Antifungal activity of propolis on different species of Candida

Propolis is a resinous material collected by bees from the buds or other parts of plants. It is known for its biological properties, having antibacterial, antifungal and healing properties. The antifungal activity of propolis was studied in sensitivity tests on 80 strains of Candida yeasts: 20 strains of Candida albicans, 20 strains of Candida tropicalis, 20 strains of Candida krusei and 15 strains of Candida guilliermondii. The yeasts showed a clear antifungal activity with the following order of sensitivity: C. albicans > C. tropicalis > C. krusei > C. guilliermondii. Patients with full dentures who used a hydroalcoholic propolis extract showed a decrease in the number of Candida.

Isolamento de fungos produtores de enzimas pectinolíticas

Soil samples collected in the campus, UNESP, Araraquara, SP, were employed to isolate and characterize fungi strains with potential pectinolytic enzymes. These enzymes have arisen great interest due to its increasing application in the food industry. Two hundred forty six strains were isolated based on the appearance of colony on PDA medium, morphology (septate mycelia, nonseptate conidiophore, black conidia, and clublike spore-bearing head), after 48 h of growth at 30°C. Strains were selected in solid medium containing pectin citrus as sole carbon source and 0.5% rutenium red. The characterization of pectinolytic production was performed in solid culture and batch fermentation medium containing pectin citrus. The enzyme pectinolytic production was evaluated at 30°C, without agitation in 100 mL of medium containing 2% pectin citrus, 0.2% ammonium sulphate, 0.2% magnesium sulphate, and 0.05% potassium phosphate. The maximum pectinolytic activity (15U/mL) was observed in the medium after Aspergillus sp CFCF-0492 growth, while Aspergillus sp CFCF-CC1 showed the higher level of the final biomass. The pectinolytic activity is more preserved when the fungi-spores were maintained in agar-Czapeck medium.

Screening for yeast with antibacterial properties from an ethanol distillery

A general screening for the expression of antibacterial activity and non-flocculating type of yeast strains from must and fermented broth of alcohol distilleries was performed. From 60 strains only Saccharomyces sp. M26 presented a inhibitory halo in Lactobacillus fermentum culture and significant reduction in the culture turbidity (71%) and specific growth rate (56%) when compared to the control. Freezing did not affect the antibacterial activity of the Saccharomyces sp. M26 extract and heating at 90°C for 20 min completely destroyed this activity. It is expected the decrease of lactic acid bacteria growth in the S. cerevisiae alcoholic fermentation should allow for better control of these bacteria in the process. © 2003 Elsevier Ltd. All rights reserved.

Cryptococcosis outbreak in psittacine birds in Brazil

An outbreak of cryptococcosis occurred in a breeding aviary in São Paulo, Brazil. Seven psittacine birds (of species Charmosyna papou, Lorius lory, Trichoglossus goldiei, Psittacula krameri and Psittacus erithacus) died of disseminated cryptococcosis. Incoordination, progressive paralysis and difficulty in flying were seen in five birds, whereas superficial lesions coincident with respiratory alterations were seen in two birds. Encapsulated yeasts suggestive of Cryptococcus sp. were seen in faecal smears stained with India ink in two cases. Histological examination of the birds showed cryptococcal cells in various tissues, including the beak, choana, sinus, lungs, air sacs, heart, liver, spleen, kidneys, intestines and central nervous system. High titres of cryptococcal antigen were observed in the serum of an affected bird. In this case, titres increased during treatment and the bird eventually died. Yeasts were isolated from the nasal mass, faeces and liver of one bird. Cryptococcus neoformans var. gattii serovar B was identified based on biochemical, physiological and serological tests. These strains were resistant (minimum inhibitory concentration 64 μ g/ml) to fluconazole. This is the first report of C. neoformans var. gattii occurring in psittacine birds in Brazil. © 2004 ISHAM.

Experimental candidosis and recovery of Candida albicans from the oral cavity of ovariectomized rats

