SciELO - Scientific Electronic Library Online

vol.33 issue1Dietary addition of curcumin favors weight gain and has antioxidant, anti-inflammatory and anticoccidial action in dairy calves author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand




Related links

  • On index processCited by Google
  • Have no similar articlesSimilars in SciELO
  • On index processSimilars in Google


Revista Colombiana de Ciencias Pecuarias

Print version ISSN 0120-0690On-line version ISSN 2256-2958

Rev Colom Cienc Pecua vol.33 no.1 Medellín Jan./Mar. 2020 

Original research article

Identification of Malassezia species as part of normal skin and ear canal microbiota in horses*

Identificación de especies de Malassezia como microbiota normal en piel y canal auditivo externo de equinos

Identificação de espécies de Malassezia como microbiota normal em pele e canal auditivo externo de cavalos

Rubiela Castañeda-Salazar1 

Diana M Rodríguez-Sandoval1 

Adriana P Pulido-Villamarín1 

Melva Y Linares-Linares1 

Olimpo J Oliver-Espinosa2 

1 Departamento de Microbiología, Facultad de Ciencias, Unidad de Investigaciones Agropecuarias (UNIDIA) - Línea de Epidemiología y Salud Animal. Pontificia Universidad Javeriana, Bogotá, Colombia.

2 Departamento de Salud Animal, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogota, Colombia.



The yeasts of the genus Malassezia are considered part of the normal skin microbiota in humans and animals. In horses, several species of the genus Malassezia have been reported in different areas of the skin and ear canal.


Isolate, characterize and identify the different species belonging to the genus Malassezia isolated from the ear canal and skin of equine patients with no dermatological lesions that were referred to the large animal clinic of veterinary teaching hospital at the National University of Colombia.


22 horses were evaluated and sampled. Eighty-two samples were obtained by swabbing either the ear canals (left and right), skin areas of prepuce, mammary gland and inguinal region. The samples were examined by cytological evaluation and were cultured on modified Dixon’s agar and phenotypic and molecular identification were performed for yeast colonies.


Fourteen yeast isolates were obtained from the 82 samples. Biochemical identification determined that 50% (n=7) were Malassezia spp., 35.7% (n=5) were identified as Candida spp. and 14.3% (n=2) as Cryptococcus spp.. Using molecular tests, the Malassezia species were M. slooffiae (28.6%) and M.nana (57.1%); only one isolate was classified as Trichosporo asahii.


M.nana and M. slooffiae were identified as part of the normal ear canal and skin microbiota in the evaluated horses. The observed prevalence of Malassezia spp. was 18.2% (n=4/22) in this study sample.

Keywords: culture; ear canal; horses; identification; Malassezia; normal microbiota; PCR; skin; yeast



Las levaduras del género Malassezia hacen parte de la microbiota normal cutánea de humanos y animales. En equinos se han reportado diferentes especies de Malassezia aisladas de varias regiones de piel y canal auditivo externo.


Aislar, caracterizar e identificar las especies del género Malassezia spp. a partir de canal auditivo externo y piel de equinos sin lesiones dermatológicas, remitidos a la Clínica de Grandes Animales de la Facultad de Medicina Veterinaria y de Zootecnia de la Universidad Nacional de Colombia.


Se evaluaron 22 equinos, a partir de los cuales se obtuvieron 82 muestras entre hisopados de canal auditivo externo (izquierdo y derecho) y diferentes regiones de piel (prepucio, glándula mamaria e ingle). Las muestras fueron procesadas mediante examen directo y cultivo en agar Dixon modificado. A partir de los aislamientos en los que se observaron colonias morfológicamente compatibles con Malassezia spp. se realizó la identificación fenotípica y molecular.


De las 82 muestras procesadas se obtuvieron 14 aislamientos de levaduras, de las cuales mediante identificación bioquímica el 50% (n=7) correspondió a Malassezia spp., el 35,7% (n=5) a Candida spp., y el 14,3% (n=2) a Cryptococcus spp. Luego mediante pruebas moleculares se identificaron las especies del género Malassezia como: M. slooffiae (28,6%) y M . nana (57,1%); y un aislamiento correspondió a Trichosporon asahii. Conclusión: Se logró identificar las especies M.nana y M. slooffiae como microbiota normal de la piel y el canal auditivo en los equinos evaluados. La prevalencia de Malassezia spp. para la población evaluada fue de 18,2% (n=4/22).

