Prevalence of adhesion Virulence factor genes, antibiogram, and pathogenicity of avian Pasteurella multocida isolate from Iran

Document Type : Original Articles

Authors

1 Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran

2 Pasteurella National Research Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Tehran, Iran

3 Department of Aerobic Bacterial Vaccine Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran

Abstract

Pasteurella multocida possesses various virulence factors, including capsule, lipopolysaccharide, fimbriae, toxins, outer membrane proteins, and adhesions. Adhesins have a crucial role in mediating colonization and invasion of the host. The aim of the present study was to identify the prevalence of adhesion factor genes and resistance/sensitivity patterns among the avian P. multocida isolates from Iran. A total of 30 isolates of P. multocida were used for this study. All isolates were obtained from the poultry cases dead from fowl cholera in the northern parts of Iran. The results of the polymerase chain reaction analysis for the frequency of virulence-associated genes showed that the genes encoding adhesins (i.e., ptfA, fimA, hsf-1, pfhA, and ompH) were found in all (100%) of the isolates. However, the frequency of two genes including tadD and toxA were 50% and 70%, respectively. Thegenotyping patterns were classified into four groups according to the virulence factors in P. multocida isolates. Genotype pattern I, which included the isolates harbouring all of the examined virulence factor genes showed the highest frequency (43.3%). Pathogenicity test showed that all of the isolates classified as genotype I were pathogen or highly pathogen in the mice model. The sensitivity of the isolates to penicillin, ampicillin, lincospectin, florfenicol, tylosin, and tiamulin was 100%. However, the sensitivity rates to flumequin, enrofloxacine, and nalidixicacid were 96.6% and 80%, respectively. The findings of the current study will be helpful to elucidate the disease process and develop an efficient multivalent local vaccine.
 

Keywords

Main Subjects


Article Title [French]

La prévalence des gènes de facteurs de Virulence de facteur d'adhérence, antibiogramme et pathogénicité dans les isolats des oiseaux de Pasteurella multocida en Iran

Abstract [French]

Les adhésions ont un rôle très important dans la création de la colonisation et l'attaque à l'hôte. En outre, leur présence au niveaux de bactéries sont souvent associées à la virulence. Les adhésions étaient le but des études récentes de génotypage virulence. Le but de la présente étude est d'identifier la prévalence des gènes de facteurs de virulence et les modèles de résistance/sensibilité dans les isolats des oiseaux de Pasteurella multocida en Iran. Un total de 30 isolats de Pasteurella multocida ont été utilisés dans la présente étude. Tous les isolats proviennent des oiseaux morts de choléra aviaire dans les régions du Nord de l'Iran. Les résultats de l'analyse PRC pour déterminer la fréquence des gènes associés à la virulence ont montré que le gènes codant des adhésions ptfA, fimA, hsf-1, pfhA et ompH ont été trouvés (100%) dans tous les isolats. En revanche, la fréquence des deux gènes de tadD et toxA était respectivement 50% et 70%. La fréquence des modèles de génotype sont classées en quatre groupes (I-IV) selon les facteurs de virulence des isolats Pasteurella multocida. Génotype modèle I comprend les isolats qui contiennent tous les gènes de facteurs de virulence testés. Ce modèle a montré la plus haute fréquence (43.3%). Des tests de pathogéncité ont montré que tous les isolats classés comme génotype I étaient aiguë ou suraiguë dans un modèle de souris. La sensibilité des isolats à Pénicilline, ampicilline, lincoseptin, florfenicol, Tiamuline et tylosine était 100% et la sensibilité envers Flemequine, Enrofloxacin et Nalidixic acid était respectivement 96% et 80%. Les résultats de cette recherche seront utile pour clarifier le processus de maladie et aussi pour produire un vaccin multivalent en utilisant des souches régionales.

Keywords [French]