The aim of this study was to analyze the development of candidosis and the recovery of C. albicans from the oral cavity of ovariectomized and sham-ovariectomized rats. One hundred aiid twenty-four rats originally negative for Candida spp. in the oral cavity were divided into two groups: ovariectomized and sham-ovariectomized. Fifty-eight ovariectomized and the same quantity of sham-ovariectomized rats were inoculated with C. albicans for the study of candidosis development and recovery of yeast. Four animals from each group were not inoculated with yeast suspension and were submitted to tongue dorsum morphologic analysis by optical and scanning electron microscopy. The development of candidosis in the tongue dorsum was observed by optical and scanning electron microscopy in the periods of 6 hr, 24 hr, 7 days and 15 days after the last inoculation. Recovery of C. albicans was performed by oral samples plating on Sabouraud agar after 1,2, 5 and 7 days and progressively at each 15-day interval until negative cultures for yeasts were obtained. The results were analyzed by Mann-Whitney and Student's t tests. The tongue dorsum of sham-ovariectomized and ovariectomized rats, not infected by Candida, presented normal aspect. Among the infected rats, the ovariectomized group showed less occurrence of candidosis lesions and lower recovery of C. albicans from the oral cavity in relation to the sham-ovariectomized group. It could be concluded that candidosis was less frequent from the oral cavities of ovariectomized rats in relation to sham-ovariectomized.

Inhibitory effect of deferoxamine on Paracoccidioides brasiliensis survival in human monocytes: Reversal by holotransferrin not by apotransferrin

The mechanisms used by Paracoccidioides brasiliensis to survive into phagocytic cells are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Thus, the objective of this paper was to investigate the role of intracellular iron in regulating the capacity of P. brasiliensis yeast cells to survive within human monocytes. Treatment of monocytes with deferoxamine, an iron chelator, suppressed the survival of yeasts in a concentration-dependent manner. The effect of deferoxamine was reversed by iron-saturated transferrin (holotransferrin) but not by nonsaturated transferrin (apotransferrin). These results strongly suggest that P. brasiliensis survival in human monocytes is iron dependent.

Interação de Paracoccidioides brasiliensis com células endoteliais

Paracoccidioidomycosis has a variety of clinical manifestations and Paracoccidioides brasiliensis, the causative agent, may infect many tissues, most importantly the lungs. Migration of pathogenic yeasts to the endothelial cell layer is considered a prerequisite for multiple organ invasion and dissemination of the fungus. In this study of the adhesion of P. brasiliensis to endothelial cells in vitro, we investigated whether this adhesion could represent a mechanism of dissemination. To this end, as well as using conventional optical microscopy, an alternative in vivo technique was developed, to detect the presence of fungal cells in umbilical cords embedded in paraffin wax. An experiment on the migration of P. brasiliensis through an endothelial cell monolayer was carried out, and the migration of yeast cells was greater, and took less time, in control wells with no cells. The fungus crossed the monolayer, but, compared to control wells, the migration-rate was about 30% lower. This shows that the monolayer only partially blocked migration of the fungus. In these experiments, we had great difficulty finding P. brasiliensis adhered to the cell monolayer, when it was examined at different times, suggesting that migration of the fungus across the endothelial layer is very fast, and cannot normally be observed in cell culture in vitro. Thus, P. brasiliensis can cross the endothelium rapidly and probably invades deeper tissue.

Chloroquine inhibits Paracoccidioides brasiliensis survival within human monocytes by limiting the availability of intracellular iron

The mechanisms used by Paracoccidioides brasiliensis (Pb 18) to survive into monocytes are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens, including P. brasiliensis, whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Chloroquine, by virtue of its basic properties, has been shown to prevent release of iron from holotransferrin by raising endocytic and lysosomal pH, and thereby interfering with normal iron metabolism. Then, in view of this, we have studied the effects of CHLOR on P. brasiliensis multiplication in human monocytes and its effect on the murine paracoccidioidomycosis. CHLOR induced human monocytes to kill P. brasiliensis. The effect of CHLOR was reversed by FeNTA, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment. CHLOR treatment of Pb 18-infected BALB/c mice significantly reduced the viable fungi recovery from lungs, during three different periods of evaluation, in a dose-dependent manner. This study demonstrates that iron is of critical importance to the survival of P. brasiliensis yeasts within human monocytes and the CHLOR treatment in vitro induces Pb 18 yeast-killing by monocytes by restricting the availability of intracellular iron. Besides, the CHLOR treatment in vivo significantly reduces the number of organisms in the lungs of Pb-infected mice protecting them from several infections. Thus, CHLOR was effective in the treatment of murine paracoccidioidomycosis, suggesting the potential use of this drug in patients' treatment.

