Fabrication and characterization of physically crosslinked alginate/chitosan-based hydrogel loaded with neomycin for the treatment of skin infections caused by Staphylococcus aureus.

Staphylococcus aureus is a pathogen widely involved in wound infection due to its ability to release several virulence factors that impair the skin healing process, as well as its mechanism of drug resistance. Herein, sodium alginate and chitosan were combined to produce a hydrogel for topical delivery of neomycin to combat S. aureus associated with skin complications. The hydrogel was formulated by combining sodium alginate (50 mg/mL) and chitosan (50 mg/mL) solutions in a ratio of 9:1 (HBase). Neomycin was added to HBase to achieve a concentration of 0.4 mg/mL (HNeo). The incorporation of neomycin into the product was confirmed by scanning electron microscopy, FTIR and TGA analysis. The hydrogels produced are homogeneous, have a high swelling capacity, and show biocompatibility using erythrocytes and fibroblasts as models. The formulations showed physicochemical and pharmacological stability for 60 days at 4 ± 2 °C. HNeo totally inhibited the growth of S. aureus after 4 h. The antimicrobial effects were confirmed using ex vivo (porcine skin) and in vivo (murine) wound infection models. Furthermore, the HNeo-treated mice showed lower severity scores than those treated with HBase. Taken together, the obtained results present a new low-cost bioproduct with promising applications in treating infected wounds.

Development and Characterization of Hydroxyethyl Cellulose-Based Gels Containing Lactobacilli Strains: Evaluation of Antimicrobial Effects in In Vitro and Ex Vivo Models.

This study aimed to develop a hydroxyethyl cellulose-based topical formulation containing probiotics and to evaluate its antimicrobial action using in vivo and ex vivo models. Initially, the antagonistic effects of Lacticaseibacillus rhamnosus ATCC 10863, Limosilactobacillus fermentum ATCC 23271, Lactiplantibacillus plantarum ATCC 8014 and Lactiplantibacillus plantarum LP-G18-A11 were analyzed against Enterococcus faecalis ATCC 29212, Klebsiella pneumoniae ATCC 700603, Staphylococcus aureus ATCC 27853 and Pseudomonas aeruginosa ATCC 2785. The best action was seen for L. plantarum LP-G18-A11, which presented high inhibition against S. aureus and P. aeruginosa. Then, lactobacilli strains were incorporated into hydroxyethyl cellulose-based gels (natrosol); however, only the LP-G18-A11-incorporated gels (5% and 3%) showed antimicrobial effects. The LP-G18-A11 gel (5%) maintained its antimicrobial effects and viability up to 14 and 90 days at 25 °C and 4 °C, respectively. In the ex vivo assay using porcine skin, the LP-G18-A11 gel (5%) significantly reduced the skin loads of S. aureus and P. aeruginosa after 24 h, while only P. aeruginosa was reduced after 72 h. Moreover, the LP-G18-A11 gel (5%) showed stability in the preliminary and accelerated assays. Taken together, the results show the antimicrobial potential of L. plantarum LP-G18-A11, which may be applied in the development of new dressings for the treatment of infected wounds.

In Silico and In Vitro Analysis of Sulforaphane Anti-Candida Activity.

Oropharyngeal candidiasis/candidosis is a common and recurrent opportunistic fungal infection. Fluconazole (FLZ), one of the most used and effective antifungal agents, has been associated with a rise of resistant Candida species in immunocompromised patients undergoing prophylactic therapy. Sulforaphane (SFN), a compound from cruciferous vegetables, is an antimicrobial with yet controversial activities and mechanisms on fungi. Herein, the in silico and antifungal activities of SFN against C. albicans were investigated. In silico analyzes for the prediction of the biological activities and oral bioavailability of SFN, its possible toxicity and pharmacokinetic parameters, as well as the estimates of its gastrointestinal absorption, permeability to the blood-brain barrier and skin, and similarities to drugs, were performed by using different software. SFN in vitro anti-Candida activities alone and in combination with fluconazole (FLZ) were determined by the broth microdilution method and the checkerboard, biofilm and hyphae formation tests. Amongst the identified probable biological activities of SFN, nine indicated an antimicrobial potential. SFN was predicted to be highly absorbable by the gastrointestinal tract, to present good oral availability, and not to be irritant and/or hepatotoxic. SFN presented antifungal activity against C. albicans and prevented both biofilm and hyphae formation by this microorganism. SFN was additive/synergistic to FLZ. Overall, the data highlights the anti-Candida activity of SFN and its potential to be used as an adjuvant therapy to FLZ in clinical settings.

