Isabel Martins Madrid

Epidemiological Findings and Laboratory Evaluation of Sporotrichosis: A Description of 103 Cases in Cats and Dogs in Southern Brazil

Sporotrichosis is a subcutaneous mycosis, which affects mainly small animals, and is considered an important public health disease. This paper describes the epidemiological and laboratory characteristics of 103 clinical cases of sporotrichosis diagnosed over a 10-year period in southern Brazil. The 92 cats and 11 dogs from eight municipalities in Rio Grande do Sul State developed especially the disseminated cutaneous and fixed cutaneous forms of the disease. Respiratory signs such as sneezing, serous nasal discharge and dyspnea were found in about 57% of the animals. The detection of Sporothrix schenckii in different clinical samples showed highest isolation in testicles (46.6%), oral cavity (45.2%) and conjunctival mucosa (38.1%). A differentiated histological pattern was found between the fixed cutaneous and disseminated cutaneous (DC) manifestations of the disease; well-organized granulomas of nodular distribution and various fungal structures prevailed in the DC form in cats. Melanin detection in S. schenckii cells by the Fontana–Masson technique was positive in 45.4% of the samples. The study revealed that the State of Rio Grande do Sul is an endemic sporotrichosis area and demonstrated the possibility of involvement of other pathways in the infection and spread of the disease. In addition, it emphasized the importance of laboratory tests for mycosis confirmation, especially in dogs that develop clinical manifestations without the presence of cutaneous lesions.

Feline Sporotrichosis in the Southern Region of Rio Grande Do Sul, Brazil: Clinical, Zoonotic and Therapeutic Aspects

Clinical cases of feline sporotrichosis, originating in the Pelotas region and diagnosed at the Laboratory of Infectious Diseases (UFPel), were studied in the period from 2002 to 2006. The animals were evaluated according to the clinical forms of the mycosis, time of lesion appearance, severity of the clinical diagnosis and evolution of cutaneous lesions throughout the treatment period. Mycological analyses, carried out through direct examination, cultivation of tissue samples and exudates of feline lesions all confirmed the diagnosis of sporotrichosis in the 15 animals under study. The cutaneous dissemination form was observed in 10 animals, of which three showed prostration, anorexia and dehydration. The zoonosis occurred in 20% of case studies, and the pet owners and one attendant at a veterinary clinic were infected, developing the fixed and disseminated cutaneous forms. The treatment of mycosis was carried out with itraconazole, 10 mg kg−1, once a day, on 12 animals. The cure of the clinical symptoms was observed on 50% of the felines. This study demonstrates a good clinical response of felines with sporotrichosis, when they were treated itraconazole and calls the attention for the incidence of human sporotrichosis on people related to the veterinary activity as well as for pet owners.

Role of melanin in the pathogenesis of cutaneous sporotrichosis.

In this study, we evaluate the clinical, histopathological and ultra structural aspects of the infection by field strain (MEL(+)) and mutated strain (MEL(-)) of Sporothrix schenckii. The development of sporotrichosis at the inoculation sites was similar for both groups of animals after the second week of the experimental period. Statistical differences appeared only at weeks 3 and 5. The pigmented isolate had greater tissue invasive capacity, promoting the formation of multifocal granulomas, whereas the albino isolate induced an enhanced inflammatory response, restraining the agent to the core of the granulomas.

