Diagnosis of Chlamydia abortus by Isolation in Cell Culture and Real Time PCR in Aborted Sheep and Goats
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Abstract
Background: Ovine enzootic abortion (OEA) caused by Chlamyidia abortus is one of the most important abortive disease in small ruminants. Diagnosis of Ovine enzootic abortion depends on the isolation and detection of the agent or its nucleic acid. The aim of the present study was to detect Chlamydia abortus using both isolation method and real-time PCR in Brucella free flocks in Iran.
Methods: Twenty-eight vaginal and conjunctival swab samples which were Chlamydia abortus seropositive, were selected from ewes and does with recent abortion. Then the samples were tested by real-time PCR and positive molecular samples were inoculated into McCoy cells.
Results: Using real-time PCR, 18 samples (64.3%) were positive and 7 (25%) of them were isolated in cell culture.
Conclusion: The present results indicate that Chlamydia can play a relatively significant role in the abortion in does and ewes in Iran. Although the isolation of Chlamydia abortus have 100% specificity, because of low sensitivity, time consuming, cost and high probability of contamination, it is not suitable for routine laboratory diagnosis. Therefore, applying real-time PCR which have high sensitivity and specificity is recommended.
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2. Ghorbanpoor M, Seed G, Razieh H. Serological Study on Enzootic Abortion of Ewes in Ahvaz, Iran. J Anim Vet Adv 2007; 6(10):1194-96.
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12. Livingstone M, Wheelhouse N, Maley SW, et al. Molecular detection of Chlamydophila abortus in post-abortion sheep at oestrus and subsequent lambing. Vet Microbiol 2009; 135(1-2):134-41.
13. Esmaeili H, Sharifi M, Gharagozloo F, et al. Infectious and non-infectious causes of abortion in Saanen and Alpine goats. J Med Bacteriol 2023; 11:1-9.
14. Pantchev A, Sting R, Bauerfeind R, et al. New real-time PCR tests for species-specific detection of Chlamydophila psittaci and Chlamydophila abortus from tissue samples. Vet J 2009; 181:145-50.
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17. Puysseleyr KD, Puysseleyr LD, Geldhof J, et al. Development and validation of a real-time pcrforChlamydia suis diagnosis in swine and humans. PlosOne 2014; 9:1-7.
18. Wilde J, Eiden J, Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol 1990; 28:1300-7.
19. Livingstone M, Entrican G, Wattegedera S, et al. Antibody responses to recombinant protein fragments of the major outer membrane protein and polymorphic outer membrane protein POMP90 in Chlamydophila abortus-infected pregnant sheep. Clin Vaccine Immunol 2005; 12 (6):770–7.
20. O’Neill LM, O’Driscoll Á, Markey B. Comparison of three commercial serological tests for the detection of Chlamydia abortus infection in ewes. Ir Vet J 2018; 71:1-9.
21. Osman KM, Ali HA, ElJakee JA, et al. Chlamydophila psittaci and Chlamydophila pecorum infections in goats and sheep in Egypt. Rev Sci Tech Off Int Epizoot 2011; 30(3):939-48.
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23. Andersen, A., Vanrompay, D., 2008. Chlamydiosis. In: Dufour-Zavala, L., Swayne, D.E., Glisson, J., Pearson, J., Reed, W., Jackwood, M. (Ed.), A laboratory manual for the isolation, identification, and characterization of avian pathogens. American Association of Avian Pathologists, Athens, pp: 65-74.
24. Amin AS. Comparison of polymerase chain reaction and cell culture for the detection of Chlamydophila species in the semen of bulls, buffalo-bulls, and rams. Vet J 2003; 166(1):86-92.
25. McElena C, Cross G. Methods of detection of Chlamydia psittaciin domesticated and wild birds. Aust Vet J 1999; 77:516-21.
26. Jenab A, Golbang N, Golbang P, et al. Diagnostic value of PCR and ELISA for Chlamydia trachomatis in a group of asymptomatic women in Isfahan, Iran. Int J fertil Steril 2009; 2:193-8.
27. Esmaeili H, Hamedi M, Madani SA. Isolation of Chlamydia spp. from ewes and does in Iran. Arch Razi Inst 2017; 72(4):249-53.
2. Ghorbanpoor M, Seed G, Razieh H. Serological Study on Enzootic Abortion of Ewes in Ahvaz, Iran. J Anim Vet Adv 2007; 6(10):1194-96.
3. Hamedi M, Esmaeili H, Madani SA et al. The frequency of abortion caused by Chlamydia abortus in aborted fetuses of sheep and goats in Iran. J Med Bacteriol 2020; 9:1-8.
4. Esmaeili H, Bolourchi M, Mokhber-Dezfouli MR, et al. Detection of Chlamydia abortus and risk factors for infection in small ruminants in Iran. Small Rumin Res 2021; 197:106339.
5. Esmaeili H, Shakeri A, Nosrati Rad Z, et al. Causes of abortion in Iranian sheep flocks and associated risk factors. Vet Res Commun 2022; 46:1227-38.