Ethyl Acetate Fraction of Punica granatum and Its Galloyl-HHDP-Glucose Compound, Alone or in Combination with Fluconazole, Have Antifungal and Antivirulence Properties against Candida spp.

Candidiasis is the most common fungal infection among immunocompromised patients. Its treatment includes the use of antifungals, which poses limitations such as toxicity and fungal resistance. Plant-derived extracts, such as Punica granatum, have been reported to have antimicrobial activity, but their antifungal effects are still unknown. We aimed to evaluate the antifungal and antiviral potential of the ethyl acetate fraction of P. granatum (PgEA) and its isolated compound galloyl-hexahydroxydiphenoyl-glucose (G-HHDP-G) against Candida spp. In silico analyses predicted the biological activity of G-HHDP-G. The minimum inhibitory concentrations (MIC) of PgEA and G-HHDP-G, and their effects on biofilm formation, preformed biofilms, and phospholipase production were determined. In silico analysis showed that G-HHDP-G has antifungal and hepatoprotective effects. An in vitro assay confirmed the antifungal effects of PgEA and G-HHDP-G, with MIC in the ranges of 31.25-250 µg/mL and 31.25 ≥ 500 µg/mL, respectively. G-HHDP-G and PgEA synergistically worked with fluconazole against planktonic cells. The substances showed antibiofilm action, alone or in combination with fluconazole, and interfered with phospholipase production. The antifungal and antibiofilm actions of PgEA and G-HHDP-G, alone or in combination with fluconazole, in addition to their effects on reducing Candida phospholipase production, identify them as promising candidates for therapeutics.

Effect of Terminalia catappa Linn. on Biofilms of Candida albicans and Candida glabrata and on Changes in Color and Roughness of Acrylic Resin.

This study aimed to investigate the effect of the n-butanol fraction of Terminalia catappa Linn., (FBuTC) on biofilm of Candida albicans and Candida glabrata, as well as changes in color and roughness of polymethyl methacrylate resin (PMMA). The susceptibility of C. albicans and C. glabrata to FBuTC was evaluated by means of the Minimum Inhibitory and Minimum Fungicidal Concentration (MIC and MFC). PMMA acrylic resin discs (N= 108) were fabricated. For the susceptibility tests, biofilms of C. albicans and C. glabrata were developed on discs for 48 h and immersed in phosphate-saline buffer solution (PBS), 1% sodium hypochlorite (SH 1%), or FBuTC at MIC, 5xMIC, or 10xMIC. For the color and roughness change tests, the discs were immersed in distilled water, SH 1%, or FBuTC in the concentrations of 0.25 mg/mL, 2.5 mg/mL, or 25.0 mg/mL. After 28 days of incubation, color change was evaluated by spectrophotometry and roughness, by using a profilometer. The biofilms were investigated by one-way ANOVA and, the color and roughness changes (two-way ANOVA and the Tukey test; α=0.05). For both MIC and MFC the value of 0.25 mg/mL of FBuTC was observed for the planktonic cells of C. albicans and C. glabrata. Exposure to FBuTC at 10xMIC had a significant effect on the biofilm of C. albicans, showing a reduction in cell counts when compared with PBS, (p=0.001). For the biofilm of C. glabrata, the MIC was sufficient for significantly reducing the cell count (p<0.001). No important changes in color and roughness of the acrylic resin were observed, even after 28 days, irrespective of the concentration of FBuTC used (p >0.05). It could be concluded that the immersion of acrylic resin for dental prosthesis in FBuTC was effective in reducing the biofilms of C. albicans and C. glabrata without evidence of change in roughness and color of this substrate.