Sporothrix brasiliensis outbreaks and the rapid emergence of feline sporotrichosis

Sporotrichosis is the main subcutaneous mycosis in Brazil, and is caused by Sporothrix schenckii and allied species. Sporothrix propagules present on soil and plant debris may be traumatically inoculated into the cutaneous/ subcutaneous tissues of the warm‐blooded host. An alternative route involves direct animal–animal and animal–human transmissions through deep scratches and bites of diseased cats. Sporotrichosis is much more common than previously appreciated with several cases emerging over the years especially in South and Southeast Brazil. We conducted an epidemiological surveillance in endemic areas of feline sporotrichosis in the southern region of Rio Grande do Sul state, Brazil. Over the last 5‐year period the number of feline sporotrichosis in Rio Grande increased from 0.75 new cases per month in 2010 to 3.33 cases per month in 2014. The wide geographic distribution of diagnosed cases highlights the dynamics of Sporothrix transmission across urban areas with high population density. Molecular identification down to species level by PCR‐RFLP of cat‐transmitted Sporothrix revealed the emergence of the clonal offshoot S. brasiliensis during feline outbreaks; this scenario is similar to the epidemics taking place in the metropolitan areas of Rio de Janeiro and São Paulo. Controlling and preventing sporotrichosis outbreaks are essential steps to managing the disease among humans and animals.

Perfil de suscetibilidade de leveduras do gênero Candida isoladas de animais ao óleo essencial de Rosmarinus officinalis L

ABSTRACT: Susceptibility profile of Candida spp. isolated from animals to the essentialoil of Rosmarinus officinalis L. In this work the antifungal activity and chemical composition ofthe Rosmarinus officinalis (rosemary) essential oil against Candida spp. were evaluated. Theessential oil was obtained by hydrodistillation of aerial parts, and analyzed by gas chromatographywith a flame ionization detector (GC/FID) and gas chromatography coupled with mass spectrometry(GC/MS). This oil was tested against fourteen isolates of Candida sp. (ten C. albicans and fourno-albicans species) by the microdillution broth assay (CLSI-M27A2). Chromatography analysesof the essential oil showed compounds, as camphor, verbenone and 1,8-cineole as majorconstituents . MIC and MFC values for C. albicans standard strains were 1.25 - 2.5 µL mL -1 and2.5 - 5.0 µL mL -1 , respectively. MIC and MFC for the non-albicans species were 1.25 – 5.0 µL mL -1 and 2.5 – 10.0 µL mL -1 , respectively. C. albicans isolates obtained from animals exhibited MIC andMFC values of 2.5 - > 10.0 µL mL

Disease progression of dermatophytic pseudomycetoma in a Persian cat

BACKGROUND Dermatophytic pseudomycetoma is a subcutaneous fungal infection by Microsporum canis. AIMS This work describes a case of dermatophytic pseudomycetoma in a Persian cat. PATIENT AND METHODS A 3-year old female Persian cat showing alopecia, scaling and ulcerated nodules throughout the body, with presence of ulcerated nodules with yellow granular discharges on the dorsum, close to the tail. Mycological and histopathological examinations were realized. RESULTS Diagnosis of dermatophytic pseudomycetoma was established. The cat was treated with griseofulvin, and surgical excision was carried out. Response to therapy was effective during the first months, during which a reduction in nodule frequency was observed. However, despite maintaining the therapy levels, the lesions relapsed and progressed to the point of causing the animals death. CONCLUSION The clinic evolution showed the progressive and recurrent character of dermatophytic pseudomycetoma.

Clinical and mycological analysis of dog's oral cavity

The oral microbiota of humans and animals is made up of a wide variety of yeasts and bacteria, but microbiota of dogs is not totally described. Although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease The aim of this study was to evaluate and correlate oral alterations with the presence of yeasts in oral cavity of female dogs. After clinical evaluation samples from healthy and from dogs with oral diseases were obtained from three different oral sites by swabs, curettes, millimeter periodontal probes and HA membrane tip in cellulose ester. Yeast identification was performed through macroscopic and microscopic colony features and biochemical tests. Dental calculus was the most prevalent occurrence in the oral cavity of 59 females. However, the isolation of yeasts was significantly higher (p < 0.05) in animals suffering from halitosis. Eleven yeast species were identified, namely: Malassezia pachydermatis, Rhodotorula spp., Candida albicans, C. catenulata, C. famata, C. guilliermondii, C. parapsilosis, C. intermedia, Trichosporon asahii, T. mucoides and Cryptococcus albidus. It could be concluded that the yeasts are part of the microbiota from the different sites of the oral cavity of the female canines studied without causing any significant alterations except halitosis.

