In vitro antifungal activity and phytochemical characterization of Eugenia uniflora, Libidibia ferrea and Psidium guajava

The aim of this study was evaluate the in vitro antifungal activity of crude extracts from Eugenia uniflora, Libidibia ferrea and Psidium guajava. The extracts were obtained by turbo-extraction using water (AQ) or acetone-water (AC-W) (7:3, v/v) as solvents and lyophilized to obtain the crude extracts (CE). The CE were characterized by UV-Vis, TLC and HPLC. The activity of CEs was investigated against clinical isolates of Candida spp. and the Minimum Inhibitory Concentration (MIC), MIC50 and MIC90 were determinated. The analysis by TLC showed that all CEs presented polyphenols (flavonoids and tannins). The CEs from E. uniflora showed higher amount of polyphenols (30.35 ± 2.15%, AC-W) and the HPLC analysis revealed the tannins in all extracts. The CEs of E. uniflora showed MIC range from 1.9 to 500.0 µg/mL, and lower values of MIC50 and MIC90 against non-albicans Candida isolates. Regarding L. ferrea and P. guajava, the results showing MIC from 3.9 to 1000.0 µg/mL (CE-AQ) against C. albicans. The results demonstrate antifungal performance from CE against various species of Candida spp., suggesting that the herbal species may be use as new potential antifungal agents. Additionally, the polyphenol content can play a pivotal role in the antifungal properties of CE.

Loop-mediated Isothermal Amplification and nested PCR of the Internal Transcribed Spacer (ITS) for Histoplasma capsulatum detection.

BACKGROUND: Histoplasmosis is a neglected disease that affects mainly immunocompromised patients, presenting a progressive dissemination pattern and a high mortality rate, mainly due to delayed diagnosis, caused by slow fungal growth in culture. Therefore, a fast, suitable and cost-effective assay is required for the diagnosis of histoplasmosis in resource-limited laboratories. This study aimed to develop and evaluate two new molecular approaches for a more cost-effective diagnosis of histoplasmosis. METHODOLOGY: Seeking a fast, suitable, sensitive, specific and low-cost molecular detection technique, we developed a new Loop-mediated Isothermal Amplification (LAMP) assay and nested PCR, both targeting the Internal Transcribed Spacer (ITS) multicopy region of Histoplasma capsulatum. The sensitivity was evaluated using 26 bone marrow and 1 whole blood specimens from patients suspected to have histoplasmosis and 5 whole blood samples from healthy subjects. All specimens were evaluated in culture, as a reference standard test, and Hcp100 nPCR, as a molecular reference test. A heparin-containing whole blood sample from a heathy subject was spiked with H. capsulatum cells and directly assayed with no previous DNA extraction. RESULTS: Both assays were able to detect down to 1 fg/µL of H. capsulatum DNA, and ITS LAMP results could also be revealed to the naked-eye by adding SYBR green to the reaction tube. In addition, both assays were able to detect all clades of Histoplasma capsulatum cryptic species complex. No cross-reaction with other fungal pathogens was presented. In comparison with Hcp100 nPCR, both assays reached 83% sensitivity and 92% specificity. Furthermore, ITS LAMP assay showed no need for DNA extraction, since it could be directly applied to crude whole blood specimens, with a limit of detection of 10 yeasts/µL. CONCLUSION: ITS LAMP and nPCR assays have the potential to be used in conjunction with culture for early diagnosis of progressive disseminated histoplasmosis, allowing earlier, appropriate treatment of the patient. The possibility of applying ITS LAMP, as a direct assay, with no DNA extraction and purification steps, makes it suitable for resource-limited laboratories. However, more studies are necessary to validate ITS LAMP and nPCR as direct assay in other types of clinical specimens.

Eumicetoma e actinomicetoma: uma breve revisão da literatura / Eumycetoma and actinomycetoma: a brief review of the literature

Micetoma é uma infecção que acomete o tecido subcutâneo após a inoculação de microrganismos na pele em locais de pequenos traumas. Caracteriza-se pela ocorrência de tumoração, associada à formação de fistulas e à drenagem de grãos. Trata-se de um grupo de infecções subcutâneas de difícil tratamento com epidemiologia bem definida, acometendo preferencialmente trabalhadores rurais do gênero masculino. Os agentes causadores podem ser fungos ou bactérias. Este artigo propõe-se à revisão dos dados recentes da epidemiologia e tratamento dessas infecções.