Palabras clave: canal auditivo; cultivo; equinos; identificación; levaduras; Malassezia; microbiota normal; PCR; piel



As leveduras do gênero Malassezia fazem parte da microbiota cutânea normal de humanos e animais. Em cavalos, diferentes espécies de Malassezia isoladas de várias regiões da pele e do canal auditivo externo foram reproduzidas.


Isolar, caracterizar e identificar as espécies do gênero Malassezia spp. do canal auditivo externo e pele eqüinos sem lesões cutâneas, referiu-se à Clínica de Grandes Animais da Faculdade de Medicina Veterinária e Zootecnia da Universidade Nacional da Colômbia.


22 equinos foram avaliadas a partir dos quais 82 amostras a partir de esfregaços do canal auditivo externo (esquerda e direita) e diferentes regiões da pele (prepúcio, glândula mamaria e virilha) foram obtidos. As amostras foram processadas por exame direto e cultura em ágar Dixon modificado. Dos isolados nos quais as colônias foram observadas morfologicamente compatíveis com Malassezia spp. identificação fenotípica e molecular foi realizada.


Das 82 amostras processadas 14 isolados de levedura, que foram obtidos por identificação bioquímica de 50% (n=7) correspondia a Malassezia spp., 35,7% (n=5) a Candida spp., e 14,3% (n=2) para Cryptococcus spp.. Em seguida, usando o teste molecular espécie Malassezia foram identificadas como M. slooffiae (28,6%) e M . nana (57,1%); e um isolamento correspondia a Trichosporon asahii.


As espécies M.nana e M. slooffiae foram identificadas como microbiota de pele normal e do canal auditivo nos equídeos avaliados. A prevalência de Malassezia spp. para a população avaliada foi 18,2% (n=4/22).

Palavras-chave: canal auditivo; cavalos; cultivo; identificação; levedura, Malassezia; microbiota normal; PCR; pele


The yeasts of the genus Malassezia are classified in the phylum basidiomycota, order Malasseziales, family Malasseziaceae (Crespo et al., 2008a; Gaitanis et al., 2012). These yeasts are considered part of the normal skin microbiota in humans and animals (Hernández, 2005). This genus is physiologically characterized for being lipid-dependent due to its incapacity to synthesize saturated fatty acids that is manifested by requiring aexogenous source of these fatty acids to grow; however, M.pachydermatis is an exception given the lack of need for lipids to grow in cultures (Giusiano, 2006). Based on these characteristics, the media commonly used are Dixon’s and Leeming, and Notman agars that allow an adequate isolation of these yeasts (Hernández, 2005; Kaneko et al., 2007).

Seventeen Malassezia species have been described so far, being lipid-dependent the following ones: M. furfur, M. globosa, M. obtusa, M. restricta, M. slooffiae, M. sympodialis, M. dermatis, M. japónica, M. yamatoensis, M. nana, M. caprae, M. equina, M. cuniculi, M. brasiliensis, M. psittaci and M. vespertilionis (Guého et al., 1996; Sugita et al., 2003; Hirai et al., 2004; Cabañes et al., 2007; Cabañes et al., 2011), with the exception of M. pachydermatis (Giusiano, 2006; Lorch et al. 2018) as already mentioned.

In animals, M. slooffiae, M. globosa, M. sympodialis and M. pachydermatis have been reported in the ear canal of dogs and cats (Pulido et al., 2010; Salah et al., 2010); M.nana in cattle with or without external otitis and also in the ear canal of healthy horses (Hirai et al., 2004; Aldrovandi et al., 2016), and M. sympodialis, M. slooffiae, M. furfur and M. pachydermatis in the ear canal of pigs (Nardoni et al., 2010).

There have been few studies on the presence of Malassezia spp. as part of the normal microbiota in horse skin; however some reports have shown them to be present in different areas of the skin surface, such as the inguinal area, back, perineum and ear canal, where species such as: M. pachydermatis, M. furfur, M. restricta, M. slooffiae, M. obtusa and M. globosa have been isolated from; in addition, M. furfur, M. restricta, M. sympodialis and M. globosa have been isolated from the axillar region (Zia et al., 2014; White et al., 2006; Crespo et al., 2002), and M. equina from the udder cleft of mares as well as from the preputial fossa of stallions and geldings (Cabañes et al., 2007; White et al., 2006).