6. Esmaeili H, Bolourchi M, Mokhber-Dezfouli MR. Seroprevalence of Chlamydia abortus infection in sheep and goats in Iran. Iran J Vet Med 2015; 9:73-7.
7. World health organization for animal health, 2018. Enzootic abortion of ewes. WOAH, France, Chapter: 3.7.5. pp. 1456-65.
8. Berri M, Rekiki A, Boumedine KS, et al. Simultaneous differential detection of Chlamydophila abortus, Chlamydophila pecorum and Coxiella burnetii from aborted ruminant's clinical samples using multiplex PCR. BMC Microbiol 2009; 9(1):130.
9. Madani A, Peighambari M, Barin A. Isolation of Chlamydophila psittaci from pet birds in Iran. Iran J Vet Res 2011; 5:95-8.
10. Soomro SA, Murray RD, Woldehiwet Z, et al. Comparisons of polymerase chain reaction and isolation in cell culture and embryonated hen’s eggs for the detection of Chlamydophila abortus in aborted ovine/caprine placenta. Vet World 2012; 2:32-9.
11. Domeika M, Savicheva A, Sokolovskiy E, et al. Guidelines for the laboratory diagnosis of Chlamydia trachomatis infections in East European coubtries. J Eur Acad Dermatol Venereol 2009; 23:1353-63.
12. Livingstone M, Wheelhouse N, Maley SW, et al. Molecular detection of Chlamydophila abortus in post-abortion sheep at oestrus and subsequent lambing. Vet Microbiol 2009; 135(1-2):134-41.
13. Esmaeili H, Sharifi M, Gharagozloo F, et al. Infectious and non-infectious causes of abortion in Saanen and Alpine goats. J Med Bacteriol 2023; 11:1-9.
14. Pantchev A, Sting R, Bauerfeind R, et al. New real-time PCR tests for species-specific detection of Chlamydophila psittaci and Chlamydophila abortus from tissue samples. Vet J 2009; 181:145-50.
15. Thrusfield, M., 2005. Veterinary epidemiology, 2nd ed, Blackwell Science, Oxford.
16. Sachse K, Vretou E, Livingstone IM, et al. Recent developments inthe laboratory diagnosisof chlamydial infections (Review). Vet Microbiol 2009; 135:2-21.
17. Puysseleyr KD, Puysseleyr LD, Geldhof J, et al. Development and validation of a real-time pcrforChlamydia suis diagnosis in swine and humans. PlosOne 2014; 9:1-7.
18. Wilde J, Eiden J, Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol 1990; 28:1300-7.
19. Livingstone M, Entrican G, Wattegedera S, et al. Antibody responses to recombinant protein fragments of the major outer membrane protein and polymorphic outer membrane protein POMP90 in Chlamydophila abortus-infected pregnant sheep. Clin Vaccine Immunol 2005; 12 (6):770–7.
20. O’Neill LM, O’Driscoll Á, Markey B. Comparison of three commercial serological tests for the detection of Chlamydia abortus infection in ewes. Ir Vet J 2018; 71:1-9.
21. Osman KM, Ali HA, ElJakee JA, et al. Chlamydophila psittaci and Chlamydophila pecorum infections in goats and sheep in Egypt. Rev Sci Tech Off Int Epizoot 2011; 30(3):939-48.
22. Vanrompay, D., 2000. Laboratory medicine: avian and exotic pets, 1st ed. WB Saunders, London.
23. Andersen, A., Vanrompay, D., 2008. Chlamydiosis. In: Dufour-Zavala, L., Swayne, D.E., Glisson, J., Pearson, J., Reed, W., Jackwood, M. (Ed.), A laboratory manual for the isolation, identification, and characterization of avian pathogens. American Association of Avian Pathologists, Athens, pp: 65-74.
24. Amin AS. Comparison of polymerase chain reaction and cell culture for the detection of Chlamydophila species in the semen of bulls, buffalo-bulls, and rams. Vet J 2003; 166(1):86-92.
25. McElena C, Cross G. Methods of detection of Chlamydia psittaciin domesticated and wild birds. Aust Vet J 1999; 77:516-21.
26. Jenab A, Golbang N, Golbang P, et al. Diagnostic value of PCR and ELISA for Chlamydia trachomatis in a group of asymptomatic women in Isfahan, Iran. Int J fertil Steril 2009; 2:193-8.
27. Esmaeili H, Hamedi M, Madani SA. Isolation of Chlamydia spp. from ewes and does in Iran. Arch Razi Inst 2017; 72(4):249-53.
| Files | ||
| Issue | Vol 12 No 4 (2024) | |
| Section | Original Articles | |
| DOI | https://doi.org/10.18502/jmb.v12i4.17003 | |
| Keywords | ||
| Abortion Cell culture Chlamydia Real-time PCR Small ruminants. | ||
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How to Cite
1.
Esmaeili H, Hamedi M, Madani SA, Barin A, Haji Agha Khiyabani F. Diagnosis of Chlamydia abortus by Isolation in Cell Culture and Real Time PCR in Aborted Sheep and Goats. J Med Bacteriol. 2024;12(4):9-16.
