Candida glabrata, geralmente comensal, surgiu como uma causa comum de infecções fúngicas graves com risco de morte. Dada a resistência crescente aos azóis, a orientação recente é utilizar as equinocandinas como a primeira escolha para o tratamento de infecções sistêmicas por C. glabrata. No entanto, esta é a primeira espécie de Candida que foi detectada com resistência significativa às equinocandinas. Esta levedura é capaz de colonizar tecidos do hospedeiro, bem como superfícies abióticas (catéteres, próteses) onde desenvolve um crescimento em multicamadas caracterizado como biofilme. A natureza da estrutura do biofilme e os atributos fisiológicos a ele conferidos resultam em uma resistência inerente a agentes antimicrobianos, impactando negativamente na saúde do paciente. Este estudo teve como objetivo a indução in vitro de resistência à anidulafungina em células planctônicas e sésseis de sete cepas sensíveis de C. glabrata, além da verificação do desenvolvimento de resistência cruzada com fluconazol. A indução de resistência foi realizada submetendo as cepas a concentrações sub-inibitórias do antifúngico. A determinação de concentração inibitória mínima através de microdiluição em caldo foi realizada previamente e posteriormente à indução de resistência e, também, para a verificação de resistência cruzada com fluconazol. O método de indução de resistência resultou em cepas fortemente resistentes à anidulafungina, com concentrações inibitórias mínimas variando de 1 a 2 μg/mL. Antes da indução da resistência, as formas planctônica e séssil das cepas eram todas sensíveis ou sensíveis dose-dependente ao fluconazol. Após a indução de resistência à anidulafungina esta sensibilidade ao fluconazol não foi mantida, tornando as cepas resistentes a este antifúngico. Clinicamente, esta resistência cruzada poderia implicar em falha terapêutica ao utilizar o fluconazol em pacientes previamente expostos a concentrações sub-inibitórias de anidulafungina por longos períodos. / Candida glabrata, usually commensal, has emerged as a common cause of serious life threatening fungal infections. Given the increasing resistance to azoles, the recent guidance is to use echinocandins as the first choice for the treatment of systemic infections by C. glabrata. However, C. glabrata is the first species of Candida that has been detected with significant resistance to echinocandins. This yeast is able to colonize host tissues as well as abiotic surfaces (catheters, prostheses) where it develops a multi-layer growth characterized as biofilm. The nature of the biofilm structure and the physiological attributes conferred on it, result in an inherent resistance to antimicrobial agents, negatively impacting the patient's health. This study aimed to induce in vitro resistance to anidulafungin in planktonic and sessile cells of seven sensitive C. glabrata strains, as well as to verify the development of cross-resistance with fluconazole. The induction of resistance was performed by subjecting the isolates to sub-inhibitory concentrations of the antifungal. The determination of minimum inhibitory concentration by broth microdilution was performed before and after induction of resistance and also for fluconazole cross-resistance verification. The resistance induction test resulted in strains strongly resistant to anidulafungin, with minimum inibitory concentrations ranging from 1 to 2 μg mL-1. Prior to induction of resistance, the planktonic and sessile forms of the strains were all sensitive or sensitive dose-dependent to fluconazole. However, after the induction of resistance to anidulafungin, this sensitivity to fluconazole was not maintained, making the strains resistant to this antifungal. Clinically, this cross-resistance could lead to therapeutic failure when using fluconazole in patients previously exposed to sub-inhibitory concentrations of anidulafungin for long periods.

