Estudo da população de Malassezia pachydermatis em otite externa canina e avaliação da sensibilidade in vitro e in vivo frente a antifúngicos.

Otite externa é uma enfermidade comumente observada em cães encaminhados a clínica veterinária e a etiologia desta doença varia em função de diversas combinações entre os fatores predisponentes, primários e perpetuantes responsáveis pela enfermidade. Este trabalho teve como objetivo pesquisar a presença da Malassezia pachydermatis em otite externa canina e avaliar a suscetibilidade in vivo e in vitro da levedura frente ao cetoconazol e tiabendazol. Para isto foi pesquisada a presença da levedura M. pachydermatis em otite externa de 168 cães encaminhados aos Hospitais de Clínicas Veterinárias da Universidade Federal do Rio Grande do Sul e da Universidade Federal de Pelotas, assim como em clínicas e canis particulares. Após identificação morfológica e bioquímica da levedura, foi realizada extração de DNA pelo método de fenol-clorofórmio de amostras selecionadas para análise pela técnica de RAPD- PCR, para verificação de heterogeneidade molecular. Foi realizada a reprodução experimental de malasseziose ótica em 14 cães inoculados com a levedura para posterior tratamento. Dois grupos de 30 cães com malasseziose ótica foram selecionados para tratamento com os produtos comerciais otológicos contendo tiabendazol e cetoconazol (Otodem plus® e Aurivet® respectivamente). Com os isolados de M. pachydermatis obtidos desses animais tratados foi realizado antifungigrama através da técnica de Microdoluição em Caldo (MC) para o tiabendazol e técnica do ETEST para o cetoconazol. Os resultados deste último foram comparados com a técnica de MC. A M. pachydermatis foi isolada em 139 (82,7%) casos de otite externa, sendo que molecularmente, a levedura apresentou diferenças, recebendo nove subdivisões a partir do primer utilizado. Os animais inoculados com a levedura desenvolveram a otite externa e o tratamento realizado com os produtos comerciais Aurivet® e Otodem plus® foi eficaz. Dos 60 animais tratados para malasseziose, 86,7% apresentaram cura clínica. A CIM do tiabendazol através da técnica de MC variou de 0,03 a >4mg/ml, com média de 3,67mg/ml. A CIM do cetoconazol, através do ETEST, variou de 0,004 a 0,75mg/ml, com média de 0,156mg/ml. A CIM do cetoconazol, através do método MC variou de 0,0009375 a 0,06mg/ml, com CIM média de 0,00815mg/ml. Através do cálculo de CIM50 e CIM90, observou-se frente ao tiabendazol, resistência em 13,7% dos isolados, sensibilidade intermediária em 47,1% e 39,2% isolados foram sensíveis. Quanto ao cetoconazol, através da técnica do ETEST, a resistência foi observada em 11,1% dos isolados, sensibilidade intermediária foi encontrada em 41,7% e 47,2% isolados foram sensíveis. Pela MC, foi observada resistência em 15,4% isolados, sensibilidade intermediária em 35,9% isolados e 48,7% foram sensíveis. As médias das CIMs observadas entre os 17 isolados testados simultaneamente frente ao cetoconazol com as duas metodologias, ETEST e MC, foi 0,103mg/ml e 0,0119mg/ml respectivamente. Nesta comparação podemos observar a concordância de resultados em apenas seis amostras (35,3%), quatro sensíveis e duas, sensibilidade intermediária. As combinações terapêuticas testadas nos animais foram eficazes no tratamento da malasseziose ótica canina, porém não houve relação entre resultado do teste in vitro e a resposta in vivo dos antifúngicos frente a levedura.

Epidemiologia de Otodectes cynotis (Acari: Sarcoptiformes) e sua associação com a infecção por Malassezia pachydermatis (Cryptococcales: cryptococcaceae) no canal auditivo de cães naturalmente infectados

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Isolation of Malassezia pachydermatis and M. sympodialis from the external ear canal of cats with and without otitis externa

The objective of this study was to determine the presence of Malassezia spp. in the external ear canal of cats with and without otitis. Forty-five animals were studied, 20 with and 25 without otitis externa (OE). Cerumen or secretion from external ear canal samples was cultured on modified Mycosel agar and sterile olive oil was added to the surface of the medium before specimen seeding. The isolates were analysed for macro- and micromorphology and identified by catalase tests and on the basis of growth on Tween 20, 40, 60 and 80. Malassezia spp. were isolated from 15 out of 20 (75%) animals with otitis and from 7 out of 25 (28%) cats without OE; the difference between the two groups was statistically significant (P <= 0.05). Malassezia pachydermatis and M. sympodialis were isolated from 60% (12/20) and 40% (8/20) of cats with otitis, respectively, with no significant difference in the frequency of isolation between the two species. In the microflora of the healthy ear canal M pachydermatis was significantly more common (6/25, 24%) than M sympodialis (1/25, 4%). The present investigation confirms that M sympodialis can also act as an actiological agent of feline OE, and if commercial veterinary laboratories do not use media with added lipids for the isolation of Malassezia spp., this might lead to false-negative results.

