Isolation and identification of molds flora in external genital tract of healthy mares


1 Agriculture Research, Education and Extention Organisation (AREEO), Ministry of Agiculture Jahad, Iran & Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran


BACKGROUND: Microorganisms are responsible for development of many diseases, which directly or indirectly affect reproductive performance in mares, like various animal species. Fungi are present at mucous membranes of reproductive tract as mycoflora in warm blooded animals. ObjectiveS: The aim of this study was isolation and identification of molds of external reproductive tract in mares. Methods: Samples were collected from genital tract of 151 mares in different ages from horse breeding farms and clubs suburb of Tehran from April to December, 2014. Age, pregnancy, insemination and mating were recorded. Samples were taken by sterile cotton swabs from 3 different sites, (1) vestibule, (2) clitoral fossa and (3) vagina and transferred under cold condition to the laboratory of Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran,Iran. Samples were inoculated in Sabouraud glucose agar supplemented with antibiotic (chloramphenicol; 0.005) and kept at 30°C for 10-15 days before being considered negative. Fungal identification relied on morphologic and physiologic features. SPSS Version 20 was used for statistical analysis of the data. Results: Totally, 666 filamentous fungi colonies were isolated. Maximum colony (CFUs) were 20 and 37 colonies for each site and horse, respectively. In total, 81.5 percent of mares and 48.3 percent of different sites were positive. The most predominant isolates were Aspergillus spp 362(54.35%), Scopulariopsis 93(13.96%), Cladosporium 35(5.25%), Penicillium 31(4.65%), Alternaria 26(3.90%), Fusarium 20(3.03%). Aspergillus took high occurrence with 362 and A. fumigatus with 156 colonies followed by Scopulariopsis 93, Cladosporium 35 and Pencillium 31. A. fumigatus had significant difference among other fungi pConclusionS: Isolated filamentous fungi could be resident flora of external reproductive tract of mares. These saprophytic pathogen fungi in predisposing conditions could be causative agents of endometritis and infertility in mares.


