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Dermatophytes isolated from pet, dogs and cats, in Lisbon, Portugal (2000-2004)

Dermatófitos isolados de animais de companhia (cão e gato), em Lisboa, Portugal (2000-2004)

F. Bernardo1, A. Lança1 M. M.Guerra 1 H. Marina Martins2*

1 CIISA-Faculdade Medicina Veterinária–Polo Universitário da Ajuda-Av. Universidade Técnica, 1300-447, Lisboa- Portugal. 2 Laboratório Nacional Investigação Veterinaria – Estrada de Benfica , 701, 1549-011, Lisboa, Portugal

Summary: Ringworm are fungi infection with a very high rel- Cladosporium spp. (17,2%) e Penicillium spp. (11,4%). Alguns evance in animal and human health. During five years (2000- destes fungos contaminantes são referidos como sendo dotados 2004), 1298 samples were analyzed, being 978 from clinical de capacidade alergizante. material (skin scrapings and ), obtained from dogs and 320 samples from cats. These samples for detection. Palavras chave: Dermatófitos, , ani- One thousand and sixty four samples were negatives for dermat- mais de companhia. ophytes (82.0%) and 588, presented contamination with sapro- phytes fungi (45.3%) . Mycological culture revealed ringworm in 146 samples of dogs (11.2%) and 89 samples in cats (6.9 %). Microsporum canis was isolated in 63.7% of infected dogs and 82.0 % of cats. The other dermatophytes isolated belong to dif- ferent species: Microsporum gypseum (5.5% in dogs and 5.6% Introduction in cats), Microsporum vanbreuseghemii (1.4% in dogs), and Mi- crosporum nanum (2.2% in cats). Trichophyton mentagrophytes was isolated in 24.0% of the positive samples from dogs and is a specific fungal diseases of the 7.9% in cats. In dogs Trichophyton ajelloi and T. terrestre were skin. Dermatophytes fungi, with an affinity for corni- found 2.0% and 3.4% respectively. In 588 samples other con- fied epidermis and adnexal structures, are common in taminants fungi were found. The most prevalent were Alternaria pet. Its incidence is related to geographic region, cli- spp. (22.6%), Candida sp. (14.5%), Cladosporium spp.(17.2%) mate and animal husbandry techniques. It is frequently and Penicillium spp. (11.4%), Some of these fungi have potential allergenic activity. found in young, stray sick or debilitated animals. Der- matophytes are fungi species belonging to three gen- Key word: Dermatophytes, pet, Microsporum, Trichophyton. era: Microsporum, Trichophyton and Epidermophyton. They can be divided into three groups on the basis of natural habitat: geophilic- dermatophytes found main- Resumo: As tinhas animais são infecções micóticas de grande relevância nosológica e com impacto em saúde Pública. Durante ly infect both humans and animals (M. gypseum, T. cinco anos (2000-2004), foram analisadas 1298 amostras, sendo terrestre) antropophilic – those found mainly in hu- 978 de pêlos e escamas cutâneas de cães e 320 de gatos. Mil e mans transmitted amongst humans, but very seldom to sessenta e quatro amostras foram negativas (82,0%) das quais animals (M. audouinii) ) and zoohpilic – those found in 588 apresentavam fungos contaminantes (45,3%). Os exames animals, but transmitted to other or humans (M. canis, micológicos revelaram dermatófitos em 146 amostras obtidas de cães (11,2%) e em 89 de gatos (6,9%). Microsporum canis foi a T. mentagrophytes) (Scott et al. 1995). Zoophilic der- espécie de maior prevalência nos cães (63,7%) e nos gatos (82,0 matophytes induce lesions which are most often quite %) . Outras espécies de dermatófitos encontrados foram: Micro- inflamatory in humans (Arreseet al., 2000) sporum gypseum (5,5% em cão e 5,6% em gato); Microsporum These fungi share the ability to utilise keratin as a vanbreuseghemii (1,4% em cão). foi iso- nutrient substrate, and the infection of keratinised tis- lado apenas em 3 casos (2,2% em gato); Trichophyton menta- grophytes 24,0% em cão e 7,9% em gato. Nos cães Trichophyton sues is also termed ringworm (Sparkes et al., 1993). ajelloi e T terrestre. foram isolados em 2,0% e 3,4% respectiva- The skin lesions that appear are variable and do not mente. Nas amostras contaminadas com fungos saprófitas (588) necessarily form a ring. There will be , usually predominaram: Alternaria spp. (22,6%), Candida sp. (14,5%), in small patches at first. There might be scratching due to itchiness (Muller et al., 1989). Dogs and cats can suffer a infection at any age, but ringworm infections is more frequent in Correspondência: [email protected] the young. (Wright, 1989; Pier et al., 1998). In adition

