Biofilm of Candida albicans: formation, regulation and resistance.

Candida albicans is the most common human fungal pathogen, causing infections that range from mucous membranes to systemic infections. The present article provides an overview of C. albicans, with the production of biofilms produced by this fungus, as well as reporting the classes of antifungals used to fight such infections, together with the resistance mechanisms to these drugs. Candida albicans is highly adaptable, enabling the transition from commensal to pathogen due to a repertoire of virulence factors. Specifically, the ability to change morphology and form biofilms is central to the pathogenesis of C. albicans. Indeed, most infections by this pathogen are associated with the formation of biofilms on surfaces of hosts or medical devices, causing high morbidity and mortality. Significantly, biofilms formed by C. albicans are inherently tolerant to antimicrobial therapy, so the susceptibility of C. albicans biofilms to current therapeutic agents remains low. Therefore, it is difficult to predict which molecules will emerge as new clinical antifungals. The biofilm formation of C. albicans has been causing impacts on susceptibility to antifungals, leading to resistance, which demonstrates the importance of research aimed at the prevention and control of these clinical microbial communities.

Ants (Hymenoptera: Formicidae) as carriers of fungi in hospital environments: an emphasis on the genera Tapinoma and Pheidole.

The aim of this study was to evaluate the presence of filamentous fungi and yeasts on the external surface of ants at hospitals. From March 2007 to February 2008, 2,899 ants were evaluated in two public hospitals in the city of Fortaleza, Ceará, in northeastern Brazil. The ants were attracted by nontoxic baits, distributed within critical and semicritical hospital areas. The fungi were identified through macro- and micromorphological analysis, biochemical profile, and growth in chromogenic medium. From this study, 5 genera and 13 species of ants were identified, from critical (8% of the collected ants) and semicritical (92%) areas, during the daytime (48%) and nighttime (52%) periods. In the mycological analysis, 75% of the ants were fungi carriers, with the species Tapinoma melanocephalum and species from the genus Pheidole having the most potential as carriers of airborne fungi (75 and 18%, respectively) and yeasts (6 and 1%, respectively). In summary, ants act as carriers of airborne fungi and yeasts, including some pathogenic species.

Antifungal activity of essential oils of Croton species from the Brazilian Caatinga biome.

AIMS: To find new antifungal agents among essential oils from Brazilian Croton species. METHODS AND RESULTS: Plant leaves were steam distilled and the obtained essential oils were analyzed by gas chromatography/mass spectroscopy. The main constituents were estragole and anethole for Croton zehntneri, methyl-eugenol and bicyclogermacrene for Croton nepetaefolius and spathulenol and bicyclogermacrene for Croton argyrophylloides. The antifungal activity of essential oils was evaluated against Candida albicans, Candida tropicalis and Microsporum canis by the agar-well diffusion method and the minimum inhibitory concentration (MIC) by the broth microdilution method. Essential oils of Croton species demonstrated better activity against M. canis. Among the three plants C. argyrophylloides showed the best results, with MIC ranging from 9 to 19 microg ml(-1). The acute administration of the essential oil up to 3 g kg(-1) by the oral route to mice was devoid of overt toxicity. CONCLUSIONS: The studied essential oils are active in vitro against the dermatophyte M. canis and present relative lack of acute toxicity in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its antifungal activity and low toxicity, the essential oils of studied Croton species are promising sources for new phytotherapeutic agents to treat dermatophytosis.

Chemical composition, toxicological aspects and antifungal activity of essential oil from Lippia sidoides Cham.

OBJECTIVES: The aims of this study were to test the essential oil from Lippia sidoides Cham. for antifungal activity, in vitro, against Candida spp. and Microsporum canis, to evaluate its acute and subchronic toxicological effects, in vivo, and to determine its chemical constituents. METHODS: The antifungal activity, in vitro, was initially evaluated by the agar-well diffusion technique, and the MIC and minimum fungicidal concentration (MFC) were determined by the broth microdilution method. The acute and subchronic toxicological effects were determined in mice and rats, respectively. The chemical composition of the essential oil was determined by gas chromatography coupled to mass spectroscopy. RESULTS: The essential oil obtained from L. sidoides was effective against all tested strains by the agar-well diffusion method. The MICs of L. sidoides essential oil for strains of M. canis ranged from 4 to 70 mg/L and the MFCs ranged from 9 to 150 mg/L. The MICs for strains of Candida spp. ranged from 620 to 2500 mg/L and the MFCs ranged from 1250 to 5000 mg/L. The main constituents of L. sidoides essential oil were thymol (59.65%), E-caryophyllene (10.60%) and p-cymene (9.08%). The acute administration of the essential oil up to 3 g/kg by the oral route to mice was devoid of overt toxicity. The 30 day oral administration of L. sidoides oil (117.95 mg/kg/day) to rats did not induce any significant histopathological, haematological or serum biochemical alterations. CONCLUSIONS: The essential oil from L. sidoides may be a promising source in the search for new antifungal drugs due to its efficacy and low toxicity.

