doi: 10.14202/vetworld.2019.331-336
Share this article on [Facebook] [LinkedIn]
Article history: Received: 06-07-2018, Accepted: 02-01-2019, Published online: 24-02-2019
Corresponding author: Nidhi Rawat
E-mail: dr_nidhirawat@yahoo.com
Citation: Rawat N, Gilhare VR, Kushwaha KK, Hattimare DD, Khan FF, Shende RK, Jolhe DK (2019) Isolation and molecular characterization of Mannheimia haemolytica and Pasteurella multocida associated with pneumonia of goats in Chhattisgarh, Veterinary World, 12(2): 331-336.Aim: The purpose of this study was to isolate and characterize the Mannheimia haemolytica and Pasteurella multocida from blood, nasal discharge, and lung tissue of pneumonic goats.
Materials and Methods: A total of 14 goats were investigated for pneumonic pasteurellosis. Of 14 goats, nasal swabs and blood samples were collected from 10 clinically diseased animals. Moreover, lung tissue and heart blood samples were collected during necropsy of four goats died with pneumonia. All the samples were processed for the isolation of M. haemolytica and P. multocida in the laboratory. Bacterial isolates were identified by cultural and biochemical characters and 16S rRNA sequence analysis. All the isolates were subjected to susceptibility testing using commonly used antimicrobials. M. haemolytica isolates were characterized by PHSSA gene detection. P. multocida isolates were characterized by KMT1 gene detection and capsule typing.
Results: On necropsy of dead goats, the pneumonia was characterized as acute fibrinous bronchopneumonia. Bacterial culture revealed the isolation of M. haemolytica (7) and P. multocida (5) of 10 clinical cases. Moreover, M. haemolytica and P. multocida were coisolated from two of the lung tissues. Furthermore, one of the other two lung tissues showed the isolation of M. haemolytica while the other showed recovery of P. multocida. Bacterial isolates were specifically identified by the 16S rRNA sequence analysis. The isolates showed reduced susceptibility to β-lactams, aminoglycosides, and fluoroquinolones . Moreover, the PHSSA and KMT1 genes were specifically detected among M. haemolytica, and P. multocida isolates, respectively. All P. multocida isolates belonged to serogroup A.
Conclusion: The present study reported an occurrence of pneumonic pasteurellosis caused by M. haemolytica and P. multocida in a goat flock.
Keywords: bronchopneumonia, goat, Mannheimia haemolytica, Pasteurella multocida, pneumonic pasteurellosis.
1. Mohamed, R.A. and Abdelsalam, E.B. (2008) A review on pneumonic pasteurellosis (respiratory mannheimiosis) with emphasis on pathogenesis, virulence mechanism and predisposing factors. Bulg. J. Vet. Med., 11(3): 139-160.
2. Marru, H.D., Anijajo, T.T. and Hassen, A.A. (2013) A study on ovine pneumonic pasteurellosis: Isolation and identification of pasteurellae and their antibiogram susceptibility pattern in Haramaya district, Eastern Hararghe, Ethiopia. BMC Vet. Res., 9(1): 239. [Crossref]
3. Falade, S. (2002) Further Pasteurella isolates from the Republic of Zambia. Trop. Vet., 20(3): 130-131. [Crossref]
4. Dowling, A., Hodgson, J.C., Schock, A., Donachie, W., Eckersall, P.D. and Mckendrick, I.J. (2002) Experimental induction of pneumonic pasteurellosis in calves by intratracheal infection with Pasteurella multocida biotype A: 3. Res. Vet. Sci., 73(1): 37-44. [Crossref]
5. Abdullah, F.F.J., Tijjani, A., Adamu, L., Chung, E.L.T., Abba, Y., Mohammed, K., Saharee, A.A., Haron, A.W., Sadiq, M.A. and Mohd, A.M.L. (2014) Pneumonic pasteurellosis in a goat. Iran. J. Vet. Med., 8(4): 293-296.