Trichosporon inkin meningitis in Northeast Brazil: first case report and review of the literature.

BACKGROUND: Trichosporon species may colonize the skin, respiratory tract and gastrointestinal tract of human beings. The yeast is recognized as etiological agent of white piedra, a superficial mycosis. Nevertheless, immunocompromised hosts may develop invasive Trichosporonosis. Central nervous system trichosporonosis is a very rare clinical manifestation. In fact, only a few cases have been published in the literature and none of them was caused by Trichosporon inkin. CASE PRESENTATION: Here we report the first clinical case of meningoencephalitis due to this species in a female previously healthy patient under corticosteroids and antibiotics therapy for several months. She was submitted to an invasive procedure to remove a left sided acoustic neuroma and further developed a cerebrospinal fistula. After some days of the procedure, she presented a predominantly and intensive occipital holocranial headache, followed by vomiting, hyporexia, weight loss, asthenia, irritability, difficulty to concentrate and rotator vertigo. The patient further developed a cerebrospinal fistula in the occipital region and was submitted to a surgical correction. After several months of clinical interventions, she was diagnosed with CNS Trichosporonosis, after Magnetic Resonance Imaging and positive microbiological cultures obtained within two different occasions (2 weeks apart). Despite the antifungal therapy with Amphotericin B and Voriconazole, the patient did not survive. CONCLUSIONS: Despite CNS Fungal infections are mostly due to Cryptococcus spp., other emergent yeasts, such as T. inkin may be considered as a likely etiological agent. This is the first case report of CNS Trichosporonosis, where species identification was performed with rDNA sequencing.

Recurrent Oral Manifestations of Paracoccidioidomycosis without Pulmonary Involvement: Case Report and Review of Literature / Manifestación Oral Recurrente de Paracoccidioidomcosis sin Compromiso Pulmonar: Reporte de Caso y Revisión de la Literatura

This article describes a case of recurrence of chronic paracoccidioidomycosis (PCM) 11 years following the initial diagnosis. The patient was a 51-year-old white Brazilian female, which had been previously diagnosed with PCM. The physical examination revealed the presence of a single crusted lesion in the upper lip and an elevated lesion with fibrous scar appearance on right buccal mucosa. Although the diagnosis of PCM, the absence of pulmonary involvement led to the biopsy of the lesion localized on the buccal mucosa and the histopathological analysis in H&E and PAS stains revealed no morphological changes suggestive of any lesion. However, the exfoliative cytology stained with Grocott-Gomori showed the presence of Paracoccidioides brasiliensis and the infection caused by this fungus was proven. Of this way, we emphasize the importance of stomatologic evaluation to the diagnosis of diseases that usually manifestssystemically. So, the correct diagnosis of oral manifestations of PCM, is essential to ensure early and safe intervention.

En este artículo se describe un caso de paracoccidioidomicosis crónica (PCM) recurrente 11 años después del diagnóstico inicial. La paciente una mujer blanca de Brasil de 51 años de edad, que había sido diagnosticada previamente con PCM. El examen físico reveló la presencia de una lesión única en el labio superior y una lesión elevada con apariencia de cicatriz fibrosa en la mucosa bucal derecha. No obstante el diagnóstico de PCM, la ausencia de compromiso pulmonar llevó a la biopsia de la lesión localizada en la mucosa bucal y el análisis histopatológico no demostró cambios morfológicos indicativos de cualquier lesión. Sin embargo, la citología exfoliativa teñida con Grocott-Gomori mostró la presencia de Paracoccidioides brasiliensis y la infección causada por este hongo fue probado. De esta manera, hacemos hincapié en la importancia de la evaluación odontológica para el diagnóstico de enfermedades que se manifiestan generalmente por vía sistémica. Por lo tanto, el diagnóstico correcto de las manifestaciones orales de PCM es esencial para asegurar la intervención temprana y segura.

Species distribution and virulence factors of Candida spp. isolated from the oral cavity of kidney transplant recipients in Brazil.

