Isolation, Molecular Characterization and Antibiotic Susceptibility Pattern of Pasteurella multocida Isolated from Cattle and Buffalo from Ahwaz, Iran

Document Type : Original Articles


1 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Graduate of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran


Pasteurella multocida is a gram-negative, opportunistic pathogen and the common inhabitant of the upper respiratory tract of domesticated and wild animal species. It is a causative agent of numerous economically important diseases worldwide, such as enzootic bronchopneumonia in ruminants and haemorrhagic septicemia in cattle and buffalo. The present study was undertaken to determine the prevalence of P. multocida carriers, PCR-serogrouping and antibiotic susceptibility status of isolates detected in cattle and buffalo in Ahvaz, Iran. Nasopharyngeal and nasal swabs were collected from 227 cattle and 174 buffaloes. The swabs were streaked on MacConkey and sheep blood agar and incubated for 24-48 h at 37 °C. The presumptive P. multocida colonies were identified based on standard biochemical testing and further confirmed by PCR. A multiplex PCR was used to determine the five pathogen serogroups (i.e., A, B, D, E, and F). Antimicrobial susceptibility of P. multocida isolates was determined using the Kirby–Bauer disc diffusion method on Mueller-Hinton agar supplemented with 5% sheep blood. Out of 401 tested samples, P. multocida was recovered from 10/227 (4.4%) and 12/174 (6.89%) cattle and buffaloes, respectively. Fifteen isolates (68.19%) belonged to serogroup A, 5 (22.72%) to serogroup D, and 2 (9.09%) isolates were untypeable. No isolate belonged to B, E, and F serogroups. All P. multocida isolates were sensitive to nitrofurantoin, florfenicol, ciprofloxacin, enrofloxacin, trimethoprim-sulfamethoxazole, oxytetracycline, and ceftriaxone. The most common finding was resistance to tylosin (90.9%), followed by resistance to oxacillin (54.54%).


Main Subjects

Article Title [French]

L'isolement, l'identification moléculaire et profil de sensibilité aux antibiotiques en Pasteurella multocida isolé des vaches et des buffles à Ahvaz en Iran

Abstract [French]

Pasteurella multocida est une bactérie gram-négative, opportuniste pathogène et flore normale des voies respiratoires supérieures des espèces animales domestiques et sauvages. Cette bactérie cause des maladies économiquement importantes partout dans le monde telles que bronchopneumonie enzootique chez les ruminants et la septicémie hémorragique chez les vaches et les buffles. L'objectif de cette étude est la détermination de la prévalence des porteurs de Pasteurella multocida chez les vaches et les buffles à Ahvaz, la détermination des sérogroupes capsulaires de bactéries et aussi l'évaluation de la sensibilité aux antibiotiques des isolats. Pour ce faire, les prélèvements nasal et rhinopharyngé ont été collectés de 227 vaches et 174 buffles. Les écouvillons ont été mises en culture dans environnement Mac et Cancan et gélose au sang de mouton et mises à incuber pendant 24-48 heures à 37 C. Colonies soupçonnées à Pasteurella multocida ont été identifiées sur la base des tests biochimiques standards et confirmées par PCR. Les multiples réactions de PCR ont été utilisées pour déterminer les cinq sérogroupes capsulaires bactérien (A, B, D, E et F). L'évaluation de la sensibilité aux antibiotiques des isolats de Pasteurella multocida a été également mise en place en utilisant la méthode de diffusion sur disque (Kirby-Bauer) sur un milieu de gélose de Mueller Hinton additionné de 5% sang de mouton. Parmi 401 échantillons, Pasteurella multocida était trouvé respectivement de 10.227 (4.4%) et 12.174 (6.89%) des vaches et des buffles. 15 isolats (68.19%) appartenait au sérogroupe A, 5 isolats (22.72%) appartenait au sérogroupe D et 2 isolats (9.09%) étaient inclassable. Aucun isolats n’appartenait au sérogroupes B, E et F. Tous les isolats de Pasteurella multocida étaient sensibles à nitrofurantoïne, Florfenicol, ciprofloxacine, enrofloxacine,Triméthoprime sulfaméthoxazole , oxytétracycline, ceftriaxone. La plus grande résistance était à la tylosine (90.9%), ensuite à oxacilline (54.54%), streptomycine (45.45%), ampicilline (27.27%) , Erythromycin (13.63%) et enfin pénicilline (9.1%).