Cytotoxicity and anti-Sporothrix brasiliensis activity of the Origanum majorana Linn. oil

The study aimed to evaluate the anti-Sporothrix sp. activity of the essential oil of Origanum majorana Linn. (marjoram), its chemical analysis, and its cytotoxic activity. A total of 18 fungal isolates of Sporothrix brasiliensis (n: 17) from humans, dogs and cats, and a standard strain of Sporothrix schenckii (n: 1) were tested using the broth microdilution technique (Clinical and Laboratory Standard Institute – CLSI M27-A3) and the results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). The MIC50 and MIC90 of itraconazole against S. brasiliensis were 2 μg/mL and 8 μg/mL, respectively, and the MFC50 and MFC90 were 2 μg/mL and >16 μg/mL, respectively, with three S. brasiliensis isolates resistant to antifungal. S. schenckii was sensitive at MIC of 1 μg/mL and MFC of 8 μg/mL. For the oil of O. majorana L., all isolates were susceptible to MIC of ≤2.25–9 mg/mL and MFC of ≤2.25–18 mg/mL. The MIC50 and MIC90 were ≤2.25 mg/mL and 4.5 mg/mL, respectively, and the MFC50/90 values were twice more than the MIC. Twenty-two compounds were identified by gas chromatography with a flame ionization detector (CG-FID) and 1,8-cineole and 4-terpineol were the majority. Through the colorimetric (MTT) assay, the toxicity was observed in 70–80% of VERO cells between 0.078 and 5 mg/mL. For the first time, the study demonstrated the satisfactory in vitro anti-Sporothrix sp. activity of marjoram oil and further studies are needed to ensure its safe and effective use.

Inhibitory effect of sodium hypochlorite and chlorhexidine digluconate in clinical isolates of Sporothrix schenckii

The susceptibility of Sporothrix schenckii isolates from clinical cases of canine, feline and human sporotrichosis, and from the environment, was evaluated with 4% sodium hypochlorite and 6.6% chlorhexidine digluconate using the broth microdilution, agar diffusion and direct exposure techniques. The minimal inhibitory concentration was smaller than 0.8% for chlorhexidine digluconate and between 8% and 4% for sodium hypochlorite. Inhibition zones were not found in agar diffusion for sodium hypochlorite, and zones averaging 1.9 mm were found for chlorhexidine digluconate. In the direct exposure test, sodium hypochlorite demonstrated best performance at 20 min of contact, as chlorhexidine digluconate presented little antimicrobial activity.

Atividade antifúngica do óleo essencial de Origanum vulgare frente a Malassezia pachydermatis

The aim of this study was to evaluate the in vitro antifungal activity of essential oil of Origanum vulgare against clinical isolates of Malassezia pachydermatis. The dried leaves of O. vulgare were purchased from a commercial distributor with certified quality and origin and referred for essential oil extraction and chromatography. The technique for in vitro testing was microdilution (CLSI M27A3) with modifications to phytochemicals and M. pachydermatis. The essential oil of O. vulgare was tested at concentrations from 28 to 0.87mg/mL in Sabouraud broth diluted with 1% of tween 80. All isolates were tested in duplicate. In the chromatographic analysis of the essential oil 12 compounds were identified, and thymol, α-terpinene, 4-terpineol were the major compounds. The MIC and the MFC of the 42 isolates of M. pachydermatis ranged from <0.87 to 7mg/mL with MIC50 and MIC90 values of 1.18 and 3.28 mg/mL, respectively. With this study it was concluded that M. pachydermatis is sensible to O. vulgare essential oil even at low concentrations. Thus, the essential oil of O. vulgare is presented as bioprospecting in the promising new drugs for the treatment of otitis and dermatitis in small animal clinic.