Betulinic Acid Prevents the Acquisition of Ciprofloxacin-Mediated Mutagenesis in Staphylococcus aureus.

The occurrence of damage on bacterial DNA (mediated by antibiotics, for example) is intimately associated with the activation of the SOS system. This pathway is related to the development of mutations that might result in the acquisition and spread of resistance and virulence factors. The inhibition of the SOS response has been highlighted as an emerging resource, in order to reduce the emergence of drug resistance and tolerance. Herein, we evaluated the ability of betulinic acid (BA), a plant-derived triterpenoid, to reduce the activation of the SOS response and its associated phenotypic alterations, induced by ciprofloxacin in Staphylococcus aureus. BA did not show antimicrobial activity against S. aureus (MIC > 5000 µg/mL), however, it (at 100 and 200 µg/mL) was able to reduce the expression of recA induced by ciprofloxacin. This effect was accompanied by an enhancement of the ciprofloxacin antimicrobial action and reduction of S. aureus cell volume (as seen by flow cytometry and fluorescence microscopy). BA could also increase the hyperpolarization of the S. aureus membrane, related to the ciprofloxacin action. Furthermore, BA inhibited the progress of tolerance and the mutagenesis induced by this drug. Taken together, these findings indicate that the betulinic acid is a promising lead molecule in the development helper drugs. These compounds may be able to reduce the S. aureus mutagenicity associated with antibiotic therapies.

Climate drivers of hospitalizations for mycoses in Brazil.

Climate can modulate human health at large spatial scales, but the influence of global, regional, and local environments remains poorly understood, especially for neglected diseases, such as mycoses. In this work, we present the correlation between climatic variables and hospitalizations for mycoses in Brazilian state capitals, evaluating the period of 2008 to 2016 at different time scales. The results indicate that climate modulates the hospitalizations for mycoses differently at annual and monthly time scales, with minimum temperature as a key climatic variable during periods of high prevalence in the 10 Brazilian capitals with the highest hospitalizations for mycoses rates. The greatest number of hospitalizations coincided with La Niña events, while a reduction was observed during El Niño events, thereby demonstrating the influence of the Pacific Interdecadal Climate Oscillation on the prevalence of mycoses in Brazil. At a regional scale, the mycoses burden in Brazil appears to respond differently to local and global climatic drivers.

Hydroalcoholic Extract and Ethyl Acetate Fraction of Bixa orellana Leaves Decrease the Inflammatory Response to Mycobacterium abscessus Subsp. massiliense.

The incidence of infections caused by rapidly growing mycobacteria (RGM), especially Mycobacterium abscessus subsp. massiliense (Mabs), is increasing worldwide. Severe infections are associated with abscess formation and strong inflammatory response. This study evaluated the antimicrobial and anti-inflammatory activities of a hydroalcoholic extract (BoHE) and ethyl acetate fraction (BoEA) of Bixa orellana leaves. Antimicrobial activity was evaluated by broth microdilution to determine the minimum inhibitory (MIC) and the minimum bactericidal (MBC) concentrations. Cytotoxicity was evaluated using erythrocytes and RAW 264.7 cells. Nitric oxide (NO) was assayed in stimulated RAW 264.7 cells, and inflammatory cell migration and acute toxicity were evaluated in a Mabs-induced peritonitis mouse model. The compounds present in BoEA were identified by high performance liquid chromatography and mass spectrometry (HPLC-MS). The MIC and MBC values were 2.34 mg/mL and 37.5 mg/mL for BoHE and 0.39 mg/mL and 6.25 mg/mL for BoEA. The extracts did not induce significant toxicity in erythrocytes and RAW 264.7 cells. High levels of NO induced by Mabs were decreased by treatment with both extracts. The anti-inflammatory activity was confirmed in vivo by significant reduction of the cell migration to the peritoneum following BoHE and BoEA pretreatment. Animals treated with BoHE or BoEA did not show signs of acute toxicity in stomach, liver, and kidney. The chemical characterization of BoEA (the most active extract) revealed that kaempferol-3-O-coumaroyl glucose is its major component. The extract of B. orellana may be effective for treating infections caused by Mabs.