Orientadores: Altair Antoninha Del Bel Cury, Renata Cunha Matheus Rodrigues-Garcia / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-06T13:46:58Z (GMT). No. of bitstreams: 1 Pereira-Cenci_Tatiana_M.pdf: 1048789 bytes, checksum: e19520a3c91fdd0a8f929e43936e3dcc (MD5) Previous issue date: 2006 / Resumo: A candidose é a infecção oral fúngica mais comum diagnosticada em humanos, com prevalência de até 67% em usuários de prótese. Embora tenha sido inicialmente associada apenas a Candida albicans, outras espécies podem ser responsáveis por mais de 50% dos casos de infecção. Ainda, fatores como presença de saliva e bactérias parecem desempenhar importante papel na colonização por Candida. Assim, este estudo objetivou verificar a influência destes fatores na a:fesão de duas espécies de Candida (Candida albicans e Candida glabrata) sobre a superfície de resinas acrílicas e reembasadores. Corpos de prova (2,5x1 ,2xO,2 cm) confeccionados com duas resinas acrílicas (convencional e de microondas) e dois reembasadores (temporário e permanente) tiveram sua rugosidade (Ra) e energia livre de superfície (ELS) mensuradas, sendo aleatoriamente divididos de acordo com a exposição aos fatores: presença ou ausência de saliva, presença ou ausência de bactérias e espécie de Candida. Os espécimes foram levados a uma câmara de fluxo utilizando-se uma bomba peristáltica para perfusão de cultura de bactérias seguida por uma das espécies de Candida, ou apenas a cultura de uma das espécies de Candida. A contagem das células de Candida aderidas foi realizada em microscópio óptico (400x). Os dados foram submetidos à análise de variância para Ra e adesão, e ao teste de Kruskal-Wallis para ELS (a=0,05). O reembasador temporário apresentou a maior Ra, seguido do permanente, enquanto as resinas acrílicas exibiram as menores rugosidades (p<0,0001). Os valores de ELS foram similares para os materiais, mas diferentes do reembasador temporário (p<0,0001). A adesão de C. albicans e C. glabrata variou de 3,2 a 564,4 e 3,2 a 1400,4 cel/mm2 respectivamente, com diferenças estatísticas (p<0,05) em alguns grupos. O reembasador temporário mostrou maiores níveis de adesão. A colonização foi diminuída pela saliva, enquanto na presença de bactérias e saliva houve aumento da adesão (p<0,05). Estes resultados sugerem que a adesão inicial das duas espécies de Candida foi fortemente afetada pela rugosidade, presença de saliva e bactérias, mas não pela energia livre de superfície / Abstract: Candida-associated stomatitis is reported in up to 67% of a population wearing dentures. Recently, disease-associated Candida species have shifted from C. albicans to norralbicans species. Since factors such as presence of saliva and oral bacteria appear to play a major role in the initial phases of yeasts adhesion, this study aimed to determine whether these factors produced differences in acrylic resins and denture liners C. albicans and C. glabrata adherence. Samples (2.5x1.2xO.2 em) of two acrylic resins (heat and microwavecured) and two denture liners (soft and hard) were prepared and had their surface free energy (SFE) and surface roughness (Ra) measured and were randomly divided according to their exposure to the following factors: saliva coating or uncoating, presence or absence of bacteria and Candida species. Specimens were assayed in a flow chamber connected to a peristaltic pump for perfusion of bacteria culture plus one of the Candida species culture or only the Candida culture (control). Adhesion was determined by count on a light microscope (400 x). Statistical analyses was performed by ANOVA (Ra and Candida species adhesion) and Kruskal-Wallis (SFE) (a=.05). Soft liner presented the roughest surface, followed by the hard liner, whereas acrylic resins exhibited the smoothest surfaces (p<.0001). The SFE values of ali materiais were similar but different from the soft liner (p<.0001). C. albicans and C. glabrata adhesion on the materiais ranged fr0m 3.2 to 564.4, and 3.2 to 1400.4 cells mm-2 respectively, with statistically signific,ant differences (p<.05) in some cases. The soft liner exhibited the highest levels of adhesion. The overall colonization was significantly decreased by saliva (p<.Oq), while bacteria increased the adhesion in the presence of saliva. These results taken together suggest that initial adhesion of Candida species was strongly affected by the surface roughness, presence of saliva and bacteria, but not by surface free energy / Mestrado / Protese Dental / Mestre em Clínica Odontológica

orionpizango@gmail.com / Case report: A 43-year-old female presented with decreased visual acuity in the right eye.“Snowball-like” retinal lesions were found in both eyes on examination. Due to a lackof improvement with intravitreal antifungal empirical treatment, vitreous culture wasperformed and Candida glabrata was isolated. The patient then received intravitreal ampho-tericin B, as well as systemic treatment with caspofungin and amphotericin B lipid complex.Discussion: Endogenous fungal endophthalmitis is a sight-threatening condition. There arefew reports of C. glabrata endogenous endophthalmitis. Treatment regimens for Candidaendophthalmitis include combinations of systemic and/or intravitreal antifungals, as wellas vitrectomy.