Although yeasts belonging to the genus Candida are frequently seen as commensals in the oral cavity, they possess virulence attributes that contribute for pathogenicity. The aims of the present study were to study the prevalence of Candida spp. isolated from the oral cavity of renal transplant recipients and to analyze strains virulence factors. We isolated a total of 70 Candida strains from 111 transplant recipients, and Candida albicans was the most prevalent species (82.86 %). Oral candidiasis was diagnosed in 14.4 % kidney transplant patients, while 11 isolates (15.7 %) corresponded to non-Candida albicans Candida (NCAC) species. C. albicans adhered to a higher extension than NCAC strains. Some isolates of Candida tropicalis were markedly adherent to human buccal epithelial cells and highly biofilm-forming strains. Regarding proteinase activity, Candida orthopsilosis was more proteolytic than Candida metapsilosis. Candida glabrata and Candida dubliniensis showed very low ability to form biofilm on polystyrene microtiter plates. We have demonstrated here diverse peculiarities of different Candida species regarding the ability to express virulence factors. This study will contribute for the understanding of the natural history and pathogenesis of yeasts belonging to the genus Candida in the oral cavity of patients who were submitted to kidney transplant and are under immunosuppressive therapies.

A 7-year survey of superficial and cutaneous mycoses in a public hospital in Natal, Northeast Brazil

In the present study, we determined the frequency of superficial and cutaneous mycoses and their etiologic agents during a 7-year period (2002-2008) in Natal, Brazil. A total of 1,717 specimens of skin, nail, and hair were collected from 1,382 patients with suspected mycoses lesions and were then subjected to direct microscopy and culture.

Rhinocerebral zygomycosis in a diabetic patient / Zigomicose rinocerebral em paciente diabética

Rhinocerebral zygomycosis is the most frequent form of fungal infection caused by members of the Zygomycetes class. A fatal case of rhinocerebral zygomycosis caused by Rhizopus (oryzae) arrhizus with histopathological and mycological diagnosis is reported in a diabetic patient.

Zigomicose rinocerebral é a forma mais frequente das infecções fúngicas causadas por membros da classe Zygomicetes. É relatado um caso fatal de zigomicose rinocerebral por Rhizopus (oryzae) arrhizus com diagnóstico histopatológico e micológico, em paciente diabética.

Evaluation of different methods for detecting methicillin resistance in staphylococcus aureus isolates in a university hospital located in the northeast of Brazil

Many methods have been described for the detection of methicillin-resistant Staphylococcus aureus (MRSA), but the heterogeneous expression of methicillin resistance affects the reliability of these methods. The aim of the present study was to evaluate some methods for detecting methicillin resistance in Staphylococcus aureus isolates in a university hospital located in the Northeast of Brazil. Among the isolates, 15 were methicillin-susceptible and 45 were methicillin-resistant, including low-level heterogeneous resistance strains. Both the 30 µg-cefoxitin disk and PBP2a test had 100 percent sensibility/specificity and appear to be good options for the detection of MRSA in the clinical laboratory.

Invasive aspergillosis associated with systemic lupus erythematosus and cardiac postoperative complication / Aspergilose invasiva associada ao lúpus eritematoso sistêmico e a complicação cardíaca pós-operatória

Aspergillus is a ubiquitous fungus which can cause a variety ofclinical syndromes. This fungus has emerged as agent of systemic infections and has therefore gained considerable public health importance. This paper describes two cases of invasive aspergillosis caused by A. fumigatus in immuno-suppressed patients and underscores the importance of early identification of Aspergillus infection associated with systemic lupus erythematosus and cardiac postoperative complications.

Aspergillus é um fungo ubíquito que pode causar uma variedade de síndromes clínicas. Este fungo tem emergido como agente de infecções sistêmicas e tem conquistado considerável importância para saúde pública. Este trabalho descreve dois casos de aspergilose invasiva causada por A. fumigatus em pacientes imunossuprimidos e relata a importância da identificação precoce em infecção por Aspergillus associada ao lúpus eritematoso sistêmico e a complicações cardíacas pós-operatórias.

Prevalence of Candida species in umbilical catheters implanted in newborns in Natal, Brazil

All the newborns with umbilical venous catheters, hospitalized in the neonatal intensive care unit at Januário Cicco Maternity Hospital in Natal, Brazil, between January, 2003 and December, 2004 were studied. The prevalence of Candida species in the tips of intra-venous catheters was assessed, as well as the coexisting exposures that the patients were subjected to. Catheter tips were cultivated in blood agar and when yeast culture occurred, the colony was subcultivated for species identification through morphologic and biochemical assays. From a total of 240 catheters, 41 were positive for yeasts, and 34 were submitted to identification. The following agents were isolated: 13 C. albicans (38 percent), 10 Candida parapsilosis (29 percent), 8 C. tropicalis (20 percent), one C. guilliermondii (3 percent), one C. famata (3 percent) and one Trichosporon spp. (3 percent). Three patients among those with positive catheter tip fungal cultures were also hemoculture positive, with the same fungal species at both sites. Among the coexisting exposures, it should be pointed out that all the patients underwent broad spectrum antibiotic therapy, used a nasogastric probe, in addition to undergoing other invasive procedures such as mechanical ventilation and umbilical catheter implantation.