Given the limited number of studies worldwide and the lack of studies on Malassezia spp. as part of the normal equine skin microbiota in tropical regions as Colombia, the main objective in this study was to isolate, identify and characterize these yeasts from different body regions of horses with no dermatological diseases.

Materials and Methods

Ethical Considerations

This research project was approved by the Bioethical committee of the science faculty of the Pontificia Universidad Javeriana by resolution No 14 on 08-10-14. It was also approved by the University Animal Care committee, by resolution C-023-14 on 01-10-14.

Sampling population

During a study period of 4 months, 22 horses, mares, geldings and stallions with ages that ranged between 26 month and 12 years, admitted to the Large Animal Clinic-Veterinary Teaching Hospital at the National University of Colombia were studied. These horses were presented with various clinical diseases none involving the skin, but these diseases did not represented a risk factor for yeast colonization of the skin or ear canal. The patients did not have any antibiotic, antimycotic nor costicosterioid treatments.


The skin of the twenty-two horses was sampled using swabs. Samples were collected from the preputial area (n=10), mammary gland (n=12), inguinal area (n=16) and both ear canals (n=44). Eighty-two samples were collected and transported in sterile tubes at room temperature to the microbiology laboratory at the Pontificia Universidad Javeriana within a two hours-range after sampling.

Sample processing

Each sample was examined directly by making a slide smears using Gram staining to identify any yeast-like structure, oval or rounded blastoconidia of different sizes, filament like structure and bacteria. All samples were cultured on modified Dixon’s agar (malt extract 36 g, Peptone 6 g, Ox bile 20 g, Tween 40-10 ml, Glycerol 2 ml, Oleic acid 2 ml, Agar 12 g, deionized water 1,000 ml) (Resusta et al., 2007), and Sabouraud agar (Oxoid Hampshire, United Kingdom) supplemented with chloramphenicol (Sigma-Aldrich - St. Louis, MO, USA), and were incubated at 32 °C during 5 days (Hernández, 2005; Ashbee 2007; Pulido et al., 2010; Cafarchia et al., 2011; Aldrovandi et al., 2016).

All colonies morphologically compatible with Malassezia spp. were plated again to obtain pure colonies and describe them macroscopically along with the different morphological characteristics such as size, texture, color, shape, as well as the margin and surface of minimum 10 colonies. In order to describe adequately the microscopic appearance, 30 cells were used to measure length and width; such measurements were made using light microcopy (software Leica Microsystems®.DM 100 LED version 2.1.0.).

The colonies were considered to be Malassezia spp. (positive to Urea), underwent a series of biochemical and physiological tests that included assimilation of lipid supplements as Cremophor-EL (Sigma-Aldrich St. Louis, MO, USA), and Tweens (Merck Darmstadt, Germany/Sigma-Aldrich St. Louis, MO, USA), growth at 37 and 40 °C, enzymatic tests such catalase and β-glucosidase (Cafarchiaand Otranto, 2004; Kindo et al., 2004; Ashbee, 2007; Crespo et al., 2008b; Guého-Kellermann et al., 2010). Additionally, phospholipase activity was evaluated using 10% egg-yolk Sabouraud agar (Oxoid Hampshire, United Kingdom) on 4 points plating manner. The results were evaluated after 21 days using the averaging of the Pz values (Cafarchia and Otranto, 2004; Coutinho, 2005; Hurtado-Suarez et al., 2016).

The colonies that resulted morphologically and biochemically compatible with Candida spp. (Urea negative) were confirmed by chromogenic Chromagar Candida® (Becton Dickinson GmbH Heidelberg, Germany).

Culture quality control and strain storage

Reference strains of M. furfur CBS 7019, M. pachydermatis CBS 1879, M. sympodialis CBS 7222, M. slooffiae CBS 7956 and M. globosa CBS 7966 were used. The control strains Candida albicans ATCC 90028, Candida krusei ATCC 6258 and Candida parapsilosis ATCC 22019 were used as positive controls for phospholipase activity. All strains, including the isolated ones, were preserved in skim milk at -20°C.