We report a rare case of Candida glabrata liver abscess and fungemia complicating severe calculus cholecystitis in a 64-year-old female patient who had no history of immunosuppression or diabetes mellitus. The patient underwent cholecystectomy, resection of liver abscess, and systemic antifungal therapy using micafungin. Copyright

Objectives: Candida glabrata is the second most common cause of oral candidiasis, second only to C. albicans. Incidence of antifungal resistance has shown a steady increase for C. glabrata. Iron has shown to modulate C. albicans pathogenesis and affect drug-susceptibility. Here, we assess the effect of iron on the growth, antifungal-susceptibility, biofilms, and cell wall of C. glabrata.Methods: Growth, minimal inhibitory concentration (MIC), and biofilm experiments were conducted using 96-well polystyrene plates. Yeast Nitrogen Base medium was used for growth experiments. Cultures of C. glabrata and C. albicans were grown over two nights in respective media containing varying iron concentrations. Rosewell Park Memorial Institute medium was used for MIC and biofilm experiments. Serial dilution was performed to obtain desired concentrations of antifungal drugs. For all experiments, growth was assessed at OD600nm over 24 hours using BioTek Synergy Multi Mode Reader. Paraformaldehyde treated cells and specific stains were used for cell wall studies. Results: Growth of C. glabrata declined significantly below 5μM iron, while C. albicans continued to grow at decreasing iron concentrations, up to 0.5μM. MIC experiments revealed 1.562μM, 1.562μM, and 4μM, as the MIC for Deferasirox, Nystatin, and Fluconazole, respectively. Drug synergy experiments revealed a 128-fold reduction in the amount of Nystatin and Fluconazole needed, with the addition of 1/8th of Deferasirox concentration. The biofilm experiments were inconclusive and the cell wall studies showed decreased levels of mannan, chitin, and an increased β-glucan exposure in high iron conditions. Conclusion: C. glabrata is more sensitive to alterations in environmental iron when compared to C. albicans. Drug synergy experiments underscore the importance of Deferasirox in lowering the MICs of Nystatin or Fluconazole. This can allow use of classical antifungals at lower doses, thereby limiting their side effects. Cell wall studies discuss the effect of iron on the virulence of the C. glabrata. / Oral Biology

Vulvovaginal candidiasis is a common gynecologic diagnosis that can be treated empirically with fluconazole. We present a patient that developed post-operative () fungemia after being empirically treated for vulvuovaginal candidiasis with fluconazole multiple times throughout the year prior to robotic total laparoscopic hysterectomy and bilateral salpingo-oophorectomy. is becoming increasingly resistant to azole antimycotic therapy. It is likely that this patient had undertreated fluconazole-resistant vulvovaginitis prior to surgery, and that the pelvic infection was the source of fungemia.