Keywords [French]

  • Pasteurella multocida
  • buffle
  • vache
  • sérogroupes
  • la sensibilité aux antibiotiques
Arashima, Y., Kumasaka, K., 2005. Pasteurellosis as zoonosis. Intern Med 44, 692-693.
Arora, A.K., Virmani, S., Jand, S.K., Oberoi, M.S., 2005. Isolation, characterization and antibiogram of Pasteurella multocida isolates from different animal species. Indian J Anim Sci 75, 749–752.
Arumugam, N.D., Ajam, N., Blackall, P.J., Asiah, N.M., Ramlan, M., Maria, J., et al., 2011. Capsular serotyping of Pasteurella multocida from various animal hosts - a comparison of phenotypic and genotypic methods. Trop Biomed 28, 55-63.
Baur, A.W., Kirby, W.M.M., Sherris, J.C., Turek, N., 1966. Antibiotic susceptibility testing by standardized single disc method. Am J Clin Pathol 45, 493-496.
Boyce, J.D., Adler, B., 2000. The capsule is a virulence determinant in the pathogenesis of Pasteurella multocida M1404 (B:2). Infect Immun 68, 3463-3468.
Caprioli, A., Busani, L., Martel, J.L., Helmuth, R., 2000. Monitoring of antibiotic resistance in bacteria of animal origin: epidemiological and microbiological methodologies. International Journal of Antimicrobial Agents 14, 295-301.
Carter, G.R., 1955. Studies on Pasteurella multocida. I. A hemagglutination test for the identification of serological types. Am J Vet Res 16, 481-484.
Davies, R.L., MacCorquodale, R., Reilly, S., 2004. Characterisation of bovine strains of Pasteurella multocida and comparison with isolates of avian, ovine and porcine origin. Vet Microbiol 99, 145-158.
Dey, S., Singh, V.P., Kumar, A.A., Sharma, B., Srivastava, S.K., Singh, N., 2007. Comparative sequence analysis of 16S rRNA gene of Pasteurella multocida serogroup B isolates from different animal species. Res Vet Sci 83, 1-4.
Dutta, T.K., Singh, V.P., Kumar, A.A., 2001. Rapid and specific diagnosis of haemorrhagic septicemia by using PCR assay. Indian J Anim Health 40, 101– 107.
Dziva, F., Christensen, H., van Leengoed, L.A., Mohan, K., Olsen, J.E., 2004. Differentiation of Pasteurella multocida isolates from cases of atrophic rhinitis in pigs from Zimbabwe by RAPD and ribotyping. Vet Microbiol 102, 117-122.
Dziva, F., Muhairwa, A.P., Bisgaard, M., Christensen, H., 2008. Diagnostic and typing options for investigating diseases associated with Pasteurella multocida. Vet Microbiol 128, 1-22.
Garcia, N., Fernandez-Garayzabal, J.F., Goyache, J., Dominguez, L., Vela, A.I., 2011. Associations between biovar and virulence factor genes in Pasteurella multocida isolates from pigs in Spain. Vet Rec 169, 362.
Haji Hajikolaei, M.R., Ghorbanpour, M., Seyfi-Abadshapouri, M.R., Rasooli, A., Jahferian, H., 2006. Occurrence of Pasteurella multocida in the Nasopharynx of Healthy Buffaloes and Their Immunity Status. B Vet I Pulway 50, 435-438.
Hanan, M.S., Riad, E.M., el-Khouly, N.A., 2000. Antibacterial efficacy and pharmacokinetic studies of ciprofloxacin on Pasteurella multocida infected rabbits. Dtsch Tierarztl Wochenschr 107, 151-155.