Ação antifúngica do resíduo líquido integral e fracionado do desfibramento da folha do sisal (Agave sisalana) sobre Malassezia pachydermatis, Tricophyton mentagrophytes e Candida albicans

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Susceptibility variation of Malassezia pachydermatis to antifungal agents according to isolate source

Malassezia pachydermatis is associated with dermatomycoses and otomycosis in dogs and cats. This study compared the susceptibility of M. pachydermatis isolates from sick (G1) and healthy (G2) animals to azole and polyene antifungals using the M27-A3 protocol. Isolates from G1 animals were less sensitive to amphotericin B, nystatin, fluconazole, clotrimazole and miconazole.

Etiologia, perfil de sensibilidade aos antimicrobianos e aspectos epidemiológicos na otite canina: estudo retrospectivo de 616 casos

A retrospective study of etiology, antimicrobial susceptibility profile and multiple drug resistance, and major epidemiological aspects were investigated in 616 cases of canine otitis. Staphylococcus beta hemolitic (26.27%), Malassezia pachydermatis (12.35%), and Pseudomonas aeruginosa (8.8%) were the most common microorganisms identified. The isolates were susceptible mainly to norfloxacin (89.62%), gentamicin (83.25%), and ofloxacin (80.16%). High occurrence of resistance of isolates was observed to neomicin (30.84%) and cephalexin (27.63%). Multiple drug resistance to three or more and five or more of antimicrobials tested was observed in 34.9% and 15.5% of isolates, respectively. The cases of canine otitis occurred predominantly in first years of age, in mixed breeds animals, at autumn season. The presence of itch, bad smell, and secretion in ear conduct were the major signs observed at clinical examination.

Produção, purificação e ação antimicrobiana do extrato fúngico do Trichosporon asahii

Pós-graduação em Microbiologia - IBILCE

Highlights in pathogenic fungal biofilms

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation of Sporothrix schenckii from the claws of domestic cats (indoor and outdoor) and in captivity in São Paulo (Brazil)

Sporotrichosis is a subcutaneous mycosis and is also a zoonosis (sapro- and anthropozoonosis). The objective of the present study was to determine the occurrence of sporotrichosis in domestic cats and in wild or exotic felines in captivity through the isolation of Sporothrix spp. from claw impressions in a culture medium. The samples included 132 felines, of which 120 (91.0 %) were domestic cats, 11 (8.3 %) were wild felines, and one (0.7 %) was an exotic felid. Twenty-one (17.5 %) were outdoor cats. Of the total, 89 (67.4 %) had contact with other animals of the same species. It was possible to isolate Sporothrix schenckii from the claws of one (0.7 %) of the felids probed; this animal exhibited generalised sporotrichosis and had infected a female veterinarian. The potential pathogenic agents Microsporum canis and Malassezia pachydermatis were isolated in 12.1 and 5.3 % of the animals, respectively. The following anemophilous fungi, which were considered to be contaminants, were also isolated: Penicillium sp. (28 or 21.2 %), Aspergillus sp. (13 or 9.8 %), Rhodotorula sp. (5 or 3.8 %), Candida sp. (5 or 3.8 %), Trichoderma sp. (1 or 0.7 %), and Acremonium sp. (1 or 0.7 %). Due to the low magnitude of occurrence (0.7 %) of Sporothrix in feline claws, the potential of the cats evaluated in this study to be sources of infection in the city of São Paulo is considerably low.

Novel microbicidal agents derived from Naja atra cardiotoxin 1 (CTX-1)