6. Shanthalingam, S., Goldy, A., Bavananthasivam, J., Subramaniam, R., Batra, S.A., Kugadas, A., Raghavan, B., Dassanayake, R.P., Jennings-Gaines, J.E., Killion, H.J., Edwards, W.H., Ramsey, J.M., Anderson, N.J., Wolff, P.L., Mansfield, K., Bruning, D. and Srikumaran, S. (2014) PCR assay detects Mannheimia haemolytica in culture-negative pneumonic lung tissues of bighorn sheep (Ovis canadensis) from outbreaks in the Western USA, 2009-2010. J. Wildl. Dis., 50(1): 1-10. [Crossref]
7. Dabo, S.M., Taylor, J.D. and Confer, A.W. (2007) Pasteurella multocida and bovine respiratory disease. Anim. Health Res. Rev., 8(2): 129-150. [Crossref] [PubMed]
8. Hassan, G.M., El-Feky, Z.A., Eissa, E.A. and Teleb, A.A. (2016) Rapid diagnosis of virulent Pasteurella multocida isolated from farm animals with clinical manifestation of pneumonia respiratory infection using 16S rDNA and KMT1 gene. Asian Pac. J. Trop. Dis., 6(1): 21-26. [Crossref]
9. Quinn, P.J., Markey, B.K., Carter, M.E., Donnelly, W.J. and Leonard, F.C. (2001) Veterinary Microbiology and Microbial Disease. 1st ed. Wiley-Blackwell, USA.
10. CLSI (Clinical and Laboratory Standards Institute). (2008) Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals-Document M31-A3. Wayne, PA, USA.
11. CLSI (Clinical and Laboratory Standards Institute). (2013) Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals; Second Informational Supplement-Document VET01-S2. Wayne, PA, USA.
12. Lane, D.J. (1991) 16S/23S rRNA sequencing. In: Stackenbrandt, E. and Goodfellow, M., editor. Nucleic Acid Techniques in Bacterial Systematics. John Wiley and Sons, Chichester, United Kingdom. p115-175.
13. Hawari, A.D., Hassawi, D.S. and 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. J. Biol. Sci., 8(7): 1251-1254. [Crossref]
14. Townsend, K.M., Frost, A.J., Lee, C.W., Papadimitriou, J.M. and Dawkins, H.J.S. (1998) Development of PCR assays for species and type specific identification of Pasteurella multocida isolates. J. Clin. Microbiol., 36(4): 1096-1100. [PubMed] [PMC]
15. Townsend, K.M., Boyce, J.D., Chung, J.Y., Frost, A.J. and Adler, B. (2001) Genetic organization of Pasteurella multocida cap loci and development of a multiplex capsular PCR typing system. J. Clin. Microbiol., 39(3): 924-929. [Crossref] [PubMed] [PMC]
16. Brogden, K.A., Lehmkuhl, H.D. and Cutlip, R.C. (1998) Pasteurella haemolytica complicated respiratory infections in sheep and goats. Vet. Res., 29(3-4): 233-254. [PubMed]
17. Galapero, J., Fernandez, S., Perez, C.J., Calle-Alonso, F., Rey, J. and Gomez, L. (2016) Identifying risk factors for ovine respiratory processes by using Bayesian networks. Small Rumin. Res., 136(3): 113-120. [Crossref]
18. Kumar, J., Dixit, S.K. and Kumar, R. (2015) Rapid detection of Mannheimia haemolytica in lung tissues of sheep and from bacterial culture. Vet. World, 8(9): 1073-1077. [Crossref] [PubMed] [PMC]
19. Black, P. and Nunn, M. (2009) Impact of Climate Change and Environmental Change on Emerging and Re-Emerging Animal Diseases and Animal Production. In: Compendium of Technical Items Presented in 77th General Session of the OIE World Assembly of Delegates, Paris, France, 24th-29th May. p15.