Comparative study of the clinical and anti-microbial efficacy of tongue cleaners

Candida species have frequently been isolated from the oral cavities of a variety of patients, such as elderly people, dentures users, immunocompromised and health patients. Yeasts may be associated with immune response and local factors such as poor oral hygiene. It was evaluated effectiveness of tongue cleaner showing which types would be preferred by patients, changes in tongue coating and in saliva yeasts counting. Thirty patients were selected and randomly distributed into three groups. This crossover blind study evaluated the effect of tongue cleaning using: a plastic and a steel tongue scraper and a nylon soft-bristle toothbrush. All patients were instructed to use the cleaners twice a day for one week (fifteen-day wash-out period). Saliva and tongue coating samples were collected from each patient from each test period, the yeasts were counted by colony forming units per mL (CFU/ mL) and the species were identified. The patients were questioned about cleaner preference. An increase in the percentage of patients with no tongue coating after scraping was observed. A reduction in the mean number of Candida species in tongue coating was observed only after nylon soft-bristle toothbrush cleaner. Candida albicans was the prevalent species. Volunteers preferred to the steel tongue scraper (60%). Tongue cleaners reduced the tongue coating and the mean number of saliva's yeasts. Degree of tongue coating favors the Candida species colonization.

Environmental isolation of black yeast-like fungi involved in human infection

The present study focuses on potential agents of chromoblastomycosis and other endemic diseases in the state of Paraná, Southern Brazil. Using a highly selective protocol for chaetothyrialean black yeasts and relatives, environmental samples from the living area of symptomatic patients were analysed. Additional strains were isolated from creosote-treated wood and hydrocarbon-polluted environments, as such polluted sites have been supposed to enhance black yeast prevalence. Isolates showed morphologies compatible with the traditional etiological agents of chromoblastomycosis, e.g. Fonsecaea pedrosoi and Phialophora verrucosa, and of agents of subcutaneous or systemic infections like Cladophialophora bantiana and Exophiala jeanselmei. Some agents of mild disease were indeed encountered. However, molecular analysis proved that most environmental strains differed from known etiologic agents of pronounced disease syndromes: they belonged to the same order, but mostly were undescribed species. Agents of chromoblastomycosis and systemic disease thus far are prevalent on the human host. The hydrocarbon-polluted environments yielded yet another spectrum of chaetothyrialean fungi. These observations are of great relevance because they allow us to distinguish between categories of opportunists, indicating possible differences in pathogenicity and virulence.

Patulina: Incidência e controle em derivados de maçã

The patulin, (4hydroxy-4furo[3,2-c]pyran(6H)-1), is a thermal resistent mycotoxin produced by several species of fungi are common in plants, mainly in derivatives and apples. Studies on the toxicity in animals have shown that mycotoxin has character teratogenic, and carcinogenic in mice immunotoxic. Its biosynthesis is well understood involving a series of reactions of condensation and oxiredução, many catalyzed by enzymes. The danger of contamination of food with patulin, warning about the need for a more rigorous control. Recent research aimed their removal and degradation as well as increase the sensitivity of the tests, making them faster and at less cost. The removal of patulin of food is made with composite adsorbents, with inconvenience to diminish the quality of the product by adsorbs other components desirable. The degradation is made with sulfur compounds, which are not allowed in food in many countries, and the growth of yeasts, such as the production of cider. Many yeasts have resistance against patulin and produce compounds capable degrade it. Here, we reviewed research on patulin with emphasis on its influence in food industry, incidence of patulin in apple juice and other foods, maximum permissible concentrations, health effects, biosynthesis, removal, degradation and most widely used methods for its detection and quantification.

Avaliação do potencial antagônico de leveduras, visando biocontrole de deterioração por penicillium expansum

Considerable losses during apple fruit storage occur due to microbiological diseases, mainly caused by Penicillium expansum, which in addition to fruit pulp deterioration produces patulin, a mycotoxin with carcinogenic and teratogenic activity. Biological control of post-harvest disease by antagonist yeasts focused on killer toxins is an appreciable alternative to the chemical fungicides, due to the low possibility of toxic residues demonstrated during fermentative processes. Twenty out of 44 yeasts (16 isolated from fruits, 10 from corn silage and 18 from laboratory anthill), showed antagonism against spores of P. expansum. The assay in solid medium pointed the strongest nutrient competition antagonism by D. hansenii strain C1 (31 mm inhibition diameter), while D. hansenii strain C7 (15 mm) showed higher antibiosis and parasitism pattern. In the following step the extracellular activity was tested performing the assay with culture supernatant in Yeast Medium agar, where C. guilliermondii P3 was more effective against conidia germination (inhibition rate of 58.15%) while P. ohmeri showed better inhibition on micelial growth (66.17%). The antibiosis showed by both yeasts could suggest probable mechanism associated with killer phenomenon, once both strains were killer positive against sensitive reference strains (S. cerevisiae NCYC 1006 and P. kluyveri CAY-15). In order to enhance the production of antifungal substance, these yeasts were cultivated with P. expansum, but the difference between culture supernatant obtained from yeasts cultivated alone and with mould was not significant (P > 0.05). The results demonstrated that the yeasts application constitute a promising tool, enhancing the biological control of P. expansum in post-harvest diseases of apple fruit.