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Candida species are the most common etiologic agent of opportunistic fungal infections. Candida albicans is the most frequent species in candidiasis, however species of non-albicans species have emerged increasingly in hospitals in which the use of azoles as a prophylactic or therapeutic is common. C. glabrata is the species most relevant in this scenario, causing infections with high morbidity and mortality due to high rates of resistance to antifungal azoles, especially fluconazole, and reduced sensitivity to amphotericin B and echinocandins. The remarkable resistance of these species to antifungal agents requires the search for new therapeutic options. An alternative is the combination of drugs with different mechanisms of action. This study aimed to evaluate the in vitro susceptibility of Candida glabrata to conventional antifungal agents (amphotericin B, ketoconazole, itraconazole, fluconazole and voriconazole), and these associations with non-antifungal drugs (chlorpromazine, linezolid, minocycline, sulfamethoxazole, tacrolimus and diphenyl diselenide). Two groups of clinical isolates of C. glabrata were evaluated. The first group consists of fluconazole-sensitive clinical isolates (FS) (n = 30) and the second, derivative of the first, fluconazole-resistant clinical isolates (FR) (n = 30). Based on protocol M27-A3 (CLSI, 2008), the checkerboard method. In isolation, MICs (Minimum Inhibitory Concentration) for fluconazole ranged from 0.25 to ≥ 64.00 μg/mL (FS) and 64.00 to 256.00 μg/mL (FR); for amphotericin B ranged from 0.06 to 0.50 μg/mL (FS) and 0.03 to 0.50 μg/ mL (FR); for itraconazole 0.50 to 8.00 μg/ mL (FS) and 1.00 to 16.00 μg/mL (FR); for ketoconazole 0.13 to 2.00 μg/mL (FS) and 0.50 to 16.00 μg/mL (FR); for voriconazole 0.13 to 4.00 μg/mL (FS) and 1.00 to 16.00 μg/mL (FR). The combinations of tacrolimus with azoles showed significant synergism rates; for the group FS tacrolimus + ketoconazole (43%), tacrolimus + itraconazole (43%), tacrolimus + voriconazole (37%); for the group FR ketoconazole + tacrolimus (77% ) tacrolimus + itraconazole (73%), tacrolimus + voriconazole (63%). Linezolid combined with ketoconazole and voriconazole showed high rates of synergism against the FR group, 76.67% and 70%, respectively. Minocycline + amphotericin B obtained 46.67% (FS) and 36.67% (FR) of synergism; chlorpromazine + amphotericin B was synergistic for 40% of the FR. Chlorpromazine combined to azoles showed high rates of antagonism for resistant group, 76.67%, 70% and 80% for ketoconazole, itraconazole and voriconazole, respectively, for the sensitive group combinations had higher rates of indifference. Sulfamethoxazole evidenced a higher percentage of indifferent interactions associated with all tested antifungals, 70% (FS), 73.34% (FR) associated to ketoconazole, 60% (FS), 43.34% (FR) associated to itraconazole, 56.67% (FS), 66.67% (FR) ins association with voriconazole, 86.66% (FS), 80% (FR) associated to amphotericin B. Synergistic (76.66%) and indifferent (23.34%) interactions were observed for diphenyl diselenide + amphotericin B combination for the sensitive group. Diphenyl diselenide + fluconazole combination demonstrated indifferent (50%) and antagonistic (40%) interactions for sensitive group, whereas synergistic interactions were observed in 10% of the isolates. The most significant associations deserve in vivo evaluation in order to verify their potential in the treatment of infections caused by C. glabrata. / Candida spp é o mais frequente agente etiológico de infecções fúngicas oportunistas. Candida albicans é a espécie mais isolada nas candidíases, no entanto espécies