Molecular identification

DNA extraction was performed using the fungi/yeast genomic DNA kit (Norgen® Thorold, ON, Canadá). The extracted DNAs were treated with RNAse (Promega, Madison, USA), 2 mg/mL at 37 °C for 4 hours (Gemmer et al., 2002), followed by amplification of region 5.8S DNAr, using primers ITS3 (5’-GCATCGATGAAGAACGCAGC-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’) (Gaitanis et al., 2002; Hernández, 2005). The amplification was done by using PCR at reaction volumes of 50 μL having 45 μL of PCR SuperMix (1.1X) (Invitrogen CA, USA), 1 μL of each one of the primers (10 pmol/μL) and 3 μL of genomic DNA (20 ng/mL). The conditions of the reaction were an initial denaturation cycle at 95°C for 5 minutes, 30 cycles at 95 °C for 1 minute, 55 °C for 1 minute and at 72 °C for 1.5 minutes, and an extension final cycle at 72 °C for 5 minutes (Gaitanis et al., 2002; Hernández, 2005). The amplification products were detected by 1.5% agarose gel electrophoresis (Promega Madison, USA) with buffer TBE 1X (Promega Madison, USA), they were stained with ethidium bromide (Invitrogen CA, USA). The gen 5.8S RNAr amplification products were sent for purification and sequencing to Macrogen® Inc (Korea). The sequences were analyzed using BLASTn ( cgi).


Cytological examination of the direct Gram stained smears showed yeasts of different forms and sizes in 17.07% (n=14) of all swab samples. The macroscopic morphology of the colonies that were identified as Malassezia spp. were cream in color, flat and small with smooth margins with diameters between 1-2.1 mm. These yeasts were isolated mainly from left ear canals (n=5) and udders (n=2). The microscopic measurements showed different sizes of cells that varied from 1.5-2.4 μm wide and 2.2-3.2 μm in length.

Biochemical identification test allowed to determine that 50% (n=7) of the yeast isolates belonged to the Malassezia genus. The species that were identified included M.pachydermatis, M.globosa, M.obtusa, M slooffiae and M.sympodialis in 14.3% (n=1) for each isolate. The remaining two isolates were only identified as belonging to the Malassezia genus and were reported as Malassezia spp. (see Table 1). However, the molecular identification tests did not agree with the biochemical tests. The molecular tests identified M.nana as the most frequent species being 57.1% (n=4) of the isolates, followed by M. slooffiae with 28.6% (n=2) of the isolates; the remaining isolate was identified as belonging to a different genus Trichosporon asahii (see Table 1). The observed agreement between the two tests was 85.7% when genus identification was tested, but only 14.3% agreement regarding species identification. The rest of the yeasts isolated were determined to be Candida spp. in 35.7% (n=5), which were isolated from the preputial and inguinal areas, and Cryptococcus spp. was isolated from ear canal in 14.3% (n=2) of the cases.

On the other hand, Malassezia spp. was only determined to be present in only 4 out of 22 horses, showing a prevalence of 18.2% in this sample.

In this present study, 28.57% (n=2) of the isolates were phospholipase positive, which was very high for M.nana and high for M. slooffiae. The remaining 57.1% (n=4) of the isolates did not present this activity.

Candida spp. was isolated in 5 samples presenting negative urease activity and grew in Sabouraud agar. When using Medio CHROMagar Candida BD Becton-Dickinson® media Candida glabrata was determined in 80% (n=4) of these isolates and Candida tropicalis in 20% (n=1) of the cases. Finally, two samples yielded yeast belonging to genus Cryptococcus, basing their classification on positive urease activity and the presence of capsule, when stained with indian ink (Resusta et al., 2007).

Due to the small numbers of isolates, it was not possible to establish any association between isolates and variables such as age, sex or breed.

Table 1 Biochemical and molecular identification of the Malassezia spp. isolates from ear canal and udder. 

EC: Ear canal U: Udder **Grow on Dixon agar at different incubation temperatures. Phospholipase activity: Very high: Pz<0,64. High: Pz between >0,64 and <1. Null: Pz =1. NG: No growth on 10% egg yolk Sabouraud agar.


Microscopic characteristics of the colonies identified as Malassezia spp. that included shape, margin, size, volume and texture are in accordance with the characteristics reported by several authors (Ashbee, 2007; Arenas, 2008; Crespo et al., 2008a; Guého-Kellermann et al., 2010; Hurtado-Suárez et al., 2016); however, there were differences in colony sizes, which could be attributed to possible limitations when measuring the organisms due to the distribution of the colonies on the agar. Regarding the microscopic characteristics based on the observed differences in the cell length, they were also partially similar to what is reported. This latter observation could also be due to limitations in the measuring as consequence of the cell distribution on the smear. These observations clearly suggest that both methodologies have a limited value when these yeasts are to be identified (Guého et al., 1996; Sugita et al., 2003; Hirai et al., 2004; Hernández, 2005; Cabañes et al., 2007; Crespo et al., 2008b).

