Isolation, identification, and monitoring of antibiotic resistance in Pasteurella multocida and Mannheimia haemolytica isolated from sheep in East Azerbaijan province, Iran

Document Type: Original Articles


1 Virus Biobank Laboratory, Razi Vaccine and Serum Research Institute, Karaj, Iran

2 Departent ofm Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, Iran Department of Research and Development, Razi Vaccine and Serum Research Institute, Karaj, Iran

3 Department of Aerobic Bacterial Vaccines, Razi Vaccine and Serum Research Institute, Karaj, Iran

4 Department of Research and Development, Razi Vaccine and Serum Research Institute, Karaj, Iran

5 Technical Director of Veterinary Vaccines, Razi Vaccine and Serum Research Institute, Karaj, Iran


The present study was carried out in order to isolate, identify, and assess the antimicrobial susceptibility of the causative agent(s) of pneumonic pasteurellosis in sheep in East Azerbaijan province, northwest of Iran. Pneumonia was detected in 320 cases, and the affected lungs were sampled in the slaughterhouse. The samples were investigated bacteriologically for the isolation of two microorganisms from the Pasteurellaceae family. Pasteurella multocida was isolated from six (1.87%) samples, while none of the lung tissues were positive for Mannheimia haemolytica. After the isolation and detection of microorganisms via cultural and morphological tests, the bacteria were identified on the basis of biochemical criteria and polymerase chain reaction (PCR) technique. Antimicrobial susceptibility testing was performed on all P. multocida isolates, using broth microdilution method. Evaluation of the minimum inhibitory concentration (MIC) of eight antimicrobial agents against the tested isolates showed that all the organisms were resistant to amoxicillin and relatively susceptible to ceftiofur. In conclusion, P. multocida was introduced as the main cause of ovine pneumonic pasteurellosis in the studied district, and the outbreak frequency significantly varied in different seasons of the year (P<0.05).

Main Subjects

Article Title [French]

Isolation, identification et contrôle de l’antibiorésistance des souches de Pasteurella multocida et Mannheimia haemolytica isolées à partir de moutons de l’Est Azarbaijan, Iran

Abstract [French]

L’objectif de cette étude était d’isoler, d’identifier et d’évaluer la susceptibilité antimicrobienne des agents responsables de la pasteurellose pneumonique chez les moutons de l’Est Azerbaïdjan, situé au nord ouest de l’Iran. Trois cent vingt (320) cas de pneumonie ont été détectés et le prélèvement des poumons affectés a été mené dans un abattoir. Tous les échantillons ont été ensuite soumis à des analyses bactériologiques et l’incidence de deux microorganismes spécifiques appartenant à la famille Pasteurellaceae a été étudiée. Selon nos analyses, six prélèvements pulmonaires étaient contaminés par Pasteurella multocida (1,87%), alors qu’aucun échantillon positif à la souche Mannheimia haemolytica n’a été détecté. Après l’isolation et la détection des microorganismes par des testes de culture et d’observation morphologiques, les souches bactériennes ont été identifiées grâce à leurs propriétés biochimiques et la méthode de réaction en chaîne par polymérase (PCR). La susceptibilité antimicrobienne de tous les isolats de P. multocida a été évaluée par la méthode de microdilution du bouillon de culture. L’évaluation de la concentration inhibitrice minimum (MIC) de 8 agents antimicrobiens vis-à-vis des isolats identifiés a révélé une résistance à l’ amoxicilline et une susceptibilité relative au ceftiofur. En conclusion, P. multocida se présente comme la cause majeure de pasteurellose pneumonique ovine dans la province de l’Est Azerbaïdjan et la fréquence cette épidémie varie de façon significative d’une saison à l’autre (P<0.05).

Keywords [French]

  • Isolation
  • Pasteurella multocida
  • Mannheimia haemolytica
  • Mouton
  • PCR
  • Résistance antimicrobienne
  • Iran

Arashima, Y., Kumasaka, K., 2005. Pasteurellosis as zoonosis. Intern Med 44, 692-693.

Azizi, S., Korani, F.S., Oryan, A., 2013. Pneumonia in slaughtered sheep in south-western Iran: pathological characteristics and aerobic bacterial aetiology. Vet Ital 49, 109-118.

Brogden, K.A., Nordholm, G., Ackermann, M., 2007. Antimicrobial activity of cathelicidins BMAP28, SMAP28, SMAP29, and PMAP23 against Pasteurella multocida is more broad-spectrum than host species specific. Vet Microbiol 119, 76-81.

Chung, J.Y., Zhang, Y., Adler, B., 1998. The capsule biosynthetic locus of Pasteurella multocida A:1. FEMS Microbiol Lett 166, 289-296.