de Candida não-albicans têm emergido de forma crescente nas unidades hospitalares em que o uso de azólicos de forma profilática ou terapêutica é frequente. C. glabrata é uma das espécies mais relevantes neste cenário, causando infecções com alta morbidade e mortalidade, devido às altas taxas de resistência frente aos antifúngicos azólicos, em especial ao fluconazol, e a reduzida sensibilidade a anfotericina B e equinocandinas. A marcante resistência desta espécie aos antifúngicos impõe a busca por novas possibilidades terapêuticas. Uma alternativa é a combinação entre fármacos com distintos mecanismos de ação. Este estudo objetivou avaliar a suscetibilidade in vitro de C. glabrata a antifúngicos convencionais (anfotericina B, fluconazol, cetoconazol, itraconazol e voriconazol) e associações destes, com fármacos e compostos não-antifúngicos (clorpromazina, linezolida, minociclina, sulfametoxazol, tacrolimus e disseleneto de difenila). Foram avaliados dois grupos de isolados clínicos de C. glabrata. O primeiro grupo composto de isolados sensíveis ao fluconazol (FS) (n=30) e o segundo, derivado do primeiro, isolados clínicos resistentes ao fluconazol (FR) (n=30). Com base no protocolo M27-A3 (CLSI, 2008), pelo método de checkerboard. Isoladamente, as CIMs (Concentrações Inibitórias Mínimas) para o fluconazol variaram de 0,25 - ≥64,00 μg/mL (FS) e 64,00 - 256,00 μg/mL (FR); para anfotericina B variaram de 0,06 - 0,50 μg/mL (FS) e 0,03 - 0,50 μg/mL (FR); para o itraconazol 0,50 - 8,00 μg/mL (FS) e 1,00 - 16,00 μg/mL (FR); para cetoconazol 0,13 - 2,00 μg/mL (FS) e 0,50 - 16,00 μg/mL (FR); para voriconazol 0,13 - 4,00 μg/mL (FS) e 1.00 - 16.00 μg/mL (FR). As combinações de tacrolimus com azólicos apresentaram consideráveis taxas de sinergismo; para o grupo FS tacrolimus + cetoconazol (43%), tacrolimus + itraconazol (43%), tacrolimus + voriconazol (37%). Para o grupo FR tacrolimus + cetoconazol (77%), tacrolimus + itraconazol (73%), tacrolimus + voriconazol (63%). Linezolida associada ao cetoconazol e ao voriconazol apresentou altas taxas de sinergismo frente ao grupo FR, 76,67% e 70%, respectivamente. Minociclina + anfotericina B obteve 46,67% (FS) e 36,67% (FR) de sinergismo; clorpromazina + anfotericina B foi sinérgica para 40% do grupo FR. Clorpromazina combinada aos azólicos apresentou altas taxas de antagonismo para o grupo resistente, 76,67%, 70% e 80% para cetoconazol, itraconazol e voriconazol, respectivamente; para o grupo sensível as combinações com clorpromazina tiveram maiores taxas de indiferença. Interações sinérgicas (76,66%) e indiferentes (23,34%) foram observadas na combinação de anfotericina B e disseleneto de difenila para o grupo sensível. A combinação de disseleneto de difenila com fluconazol apresentou indiferença (50%) e antagonismo (40%) para o grupo sensível, 10% dos isolados apresentaram sinergismo. Sulfametoxazol apresentou um maior porcentual de interações indiferentes, associado a todos os antifúngicos testados frente aos dois grupos, 70% (FS), 73,34% (FR) associado ao cetoconazol, 60% (FS), 43,34% (FR) associado ao itraconazol, 56,67% (FS), 66,67% (FR) na associação com voriconazol, 86,66% (FS), 80% (FR) associado à anfotericina B. As associações de maior relevância merecem avaliação in vivo, a fim de verificar o potencial das mesmas no tratamento de infecções por Candida glabrata.