The Malassezia isolates were determined to be M. pachydermatis, M. globosa, M. obtusa, M. slooffiae and M. sympodialis with prevalence of 14.3% (n=1) for each one of them. These species have also been reported by other studies to be commonly isolated from normal equine skin (White et al., 2006; Crespo et al., 2002); however, isolation of M. sympodialis and M.pachydermatis from mammary gland contrasts with the reports by White (2005) and White et al.(2006), who isolated M. slooffiae and/or M.equina from this same anatomical site of healthy mares. According to biochemical test results, the Malassezia species was not identified (2.43%) in two isolates, because their results did not coincide with what has been reported in the literature for the different species (Guého et al., 1996).

The identification of one of the isolates as M. pachydermatis was based on its ability to grow on Sabouraud Agar at 32 °C, and to be able to assimilate all the Tweens (Guého-Kellermann et al., 2010; Cafarchia et al., 2011). Despite the fact that M. globosa and M. obtusa share similar characteristics such as their incapacity to grow at 40 °C and the ability to assimilate tweens, the β- glucosidasa test allowed to differentiate one from the other due to the lack of the enzyme by M. globosa (Hernández, 2005; Giusiano, 2006). In a similar manner, M. sympodialis was differentiated from M. slooffiae because it has the β- glucosidase enzyme, and also due to the capacity to assimilate Tweens 40, 60 and 80, while M. slooffiae assimilates Tweens 20, 40 and 60, according to the biochemical characteristics reported, it was possible to determine the species of the isolates (Guého et al., 1996; Sugita et al., 2003; Hirai et al., 2004; Giusiano, 2006; Kaneko et al., 2007; Salah et al., 2010).

In order to identify M.pachydermatis, the phospholipase activity was evaluated given that it has been reported to be present in higher levels in either isolates from healthy skin or with skin lesions. This previous test and the biochemical characteristic allowed identifying this species (Cafarchia & Otranto, 2004; Juntachai et al., 2009; Pini & Faggi, 2011; Ortiz et al., 2013). In the case of M. obtusa, M. slooffiae, M. globosa and M. restricta, it has been reported that they do not show phospholipase activity in healthy animals (Pini & Faggi 2011). In the present study, M. slooffiae and M.globosa showed phospholipase activity in contrast to what has been reported. The presence of this activity could not be regarded as a virulence factor given that they came from healthy equine skin samples.

M. slooffiae, M. obtusa, M. globosa, M. pachydermatis and M. sympodialis were the isolated species from the healthy skin in horses using phenotypical and biochemical tests. These findings are similar to several studies that have reported them as the most common species isolated as normal microbiota of the skin of healthy horses (Crespo et al., 2002; Giusiano, 2006; White et al., 2006).

It is important to highlight that the phenotypical and the molecular identification methods agreed in only one isolate. The molecular identification technique was able to identify the species of the two isolates that were classified only as Malassezia phenotype classification. These low agreement level could be due to the variations in the results of the biochemical test as seen in the isolates from subject 2 (see Table 1), suggesting the need to use molecular techniques to identify the Malassezia species (Gaitanis et al., 2006; Ko et al., 2011; Zia et al., 2015).

Based on the molecular tests used, M.nana (80%) was the most frequent species isolated from the ear canal of the horses in the study; it was followed in occurrence by M. slooffiae (20%). Similar findings were reported by Aldrovandi et al. (2016) and Crespo et al. (2002). They reported the same species from this anatomical site, but it is different from the findings by Shokri (2016), who isolated M.pachydermatis from the ear canal. An interesting finding was that Malassezia spp. was only isolated from ear canals and udders; however, it was not isolated from the other anatomical sample sites despite reports indicating that it has been isolated from those areas in other studies (Crespo et al., 2002; Paterson 2002; White et al., 2006; Shokri, 2016).

The frequency of isolation observed in this study was 18.2% and only one species for each horse was detected. Comparing with other studies, this frequency was lower than the reported by Crespo et al.(2002), White et al., (2006) and Shokri (2016), that found prevalences of 34,9, 54 and 60%, respectively, in healthy horses.