85 Bernardo, F. et al. RPCV (2005) 100 (553-554) 85-88 to age, risk factors include poor nutrition, high density level. Identification was performed by microscopic of animals, poor management and lack of an adequate examination of a wet mount, using lactophenol cotton quarantine period for infected pets (Olivares et al. blue solution (LPCB -Merck 1. 13741.0100). Fungi 2003). The dermatophytes are transmitted by contact were identified according to Rebell and Taplin (1974), with infected hair, fomites (clippers, brushes) or from Badillet et al.(1987), Badillet (1991), Mackenzie and the environment (spores in soil). Microsporum canis Philpot, (1981) and Guaguère and Prelaud (1999). is considered highly contagious and potentialy patho- genic for people. Cats are considered the reservoirs of Results M. canis (Muller et al. 1989). Dogs and cats harbor many saprophytic moulds and Of the 1298 samples of skin scrapings and hair, 978 on there haircoats and skin. The most common were obtained from dogs and 320 from cats (Table 1). of these fungi isolated are species of Alternaria, As- A total of fourteen genera of fungi was isolated. Twelve pergillus, Cladosporium, Mucor, Penicillium, Rhizo- of the genera are regarded as saprophytes. Two fungal pus, Candida and Mallassezia. Most of these sapro- genera are commomly considered to be pathogenic phytic isolates represent transient contamination by (Microsporum and Trichophyton spp.). airborn fungi or by fungi in soil; some of them have potential allergenic activity (Aho, 1983). Table 1. Frequence of positive clinical material from dogs and In Portugal, little information is available concerning cats (2000-2004). the dermatophytes isolated from dogs and cats (Dias Nº of samples Positives % and Martins, 1981; Bernardo et al.1989). This study Dogs 978 146 14.9 was aimed to assess the frequency of dermatophytes and others fungi contaminations among of the skin of Cats 320 89 27.8 the pets (dogs and cats). Total 1298 235 18.1

In dogs, Microsporum canis was isolated in 93 Material and methods clinical material (63.7 %) (Table 2). The other der- matophytes isolated from dogs belonged to diverse Clinical material species: Microsporum gypseum (5.5%), Trichophyton mentagrophytes was isolated in 24.0%, Trichophyton During five years (2000-2004), 1298 samples of skin terrestre in 3.4% and T. ajelloi in 0.7 %. Of 89 cats, scrapings and hair, obtained from 978 dogs and from Microsporum canis was isolated in 73 clinical mate- 320 cats was examined for dermatophytes infection. rial (82.0%), Microsporum gypseum in 5.6% and Mi- All animals presented suspected skin disease (lesions crosporum nanum were isolated only in two occasions are round with embossed edges, and occurring in any (2.2%) (Table 2). part of the body, but mainly in tail and front paws) Table 2. Frequence of dermatophytes isolated in positives dogs Direct microscopic examination and cats Dermatophytes Dogs (n+= 144) Cats (n+= 90) The hair, scales and claw material was cleared by Microsporum canis 93 (63.7 %) 73 (82.0 %) placing the material in lactophenol solution on a mi- croscope slide. A coverslip was added, and the slide M. cookei 0 1 (1.1 %) was allowed to stand for 30 min at temperature room. M. gypseum 8 (5.5 %) 5 (5.6 %) Preparation were observed for microscope examina- M. nanum 2 (1.4%) 2 (2.2 %) tion, at 200 and 400 x. M .persicolor 0 1 (1.1%) M. vangreuseghemii 2 (1.4 %) 0 Fungal culture Trichophyton ajelloi 1 (0.7 %) 0 For isolation of fungi Mycobiotic agar (Difco Ref. T. terrestre 5 (3.4 %) 0 268940) and Sabouraud dextrose agar (Oxoid, Ref. T. mentagrophytes 35 (24.0%) 7 (7.9%) CM-41) were used added with antibacterial agents- chloramphenicol (C-0378- Sigma), or a combination of The most frequent fungal isolates (contaminants) penicillin (P 4399-Sigma), streptomycin (S 6501-Sig- were Alternaria spp. (22.6%), Candida spp.(14.5%), ma) and cycloheximide (C 7698-Sigma)- a substance Clasdoporium spp (17.2%) and Penicillium spp that growth of fast growing fungi,. The clinical mate- (11.4%) (Table3). spp. sp, rial were inoculated into McCartney flasks with the Malassesia pachydermatis, Trychosporum cutaneum, culture media, incubated at 25ºC, then examined daily Scopularipsis sp., Trichoderma spp. Rhizopus spp. and for 21 days. Saprophytic fungi were identified to genus Phoma spp were significantly isolated less frequently level and the dermatophytes were identified to species (Table 3).

