The Evaluation of Brucella Spp. Isolation Rates in Ruminant Abortion Cases by Using Different Selective Media

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The aim of this study is to evaluate the success of Brucella spp. isolation in ruminant abortion cases by using different selective media. To this end, 58 samples from ruminant abortion cases were utilized. 4 selective media; namely, Farrell Medium (FM), CITA Medium (CM), Modified Thayer Martin (MTM) and Jones & Morgan (JM) were preferred for isolation. In addition to these, one medium with antibiotics was used to extend the range of the results. Suspensions prepared from organ and fetal stomach contents were inoculated to media plates and incubated at 37C° for 5-8 days in 5-10% CO2 condition. Conventional biotyping method was used to identify Brucella isolates within the level of species and biovar. MTM (67.2%) and Farrell (65.5%) outperformed the other media with regards to isolation rate. However, regarding the inhibition ability against contaminant microrganisms, Farrell (86.2%) and CITA (72%) have the highest and second highest percentages respectively. The media’s inhibition ability was examined in the samples in which Brucella spp. isolation occurred to be able to investigate the correlations between isolation and inhibition. Lower isolation percentage was observed in the samples in which the media displayed the lowest inhibition ability against contaminants. In this context, using two different selective media with high inhibition ability against contaminants may be recommended to enhance the isolation rate. Moreover, the components stimulating the growth of Brucella strains might be added to the media to obtain better results.

1. Godfroid J., Cloeckaert A., Liautard J.P., Kohler S., Fretin D., Walravens K., Garin-Bastuji B., Letesson J.J. (2005). From the discovery of the Malta fever’s agent to the discovery of a marine mammal reservoir, brusellosis has continously been a re-emerging zonosis. Vet. Res. 36, 313–326. PMid:15845228

2. Songer, J.G., K.W. Post. (2012). Brusella cinsi. [Brusella genus]. In: O. Ang, Y. Ozgur, (Eds.), Veteriner Hekimlik Mikrobiyolojisi-Hayvan Hastalığı Etkeni Olan Bakteriler ve Mantarlar. [Veterinary Microbiology: Bacterial and Fungal Agents of Animal Disease] (pp. 200-207). Istanbul, Turkey: Nobel Press. [in Turkish]

3. Yumuk Z., O’Callaghan D. (2012). Brucellosis in Turkey-an overview. Int. J. Infect. Dis. 16 (4): 228-35. PMid:22333223

4. Pappas G., Panagopoulou P., Chistou L., Akritidis N. (2006). Brucella as a biological weapon. Cell. Mol. Life. Sci. 63, 2229-2236. PMid:16964579

5. Seleem M.N., Boyle S.M., Sriranganathan N. (2010). Brucellosis: A re-emerging zoonosis. Vet. Microbiol. 140 (3-4): 392-398. PMid:19604656

6. Aydın N. (1997). Gram negatif küçük çomaklar -Brusella infeksiyonları. [Small gram negative cocci - Brucella infections]. In: M. Arda, A. Minbay, N. Leleoglu, N. Aydın, M. Kahraman, O. Akay, K.S. Diker, Özel mikrobiyoloji kitabı (pp. 110-124). Ankara, Türkiye: Medisan Yayınevi. [The Microbiology Book (pp.110-124). Ankara, Turkey: Medisan Press]. [in Turkish]

7. Doganay M., Aygen B. (2003). Human brucellosis: an overview. Int. J. Infect. Dis. 7, 173-182.

8. Perez-Sancho M., Garcia-Seco T., Dominguez L., Alvarez J. (2015). Control of animal brucellosis, The most effective tool to prevent human brucellosis. [accessed on 09.24.2016].

9. World Health Organisation. (2004). Laboratory safety manual, Third edition. Geneva.

10. OIE. Terrestrial Manual Chapter 2.7.2. (2009). Caprine and ovine brucellosis.

11. OIE. Terrestrial Manual Chapter 2.4.3. (2012). Bovine brucellosis.

12. Godfroid, J., Scholz, H.C., Barbier, T., Nicolas, C., Wattiau, P., et al. (2011). Brucellosis at the animal/ecosystem/human interface at the beginning of the 21st century. Prev Vet. Med. 102, 118-131. PMid:21571380

13. Krstevski K., Naletoski I., Mitrov D., Mrenoshki S., Cvetkovikj I., Janevski A., Dodovski A., Djadjovski I. (2015). Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples. Mac Vet Rev. 38 (2): 223-232.

14. Marin, C.M., Alabart, J.L., Blasco, J.M. (1996). Effect of antibioctics contained in two Brucella selective media on growth of Brucella abortus, B. melitensis, and B. ovis. J. Clin. Microbiol. 34 (2): 426-428. PMid:8789029 PMCid:PMC228811

15. Stack, J.A., Harrıson, M., Perrett, L.L. (2002). Evaluation of a selective medium for Brucella isolation using natamycin. J. Appl. Microbiol. 92, 724–728. PMid:11966913

16. Hornsby, R.L., Jensen, A.E., Olsen, S.C., Thoen, C.C. (2000). Selective media for isolation of Brucella abortus strain RB51. Vet. Microbiol. 73, 51-60.

17. Marin, C.M., Jimenez De Bagues, M.P., Barberan, M., Blasco, J.M.(1996). Comparison of two selective media for the isolation of Brucella melitensis from naturally infected sheep and goats. Vet. Res. 138, 409-411.

