Phylogenetic and molecular analysis based on genes 16S-rRNA, OMPA and POMP to identify Chlamydia abortus infection occurrence at the milk samples of goats and sheep in west Azerbaijan of Iran
Abstract
Background and Objectives: Enzootic abortion in sheep and goats, also called ovine enzootic abortion (OEA) or enzootic abortion of ewes (EAE), is caused by Chlamydia abortus. The disease has a major economic impact as it represents the most important cause of lamb loss in sheep in parts of Europe, North America and Africa. This serious and potentially life-threatening zoonosis can also affect pregnant women after contact with lambing ewes, leading to severe febrile illness in pregnancy and loss of the foetus.
Materials and Methods: The present study was conducted to the Phylogenetic and Molecular Analysis based on Genes 16S-rRNA, OmpA and POMP of C. abortus in milk samples collected from sheep and goats in West Azerbaijan province, Iran. During 2018, a total number of 360 milk samples were collected from sheep (n = 180) and goats (n = 180) of different regions of the province. All milk samples were subjected to DNA extraction and examined by PCR.
Results: Among 360 milk samples collected from sheep and goats, 31 (8.611%; 95% CI=6.13-11.96) were positive for Chlamydia spp. The helicase, 16S-rRNA and ompA genes were examined and resulted in 8, 31, 31 of positive samples respectively. The accession numbers have been deposited in GenBank (NCBI) (MT367602 and MT367603).
Conclusion: Phylogenetic analysis based on the gene of helicase showed that most of the isolates shared similarity > 99.97%.
2. Ghorbanpoor M, Goraninejad S, Heydari R. Serological study on enzootic abortion of ewes in Ahvaz, Iran. J Anim Vet Adv 2007; 6: 1194-1196.
3. Barati S, Moori-Bakhtiari N, Najafabadi MG, Momtaz H, Shokuhizadeh L. The role of zoonotic Chlamydial agents in ruminants abortion. Iran J Microbiol 2017; 9: 288-294.
4. Siarkou V, Lambropoulos AF, Chrisafi S, Kotsis A, Papadopoulos O. Subspecies variation in Greek strains of Chlamydophila abortus. Vet Microbiol 2002; 85: 145-157.
5. Reinhold P, Jaeger J, Liebler-Tenorio E, Berndt A, Bachmann R, Schubert E, et al. Impact of latent infections with Chlamydophila species in young cattle. Vet J 2008; 175: 202-211.
6. Berri M, Bernard F, Lecu A, Ollivet-Courtois F, Rodolakis A. Molecular characterisation and ovine live vaccine 1B evaluation toward a Chlamydophila abortus strain isolated from springbok antelope abortion. Vet Microbiol 2004;103:231-240.
7. Berri M, Rekiki A, Boumedine KS, Rodolakis A. Simultaneous differential detection of Chlamydophila abortus, Chlamydophila pecorum and Coxiella burnetii from aborted ruminant's clinical samples using multiplex PCR. BMC Microbiol 2009;9:130.
8. Greco G, Totaro M, Madio A, Tarsitano E, Fasanella A, Lucifora G, et al. Detection of Chlamydophila abortus in sheep and goat flocks in southern Italy by PCR using four different primer sets. Vet Res Commun 2005;29 Suppl 1:107-115.
9. Güler L, Hadimli H, Erganiş O, Ateş M, Ok Ü, Gündüz K. Field evaluation of a PCR for the diagnosis of Chlamydial abortion in sheep. Vet Rec 2006;159:742-745.
10. White CH, Bishop JR, Morgan DM. Microbiological methods for dairy products. In: Marshall, R. T. (ed.), Standard Methods for the Examination of Dairy Products. Washington, DC 1993:287-308.
11. Cetinkaya B, Karahan M, Atil E, Kalin R, De Baere T, Vaneechoutte M. Identification of Corynebacterium pseudotuberculosis isolates from sheep and goats by PCR. Vet Microbiol 2002;88:75-83.