86 Bernardo, F. et al. RPCV (2005) 100 (553-554) 85-88

Table 3. Contaminant mould isolated from clinical material Arrese, J.E., Martalo, O. Pierard-Franchimont, C. and Pierard, (n=588) G.E. (2000). Urban and rural mycozoonoses. Rev. Med. Liege, 55, 998-1002. Organisms N % Babillet, G. , Biévre, C. and Guého, É. (1987). Champignons Aspergillus spp. 44 (7.5) contaminants des cultures-Champignons opportunistes. Atlas clinique et biologique.Tome I. Éd. Varia, Paris, Alternaria spp. 133 (22.6) France pp. 131. Candida spp. 85 (14.5) Badillet, G. (1991). Dermatophyties et Dermatophytes- Atlas Chrysosporium sp 28 (4.8) clinique et biologique. 3ème Ed. Éditions Varia. Paris, France, pp.303. Cladosporium spp 101(17.2) Bernardo, F., Martins, H. M. and Mendes, A. M. (1989). Rastreio pachydermatis 37 (6.3) de dermatófitos em animais de companhia em Portugal. Rep. Trab. LNIV, XXI, 83-88. Penicillium spp 67 (11.4) Cabanes, F. J., Abarca, M. L., Bragulat, M.R. and Castella, G. Phoma sp. 5(0.9) (1996). Seasonal study of the fungal biota of the fur of Rhizopus nigricans 8 (1.4) dogs. Mycopathologia, 133, 1, 1-7. Cabanes, F. J., Abarca, M. L. and Bragulat, M. R. (1997). Der- Scopulariopsis spp. 3 (0.5) matophytes isolated from domestic animals in Barcelona, Trichoderma sp. 1 (0.2) Spain. Mycopathologia, 137, 2, 107-113. Caretta, G., Mancianti, F. and Ajello, L. (1989). Dermatophytes Trichosporum cutaneum 9 (1.5) and Keratinophilic fungi in cats and dogs. Mycoses, 32, Sporothrix schenchii 1 (0.2) 12, 620-626. Dias, M. C. and Martins, M. L. (1981). Dermatomicose em canídeo. Rep. Trab. INV. XIII, 39-42. Discussion Guaguère, É. and Prélaud, P. (1999). Guia Practica de Derma- tología Felina. Ed. Merial, pp 28.7. Paris, France. In Portugal, dermatophytosis are a common infec- Kaplan, W. and Ivens, M. (1961). Observations on the seasonal tious skin diseases in dogs and cats, specially during variation in incidence of ringworm in dos and cats in the Spring and Autumn. The clinical signs of dermato- United States. Sabouraudia, 1, 91-94. Mignon, B. R. and Losson, B. J. (1997). Prevalence and charac- phytosis are extremely variable, and the owners are terization of Microsporum canis carriage in cats. J. Med many times unaware that their pet is infected until they Vet Mycol. 35, 4, 149-256. themselves contract the infection. Most cases of der- Mackenzie, D. W. R. and Philpot, C. M. (1981). Isolation and matophytosis in pet are caused by Microsporum canis identification of ringworm fungi. Public Health Laboratory (Bernardo et al., 1989; Muller et al.1989). According service. Monograph Series, nº 15. H. M. Statonary Office, London, U.K. to Kaplan and Ivens (1961) cats are considered to be Moriello. K. A. and DeBoer, D. J. (1991). Fungal flora of the coat reservoirs of infection. of pet cats. Am. J. Vet. Res. 52, 602-606. In this study we characterized dermatophytes and Moriello, K. A. and Deboer, D. J. (1991a). Fungal flora of the contaminant fungal flora from pets with clinical signs. haircoat of cats with and without dermatophytosis. J. Med. Previous studies of Mignon and Losson, (1997), Vet. Mycol. 29, 5, 285-292. Cabanes et al., (1997), Morielo and DeBoer (1991), Muller, G. H., Kirk, R. W. and Scott, D. W. (1989). Small Animal Dermatology, 4 th Ed. Philadelphia, W. B. Saunders Co, conducted on cats indicate, that M. canis can be fre- Paris, p. 295-346. quently isolated and in most other studies of canine and Olivares, C. R. A. (2003). Ringworm infection in dogs and cats. feline dermatophytosis and this organism was isolated Intern. Veter. Information Service, Ithaca NY. with a higher prevalence from cats than from dogs (Ka- Pier, Ac. and Moriello, K. A. (1998). Parasitic relations hip plan and Ivens, (1961). M. gypseum M. vanbreuseg- beteween Microsporum canis and the cat. Med Mycol. 36, hemii and M. nanum were also reported by Morielo and 271-275. Rebell, G. and Taplin, D. (1974). Dermatophytes, Their recog- DeBoer (1991a), in one cat. Sparkes et al.,1993, and nition and Identification. Ed. University of Miami Press. Cabanes et al., 1996 isolate T. mentagrophytes from Florida, USA, pp124. dogs. T. terrestre has occasionally been reported to Scott, D.W., Miller, W. H. and Griffin, C. E. (1995). Small cause lesions in dogs and cat (Caretta et al., 1989). Animal Dermatology. 5 th Ed. W.B. Saunders Company. Concerning the contaminant fungal flora, Morielo London,U.K. p. 329-391. Sparkes, A. H., Gruffydd-Jones, T. J., Shaw, S. E., Wright, A. I. and DeBoer, (1991), found Aspergillus, Alternaria, and Stokes, C. R. (1993). Epidemilogical and diagnostic Cladosporium and Penicillium spp.. These moulds features of canine and feline dermatophytosis in the United are commonly found in the environment, particularly Kingdom from 1956 to 1991. The Vet. Record, 17, 57-61. on decaying plant debris. Our findings were similar to Wright, Al. (1989). Ringworm in dogs and cats. J. Small Animal. those reported by these authors. Pract. 30, 242-249.

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