In vitro susceptibility of oral Candida albicans strains to different pH levels and calcium hydroxide saturated aqueous solution

Candida albicans is present in the oral cavity and in the whole digestive tract of humans and other animals, being frequently related to endodontic treatment failure. The present study determined the incidence of C. albicans in the oral cavity and the susceptibility of isolates to different pH values and saturated calcium hydroxide aqueous solution at pH 12.5. Sixty-five patients attending the Endodontic Clinic at the Sagrado Coração University participated in the study. The collected samples were cultivated in selective media for C. albicans and the isolates were tested in terms of resistance to both alkaline pH and saturated aqueous solution of calcium hydroxide. In relation to time variables, yeast viability was assessed by the Sabouraud's agar culture and fluorescein diacetate and ethidium bromide fluorescent staining method. Results from the different pHs and experimental times, including those from different techniques measuring fungal viability, were compared using the chi-square and Fisher's exact tests (α=0.05). The yeasts became completely inviable after 48 h of contact with the calcium hydroxide solution. On the other hand, when exposed to the alkaline culture broth, the yeasts were found to be viable at pHs 9.5 and 10.5 for up to 7 days. In conclusion, C. albicans can only be completely inhibited by direct contact with saturated calcium hydroxide aqueous solution after 48 h of exposure.

Effects of probiotic bacteria on Candida presence and IgA anti-Candida in the oral cavity of elderly

Imbalance in the resident microbiota may promote the growth of opportunistic microorganisms, such as yeasts of Candida genus and the development of diseases, especially in aged people. This study evaluated whether the consumption of the probiotic Yakult LB® (Lactobacillus casei and Bifidobacterium breve) was able to influence on the specific immunological response against Candida and on the presence of these yeasts in the oral cavity of 42 healthy aged individuals. Saliva samples were collected before and after the probiotic use for 30 days, 3 times a week. The samples were plated in Dextrose Saboraud Agar with chloramphenicol, the colony-forming units (CFU/mL) were counted and the Candida species were identified. Anti-Candida IgA analysis was conducted using the ELISA technique. ANOVA and Student's t-test were used for normally distributed data and the Wilcoxon test was used for data with non-normal distribution (α=0.05). The results showed a statistically significant reduction (p<0.05) in Candida prevalence (from 92.9% to 85.7%), in CFU/mL counts of Candida and in the number of non-albicans species after consumption of the probiotic. Immunological analysis demonstrated a significant increase (p<0.05) in anti-Candida IgA levels. In conclusion, probiotic bacteria reduced Candida numbers in the oral cavity of the elderly and increased specific secretory immune response against these yeasts, suggesting its possible use in controlling oral candidosis.

Estudo da mcrobiota fúngica gastrintestinal de morcegos (Mammalia, Chiroptera) da região noroeste do estado de São Paulo: Potencial zoonótico

Bats are hosts of a rich diversity of microorganisms. Many studies indicate a close link between bats and fungi with pathogenic potential, especially for living in environments such as caves, caverns and hollow trees, favorable to the maintenance and spread of fungi. The objective was to study the gastrointestinal mycoflora of bats. Of the 98 samples belonging to 11 species of bats coming from 15 studied cities, 20% of the species were Carollia perspicillata, 19% Artibeus lituratus, 17% Molossus rufus, 13% Glossophaga soricina, 9% Nyctinomops macrotis, 8% Molossus molossus, 7% Desmodus rotundus, 2% Lasiurus ega and 1% Eptesicus furinalis, Myotis nigricans and Tadarida brasiliensis. The genus Aspergillus sp. was isolated from 29% of the samples, followed by 6% Microsporum sp. and Penicillium sp. 4% Trichophyton sp. and zygomycetes and 2% Fusarium sp. Of yeast species, 14% were from Rhodotorula sp., 10% Candida sp. and 2% Cryptococcus sp., 22% of isolates remained unidentified. All 82 cultures of organs were negative for Histoplasma capsulatum. There was a statistically significant association between the results of microbiological culture and bat species (p < 0.05). We conclude that the bats can act as disperser agents of fungi with pathogenic potential, although other studies should be performed to establish strategies to identify the main factors correlated with the growth and spread of microorganisms in nature and implication of bats in the epidemiological cycle.