Morphometric and genetic characterization of Oestrus ovis in sheep in different climatic regions of Iran

Document Type : Research Article


Department of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.


Oestrosis is nasal myiasis, which results from infestation with the larvae of flies that belong to the genus Oestrus. Nasal discharge and sneezing are the most common clinical signs in infected animals. Myiasis larvae were collected from sheep in different climatic regions of Iran. Morphological identification of the larvae was made based on the diagnostic keys. The species was confirmed by PCR amplifying the partial fragment (610 bp) of the mtCOI gene. Genetic distance was assessed in COI sequences, and a phylogenetic tree was drawn. Sequencing showed no difference in the partial COI gene among the Iranian isolates, and this gene had a high similarity with the sequences of O. ovis isolates from Iraq, Bosnia and Herzegovina, and Croatia. The present study provided the first molecular dataset for O. ovis species, which is crucial for the phylogenetic relationships assessment and the molecular identification of these parasites. 


1. Horak I. Parasites of domestic and wild animals in South Africa. XLVI 1-Oestrid fly larvae of sheep, goats, springbok, and black wildebeest in the Eastern Cape Province. Onderstepoort J Vet Res. 2005; 72(4): 315–20. Doi:10.4102/ojvr.v72i4.188.
2. Hanan BA. Seasonal prevalence of Oestrus ovis L.(Diptera: Oestridae) larvae in infested sheep in Jazan Region, Saudi Arabia. J Parasitol Vector Biol. 2013; 5(5): 66-71. Doi: 10.5897/JPVB2013.0112.
3. Angulo-Valadez CE, Scholl PJ, Cepeda-Palacios R, Jacquiet P, Dorchies P. Nasal bots a fascinating world! Vet Parasitol. 2010; 174(1-2): 19–25. Doi:10.1016/j.vetpar.2010.08.011 .
4. Dorchies PH, Tabouret G, Hoste H, Jacquiet PH. Oestrinae host-parasite interactions. In: Colwell DD, Hall MJR, Scholl PJ. (éd.): The Oestrid flies: biology, host-parasite relationships, impact and management, CABI, UK. 2006; 191-200. Doi: 10.1016/j.vetpar.2010.08.011.
 5. Ogo NI, Onovoh E, Okubanjo OO, Galindo RC, Lastra JMP. Molecular identification of Cordylobia anthropophaga Blanchard (Diptera: Calliphoridae) larvae collected from dogs (Canis familiaris) in Jos South, Plateau State, Nigeria. Onderstepoort Vet Res. 2012; 79(1):1-4. Doi:10.4102/ojvr.v79i1.349.
6. Otranto D, Traversa D. Molecular evidence indicating that Przhevalskiana silenus, Przhevalskiana aegagri, and Przhevalskiana crossii (Diptera, Oestridae) are one species. Acta Parasitol. 2004; 2(49): 173–6. Doi:
7. Otranto D, Colwell DD, Traversa D, Stevens JR. Species identification of Hypoderma (Diptera, Oestridae) affecting domestic and wild ruminants by Scanning Electron Microscopy (SEM) and molecular characterization of the cytochrome oxidase I (COI) gene. Med Vet Entomol. 2003; 17: 316–25. Doi:10.1046/j.1365-2915.2003.00446.x.
8. Otranto D, Traversa  D, Colwell DD, Guan G, Giangaspero A, Boulard C, Yin H. A third species of Hypoderma (Diptera, Oestridae) affecting cattle and yaks in China: molecular and morphological evidence. J Parasitol. 2004; 90(5): 958–65. Doi:10.1645/GE-232R.
9. Lunt DH, Zhang DX, Szymura JM, Hewitt GM. The insect cytochrome oxidase I gene: evolutionary patterns and conserved primers for phylogenetic studies. Insect Mol Biol. 1996; 5(3): 153–65. Doi:10.1111/j.1365-2583.1996.tb00049.x.