12. Daibata M, Nemoto Y, Togitani K, Fukushima A, Ueno H, Ouchi K, et al. Absence of Chlamydia psittaci in ocular adnexal lymphoma from Japanese patients. Br J Haematol 2006; 132: 651-652.
13. Zhaocai Li, Xiaoan Cao, Baoquan Fu, Yilin Chao, Jinshan Cai, Jizhang Zhou. Identification and characterization of Chlamydia abortus isolates from yaks in Qinghai, China. Biomed Res Int 2015;2015: 658519.
14. Longbottom D, Coulter L. Animal Chlamydioses and zoonotic implications. J Comp Pathol 2003;128:217-244.
15. Borel N, Thoma R, Spaeni P, Weilenmann R, Teankum K, Brugnera E, et al. Chlamydia-related abortions in cattle from Graubunden, Switzerland. Vet Pathol 2006;43:702-708.
16. Szymanska-Czerwinska M, Niemczuk K, Galinska EM. Serological and nested PCR survey to determine the occurrence of Chlamydia infections in the Polish cattle population. Ann Agric Environ Med 2013;20:682-686.
17. Arif E, Saeed N, Shwan KR. Isolation and identification of Chlamydia abortus from aborted ewes in Sulaimani Province, Northern Iraq. Pol J Microbiol 2020; 69:1-7.
18. DeGraves FJ, Gao D, Hehnen H-R, Schlapp T, Kaltenboeck B. Quantitative detection of Chlamydia psittaci and C. pecorum by high-sensitivity real-time PCR reveals high prevalence of vaginal infection in cattle. J Clin Microbiol 2003;41:1726-1729.
19. Creelman JL, McCullough SJ. Evaluation of strain-specific primer sequences from an abortifacient strain of ovine Chlamydophila abortus (Chlamydia psittaci) for the detection of EAE by PCR. FEMS Microbiol Lett 2000;190:103-108.
20. Marsilio F, Martino B, Francesco C, Meridiani I. Diagnosis of ovine Chlamydial abortions by PCR-RFLP performed on vaginal swabs. Vet Res Commun 2005;29 Suppl 1:99-106.
21. Beeckman DSA, Vanrompay DCG. Zoonotic Chlamydophila psittaci infections from a clinical perspective. Clin Microbiol Infect 2009; 15: 11-17.
22. Laroucau K, Souriau A, Rodolakis A. Improved sensitivity of PCR for Chlamydophila using pmp genes. Vet Microbiol 2001;82:155-164.
23. Taban BM, Ben U, Aytac S. Rapid detection of Salmonella in milk by combined immunomagnetic separation-polymerase chain reaction assay. J Dairy Sci 2009; 92: 2382-2388.
24. Ghorbanpoor M, Bakhtiari N-M, Mayahi M, Moridveisi H. Detection of Chlamydophila psittaci from pigeons by polymerase chain reaction in Ahvaz. Iran J Microbiol 2015;7:18-22.
25. Jee J, DeGraves F, Kim T, Kaltenboeck B. High prevalence of natural Chlamydophila spp. infection in calves. J Clin Microbiol 2004; 42:5664-5672.
26. Pinheiro Junior JW, Mota RA, Piatti RM, Oliveira AAdF, Silva AMd, Abreu SRdO, et al. Seroprevalence of antibodies to Chlamydophila abortus in ovine in the State of Alagoas, Brazil. Braz J Microbiol 2010;41:358-364.
Files | ||
Issue | Vol 13 No 4 (2021) | |
Section | Original Article(s) | |
DOI | https://doi.org/10.18502/ijm.v13i4.6972 | |
Keywords | ||
Chlamydia; Nested polymerase chain reaction; Helicase; Outer membrane protein A (OmpA); Polymorphic outer membrane protein (POMP) |
Rights and permissions | |
![]() |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |