens. The results showed a variation in pathogenicity between the strains tested, namely: serotype A:1 strains caused 80% mortality, in contrast to 20% mortality by type D strains. Pathological findings included severe congestion of the entire embryo, haemorrhaging of the skin, feather follicles and toe, and ecchymotic haemorrhages on the liver of the inoculated embryos. The observations in this study indicate that P. multocida serogroup A could be highly pathogenic for mature chickens and therefore might be a cause of considerable economic losses in commercial production. A total of 10 isolates were subjected to antimicrobial susceptibility to determine the minimal inhibitory concentration of 7 antimicrobials. All isolates were susceptible to ciprofloxacin, florfenicol, streptomycin and sulphamethoxazol with trimethoprim and with varying degrees of sensitivity to the other agents.