During my PhD course, I focused my research on antimicrobial peptides (AMPs), in particular on the aspects of their computational design and development. This work led to the development of a new family of AMPs that I designed, starting from the amino acid sequence of a snake venom toxin, the cardiotoxin 1 (CTX-1) of Naja atra. Naja atra atra cardiotoxin 1, produced by Chinese cobra snakes belonging to Elapidae family, is included in the three-finger toxin family and exerts high cytotoxicity and antimicrobial activity too. This toxin family is characterized by specific folding of three beta-sheet loops (“fingers”) extending from the central core and by four conserved disulfide bridges. Using as template the first loop of this toxin, different sequences of 20 amino acids linear cationic peptides have been designed in order to avoid toxic effects but to maintain and strengthen the antimicrobial activity. As a result, the sequence NCP-0 (Naja Cardiotoxin Peptide-0) was designed as ancestor and subsequently other 4 variant sequences of NCP0 were developed. These variant sequences have shown microbicidal activity towards a panel of reference strains of Gram-positive and Gram-negative bacteria, fungi and an enveloped virus. In particular, the sequence designed as NCP-3 (Naja Cardiotoxin Peptide-3) and its variants NCP-3a and NCP-3b have shown the best antimicrobial activity together with low cytotoxicity against eukaryotic cells and low hemolytic activity. Bactericidal activity has been demonstrated by minimum bactericidal concentration (MBC) assay at values below 10 μg/ml for Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii ( clinical isolates), Moraxella catharralis ATCC 25238, MRSA ATCC 43400, while towards Staphylococcus aureus ATCC 25923, Enterococcus hirae ATCC 10541 and Streptococcus agalactiae ATCC 13813 the bactericidal activity was demonstrated even below 1.6 μg/ml concentration. This potent antimicrobial activity was confirmed even for unicellular fungi Candida albicans, Candida glabrata and Malassezia pachydermatis (MBC 32.26-6.4 μg/ml), and also against the fast-growing mycobacteria Mycobacterium smegmatis DSMZ 43756 and Mycobacterium fortuitum DSMZ 46621 (MBC 100 μg/ml). Moreover, NCP-3 has shown a virucidal activity on the enveloped virus Bovine Herpesvirus 1 (BoHV1) belonging to herpesviridae family. The bactericidal activity is maintained in a high salt concentration (125 and 250 mM NaCl) medium and PB +20% Mueller Hinton Medium for E. coli, MRSA and Pseudomonas aeruginosa reference strains. Considering these in vitro obtained data, we propose NCP-3 and its variants NCP-3a and NCP-3b as promising antimicrobial candidates. For this reason, the whole novel AMPs family has been protected by a national patent (n°102015000015951).

Ação de extratos aquoso e alcoólico de própolis sobre Malassezia spp isoladas do conduto auditivo de cães

Pós-graduação em Medicina Veterinária - FMVZ

Malassezia spp. in acoustic meatus of bats (Molossus molossus) of the Amazon region, Brazil

The yeasts of the Malassezia genus are opportunistic microorganisms and can cause human and animal infections. They are commonly isolated from the skin and auricular canal of mammalians, mainly dogs and cats. The present study was aimed to isolate Malassezia spp. from the acoustic meatus of bats (Molossus molossus) in the Montenegro region, `` Rondonia ``, Brazil. From a total of 30 bats studied Malassezia spp. were isolated in 24 (80%) animals, the breakdown by species being as follows (one Malassezia sp. per bat, N=24): 15 (62.5%) M. pachydermatis, 5 (20.8%) M. furfur, 3 (12.5%) M. globosa and 1 (4.2%) M. sympodialis. This study establishes a new host and anatomic place for Malassezia spp., as it presents the first report ever of the isolation of this genus of yeasts in the acoustic meatus of bats.

Dermatopatias em cães: Revisão de 257 casos

The purpose of this study was to collect data on skin diseases in dogs evaluated at the Veterinary Hospital of the Northern Parana State University (UENP), in Bandeirantes, Parana, Brazil. Dermatological conditions amounted to 31.38% of all consultations in dogs, and the most common skin conditions observed were, from the most to the least common: bacterial, parasitic, fungal, immune, and keratinization disorders; general skin diseases and psychogenic conditions. The highest scoring diseases, in number of occurrences were, in descending order: Superficial folliculitis, demodicidosis, dermatophytosis, bacterial otitis, deep folliculitis, dry dekeratinization, allergic dermatitis caused by flea bites, fungal ear infections, scabies, impetigo, malassezia dermatitis, parasitical otitis, pad dermatitis, and atopic dermatitis.

Frequência de dermatopatias infecciosas, parasitárias e neoplásicas em cães na região de Garça, São Paulo - Brasil

Pós-graduação em Medicina Veterinária - FMVZ

From emotional suppression to regulated empathy : the changing face of control in the CES

Between the 1970s and the 1990s the level and type of emotionality in the Commonwealth Employment Service (the Australian national employment service) altered. Within a context of changing economic conditions and concomitant work intensification, it is argued that untenable working conditions resulted in new recruits adopting a coping strategy that led to the use rather than the suppression of emotions. The use of emotions provided workers with job satisfaction and greater control over service interactions. Management subsequently commandeered the use of emotions to complement the introduction of private sector management techniques and service delivery reforms, regaining control over worker-client interactions.