Foram estudados, entre janeiro de 2003 a dezembro de 2004, todos os recém-nascidos internados na UTI neonatal da Maternidade Escola Januário Cicco, portadores de cateter venoso umbilical. Avaliou-se a prevalência de espécies de Candida nas pontas de cateteres intra-venosos dos neonatos, e os fatores associados a que os pacientes estavam submetidos. As pontas de cateter foram cultivadas em ágar sangue, e, quando houve crescimento de colônia leveduriforme, esta foi subcultivada para a realização de provas morfológicas e bioquímicas para identificar a espécie. Do total de 240 cateteres analisados, 41 apresentaram crescimento de leveduras, sendo 34 encaminhados para identificação, observando-se os seguintes agentes: 13 C. albicans (38 por cento), 10 Candida parapsilosis (29 por cento), 8 C. tropicalis (20 por cento), 1 C. guilliermondii (3 por cento), 1 C. famata (3 por cento) e 1 Trichosporon spp. (3 por cento). Dos pacientes que apresentaram cultura de ponta de cateter positiva para fungos, três tiveram hemocultura positiva com a mesma espécie em ambos os sítios. Entre os fatores de risco destaca-se que todos os pacientes faziam uso de antibioticoterapia de amplo espectro, portavam sonda nasogástrica, além de outros procedimentos invasivos, como ventilação mecânica e cateter umbilical.

Candidíase vulvovaginal: sintomatologia, fatores de risco e colonização anal concomitante / Vulvovaginal candidiasis: symptomatology, risk factors and concomitant anal colonization

OBJETIVO: analisar pacientes com candidíase vulvovaginal quanto a sintomatologia, fatores de risco e resultados da cultura anal, identificar a freqüência de Candida albicans e não C. albicans e correlacionar as colonizações anal e vaginal. MÉTODOS: foram incluídas 99 pacientes com suspeita clínica de candidiase vulvovaginal, procedentes de Natal, RN, atendidas entre maio de 2003 e maio de 2005, perfazendo-se o total de 294 coletas. O material clínico, colhido por zaragatoas, foi semeado em CHROMagar Candida®. As leveduras foram identificadas pelo método clássico, além da prova de crescimento a 42 e 45°C e da prova do caldo Sabouraud hipertônico. A sintomatologia, fatores de risco e colonização anal foram analisados de acordo com a positividade ou negatividade para Candida spp. As culturas positivas para C. albicans nos dois sítios foram comparadas com outros resultados encontrados. Para análise estatística utilizou-se o teste do chi2, com correção de Yates e o teste exato de Fisher. RESULTADOS: a espécie mais frequente foi C. albicans em 69 por cento dos casos. Uso de roupas íntimas justas e/ou sintéticas, presença de doenças alérgicas, ocorrência de prurido, leucorréia e hiperemia apresentaram associação com a positividade vaginal para Candida spp. A chance de uma paciente com colonização anal positiva de apresentar positividade vaginal concomitante foi 2,8 e 4,9 vezes maior, respectivamente, para Candida spp e C. albicans. A chance de uma paciente com cultura anal positiva para C. albicans de apresentar resultado vaginal positivo foi 3,7 vezes maior quando comparada a espécies não C. albicans. CONCLUSÕES: C. albicans foi a espécie mais comum, tendo sido observada associação da positividade vaginal para Candida spp com uso de roupas justas e/ou sintéticas, doenças alérgicas, prurido, leucorréia e eritema (p<0,05). A positividade anal concomitante com a vaginal foi significativa, sugerindo uma possível contaminação vaginal a partir do ânus.