Blue, M.G. (1983) Mycotic invasion of the mare’s uterus. Vet Rec. 113: 31-2.
Barnes, P.D., Marr, K.A. (2006) Aspergillosis: spectrum of disease, diagnosis, and treatment. Infect Dis Clin North Am. 20: 545-61.
Bouziane, H., Latge, J.P., Fitting, C., Mecheri, S., Lelong, M., David, B. (2005) Comparison of the allergenic potency of spores and mycelium of Cladosporium. Allergol Immunopathol (Madr). 33: 125-30.
Chengappa, M.M., Madux, R.I., Grer, S.C.  (1984) Isolation and identification of yeasts and yeastlike organisms from clinical veterinary sources. J Clin Microbiol. 19: 427-428.
Cleff, M.B., Lima, A.P., Faria, R.O., Meinerz, A.R.M., Antunes, T.A., Araujo, F.B. (2005) Isolation of Candida spp from vaginal microbiota of healthy canine females during estrous cycle. Br J Microbiol. 36: 201-204.
Couto, M.A., Hughes, J.P. (1993) Sexually transmitted (venereal) disease of horses. In: Equine Reproduction. McKinnon, A.O., Voss, J.L. (eds.).  (1st ed.) Philadelphia, USA. Lea & Febiger. p. 845-54.
Cuenca-Estrella, M., Gomez-Lopez, A., Mellado, E., Buitrago, M.J., Monzo´n, A., Rodriguez-Tudela, J., Unidad, L. (2003) Scopulariopsis brevicaulis, a fungal pathogen resistant to broad-spectrum antifungal agents. Antimicrobial agents and Chemotherapy. 47: 2339-2341.
Giles, R.C., Donahue, J.M., Hong, B., Tuttle, P.A., Petrites-Murphy, M.B., Poonacha, K.B. (1993) Causes of abortion, stillbirth, and perinatal death in horses: 3527 cases (1986 -1991). J Am Vet Med Ass. 203: 1170 -5.
Hong, C.B., Donahue, J.M., Giles, J.r.R.C., Petrites-Murphy, M.B., Poonacha, K.B., Roberts, A.W. (1993) Etiology and pathology of equineplacentitis. J Vet Diagn Invest. 5: 56-63.
de Hoog, G.S, Guarro, J., Gené, J., Figueras, M.J. (2000) Atlas of Clinical Fungi. (2nd ed.) Utrecht, Centraal Bureau Voor Schimmel Cultures. Utrecht, Netherlands.
Fidel, J.R., Cutright, J., Steele, C. (2000) Effects of reproductive hormones on experimental vaginal candidioasis. Infect Immunol. 68: 651-7.
Ismail, S., Chavoshi, M., Hussaini, J., Malekshaabani, Sh. (2014) Uterine Mycotic Flora in Healthy Buffaloes and Buffaloes with Reproductive Disorders. Life Science Journal. 11: 400-402.
Khosravi, A.R., Nikaein, D., Sharifzadeh, A., Gharagozlou, F. (2014) Ocular fungal flora from healthy horses in Iran. Science Direct J Medical Mycology. 24: 29-33.
Kontoyiannis, D.P., Lewis, R.E., May, G.S., Osherov, N., Rinaldi, M.G. (2002) Aspergillus nidulans is frequently resistant to amphotericin B. Mycoses. 45: 406-7.
Kwon-Chung, K.J., Bennett, J.E. (1992) Infections due to miscellaneous molds. Medical mycology. p. 733-767.
Laing, J.A., Bringley Morgan, W.J., Wagner, W.C. (1988) Fertility and Infertility in Veterinary Practice. (4th ed.) Bailliere Tindall. London, UK.
Lyke, K.E., Miller, N.S., Towne, L., Merz, W.G. (2001) A case of cutaneous ulcerative alternariosis: rare association with diabetes mellitus and unusual failure of itraconazole treatment. Clin Infect Dis. 32: 1178-1187.
Nema, H.V., Ahuja, O.P., Bal, A., Mohapatra, L.N. (1966) Mycotic flora of the conjunctiva. Am J Ophthalmol. 62: 968- 970.
O’Bryan, T.A. (2005) Pseudallescheriasis in the 21st century. Expert Rev Anti Infect Ther. 3: 765-73.
Ogawa, H., Fujimura, M., Amaike, S., Matsumoto, Y., Kitagawa, M., Matsuda, T. (1997) Eosinophilic pneumonia caused by Alternaria alternata.Allergy. 52: 1005-1008.
Pastor, F.J., Guarro, J.  (2008) Alternaria infections: laboratory diagnosis and relevant clinical features. Clin Microbiol Infect Aug. 14: 734-46.
Pugh, D.G., Martin, M.T., Shull, J.W., Bowen, J.M. (1986) Endometrial candidiasis in five mares. J Equine Vet Sci. 6: 40-43.
Raper, K.B., Fennell, D.I. (1965) The Genus Aspergillus, Williams and Wilkins, Baltimore.  (1st ed.) New York, USA.
Rotaa, A., Calicchioa, E.,Nardonib, S., Fratinib, F., Ebanib, V.V., Sgorbinia, M., Panzania, D., Camilloa, F., Manciantib, F. (2011) Presence and distribution of fungi and bacteria in the reproductive tract of healthy stallions. Theriogenology. 76: 464-470.
Rozanski, P., Slaska, B., Rozanska, D. (2013) The current status of prevalence of yeast-like fungi in the environment of horses bred in Poland. Ann Anim Sci. 2: 365-374.
Sobel, J.D., Faro, S., Force, R., Foxman, B., Ledger, W.J., Nviriesy, P.R. (1998) Vulvovaginal candidiasis: epidemiologic, diagnostic, and therapeutic considerations. Am J Obstet Gynecol. 178: 203-11.
Sobel, J.D., Chaim, M.D. (1996) Update on treatment of vulvovaginal candidiasis. Rev Iberoam Micol. 13: 44-6.
Soeria-Atmadja, D., Onell, A., Borga, A. (2010) IgE sensitization to fungi mirrors fungal phylogenetic systematics. J Allergy Clin Immunol. 125: 1379-86.
Shokri, Sh., Khosravi, A.R., Sharifzadeh, A., Tootian, Z. (2010) Isolation and identification of yeast flora from genital tract in healthy female camels. Vet Microbiol. 144: 183-186.
Shugar, M.A., Montgomery, W.W., Hyslop, N.E. (1981) Alternaria sinusitis. Ann Otol Rhinol Laryngol. 90: 251-254.
Summerbell, R.C., Kane, J., Krajden, S. (1989) Onychomycosis, tinea pedis and tinea manuum caused by non-dermatophytic filamentous fungi. Mycoses. 32: 609-619.
Swerczek, T.W., Dennis, S.M. (2007) Fungal abortion. In: Current Therapy in Large Animal Theriogenology. (2nd ed.). Youngquist, R.S., Threrfall, W.R. (eds.). Saunders Elsevier. Philadelphia, USA. p. 188-9.
Talebkhan Garoussi, M., Khosravi, A.R., Havareshti, P. (2007) Mycoflora of cervicovaginal fluids in dairy cows with or without reproductive disorders. Mycopathologia. 164: 97-100.
Sutton, D.A., Sanche, S.E., Revankar, S.G., Fothergill, A.W., Rinaldi, M.G.(1999) In vitro amphotericin B resistance in clinical isolates of Aspergillus terreus, with a head-to-head comparison to voriconazole. J Clin Microbiol. 37: 2343-5.
Tosti, A., Piraccini, B.M., Stinchi, C., Lorenzi, S. (1996) Onychomycosis due to Scopulariopsis brevicaulis: clinical features and response to systemic antifungals. Br J Dermatol. 135: 799-802.
Traboulsi, R.S., Kattar, M.M., Dbouni, O., Araj, G.F., Kanj, S.S. (2007) Fatal brain infection caused by Aspergillus glaucus in an immunocompetent patient identified by sequencing of the ribosomal 18S-28S internal transcribed spacer.  26: 747-50.
Verma, S., Katoch, R.C., Jand, S.K., Sharm, B.M., Nigam, P. (1999) Mycobiotic flora of female genitalia of buffaloes and cows with reproductive disorders. Vet Res Commun. 23: 337-41.
Walsh, T.J., Groll, A.H. (2001) Overview: non-fumigatus species of Aspergillus: perspectives on emerging pathogens in immunocompromised hosts. Curr OpinInvestig Drugs. 2: 1366-7.
Walsh, T.J., Petraitis, V., Petraitiene, R. (2003) Experimental pulmonary aspergillosis due to Aspergillus terreus: pathogenesis and treatment of an emerging fungal pathogen resistant to amphotericin B. J Infect Dis. 188: 305-19.
Williamson, P., Munyua, S., Martin, R., Penhale, W.J. (1987) Dynamics of the acute uterine response to infection, endotoxin infusion and physical manipulation of the reproductive tract in the mare. J  Reprod Fertil. 35: 317-25.
Yu, H.(1965) Studies on fungi of the normal skin. Hifuka kiyo. Acta dermatologica.  60: 126-174.
Zafracas, A.M. (1975) Candida infection of the genital tract in thoroughbred mare. J Reprod Fertil Suppl. 23: 349 -51.