Le goéland leucophée endémique dans la région méditerranéenne française. Son mycobiote intestinal n'a jamais été étudié. Ce travail visait à décrire le rôle de ces oiseaux comme réservoir et disséminateur de champignons pathogènes pour l’homme. Nous avons collecté 177 guano de goélands dans cinq sites sur le littoral méditerranéen français; La Grande-Motte, Palavas-les-Flots, Pierre-Blanche, Frioul and Riou archipels. Nous avons identifié dix-sept espèces de levure; les plus fréquentes étant Candida krusei, Galactomyces geotrichum, C. glabrata et C. albicans. On notait d’une part une augmentation de la fréquence des espèces anthropiques de levures C. glabrata et C. albicans avec l’anthropisation des biotopes des colonies de goélands dont d’isolats résistants aux antifongiques. Nous avons analysé les communautés de champignons filamenteux aérocontaminants isolés à partir des mêmes échantillons. Nous avons identifié 16 genres de champignons filamenteux. la faible diversité et abondance de champignons filamenteux dans les zones urbaines par rapport aux suburbains ou à un environnement peu affecté par l'anthropisation et l’association claire entre certaines espèces fongiques et des environnements particuliers. nous avons analysé la génétique des populations de la levure C. glabrata. Nous avons typé par MLVA, 111 isolats de goélands et 79 isolats collectés chez des patients des hôpitaux de Nîmes, Montpellier et Marseille. Nous avons observé une diversité génétique similaire entre les populations de C. glabrata isolées chez le goéland ou chez l’homme. Les isolats de C. glabrata résistants au fluconazole se distribuaient uniformément dans les deux populations. / The yellow-legged gull is endemic in the French Mediterranean area. Their gut mycobiota has never been studied. This work aimed to describe their role in the spreading of potentially human pathogenic fungi with antifungal resistance. Therefore, we sampled 177 yellow-legged gull’s faecal samples in five sites along the Mediterranean littoral South of France; La Grande-Motte, Palavas-les-Flots, Pierre-Blanche, Frioul and Riou archipelagos. We identified seventeen yeast species; the most frequent were Candida krusei, Galactomyces geotrichum, C. glabrata, C. albicans and Saccharomyces cerevisiae. The frequency of the anthropic yeast species C. glabrata and C. albicans increased with the synanthropy of the gull’s colonies and antifungal resistance was found in each of the five most frequent yeast species. We further analyzed the airborne filamentous fungi species isolated from the same sample cultures. We identified 35 filamentous fungi species in 16 genera including 35 species. Both fungal diversity and abundance were low in urban area when compared to suburban ecocline or environments that were little affected by anthropogenic impact and particular fungal species were clearly associated with distinct environments. Finally, we analyzed the population genetic of the human pathogenic yeast C. glabrata, which were isolated from gulls (111 isolates) and from patients (79 isolates) in Nimes, Montpellier and Marseille hospitals, via MLVA analysis. We found that the C. glabrata populations isolated from gulls or humans shared a similar genetic diversity. Antifungal-resistant C. glabrata isolates were evenly distributed in both gull and human populations.

O uso de antissépticos complementa a higienização bucal reduzindo a microbiota e, consequentemente minimizando a colonização, a formação de biofilme e, assim promovendo a saúde bucal. Diante das opções de antissépticos bucais e produtos naturais, faz-se necessária a análise microbiológica da eficácia desses produtos e suas implicações no controle do biofilme. Neste sentido, têm-se como objetivos: determinar a diluição inibitória máxima (DIMax) de antissépticos bucais (Listerine®, Colgate Plax® Tea Fresh, Periogard®) e o óleo de melaleuca sobre as cepas de Candida albicans e Candida glabrata, clínicas e padrão; quantificar as unidades formadoras de colônias por tubo traqueal (UFC/TT) das leveduras em TTs revestidos com os respectivos produtos e, analisar a formação de biofilme em fragmentos de tubos revestidos ou não com antissépticos e óleo de melaleuca por C. glabrata. Trata-se de um estudo de natureza laboratorial, in vitro, realizado com cepas clinicas e padrão e subsidiado em métodos clássicos da microbiologia para o processamento das avaliações