Candida spp. was also isolated from some of the study subjects. This yeast is considered as part of the normal microbiota of both the skin of horses and its environment (Sgorbini et al., 2008; Cafarchia et al., 2013; Różański et al., 2013). This genus may produce opportunistic infections, despite being part of the normal microbiota, in patients with compromise of their immunological status or alteration of the normal microbiota (Cafarchia et al., 2013). It has been also reported that Cryptococcus spp. has been detected in the environment where horses are kept, which may explain why it was isolated in this study (Różański et al., 2013); however, it has been implicated in mycotic pneumonia in horses (Higgins & Pusterla, 2006).

Malassezia spp. is considered a normal inhabitant of the horse skin, but it has the potential to become a pathogen in cases of abnormal skin microenvironment or cutaneous infections (Paterson, 2002). In conclusion, the prevalence of Malassezia spp. in the studied population was 18.2% (4/22), and the species M.nana and M. slooffiae were identified as part of the skin microbiota in these horses using molecular tests. This is the first report that identifies Malassezia spp. as part of normal horse skin microbiota in Colombia.


To Adriana Marcela Celis Ramírez, from Universidad de Los Andes who provided the reference strains.


Aldrovandi AL, Osugui L, Coutinho SDA. Is Malassezia nana the main species in horses’ ear canal microbiome?. Braz J Microbiol 2016; 47:770-774. 10.1016/j.bjm.2016.04.017 [ Links ]

Arenas R. Micología Médica Ilustrada. 5th ed. México: Mc Graw Hill; 2008. ISBN: 978-607- 15-1125-6 [ Links ]

Ashbee H. Update on the genus Malassezia. Med Mycol 2007; 45:287-303. 10.1080/13693780701191373 [ Links ]

Cabañes J, Theelen B, Castellá G, Boekhout T. Two new lipi-dependent Malassezia species from domestic animals. FEMS Yeast Res 2007; 7(6):1064-1076. 10.1111/j.1567-1364.2007.00217.x [ Links ]

Cabañes F, Vega S, Castellá G. Malassezia cuniculi sp. nov., a novel yeast species isolated from rabbit skin. Med Mycol 2011; 49:40-48. ]

Cafarchia C & Otranto D. Association between phospholipase production by Malassezia pachydermatis and skin lesions. J Clin Microbiol 2004; 42(10):4868-4869. 10.1128/JCM.42.10.4868-4869.2004 [ Links ]

Cafarchia C, Gasser RB, Figueredo LA, Latrofa MF, Otranto D. Advances in the identification of Malassezia. Mol Cell Probe 2011; 25:1-7. 10.1016/j.mcp.2010.12.003 [ Links ]

Cafarchia C, Figueredo L, Otranto D. Fungal diseases of horses. Vet Microbiol 2013; 167:215- 34. 10.1016/j.vetmic.2013.01.015 [ Links ]

Coutinho S. Malassezia pachydermatis: enzymes production in isolates from external ear canal of dogs with and without otitis. Arq Bras Med Vet Zootec 2005; 57(2):149-153. ]

Crespo M, Abarca M, Cabañes F. Occurrence of Malassezia spp. in horses and domestic ruminants. Mycoses 2002; 45:333-337. ]

Crespo V, Crespo M, Gómez E. Diagnóstico de laboratorio de las levaduras del género Malassezia. Piel 2008a; 23(10):570-576. 10.1016/S0213-9251(08)75801-5 [ Links ]

Crespo V, Gómez E, Crespo M. La pitiriasis versicolor y las levaduras del género Malassezia. Actas Dermosifiliogr 2008b; 99:764-771. 10.1016/S0001-7310(08)74956-1 [ Links ]

Gaitanis G, Prokopios M, Hantschke M, Bassukas I, Velegrakid A. The Malassezia Genus in Skin and Systemic Diseases. Clin Microbiol Rev 2012; 25:106-141. 10.1128/CMR.00021-11 [ Links ]

Gaitanis G, Robert V, Velegraki A. Verifiable single nucleotide polymorphisms of the internal transcribed spacer 2 region for the identification of 11 Malassezia species. J Dermatol Sci 2006; 43:214-217. 10.1016/j.jdermsci.2006.03.013 [ Links ]

