Over a one year period (November 2000-December 2001), clinical specimens from 189 dogs and 38 cats, from the city of Fortaleza, Ceará, Brazil, were examined at the Specialized Medical Mycology Center at the Federal University of Ceará to detect animals with dermatophytoses. The mycological analyses were conducted by direct microscopy and by fungal culture on Sabouraud agar, Sabouraud chloramphenicol agar and Mycosel agar. Dermatophytes were isolated from 27 of the 189 (14.3%) canine specimens and 14 of the 38 (36.8%) feline specimens. The identified dermatophytes were Microsporum canis (95%), M. gypseum (2.5%) and Trichophyton mentagrophytes var. mentagrophytes (2.5%). Microsporum canis was the most common species isolated (92.6% and 100%, for dogs and cats respectively). The percentage of positive direct microscopic examinations of clinical specimens and positive cultures was 61%. There was a high proportion of positive cultures from cats less than 1 year of age, but in dogs no significant differences were detected. There were no significant differences between the sexes. Dermatophytes were more frequently isolated in March, April and May, but no significant differences were detected in the seasonal distribution of canine and feline dermatophytoses.

Knowledge of epidemiological and mycological characteristics of onychomycosis has been noted by many authors as being an important tool for control of these fungal infections. This study seeks to improve knowledge of onychomycosis epidemiology and mycological features. Samples were taken from infected fingernails and toenailsOnychomycosis is a denomination used to describe nail infection usually caused by dermatophytes, yeast, and non-dermatophytic moulds (Mercantini et al. 1996, Weitzman & Summerbell 1996. These fungi may cause onychomycosis particularly as secondary invaders after damage by trauma or disease (Haneke 1991, Elewski 1998.Onychomycosis affects approximately 5% of the population worldwide (Murray & Dawber 2002) and represents around 30% of all superficial mycotic infection (Migdley et al. 1994) and 50% of nail disorders (Drake et al. 1996, Ghannoum et al. 2000.Dermatophytes are responsible for nearly 90% of toenail onychomycosis and at least 50% of fingernail infections (Elewski 1998). Candida species, particularly C. albicans, prevail in fingernail infections (Lopes et al. 1999, Pontes et al. 2002. Non-dermatophytic moulds are rare, but a number of species, such as Fusarim spp., Scytalidium spp., and Acremonium spp. have also been described as etiological agents of onychomycosis (Migdley et al. 1994, Tosti et al. 2000, Pontes et al. 2002.The epidemiology of onychomycosis has been well studied in some countries, but few data are available in tropical countries (Kam et al. 1997). In addition, research on this theme is poorly exploited in Northeast Brazil. This study, therefore, seeks to improve knowledge of the epidemiology and the mycological features of onychomycosis. Specimen collection and processing -The specimens were obtained from clinically abnormal nails, by a vigorous scraping of the nail bed, the underside of the nail plate and the hyponychyum, after cleaning the affected areas with 80% ethanol. The samples of each patient were placed in separate sterile Petri dish and transported to Medical Mycology Specialized Center. Scales scraped from the nails were analyzed for fungal elements, such as hyphae or blastoconidia, by direct microscopy examination, in potassium hydroxide (30%). For fungal cultures, all samples were inoculated on each of three isolation media (i) Sabouraud glucose agar (SGA; Difco Laboratories, Detroit, MI), (ii) SGA with 5% chloramphenicol, and (iii) Mycosel agar (Sanofi, France). The culture tubes were incubated at 28°C and examined daily for one month. Specimens from the lesions were repeatedly collected three times when it was observed growth of a nondermatophyte alone from a specimen that has tested positive for fungi on direct microscopy.Strain identification -The yeast isolates were identified according to morphological characteristics and the biochemical profile. To determine yeast micromorphology, cornmeal-Tween 80 agar plates were streaked and stabbed with a 48-h-old yeast colony, covered with a sterile cover-