Identification of specific antibodies against the Ag85C-MPT51-HspX fusion protein (CMX) for serological screening of tuberculosis in endemic area.

OBJECTIVES: Development of new tools for rapid and accurate diagnosis of tuberculosis (TB) is considered a strategy for controlling the disease. The recombinant CMX fusion protein is composed of immunodominant epitopes of the Ag85C (Rv0129c), MPT51 (Rv3803c) and the entire HspX (Rv2031c) proteins from Mycobacterium tuberculosis H37Rv (Mtb). The aim of this study was to evaluate the applicability of a test using the CMX protein in individuals suspected of TB. METHODS: Indirect ELISA was used to measure serum anti-CMX IgM and IgG in individuals with pulmonary TB. RESULTS: Patients with pulmonary TB had higher titers of IgM (OD = 0.502 ± 0.281) than healthy controls (OD = 0.200 ± 0.125). The cutoff for IgM-ELISA was determined using ROC curve analyzes (AUC = 0.868) with a sensitivity of 80.1% and a specificity of 78.2%. Patients with pulmonary TB also had higher titers of IgG (OD = 0.525 ± 0.391) than healthy controls (OD = 0.215 ± 0.077). The cutoff for IgG-ELISA was determined using ROC curve analyzes (AUC = 0.864) with a sensitivity of 81.7% and a specificity of 74.7%. CONCLUSION: The results suggest that the recombinant protein CMX can be used in a serological test to complement the screening of individuals suspected of having active pulmonary TB.

Phytochemical Characterization of Terminalia catappa Linn. Extracts and Their antifungal Activities against Candida spp.

Terminalia catappa Linn bark is used to treat dysentery by various populations in Southeast Asian countries, and its leaves have also been used in traditional medicine to treat hepatitis in India and the Philippines. Here, the antifungal actions of crude hydro-alcoholic extract (TcHE) and fractions from T. catappa leaves were assessed via the agar diffusion and microdilution tests on Candida reference strains and clinical isolates from patients with acquired immunodeficiency syndrome (AIDS). Additionally, the potential cytotoxic effects of TcHE were assessed on cultured human peripheral blood mononuclear cells (PBMC). T. catappa fractions and sub-fractions were analyzed by gas chromatography coupled to mass spectrometry with electron impact (GC/MS/EI), high-performance liquid chromatography coupled to mass spectrometry "electrospray" ionization in positive mode (HPLC/MS/MS/ESI+) and hydrogen nuclear magnetic resonance (1HNMR). TcHE and its fractions were able to inhibit the growth of all tested Candida strains with the n-butanol (FBuOH) fraction presenting the best antifungal activity. Testing of different FBuOH sub-fractions (SF) showed that SF10 was the most active against Candida spp. Fractioning of SF10 demonstrated that 5 out of its 15 sub-fractions were active against Candida spp., with SF10.5 presenting the highest activity. Chemical analysis of SF10 detected hydrolysable tannins (punicalin, punicalagin), gallic acid and flavonoid C-glycosides. Overall, the results showed that T. catappa L. leaf extract, fractions and sub-fractions were antifungal against Candida spp. and may be useful to treat diseases caused by this fungus.

Prevalence and production of enzymes by Candida isolates from vaginal secretion samples

A candidíase vulvovaginal (VVC) ocupa o segundo lugar entre as vaginites, sendo Candida albicans responsável pela maioria dos episódios sintomáticos de VVC. Entretanto, um fato a ser considerado é o aumento de infecções por espécies de C. não albicans, o que contribui para as elevadas taxas de recidiva e resistência. Os objetivos deste estudo foram isolar Candida spp. de amostras de secreção vaginal, determinar a prevalência das espécies e verificar a produção de enzimashidrolíticas (proteases, fosfolipases, hemolisinas, catalases e gelatinases) nos isolados clínicos. Foram analisadas 144 amostras de pacientes atendidas em um hospital público em São Luís-MA.A produção de enzimas hidrolíticas foi conduzida em triplicata usando-se métodos específicos. A análise estatística foi realizada por meio dos testes c2, Kruskal-Wallis e Student-Newman-Keuh. Apresentaram cultura positiva para Candida spp. 90 pacientes. A principal espécie isolada (43,3por cento) foi Candida parapsilosis e 43 pacientes (47,8por cento) apresentaram manifestações clínicas sugestivas de VVC.al quando em condições apropriadas.