Gaitanis G, Velegraki A, Frangoulis E, Mitroussia A, Tsigonia A. Identification of Malassezia species from patient skin scales by PCR-RFLP. Clin Microbiol Infec 2002; 8:162-173. ]

Gemmer C, De Angelis Y, Theelen B, Boekhout T, Dawson T. Fast, noninvasive method for molecular detection and differentiation of Malassezia yeast species on human skin and application of the method to dandruff microbiology. J Clin Microbiol 2002; 40(9):3350-3357. 10.1128/jcm.40.9.3350-3357.2002 [ Links ]

Giusiano G. Malassezia: Estado del conocimiento y perspectivas en su estudio. Rev Argent Microbiol 2006; 38:41-48. Available from: ]

Guého-Kellermann E, Boekhout T, Begerow D. Biodiversity, Phylogeny and Ultrastructure. In: Boekhout T, Guého E, Mayser P, Velegraki A, editors. Malassezia and the skin. Science and Clinical Practice. Germany: Springer-Berlin, Heidelberg, 2010. p. 17-63. ISBN: 978-3-642- 03616-3 [ Links ]

Guého E, Midgley G, Guillot J. The genus Malassezia with description of four new species. Anton Leeuw Int JG 1996; 69:337-355. Avaliable from: ]

Hernández J. Caracterización molecular de especies del género Malassezia. Tesis de Doctorado. 2005. Facultad de Veterinaria, Universidad Autónoma de Barcelona, España. 2005. Available from: ]

Higgins JC, Pusterla N. Fungal Pneumonia in Horses. Clin Tech Equine Pract 2006; 5:218- 224. ]

Hirai A, Kano R, Makimura K, Duarte E, Hamdam J et al. Malassezia nana, a novel lipid-dependent yeast species isolated from animals. Int J Syst Evol Micr 2004; 54:623-627. 10.1099/ijs.0.02776-0 [ Links ]

Hurtado-Suárez A, Pulido-Villamarín A, Linares-Linares M, Suárez-Fernández L, Castañeda-Salazar R, Rodríguez-Bocanegra MX. Caracterización fenotípica de aislamientos de Malassezia spp. de origen canino. Rev MVZ Córdoba 2016; 21(3):5535-5546. ]

Juntachai W, Oura T, Murayama SY & Kajiwara S. The lipolytic enzymes activities of Malassezia species. Med Mycol 2009; 47:477-484. 10.1080/13693780802314825 [ Links ]

Kaneko T, Makimura k, Abe M, Shiota R, Nakamura Y, Kano R, Hasegawa A, Sugita T, Shibuya S, Watanabe S, Yamaguchi H, Abe S, Okamura N. Revised Culture-Based System for Identification of Malassezia Species. J Clin Microbiol 2007; 45(11):3737-3742. 10.1128/JCM.01243-07 [ Links ]

Kindo AJ, Sophia SKC, Kalyani J, Anandan S. Identification of Malassezia species. Indian J Med Microbi 2004; 22(3):179-181. Available from: ]

Ko, JH; Lee, YW; Choe, YB and Ahn, KJ. Epidemiologic study of Malassezia yeast in patients with Malassezia folliculitis by 26S rDNA PCR-RFLP analysis. Ann Dermatol 2011; 23(2):177-184. 10.5021/ad.2011.23.2.177 [ Links ]

Lorch JM, Palmer JM, Vanderwolf KJ, Schmidt KZ, Verant ML, Weller TJ, Blehert DS. Malasseszia vespert ilionis sp. nov.: a new cold-tolerant species of yeast isolated from bast. Persoonia. Mol. Phylog. Evol. Fungi 2018; 41(1): 56-70 ]

Nardoni S, Merildi V, Frangioni S, Ariti G, Verin R, Vannucci P, Mancianti F. Isolation and characterization of Malassezia spp. in healty swine of different breeds. Vet Microbiol 2010; 141:155-158. 10.1016/j.vetmic.2009.07.033 [ Links ]

NCBI. National Center for Biotechnology Information. Basic Local Alignment Search Tool. [Access date: January 10th, 2014]. Available from: Available from: . [ Links ]

Ortiz G, Carmen M, Alfonso J. Carrillo M, Paya M. Producción de fosfolipasa y proteinasa en cepas de Malassezia pachydermatis aisladas de perros con otitis y sin otitis. Rev Iberoam Micol 2013; 30(4):235-238. ]