  • Pasteurella multocida
  • facteurs d’adhésion
  • antibiogramme
  • pathogénicité
  • choléra aviaire
Atashpaz, S., Shayegh, J., Hejazi, M.S., 2009. Rapid virulence typing of Pasteurella multocida by multiplex PCR. Res Vet Sci 87, 355-357.
Bethe, A., Wieler, L.H., Selbitz, H.J., Ewers, C., 2009. Genetic diversity of porcine Pasteurella multocida strains from the respiratory tract of healthy and diseased swine. Vet Microbiol 139, 97-105.
Blackall, P.J., Pahoff, J.L., Bowles, R., 1997. Phenotypic characterisation of Pasteurella multocida isolates from Australian pigs. Vet Microbiol 57, 355-360.
Christensen, H., Angen, O., Olsen, J.E., Bisgaard, M., 2004. Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium. Microbiology 150, 1757-1767.
Davies, R.L., 2004. Genetic diversity among Pasteurella multocida strains of avian, bovine, ovine and porcine origin from England and Wales by comparative sequence analysis of the 16S rRNA gene. Microbiology 150, 4199-4210.
Davies, R.L., MacCorquodale, R., Baillie, S., Caffrey, B., 2003. Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis. J Med Microbiol 52, 59-67.
Doughty, S.W., Ruffolo, C.G., Adler, B., 2000. The type 4 fimbrial subunit gene of Pasteurella multocida. Vet Microbiol 72, 79-90.
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.
Fegan, N., Blackall, P.J., Pahoff, J.L., 1995. Phenotypic characterisation of Pasteurella multocida isolates from Australian poultry. Vet Microbiol 47, 281-286.
Ferreira, T.S.P., Felizardo, M.R., Sena de Gobbi, D., #xe9, Dirani, b., Gomes, C.R., et al., 2012. Virulence Genes and Antimicrobial Resistance Profiles of Pasteurella multocida Strains Isolated from Rabbits in Brazil. Sci World J 2012, 6.
Furian, T.Q., Borges, K.A., Rocha, S.L.S., Rodrigues, E.E., Nascimento, V.P.d., Salle, C.T.P., et al., 2013. Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR. Pesquisa Vet Brasil 33, 177-182.
Harper, M., Boyce, J.D., Adler, B., 2006. Pasteurella multocida pathogenesis: 125 years after Pasteur. FEMS Microbiol Lett 265, 1-10.
Hatfaludi, T., Al-Hasani, K., Boyce, J.D., Adler, B., 2010. Outer membrane proteins of Pasteurella multocida. Vet Microbiol 144, 1-17.
Katsuda, K., Hoshinoo, K., Ueno, Y., Kohmoto, M., Mikami, O., 2013. Virulence genes and antimicrobial susceptibility in Pasteurella multocida isolates from calves. Vet Microbiol 167, 737-741.
Kehrenberg, C., Schwarz, S., 2001. Occurrence and linkage of genes coding for resistance to sulfonamides, streptomycin and chloramphenicol in bacteria of the genera Pasteurella and Mannheimia. FEMS Microbiol Lett 205, 283-290.
May, B.J., Zhang, Q., Li, L.L., Paustian, M.L., Whittam, T.S., Kapur, V., 2001. Complete genomic sequence of Pasteurella multocida, Pm70. Proc Natl Acad Sci U S A 98, 3460-3465.
Portis, E., Lindeman, C., Johansen, L., Stoltman, G., 2012. A ten-year (2000-2009) study of antimicrobial susceptibility of bacteria that cause bovine respiratory disease complex--Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni--in the United States and Canada. J Vet Diagn Invest 24, 932-944.
Rigobelo, E.C., Blackall, P.J., Maluta, R.P., de Avila, F.A., 2013. Identification and antimicrobial susceptibility patterns of Pasteurella multocida isolated from chickens and japanese quails in Brazil. Braz J Microbiol 44, 161-164.
Sarangi, L.N., Priyadarshini, A., Kumar, S., Thomas, P., Gupta, S.K., Nagaleekar, V.K., et al., 2014. Virulence Genotyping of Pasteurella multocida Isolated from Multiple Hosts from India. Sci World J 2014, 10.
Sarangi, L.N., Thomas, P., Gupta, S.K., Priyadarshini, A., Kumar, S., Nagaleekar, V.K., et al., 2015. Virulence gene profiling and antibiotic resistance pattern of Indian isolates of Pasteurella multocida of small ruminant origin. Comp Immunol Microbiol Infect Dis 38, 33-39.
Sellyei, B., Banyai, K., Magyar, T., 2010. Characterization of the ptfA gene of avian Pasteurella multocida strains by allele-specific polymerase chain reaction. J Vet Diagn Invest 22, 607-610.
Shayegh, J., Atashpaz, S., Hejazi, M.S., 2008. Virulence genes profile and typing of ovine Pasteurella multocida. Asian J Anim Vet Adv 3, 206-213.
Tang, X., Zhao, Z., Hu, J., Wu, B., Cai, X., He, Q., et al., 2009. Isolation, antimicrobial resistance, and virulence genes of Pasteurella multocida strains from swine in China. J Clin Microbiol 47, 951-958.
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., O'Boyle, D., Thi Phan, T., Xuan Hanh, T., Wijewardana, T.G., Wilkie, I., et al., 1998. Acute septicaemic pasteurellosis in Vietnamese pigs. Vet Microbiol 63, 205-215.
Varga, Z., Volokhov, D.V., Stipkovits, L., Thuma, A., Sellyei, B., Magyar, T., 2013. Characterization of Pasteurella multocida strains isolated from geese. Vet Microbiol 163, 149-156.
Vasfi Marandi, M., Mittal, K.R., 1997. Role of outer membrane protein H (OmpH)- and OmpA-specific monoclonal antibodies from hybridoma tumors in protection of mice against Pasteurella multocida. Infect Immun 65, 4502-4508.
Verma, S., Sharma, M., Katoch, S., Verma, L., Kumar, S., Dogra, V., et al., 2013. Profiling of virulence associated genes of Pasteurella multocida isolated from cattle. Vet Res Commun 37, 83-89.
Wilkie, I.W., Grimes, S.E., O’Boyle, D., Frost, A.J., 2000. The virulence and protective efficacy for chickens of Pasteurella multocida administered by different routes. Vet Microbiol 72, 57-68.
Yoshimura, H., Ishimaru, M., Endoh, Y.S., Kojima, A., 2001. Antimicrobial susceptibility of Pasteurella multocida isolated from cattle and pigs. J Vet Med B Infect Dis Vet Public Health 48, 555-560.