A correlação entre a presença de espécies de Candida e a presença ou ausência de sintomas não foi estatisticamente significante (x igual 3,22 , p igual 0,073). Verificou-se a expressão de enzimas pelos isolados de Candida spp. nas seguintes porcentagens: hemolisinas (80por cento), fosfolipases (8,9por cento), proteinases (17,8por cento), catalases (64,4por cento) e gelatinases (10,0por cento). Diferenças estatisticamentesignificativas foram observadas entre os isolados em relação à expressão de fosfolipases (p igual 0,0005), sendo C. albicans a espécie mais produtora, e de catalases (p igual 0,0045), em que C. parapsilosis e C. albicans foram as espécies com expressão mais elevada. Este estudo mostrou uma predominânciade espécies não albicans em amostras vulvovaginais e a maioria dos isolados revelou-se produtora de hemolisinas e catalases, o que pode contribuir para a virulência das linhagens de Candida e para o desenvolvimento de infecção vaginal quando em condições apropriadas.

Prevalência e produção de exoenzimas por espécies de Candida provenientes da mucosa bucal de pacientes com AIDS e indivíduos hígidos / Prevalence and exoenzyme production by Candida species from the oral mucosa of AIDS patients and healthy individuals

Infecções por leveduras do gênero Candida têm gerado elevados índices de morbidade e mortalidade, longo período de permanência em hospitais, dificuldade e alto custo de tratamento.Indivíduos imunodeprimidos como os portadores da Síndrome da Imunodeficiência Adquirida (Aids) apresentam grande suscetibilidade a desenvolver essas infecções em razão do baixo número de linfócitos T-CD4, menor que 200 cel/mm3. Candida albicans é a espécie mais estudadae está relacionada com os processos de colonização e patogenicidade na boca do homem. Essa característica é decorrente, entre outros fatores, da produção de exoenzimas facilitadoras dainteração do fungo com as células do hospedeiro. Este estudo verificou a produção das exoenzimas proteinase, fosfolipase, gelatinase e hemolisina de amostras bucais de Candida isoladas de 49 pacientes com Aids (grupo teste) e de 26 indivíduos hígidos (grupo controle). C. albicans foi a espécie mais prevalente no grupo teste (59,2por cento) e Candida parapsilosis (53,8por cento) no grupo controle. C. albicans apresentou resultados significativos para a produção de proteinase (ambos os grupos) e fosfolipase no grupo teste. Já as espécies de Candida não albicans (CNA) apresentaram resultadosaltamente significativos para fosfolipase no grupo controle. Em relação às enzimas gelatinase e hemolisina, não foram encontradas diferenças significantes entre C. albicans e espécies CNA. Por fim, não foi encontrada diferença estatística na produção de exoenzimas quando foi comparado o grupo teste com o grupo controle.

Detecção de betalactamases de espectro estendido (ESBL) em isolados de Escherichia coli uropatogênicas (UPECS) oriundos de pacientes da comunidade / Detection of extended spectrum beta-lactamases (ESBLS) in isolates of uropathogenic Escherichia coli (UPECs) from patients in the community