10. Hebert PD, Stoeckle MY, Zemlak TS, Francis CM. Identification of birds through DNA Barcodes. PLoS Biol. 2004; 2(10):e312. Doi:10.1371/journal.pbio.0020312. 
11. Moreno V, Romero-Fernández I, Marchal JA, Beltrán M, Granados JE, Habela MA, Tamadon A, Rakhshandehroo E, Sarasa M, Pérez JM, Sánchez A. Molecular characterization of bot flies, Oestrus spp (Diptera, Oestridae), from domestic and wild Bovidae hosts. Vet Parasitol. 2015; 212(3-4): 473-7. Doi:10.1016/j.vetpar.2015.08.002. 
12. Cavallero S, Pombi M, Perrone V, Milardi GL, D’Amelio S, Giuliani C, Gabrielli S. Gasterophilus intestinalis (Diptera: Oestridae) in the diaphragmatic muscle: An unusual finding. Vet Parasitol. 2017; 237: 117-21. Doi:10.1016/j.vetpar.2015.08.002. Doi:10.1016/j.vetpar.2017.02.030.
13. Rakhshandehroo E, Razavi SM, Farzaneh R, Esmailnejad A, Asadpour M, Shams S. Phylogenetic analysis of goat warble fly (Przhevalskiana silenus) based on mitochondrial COI gene. J Parasit Dis. 2019; 43(2): 304-7. Doi:10.1007/s12639-019-01093-8.
14. Li XY, Pape T, Zhang D. Gasterophilus flavipes (Oestridae: Gasterophilinae) A horse stomach bot fly brought back from oblivion with morphological and molecular evidence. PLoS One. 2019;14(8):e0220820. Doi:10.1371/journal.pone.0220820.
 15. Zumpt, F. Myiasis in Man and Animals in the Old World: A Textbook for Physicians, Veterinarians, and Zoologists; 1965.
16. Otranto D, Milillo P, Traversa D, Colwell DD. Morphological variability and genetic identity in Rhinoestrus spp. Causing horse nasal myiasis. Med Vet Entomol. 2005; 19(1): 96–100. Doi:10.1111/j.0269-283X.2005.00553.x.
17. Moreno V, Romero-Fernández I, Marchal JA, Beltrán M, Granados JE, Habela MA, Tamadon A, Rakhshandehroo E, Sarasa M, Pérez JM, Sánchez A. Molecular characterization of bot flies, Oestrus spp., (Diptera, Oestridae) from domestic and wild Bovidae hosts. Vet Parsitol. 2015; 212(3-4): 473–77. Doi:10.1016/j.vetpar.2015.08.002.
18. Pape T. Phylogeny of Oestridae (Insecta: Diptera). Syst Entomol. 2001; 26(2):
19. Guo Y, Cai J, Chang Y, Li X, Liu Q, Wang X., Wang X, Zhong M, Wen J, Wang J. Identification of forensically important sarcophagid flies (Diptera: Sarcophagidae) in China, based on COI and 16S rDNA gene sequences. J Forensic Sci. 2011; 56(6): 1534-40. Doi:10.1111/j.1556-4029.2011.01882.x.
20. Tan X, Lu H, Sun Y, Chen X, Wang D, Jia F, Zhang K. Expanding the materials space of DNA via organic-phase ring-opening metathesis polymerization. Chem. 2019; 5(6): 1584-96. Doi:10.1016/j.chempr.2019.03.023.
21. Banafshi O, Hanafi-Bojd AA, Karimi M, Faghihi F, Beik-Mohammadi M, Gholami S, Javaherizadeh S, Edalat H, Vatandoost H, Telmadarraiy Z. Tick ectoparasites of animals in borderline of Iran-Iraq and their role on disease transmission. J Arthropod Borne Dis. 2018; 12(3): 252. Doi:10.18502/jad.v12i3.77.