Hunt, M.L., Adler, B., Townsend, K.M., 2000. The molecular biology of Pasteurella multocida. Veterinary Microbiology 72, 3-25.
Imran, M., Irshad, M., Shahid, M.A., Ashraf, M., 2007. Studies on the Carrier Status of Pasteurella multocida in Healthy Cattle and Buffalo in District Faisalabad. Int J Dairy Sci 2, 398- 400.
Kaan, O., Serpil, K.K., Tayfun, C., 2010. Frequency and antibiotic susceptibility of Pasteurella multocida and Mannheimia haemolytica isolates from nasal cavities of cattle. Turk J Vet Anim Sci 34, 91-94.
Khamesipour, F., Momtaz, H., Azhdary Mamoreh, M., 2014. Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran. Frontiers in Microbiology 5.
Kroemer, S., Galland, D., Guerin-Faublee, V., Giboin, H., Woehrle-Fontaine, F., 2012. Survey of marbofloxacin susceptibility of bacteria isolated from cattle with respiratory disease and mastitis in Europe. Vet Rec 170, 53.
Markey, B., Leonard, F., Archambault, M., Cullinane, A., Maguire, D., 2013. Clinical Veterinary Microbiology E-Book, Elsevier Health Sciences.
Mohamed, M.A., Mohamed, M.W., Ahmed, A.I., Ibrahim, A.A., Ahmed, M.S., 2012. Pasteurella multocida in backyard chickens in Upper Egypt: incidence with polymerase chain reaction analysis for capsule type, virulence in chicken embryos and antimicrobial resistance. Vet Ital 48, 77-86.
Mustafa, A.A., Ghalib, H.W., Shigidi, M.T., 1978. Carrier rate of Pasteurella multocida in a cattle herd associated with an outbreak of haemorrhagic septicaemia in the Sudan. Br Vet J 134, 375-378.
Naz, S., Hanif, A., Maqbool, A., Ahmed, S., Muhamm, K., 2012. Isolation, characterization and monitoring of antibiotic Resistance in Pasteurella multocida isolates from buffalo (Bubalus Bubalis) herds around lahore. J Anim Plant Sci 22, 242-245.
Rimler, R.B., Rhoades, K.R., 1987. Serogroup F, a new capsule serogroup of Pasteurella multocida. J Clin Microbiol 25, 615-618.
Shayegh, J., Atashpaz, S., Zahraei Salehi, T., Saeid Hejazi, M., 2010. Potential of Pasteurella multocida isolated from healthy and diseased cattle and buffaloes in induction of diseases. B Vet I Pulawy 54, 299-304.
Shayegh, J., Mikaili, P.J., Sharaf, D., Rastgu, A., 2009. Antimicrobal resistance evaluation of Iranian ovine and bovine Pasteurella multocida. Anim Vet Adv 8, 1753-1756.
Shivachandra, S.B., Kumar, A.A., Biswas, A., Ramakrishnan, M.A., Singh, V.P., Srivastava, S.K., 2004. Antibiotic Sensitivity Patterns among Indian Strains of Avian Pasteurella multocida. Tropical Animal Health and Production 36, 743-750.
Townsend, K.M., Boyce, J.D., Chung, J.Y., Frost, A.J., Adler, B., 2001. Genetic organization of Pasteurella multocida cap Loci and development of a multiplex capsular PCR typing system. J Clin Microbiol 39, 924-929.
Townsend, K.M., Frost, A.J., Lee, C.W., Papadimitriou, J.M., Dawkins, H.J., 1998. Development of PCR assays for species- and type-specific identification of Pasteurella multocida isolates. J Clin Microbiol 36, 1096-1100.