A produção de betalactamases de espectro estendido (ESBL) consiste em um importante mecanismo de resistência aos antimicrobianos betalactâmicos. O vasto uso desses antibióticos no tratamento das infecções do trato urinário (ITU) tem agravado ainda mais esta problemática. As bactérias do tipo UPEC são consideradas as principais responsáveis por este tipo de infecção. Estudos para acompreensão da diversidade dessas enzimas no Brasil são escassos e, além disso, muitos deles são direcionados à pesquisa de isolados de origem hospitalar. Os objetivos do presente estudo foram detectar os genes que codificam as betalactamases TEM, CTX-M e SHV por PCR multiplexe identificar os isolados que as produzem por meio do método do disco aproximação. Do total dos isolados analisados, 12,8por cento apresentaram o fenótipo positivo para ESBL, enquanto 65,8por cento amplificaram sequências homólogas para os genes de betalactamases. Destes, os predominantes foram blaTEM e blaCTX-M, com 42,7por cento e 18,8por cento, respectivamente. Dois isolados não amplificaram para nenhum dos genes em estudo, apesar de terem expressado a enzima no teste fenotípico. Os resultados obtidos neste trabalho ressaltam a necessidade de mais estudos que visem à caracterizaçãomolecular para elucidar a disseminação desse mecanismo de resistência entre bactérias oriundas de pacientes da comunidade.

Prevalência de vaginose bacteriana em pacientes que realizaram bacterioscopia de secreção vaginal em laboratório de saúde pública / Prevalence of bacterial vaginosis in patients who vaginal bacterioscopy implemented in a public health laboratory

JUSTIFICATIVA E OBJETIVOS: Vaginose bacteriana (VB) é caracterizada pela alteração na microflora da vagina, com redução e/ou ausência de espécies de Lactobacillus. É a causa mais comumde infecções vaginais em mulheres na idade reprodutiva e frequentemente está associada a corrimentos vaginais. O objetivo deste estudo foi verificar a prevalência de VB em pacientes ambulatoriais e leucocitose, relacionando com a presença de outros micro-organismos. MÉTODO: Foram analisados 765 laudos provenientes de pacientes ambulatoriais do Laboratório Central de Saúde Pública de São Luís, no período de janeiro de 2006 a março de 2009. RESULTADOS: Foram baseados na bacterioscopia de esfregaços de secreções vaginais corados pelo Gram e no exame direto à fresco para a pesquisa de Candida spp. e Trichomonas vaginalis. Quando analisada a prevalência global de micro-organismos, obteve-se 39% dos casos positivos. Gardnerella vaginallis foi o micro-organismo mais prevalente, seguido de Candida spp., Mobiluncusspp e Trichomonas vaginalis. O número de leucócitos variou, embora a presença de poucos leucócitos tenha sido maisfrequente. CONCLUSÃO: A prevalência de VB nas pacientes atendidas no período estudado foi maior na faixa etária de menores de 20 anos com aproximadamente 39% das amostras positivas para presença de micro-organismos potencialmente patogênicos.

BACKGROUND AND OBJECTIVES: Bacterial vaginosis (BV) is characterized by alterations in the microflora of the vagina,reducing and/or absence of species of Lactobacillus. It is the most common cause of vaginal infections in women of reproductive age and often is associated with vaginal discharge. The aim ofthis study was to assess the prevalence of BV in outpatients and leukocytosis, relating to the presence of other microorganisms. METHOD: There were analyzed 765 cases of Laboratório Central de Saúde Pública de São Luís, during January 2006 to March 2009. RESULTS: Were based in bacterioscopy of vaginal smears coloredby Gram and the wet mount for the research of Candida spp. and Trichomonas vaginalis. The global prevalence of microorganisms was 39%. Gardnerella vaginallis was the most prevalent microorganism, followed by Candida spp., Mobiluncus spp and Trichomonas vaginalis. The number of leukocytes varied, although the leukopenia was more frequent. CONCLUSION: It can be concluded that the prevalence of BV in patients treated during the period studied was higher in theage group younger than 20 years with approximately 39% of samples positive for the presence of potentially pathogenic microorganisms

Diversity and prevalence of airborne fungi isolated from São Luis, Northeast Brazil