Glucose improves the in vitro viability of Microsporum canis and Trichophyton mentagrophytes var. mentagrophytes.

We describe simple and cost-effective methods using carbohydrates to improve the in vitro viability of dermatophytes. Glucose and sucrose in different concentrations (3, 6, 9 and 12%) were used to maintain fifteen strains of M. canis and T. mentagrophytes var. mentagrophytes at 4 and -20 degrees C. The strains were phenotypically analyzed before storage and reevaluated at 1, 3, 6 and 9 months. At 1 and 3 months, any alterations in the viability or phenotype pattern of the stored strains were noted. At 6 months, both dermatophytes were 100% viable, when preserved in glucose (3, 6, 9 and 12%) at -20 degrees C. All T. mentagrophytes strains were also viable in sucrose (12%), at 4 degrees C and -20 degrees C. However, sucrose failed to improve the viability of M. canis at both temperatures. At 9 months, the higher viabilities without pleomorphism were seen for both dermatophytes preserved in glucose (9 and 12%) at -20 degrees C.

Identification and antimicrobial susceptibility of bacteria isolated from corneal ulcers of dogs

A total of 22 clinical specimens were obtained from 19 dogs with corneal ulcer (16 unilateral and three bilateral) for isolation and antimicrobial susceptibility evaluation of the isolated bacteria. Bacterial growth was observed in 100 percent of the samples (n=22). Staphylococcus intermedius was the predominant species (35.5 percent), followed by Corynebacterium xerosis (19.3 percent). Gentamicin, ciprofloxacin, chloramphenicol and tobramycin had a high efficacy against all of the isolated bacteria. The results evidenced that 80.7 percent of the isolates were Gram positive cocci and Gram positive bacilli, and that those microorganisms were sensitive to gentamicin, ciprofloxacin, chloramphenicol and tobramycin.

Utilizaram-se 22 amostras de material, obtidas de 19 cães com úlcera de córnea, sendo 16 unilaterais e três bilaterais, para isolamento e avaliação da susceptibilidade antimicrobiana das bactérias isoladas. Observou-se crescimento bacteriano em 100 por cento das amostras (n=22). A espécie predominante foi Staphylococcus intermedius (35,5 por cento) seguido de Corynebacterium xerosis (19,3 por cento). Gentamicina, ciprofloxacina, cloranfenicol e tobramicina apresentaram alta eficácia contra todas as bactérias isoladas. Os resultados evidenciam que 80,7 por cento dos isolados foram cocos e bacilos Gram positivos e que estes microrganismos foram sensíveis à gentamicina, ciprofloxacina, cloranfenicol e tobramicina.

Higher incidence of Malassezia pachydermatis in the eyes of dogs with corneal ulcer than in healthy dogs.

Malassezia pachydermatis is usually associated with otitis and dermatitis in dogs but it can also cause diseases in other species, including humans. In a human neonatal intensive care unit, M. pachydermatis was isolated from an infant's ocular discharge. Therefore, the aim of this study was to ascertain the presence of Malassezia spp. and its possible consequences in dogs' eyes. This research included 19 dogs with unilateral or bilateral corneal ulcers and 60 healthy dogs. A total of 158 clinical specimens from both the groups were obtained from the conjunctival sac of each eye by a calibrated platinum loop. The samples were placed on Dixon and blood agar, incubated at 35 degrees C, and examined daily for 15 days. Then, the strains were subcultured on Sabouraud agar. Of 22 clinical specimens collected from the eyes with corneal ulcers, five cultures (23%) were positive for M. pachydermatis. Of 16 samples collected from the contralateral healthy eye, cultures were positive in three samples (19%). Three animals had unilateral corneal ulcer and positive cultures for M. pachydermatis in both the eyes. Two dogs had unilateral corneal ulcer and positive cultures for M. pachydermatis in the same eye. However, from the 120 samples of 60 healthy dogs, only four clinical specimens (3%) had positive cultures for M. pachydermatis. The findings of M. pachydermatis, in a considerable percentage of clinical specimens from dogs with corneal ulcer, suggest its possible role at least as an aggravating factor in the pathophysiology of this disease.

Evaluation of Microsporum canis in different methods of storage.

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.