22. Kettle PR. A study on the sheep botfly, Oestrus ovis (Diptera: Oestridae) in New Zealand. N Z Entomol. 1973. 5(2): 185-191. Doi:10.1080/00779962.1973.9722998.
23. Metwally DM, Albasyouni SA, Barakat IAH, Al-Turaiki IM, Almuhanna AM, Bashir MA, Baghdadi HB, El-Khadragy MF, Alajmi RA. Prevalence Rate and Molecular Characteristics of Oestrus ovis L.(Diptera, Oestridae) in Sheep and goats from Riyadh, Saudi Arabia. Animals. 2021; 11(3): 689. Doi:10.3390/ani11030689.
24. Negm-Eldin MM, Elmadawy RS, Hanan GM. Oestrus ovis larval infestation among sheep and goats of Green Mountain areas in Libya. J Adv Vet Anim Res. 2015; 2(4): 382-7. Doi:10.5455/javar.2015.b105.
 25. Bosly, AH. Molecular identification studies on Oestrus ovis L. (Diptera: Oestridae) larvae-infested sheep in Jazan region, Saudi Arabia. Indian J Anim Res. 2018; 52 (1): 105-10. Doi:10.18805/ijar.v0iOF.8487.
26. Otranto D, Stevens JR. Molecular approaches to the study of myiasis-causing larvae. Int J Parasitol. 2002; 32(11): 1345-60. Doi:10.1016/S0020-7519(02)00095-4.
27. Ogo, NI, Onovoh E, Okubanjo OO, Galindo RC, Lastra JMP. Molecular identification of Cordylobia anthropophaga Blanchard (Diptera: Calliphoridae) larvae collected from dogs (Canis familiaris) in Jos South, Plateau State, Nigeria. Onderstepoort J Vet Res. 2012; 79(1): 01-04. Doi:10.4102/ojvr.v79i1.349
28. Ames C, Turner B, Daniel B. The use of mitochondrial cytochrome oxidase I gene (COI) to differentiate two UK blowfly species–Calliphora vicina and Calliphora vomitoria. Forensic Sci Int. 2006; 164(2): 179-82. Doi: 10.1016/j.forsciint.2006.01.005 .
29. Hall M, Adams Z, Wyatt N, Testa J, Edge W, Nikolausz M, Farkas R, Ready P. Morphological and mitochondrial DNA characters for identification and phylogenetic analysis of the myiasis causing flesh fly Wohlfahrtia magnifica and its relatives, with a description of Wohlfahrtia monegrosensis sp. n. Wyatt & Hall. Med Vet Entomol. 2009; 23(s1): 59-71. Doi: 10.1016/j.forsciint.2006.01.005.
31. Wang Y, Liu GH, Li JY, Xu MJ, Ye YG, Zhou DH, Song HQ, Lin RQ, Zhu XQ. Genetic variability among Trichuris ovis isolates from different hosts in Guangdong Province, China revealed by sequences of three mitochondrial genes. mtDNA. 2013; 24(1): 50-54. Doi: 10.3109/19401736.2012.710210.
32. Aleix-Mata G, Peréz JM, Sánchez A.  The complete mitochondrial genome of Oestrus ovis (Linnaeus, 1758)(Diptera: Oestridae). MitDNA Part B. 2021; 6(7):1847-48. Doi:10.1080/23802359.2021.1934174.
33. Hall MJ, Smith KG. Diptera causing myiasis in man. Medical insects and arachnids, Springer. 1993; 429-69. Doi: 10.1007/978-94-011-1554-4_12.
34. Blacket MJ, Semeraro L, Malipatil MB. Barcoding Queensland fruit flies (Bactrocera tryoni): impediments and improvements. Mol Ecol Resour. 2012; 12(3): 428-36. Doi: 10.1111/j.1755-0998.2012.03124.x.
35. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase Subunit 1 from diverse metazoan invertebrates. Mol Marine Biol Biotechnol. 1994; 3(5): 294-9. PMID: 7881515.