Airborne fungi are considerate the main agents related to allergy in patients that suffer from asthma and rinitis. The knowledge about the prevalence of these microorganisms and theis seasonal variation can be useful for human health by improving the diagnosis and treatment of respiratory allergic disease provoked by their inhalation. The present study has isolated and identified airborne funfi from São Luis city investigating their prevalence, biodiversity and seasonal variation. Samples were taken by the Petri plate gravitacional method in six urban areas. A total of 2993 fungal colonies were counted, 67,9% in the dry season and 32,1% in the rainy season. Statistical analysis showed that there are significant diference (p=0,0121) between the averages of spores in dry and rainy periods. Relative humidity, rainfall precipitation and wind speed can have influenced the diversity and concentration of fungal spores presents in the air. Among 24 genera isolated, the most prevalent ones were Aspergillus (38,7)%, Fusarium (13,90%), Curvularia (11,18%) and Penicillium (10,88%), major triggers of allergic process worldwide recognized. The data show the presence of a large number of airborne fungi spores in São Luis and all genera found are opportunistic and they may cause disease in susceptible patients.

In vitro differential activity of phospholipases and acid proteinases of clinical isolates of Candida / Atividade diferencial in vitro de fosfolipases e proteinases ácidas de isolados clínicos de Candida

INTRODUCTION: Candida yeasts are commensals; however, if the balance of normal flora is disrupted or the immune defenses are compromised, Candida species can cause disease manifestations. Several attributes contribute to the virulence and pathogenicity of Candida, including the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate the in vitro activity of phospholipases and acid proteinases in clinical isolates of Candida spp. METHODS: Eighty-two isolates from hospitalized patients collected from various sites of origin were analyzed. Phospholipase production was performed in egg yolk medium and the production of proteinase was verified in a medium containing bovine serum albumin. The study was performed in triplicate. RESULTS: Fifty-six (68.3 percent) of isolates tested were phospholipase positive and 16 (44.4 percent) were positive for proteinase activity. C. tropicalis was the species with the highest number of positive isolates for phospholipase (91.7 percent). Statistically significant differences were observed in relation to production of phospholipases among species (p<0,0001) and among the strains from different sites of origin (p=0.014). Regarding the production of acid protease, the isolates of C. parapsilosis tested presented a larger number of producers (69.2 percent). Among the species analyzed, the percentage of protease producing isolates did not differ statistically (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSIONS: The majority of C. non-albicans and all C. albicans isolates were great producers of hydrolytic enzymes and, consequently, might be able to cause infection under favorable conditions.

INTRODUÇÃO: Candida são leveduras comensais, porém, se o equilíbrio da flora normal for interrompido ou as defesas imunitárias estiverem comprometidas, espécies de Candida podem causar manifestações de doença. Vários atributos contribuem na virulência e patogenicidade de Candida, inclusive a produção de enzimas extracelulares hidrolíticas, especialmente fosfolipases e proteinases. O objetivo deste estudo foi verificar a atividade in vitro de fosfolipases e proteinases ácidas em isolados clínicos de Candida spp. MÉTODOS: Oitenta e dois isolados provenientes de pacientes hospitalizados coletados a partir de sítios de origem diversos foram analisados. A produção de fosfolipase foi verificada em meio egg yolk e a de proteinase em meio contendo soro albumina bovina. O estudo foi feito em triplicata. RESULTADOS: Cinquenta e seis (68,3 por cento) dos isolados testados apresentaram atividade de fosfolipase positiva e 16 (44,4 por cento) foram positivos para atividade de proteinase. C. tropicalis foi a espécie que apresentou o maior número de isolados positivos para fosfolipases (91,7 por cento). Diferenças estatisticamente significantes em relação à produção de fosfolipases entre as espécies e entre as cepas provenientes de diferentes sítios de origem foram detectadas. Quanto à produção de proteinases ácidas, os isolados de C. parapsilosis testados foram os maiores produtores (69,2 por cento). Entre as espécies analisadas, a porcentagem de produção de proteinase entre os isolados não diferiu estatisticamente (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSÕES: A maioria dos isolados de C. não-albicans, assim como os de C. albicans, foram grandes produtores de enzimas hidrolíticas e, consequentemente, podem ser capazes de causar infecção em condições adequadas.