20. Sadeghian, S., Dezfouli, M.R.M., Kojouri, G.A., Bazargani, T.T. and Tavasoli, A. (2011) Pasteurella multocida pneumonic infection in goat: Hematological, biochemical, clinical and pathological studies. Small Rumin. Res., 100(2-3): 189-194. [Crossref]
21. Akloul, K. and Menoueri, M.N. (2016) Pneumonia in Algerian Ouled Djellal sheep: Bacteriological study and macroscopic aspect of lung lesions. Afr. J. Microbiol. Res., 10(40): 1685-1693. [Crossref]
22. Dassanayake, R.P., Shanthalingam, S., Herndon, C.N., Subramaniam, R., Lawrence, P.K., Bavananthasivam, J., Cassirer, E.F., Haldorson, G.J., Foreyt, W.J., Rurangirwa, F.R., Knowles, D.P., Besser, T.E. and Srikumaran, S. (2010) Mycoplasma ovipneumoniae can predispose bighorn sheep to fatal Mannheimia haemolytica pneumonia. Vet. Microbiol., 145(3-4): 354-359. [Crossref] [PubMed]
23. Jakab, G.J. (1982) Viral-bacterial interactions in pulmonary infection. Adv. Vet. Sci. Comp. Med., 26: 155-171. [PubMed]
24. Dassanayake, R.P., Shanthalingam, S., Subramaniam, R., Herndon, C.N., Bavananthasivam, J., Haldorson, G.J., Foreyt, W.J., Evermann, J.F., Herrmann-Hoesing, L.M., Knowles, D.P. and Srikumaran, S. (2013) Role of Bibersteinia trehalosi, respiratory syncytial virus, and parainfluenza-3 virus in bighorn sheep pneumonia. Vet. Microbiol., 162(1): 166-172. [Crossref] [PubMed]
25. Kumar, P., Singh, V.P., Agrawal, R.K. and Singh, S. (2009) Identification of Pasteurella multocida isolates of ruminant origin using polymerase chain reaction and their antibiogram study. Trop. Anim. Health Prod., 41(4): 573-578. [Crossref] [PubMed]
26. Sarangi, L.N., Thomas, P., Gupta, S.K., Priyadarshini, A., Kumar, S., Nagaleekar, V.K., Kumar, A. and Singh, V.P. (2015) Virulence gene profiling and antibiotic resistance pattern of Indian isolates of Pasteurella multocida of small ruminant origin. Comp. Immunol. Microbiol. Infect. Dis., 38(1): 33-39. [Crossref] [PubMed]
27. Gonzalez, C., Murtaugh, M.P. and Maheswaran, S.K. (1991) Genomic distribution of a serotype 1-specific antigen coding DNA fragment of Pasteurella haemolytica. J. Vet. Med. B Infect. Dis. Vet. Public Health, 38(8): 599-609. [Crossref]
28. Prabhakar, T.G., Prabhakar, P. and Venkadabadi (2010) Molecular characterization of Pasteurella multocida isolated from an incidence of sheep pasteurellosis in Karamadai hill tract of Tamil Nadu. Tamilnadu J. Vet. Anim. Sci., 6(2): 81-87.
29. Harper, M., Boyce, J.D. and Adler, B. (2006) Pasteurella multocida pathogenesis: 125 years after Pasteur. FEMS Microbiol. Lett., 265(1): 1-10. [Crossref]
30. Harmon, B.G., Glisson, J.R., Latimer, K.S., Steffens, W.L. and Nunnally, J.C. (1991) Resistance of Pasteurella multocida A: 3, 4 to phagocytosis by turkey macrophages and heterophils. Am. J. Vet. Res., 52(9): 1507-1511. [PubMed]
31. Aski, H.S. and Tabatabaei, M. (2016) Occurrence of virulence-associated genes in Pasteurella multocida isolates obtained from different hosts. Microb. Pathog., 96(7): 52-57. [Crossref] [PubMed]
32. Singh, F., Sonawane, G.G. and Meena, R.K. (2018) Molecular detection of virulent Mannheimia haemolytica and Pasteurella multocida in lung tissues of pneumonic sheep from semiarid tropics, Rajasthan, India. Turk. J. Vet. Anim. Sci., 42(6): 556-561. [Crossref]
33. Singh, F., Sonawane, G.G. and Meena, R.K. (2019) Pathology, isolation and characterisation of virulent and diverse Mannheimia haemolytica and Pasteurella multocida associated with fatal pneumonia in sheep, Rajasthan, India. Comp. Clin. Path., doi.org/10.1007/s00580-019-02916-z. [Crossref]
34. Tabatabaei, M. and Abdollahi, F. (2018) Isolation and identification of Mannheimia haemolytica by culture and polymerase chain reaction from sheep's pulmonary samples in Shiraz, Iran. Vet. World, 11(5): 636-641. [Crossref] [PubMed] [PMC]