Paterson S. Identification of Malassezia from a horse’s skin case report. Equine Vet. Educ 2002; 14(3):121-125. ]

Pini G, Faggi E. Extracellular phospholipase activity of Malassezia strains isolated from individuals with and without dermatological disease. Rev Iberoam Micol 2011; 28(4):179-182. 10.1016/j.riam.2011.05.002. [ Links ]

Pulido A, Castañeda R, Linares M, Mercado M. Diagnóstico clínico-microbiológico de otitis externa en caninos de Bogotá - Colombia. Rev MVZ Córdoba 2010; 15(3):2215-2222. ]

Resusta A, Sánchez A, Gil J. Fundamentos básicos para el diagnóstico micológico. Rev Iberoam Micol 2007. ISBN: 978-84-611-8776-8. Available from: ]

Różański P, Ślaska B, Różańska D. Current status of prevalence of yeast-like fungi in the environment of horses bred in Poland. Ann Anim Sci 2013; 13(2): 365-374. ]

Salah BI, Makni F, Cheikhrouhou F, Neji S, Sellami H, Ayadi A. Les levures du genere Malassezia: pathologie, milieux d’isolement et d’identification. Malassezia species: Pathology, isolation and identification media. J Mycol Med 2010; 20:53-60. 10.1016/j.mycmed.2009.11.006 [ Links ]

Sgorbini M, Barsotti G, Nardoni S, Mancianti F, Rossi S, Corazza M. Fungal Flora of Normal Eyes in Healthy Newborn Foals Living in the Same Stud Farm in Italy. J Equine Vet Sci 2008; 28(9):540-543. 10.1016/j.jevs.2008.07.018 [ Links ]

Shokri H. Occurrence and distribution of Malassezia species on skin and external ear canal of horses. Mycoses 2016; 59:28-33. 10.1111/myc.12430 [ Links ]

Sugita T, Takashima M, Kodama M, Tsuboi R, Nishikawa A. Description of a new yeast species Malassezia japonica and its detection in patients with atopic dermatitis and healthy subjects. J Clin Microbiol 2003; 41:4695-4699. 10.1128/JCM.41.10.4695-4699.2003 [ Links ]

White SD. Equine Bacterial and Fungal Diseases: A Diagnostic and Therapeutic Update. Clin Tech Equine Pract 2005; 4:302-310. ]

White, SD, Vandenabeele, S.I., Drazenovich, N.L, Foley, J.E. Malassezia species isolated from the intermammary and preputial fossa areas of horses. J Vet Intern Med 2006; 20:395- 398. 10.1892/0891-6640(2006)20[395:msifti];2 [ Links ]

Zia M, Mirhendi H, Toghyani M. Detection and identification of Malassezia species in domestic animals and aquatic birds by PCR-RFLP. Iran J Vet Res 2015; 16(1):36-41. PMID: 27175148. Available from: ]

* To cite this article: Castañeda-Salazar R, Rodríguez-Sandoval DM, Pulido-Villamarín AP, Linares-Linares MY, Oliver-Espinosa OJ. Identification of Malassezia species as part of normal skin and ear canal microbiota in horses. Rev Colomb Cienc Pecu 2020; 33(1): 5-15. DOI:

Funding This work was supported by the Pontificia Universidad Javeriana. Office of Research. University research grant. Research institutes. [ID PROY 000006472]. This report is part of the research project: “Isolation and phenotypical identification of Malassezia spp. yeast in pigs, horses and cattle”.

Received: March 12, 2018; Accepted: March 18, 2019

Conflicts of interest

The authors declare they have no conflicts of interest with regard to the work presented in this report

Author contributions

Rubiela Castañeda-Salazar: who was responsible for the design or conception of the study, administered the project, coordinated the research, process the samples and wrote the paper. Diana M Rodriguez-Sandoval: who was responsible for processing the samples and collected the data; participated in the writing of the paper. Adriana P Pulido-Villamarín: who helped to process the samples, administered the project, and participated in the writing of the paper. Melva Y Linares-Linares: who helped to process the samples and participated in the writing of the paper. Olimpo J Oliver-Espinosa: who took the samples, participated in the writing of the paper and reviewed and did a critical reading and editing of the paper.

Creative Commons License This is an open-access article distributed under the terms of the Creative Commons Attribution License