18. Vicente, A.F., Antunes, J.M., Lara, G.H., Mioni, M.S.R., Allendorf, S.D., et al. (2014). Evaluation of three formulations of culture media for isolation of Brucella spp. regarding their ability to inhibit the growth of contaminating organisms. Biomed. Res. Int. 702072. PMid:24949466 PMCid:PMC4052881

19. Pappas, G. (2010). The changing Brucella ecology: novel reservoirs, new threats. Int. J. Antimicrob. Ag. 365, 8-11. PMid:20696557

20. Farrell, I.D. (1974). The development of a new selective medium for the isolation of Brucella abortus from contaminated sources. Res. Vet. Sci. 16, 280-286. PMid:4369280

21. De Miguel, M.M., Marin, C.M., Munoz, P.M., Dieste, L., Grillo, M.J., Blasco, J.M. (2011). Development of a selective culture medium for primary isolation of the main Brucella species. J. Clin. Microbiol. 49(4): 1458-1463. PMid:21270216 PMCid:PMC3122841

22. Jones, L.M., Morgan, W.W. (1958). A preliminary report on a selective medium for the culture of Brucella, including fastidous types. Bull. World. Health. Organ. 19 (1): 200-203. PMid:13585070 PMCid:PMC2537688

23. Drancourt, M., Raoult, D. (2007). Cost-effectiveness of blood agar for isolation of Mycobacteria. Plos. Negl. Trop. D. 1, 83. PMid:18060087 PMCid:PMC2100370

24. Martin, W.K., Mattick, K.L., Harrison, M., Humphrey, T.J. (2002). Evaluation of selective media for Campylobacter isolation when cycloheximide is replaced with amphotericin B. Lett. Appl. Microbiol. 34, 124–129. PMid:11849508

25. Alton, G.G., Jones, L.M., Angus, R.D., Verger, J.M. (1988). Techniques for the brucellosis laboratory. Paris, France: Institut National de la Recherche Agromique-INRA.

26. International Organisation for Standartization. ISO/TS Technical Spefication 11133-1. (2009). Microbiology of food and animal feeding guidelines on preparation and production of culture media. Geneva, Switzerland: ISO.

27. Her, M., Choa, D.H., Kang, S.I. (2010). The development of a selective medium for the Brucella abortus strains and its comparison with the currently recommended and used medium. Diagn. Microbiol. Infect. Dis. 67, 15–21. PMid:20385349

28. Jones, L.M., Dubray, G., Marly, J. (1975). Comparison of methods of diagnosis of Brucella ovis infection of rams. INRA editions. 6 (1): 11-22.

29. O’ Grady, D., Byrne, W., Kelleher, P., O’callahan, H., Kenny, K., Heneghan, T., Power, S., Egan, J, Ryan, F. (2014). A comparative assessment of culture and serology in the diagnosis. Vet. J. 199, 370-375. PMid:24507882

30. Ferreira, A.C., Almendra, C., Cardoso, R., Pereira, M.S., Pereira, A.B., Luikart, G., Correa De Sa, M.I. (2012). Development and evaluation of a selective medium for Brucella suis. Res. Vet. Sci. 93, 565-567. PMid:21968103

31. Nardi Junior, G., Megid, J., Vicente, A.F. (2015). Comparison of Brucella agar, CITA and Farrell media for selective isolation of Brucella abortus from semen of bovine bulls. Afr. J. Microbiol. Res. 9 (9): 617-620.

32. Fatolahzadeh, B., Maleknejad, P., Hejazi, M.J., Pyri, H. (2009). Development and evaluation of TUMS medium, a novel biphasic culture medium for isolation of Brucella spp. from patients. Iran. J. Microbiol.1 (2): 21-25.

33. Keppie, J., Williams, A., Witt, K., Smith, H. (1965). The role of erythritol in tissue localization of the Brucellae. Brit. J. Exp. Pathol. 46, 104–108. PMid:14295553 PMCid:PMC2093692

34. Garcia-Lobo, J.M., Sangari Garcia, J.F. (2005). Erythritol metobolism and virulence in Brucella. In: I. Lopez-Goni, I. Moriyon (Eds.), Brucella, Molecular and Celular Biology, (pp.223-236). Spain: Taylor & Francis.

35. Seleem, M.N., Boyle, S.M., Srıranganathan N. (2008). Brucella: A pathogen without classic virulence genes. Vet. Microbiol. 129, 1-14. PMid:18226477

36. Poester, F.P., Samartino, L.E., Santos, R.L. (2013). Pathogenesis and pathology of Brucellosis in livestock. Revue scientifique et technique (International Office of Epizootics). 32 (1): 105-115. PMid:23837369

37. Sperry, J.F., Robertson, D.C. (1975). Inhibition of Growth by Erythritol Catabolism in Brucella abortus. J. Bacteriol. 124 (1): 391-397. PMid:170249 PMCid:PMC235907

Macedonian Veterinary Review

The Journal of the Faculty of Veterinary Medicine-Skopje at the Ss. Cyril and Methodius University in Skopje

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CiteScore 2017: 0.32

SCImago Journal Rank (SJR) 2017: 0.195
Source Normalized Impact per Paper (SNIP) 2017: 0.387


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