propostas. Para determinar a DIMax, realizou-se a diluição dupla seriada (1/4 a 1/4096) dos antissépticos e óleo de melaleuca respectivamente, sendo as placas incubadas a 37°C por 24 horas. Considerou-se DIMax a maior diluição capaz de inibir o crescimento de todas as cepas avaliadas. Na formação de biofilme foram empregadas duas técnicas: determinação das UFC/TT e microscopia eletrônica de varredura (MEV). Na aleatorização das cepas C. glabrata utilizou-se a análise estatística pelo modelo de regressão logístico multinomial. A partir da análise dos resultados observou-se que o Listerine® apresentou a menor ação inibitória na DIMax de 1/4, óleo de melaleuca (1/16), Colgate Plax® Tea Fresh (1/64) e o Periogard® (1/128). Em termos de formação de biofilme, o tubo revestido com Colgate Plax® Tea Fresh apresentou diferença no teste de comparações múltiplas (p=0,0031), com atividade antibiofilme em todas as cepas de C. glabrata, com exceção de um isolado clínico. As fotomicrografias revelaram reprodução por brotamento presente no TT revestido com óleo de melaleuca, lise celular na ação do Periogard® e, os TTs revestidos com Colgate Plax® Tea Fresh não apresentaram biofilme, exceto na cepa 33. A formação de biofilme por células leveduriformes foi significativa apresentando-se de forma diversificada nos diferentes tubos revestidos. Estudos adicionais sobre Candida spp. em tubos traqueais são recomendáveis em pacientes, com e sem pneumonia, submetidos à ventilação mecânica / Introduction: The use of oral antiseptics is increasingly common, as a complement to the regular oral hygiene by reducing the oral microbiota, biofilm formation and thereby promoting oral health. Facing of several oral antiseptics and natural products, it is necessary microbiological analysis of the effectiveness of these antiseptics and in its implications in the control of biofilm. Objective: The aims of the study were to determine the maximum inhibitory dilution (MID) oral antiseptics (Listerine®, Colgate® Plax, Fresh Tea and Periogard®) and tea tree oil of clinical and standard strains of Candida spp. and Candida glabrata; colony forming units assay (CFU/TT) of yeast in tracheostomy tubes (TT) coated with some products; and analyze the biofilm formed in fragments of tubes coated or not with oral antiseptics and tea tree oil for Candida glabrata. Methods: This is a laboratory investigation, in vitro study performed with clinical and standard strains and subsidized of classical microbiology methods for processing the proposals reviews. To determine the MID, serial dilution was carried out (1/4 a 1/2048) of oral antiseptics and tea tree oil respectively, and the plates were incubated at 37°C for 24h. The MID was considered the highest dilution capable of inhibiting the growth of all strains tested. In biofilm formation two techniques were employed: determination of colony forming units (CFU/TT) and scanning electron microscopy (SEM). The statistical analysis by multinomial logistic regression model was used to randomization strains of C. glabrata. Results: Listerine® showed the worst performance, MID (1/4), tea tree oil (1/16), Colgate Plax® Fresh Tea (1/64) and Periogard® (1/128). In terms of biofilm formation, the multiple comparisons test presented differences (p=0.0031) for tube coated Colgate Plax® Tea Fresh with antibiofilm activity in all strains of C. glabrata, except for one clinical isolate. The photomicrographs revealed reproduction by budding in the TT coated with tea tree oil, cell lysis in action of Periogard®, and the Colgate Plax® Fresh Tea coated tube produced no visible colony, except with strain 33. Conclusion: Biofilm formation by yeast was significant presenting diverse in different coated tubes. Additional studies of Candida spp. in tracheostomy tubes are recommended in patients with and without pneumonia, undergoing mechanical ventilation