Dar, L.M., Darzi, M.M., Mir, M.S., Kamil, S.A., Rashid, A., Abdullah, S., 2013. Prevalence of lung affections in sheep in northern temperate regions of India: A postmortem study. Small Ruminant Research 110, 57-61.

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.

Demissie, T., Dawo, F., Sisay, T., 2014. Biochemical and antigenic characterization of Mannheimia, Pasteurella and Mycoplasma species from naturally infected pneumonic sheep and goats, Bishoftu, Ethiopia. African Journal of Basic & Applied Science 6, 198-204.

Ebrahimi, A., Halajifar, M., Ataei, S., Lotfalian, S., 2010. Investigation of Pasteurella multocida in sheep pneumonic lesions of Kashan abattoir. Journal of Animal Health and Disease 4, 29-32.

Ewers, C., Lubke-Becker, A., Bethe, A., Kiebling, S., Filter, M., Wieler, L.H., 2006. Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status. Vet Microbiol 114, 304-317.

Ferreira, T.S., Felizardo, M.R., Senade Gobbi, D.D., Gomes, C.R., Nogueira Filsner, P.H., Moreno, M., 2012. Virulence genes and antimicrobial resistance profiles of Pasteurella multocida strains isolated from rabbits in Brazil. Science World Journal, 1-6.

Guler, L., Gunduz, K., Sarishahin, A.S., 2013. Capsular typing and antimicrobial susceptibility of Pasteurella multocida isolated from different hosts. Kafkas University Veterinary Fakultesi Dergisi 19, 843-849.

Hawari, A.D., Hassawi, D.S., Sweiss, M., 2008. Isolation and Identification of Mannheimia haemolytica and Pasteurella multocida in Sheep and Goats using Biochemical Tests and Random Amplified Polymorphic DNA (RAPD) Analysis. Journal of Biological Sciences 8, 1251-1254.

Jabeen, A., Khattak, M., Munir, S., Jamal, Q., Hussain, M., 2013. Antibiotic susceptibility and molecular analysis of bacterial pathogen Pasteurella multocida isolated from cattle. Journal of Applied Pharmaceutic Science 3, 106-110.

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.

Kumar, A.A., Shivachandra, S.B., Biswas, A., Singh, V.P., Singh, V.P., Srivastava, S.K., 2004. Prevalent serotypes of Pasteurella multocida isolated from different animal and avian species in India. Vet Res Commun 28, 657-667.

Lichtensteiger, C.A., Steenbergen, S.M., Lee, R.M., Polson, D.D., Vimr, E.R., 1996. Direct PCR analysis for toxigenic Pasteurella multocida. J Clin Microbiol 34, 3035-3039.

Lion, C., Conroy, M.C., Carpentier, A.M., Lozniewski, A., 2006. Antimicrobial susceptibilities of Pasteurella strains isolated from humans. Int J Antimicrob Agents 27, 290-293.

Odugbo, M.O., Odama, L.E., Umoh, J.U., Lamorde, A.G., 2006. Pasteurella multocida pneumonic infection in sheep:

Prevalence, clinical and pathological studies. Small Ruminant Research 66, 273-277.

Safaee, S., Weiser, G.C., Cassirer, E.F., Ramey, R.R., Kelley, S.T., 2006. Microbial diversity in bighorn sheep revealed by culture-independent methods. J Wildl Dis 42, 545-555.

Shayegh, J., Dolgari Sharaf, J., Mikaili, P., Namvar, H., 2009. Pheno- and genotyping of Pasteurella Multocida isolated from goat in Iran. African Journal of Biotechnology 8, 3707-3710.

Songer, J.G., Post, K.W., 2004. Veterinary Microbiology - Elsevieron VitalSource: Bacterial and Fungal Agents of Animal Disease, Elsevier Health Sciences.

Tabatabaei, M., Liu, Z., Finucane, A., Parton, R., Coote, J., 2002. Protective immunity conferred by attenuated aroA derivatives of Pasteurella multocida B:2 strains in a mouse model of hemorrhagic septicemia. Infect Immun 70, 3355-3362.

Tehrani, A.A., Ras, M.B., Niazy, H., 2004. Isolation and identification of Pasteurella Haemolytica biotype A from sheep in Uremia, Iran. Iranian Journal of Veterinary Research 5, 105-109.

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.

Weiser, G.C., DeLong, W.J., Paz, J.L., Shafii, B., Price, W.J., Ward, A.C., 2003. Characterization of Pasteurella multocida associated with pneumonia in bighorn sheep. J Wildl Dis 39, 536-544.

Wilson, B.A., Ho, M., 2013. Pasteurella multocida: from zoonosis to cellular microbiology. Clin Microbiol Rev 26, 631-655.

Zamri-Saad, M., Effendy, A.W.M., Maswati, M.A., Salim, N., Sheikh-Omar, A.R., 1996. The goat as a model for studies of pneumonic pasteurellosis caused by Pasteu