The aims of this study were to test the antifungal activity, toxicity and chemical composition of essential oil from C. sativum L. fruits. The essential oil, obtained by hydro-distillation, was analyzed by gas chromatography/mass spectroscopy. Linalool was the main constituent (58.22%). The oil was considered bioactive, showing an LC50 value of 23 µg/mL in the Artemia salina lethality test. The antifungal activity was evaluated against Microsporum canis and Candida spp. by the agar-well diffusion method and the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) were established by the broth microdilution method. The essential oil induced growth inhibition zones of 28 ± 5.42 and 9.25 ± 0.5 for M. canis and Candida spp. respectively. The MICs and MFCs for M. canis strains ranged from 78 to 620 and 150 to 1,250 µg/mL, and the MICs and MFCs for Candida spp strains ranged from 310 to 620 and 620 to 1,250 µg/mL, respectively. C. sativum essential oil is active in vitro against M. canis and Candida spp. demonstrating good antifungal activity.

Crotalicidin (Ctn), a 34-residue cathelicidin from a South American rattlesnake, and its fragment (Ctn[15-34]) have shown anti-infective and cytotoxic activities against Gram-negative bacteria and certain tumor lines, respectively. The extent of such effects has been related to physicochemical characteristics such as helicity and hydrophobicity. We now report the anti-fungal activity of Ctn and its fragments (Ctn[1-14]) and (Ctn[15-34]). MIC determination and luminescent cell viability assays were used to evaluate the anti-infective activity of Ctn and its fragments (Ctn[1-14]) and (Ctn[15-34]) as anti-fungal agents against opportunistic yeast and dermatophytes. Cytotoxicity towards healthy eukaryotic cells was assessed in vitro with healthy human kidney-2 (HK-2) cells and erythrocytes. The checkerboard technique was performed to estimate the effects of combining either one of the peptides with amphotericin B. Ctn was the most active peptide against dermatophytes and also the most toxic to healthy eukaryotic cells. Fragments Ctn[1-14] and Ctn[15-35] lost activity against dermatophytes, but became more active against pathogenic yeasts, including several Candida species, both clinical isolates and standard strains, with MICs as low as 5 μm. Interestingly, the two peptide fragments were less cytotoxic to healthy HK-2 cells and less hemolytic to human erythrocytes than the standard-of-care amphotericin B. Also noteworthy was the synergy between Ctn peptides and amphotericin B, with consequent reduction in MICs of both drug and peptides. Altogether, Ctn and its fragments, particularly Ctn[15-34], are promising leads, either alone or in combined regimen with amphotericin B, for the treatment of fungal diseases.

Overall, Ctn[15-34] displays a synergistic antifungal activity with AMB, an effect that can be further developed into a multi-target therapeutic option with other antimycotics currently in use.

The main objective of this investigation was to evaluate different methods of storage for Microsporum canis based on materials and equipment that are readily available in developing countries. We tested 32 strains of M. canis at - 20 degrees C in potato dextrose agar (PDA) in its plain condition, or amended with 10% dimethyl sulfoxide or with 10% glycerol. In addition, we tested 25 degrees C storage of isolates in plain saline (0.9% NaCl) and in saline covered with a mineral-oil layer. After 9 months of storage, none of the M. canis strains frozen in PDA supplemented with glycerol survived, while only 16 and 6%, respectively, of the isolates in plain and DMSO medium lost viability. Nine month's storage in saline with or without mineral oil increased the amount of pleomorphic development of sterile hyphae; this phenomenon occurred at a significantly higher level than was seen in isolates stored at -20 degrees C. The physiological characteristics of M. canis were not affected by the different storage tests. The results suggest that, in order to ensure optimal viability, purity and pristine isolate condition, each M. canis isolate maintained should be held in at least two methods of storage, namely, PDA at -20 degrees C and saline with a mineral-oil layer at 25 degrees C.