O uso de antissépticos complementa a higienização bucal reduzindo a microbiota e, consequentemente minimizando a colonização, a formação de biofilme e, assim promovendo a saúde bucal. Diante das opções de antissépticos bucais e produtos naturais, faz-se necessária a análise microbiológica da eficácia desses produtos e suas implicações no controle do biofilme. Neste sentido, têm-se como objetivos: determinar a diluição inibitória máxima (DIMax) de antissépticos bucais (Listerine®, Colgate Plax® Tea Fresh, Periogard®) e o óleo de melaleuca sobre as cepas de Candida albicans e Candida glabrata, clínicas e padrão; quantificar as unidades formadoras de colônias por tubo traqueal (UFC/TT) das leveduras em TTs revestidos com os respectivos produtos e, analisar a formação de biofilme em fragmentos de tubos revestidos ou não com antissépticos e óleo de melaleuca por C. glabrata. Trata-se de um estudo de natureza laboratorial, in vitro, realizado com cepas clinicas e padrão e subsidiado em métodos clássicos da microbiologia para o processamento das avaliações propostas. Para determinar a DIMax, realizou-se a diluição dupla seriada (1/4 a 1/4096) dos antissépticos e óleo de melaleuca respectivamente, sendo as placas incubadas a 37°C por 24 horas. Considerou-se DIMax a maior diluição capaz de inibir o crescimento de todas as cepas avaliadas. Na formação de biofilme foram empregadas duas técnicas: determinação das UFC/TT e microscopia eletrônica de varredura (MEV). Na aleatorização das cepas C. glabrata utilizou-se a análise estatística pelo modelo de regressão logístico multinomial. A partir da análise dos resultados observou-se que o Listerine® apresentou a menor ação inibitória na DIMax de 1/4, óleo de melaleuca (1/16), Colgate Plax® Tea Fresh (1/64) e o Periogard® (1/128). Em termos de formação de biofilme, o tubo revestido com Colgate Plax® Tea Fresh apresentou diferença no teste de comparações múltiplas (p=0,0031), com atividade antibiofilme em todas as cepas de C. glabrata, com exceção de um isolado clínico. As fotomicrografias revelaram reprodução por brotamento presente no TT revestido com óleo de melaleuca, lise celular na ação do Periogard® e, os TTs revestidos com Colgate Plax® Tea Fresh não apresentaram biofilme, exceto na cepa 33. A formação de biofilme por células leveduriformes foi significativa apresentando-se de forma diversificada nos diferentes tubos revestidos. Estudos adicionais sobre Candida spp. em tubos traqueais são recomendáveis em pacientes, com e sem pneumonia, submetidos à ventilação mecânica / Introduction: The use of oral antiseptics is increasingly common, as a complement to the regular oral hygiene by reducing the oral microbiota, biofilm formation and thereby promoting oral health. Facing of several oral antiseptics and natural products, it is necessary microbiological analysis of the effectiveness of these antiseptics and in its implications in the control of biofilm. Objective: The aims of the study were to determine the maximum inhibitory dilution (MID) oral antiseptics (Listerine®, Colgate® Plax, Fresh Tea and Periogard®) and tea tree oil of clinical and standard strains of Candida spp. and Candida glabrata; colony forming units assay (CFU/TT) of yeast in tracheostomy tubes (TT) coated with some products; and analyze the biofilm formed in fragments of tubes coated or not with oral antiseptics and tea tree oil for Candida glabrata. Methods: This is a laboratory investigation, in vitro study performed with clinical and standard strains and subsidized of classical microbiology methods for processing the proposals reviews. To determine the MID, serial dilution was carried out (1/4 a 1/2048) of oral antiseptics and tea tree oil respectively, and the plates were incubated at 37°C for 24h. The MID was considered the highest dilution capable of inhibiting the growth of all strains tested. In biofilm formation two techniques were employed: determination of colony forming units (CFU/TT) and scanning electron microscopy (SEM). The statistical analysis by multinomial logistic regression model was used to randomization strains of C. glabrata. Results: Listerine® showed the worst performance, MID (1/4), tea tree oil (1/16), Colgate Plax® Fresh Tea (1/64) and Periogard® (1/128). In terms of biofilm formation, the multiple comparisons test presented differences (p=0.0031) for tube coated Colgate Plax® Tea Fresh with antibiofilm activity in all strains of C. glabrata, except for one clinical isolate. The photomicrographs revealed reproduction by budding in the TT coated with tea tree oil, cell lysis in action of Periogard®, and the Colgate Plax® Fresh Tea coated tube produced no visible colony, except with strain 33. Conclusion: Biofilm formation by yeast was significant presenting diverse in different coated tubes. Additional studies of Candida spp. in tracheostomy tubes are recommended in patients with and without pneumonia, undergoing mechanical ventilation