PURPOSE: to analyze patients with vulvovaginal candidiasis with respect to risk factors, symptomatology and results of anal culture, to identify the frequency of species of Candida albicans and non-C. albicans, and to correlate anal and vaginal colonization. METHODS: a total of 99 patients were included with suspected vulvovaginal candidiasis, from Natal, Brazil, between May 2003 and May 2005, totalling 294 collections. The clinical material, obtained by vaginal and anal swabs, was seeded on CHROMagar Candida®. The yeasts were identified using the classic method, in addition to the growth test at 42° and 45°C and the Hypertonic Saboraud broth test. Symptomatology, risk factors and anal colonization were assessed according to positive or negative culture for Candida spp. The cultures positive for C. albicans at the two sites were compared with other results encountered. YatesÆ chi2 test and FisherÆs exact test were used for statistical analysis. RESULTS: the most frequent was C. albicans in 69 percent of the cases. Wearing tight and/or synthetic underclothing, the presence of allergic diseases, the occurrence of itching, leukorrhea and hyperemia showed a significant association with positive culture for Candida spp in the vagina. The chance of a patient with positive anal colonization to present with concomitant positive vaginal colonization was 2.8 and 4.9 times greater for Candida spp and C. albicans, respectively. The risk of a patient with anal culture positive for C. albicans to present with positive vaginal colonization was 3.7 times greater when compared to non-C. albicans species. CONCLUSIONS: the most common species was C. albicans, and a relevant association between vaginal cultures positive for Candida spp and the use of tight and/or synthetic underclothing, allergic diseases, the occurrence of pruritus, leukorrhea and erythema was observed (p<0.05). Positive anal colonization concomitant with vaginal colonization was significant,suggesting...

Candidíase vulvovaginal: uma revisão de literatura / Vulvovaginal candidiasis: a review

A candidíase vulvovaginal é a infecção da mucosa vaginal causada por fungos do gênero Candida, colonizantes habituais do trato gastrintestinal, que podem, em condições especiais, tornar-se patogênicos. É uma das causas mais comuns de vaginite, com incidência aproximada de 25 porcento, que acomete cerca de 75 porcento das mulheres, pelo menos uma vez na vida e se caracteriza por leucorréia, prurido intenso, hiperemia vulvar, disúria e dispareunia. Vários fatores predisponentes estão relacionados com a ocorrência da candidíase vulvovaginal: gravidez, diabetes mellitus, uso de anticoncepcionais hormonais, terapia de reposição hormonal, antibióticos e corticóides, doenças imunossupressoras e outros. Para haver infeção é necessário um estímulo estrogênico alto, razão pela qual sua ocorrência é mais comum durante o menacme e na mulher climatérica sob terapia de reposição hormonal, sendo o processo de adesão do fungo primordial para que ela se estabeleça. Entre as mulheres que apresentam candidíase vulvovaginal, cerca de 5 porcento desenvolve a forma recorrente, definida por pelo menos quatro episódios de vaginite por cândida sp no intervalo de um ano. O diagnóstico é essencialmente clínico, podendo ser corroborado por exame microscópico e excepcionalmente por cultura. O tratamento pode ser tópico e/ou sistêmico, sendo os imidazóis as drogas mais indicadas

Candida dubliniensis identification in Brazilian yeast stock collection

We investigated the presence of Candida dubliniensis among isolates previously identified as Candida albicans and maintained in a yeast stock collection from 1994 to 2000. All isolates were serotyped and further evaluated for antifungal susceptibility profile. After doing a screening test for C. dubliniensis isolates based on the capability of colonies to grow at 42C, its final identification was obtained by randomly amplified polymorphic DNA (RAPD) analysis using three different primers. A total of 46 out of 548 screened isolates did not exhibit growth at 42C and were further genotyped by RAPD. Eleven isolates were identified as C. dubliniensis with RAPD analysis. Regarding serotypes, 81.5 percent of C. albicans and all C. dubliniensis isolates belonged to serotype A. Of note, 9 out of 11 C. dubliniensis isolates were obtained from patients with acquired immunodeficiency syndrome (Aids) and all of them were susceptible to azoles and amphotericin B. We found 17 (3 percent) C. albicans isolates that were dose-dependent susceptibility or resistant to azoles. In conclusion, we found a low rate of C. dubliniensis isolates among stock cultures of yeasts previously identified as C. albicans. Most of these isolates were recovered from oral samples of Aids patients and exhibited high susceptibility to amphotericin B and azoles. C. albicans serotype A susceptible to all antifungal drugs is the major phenotype found in our stock culture

Multicenter Brazilian study of oral Candida species isolated from Aids aatients

Oropharyngeal candidiasis continues to be considered the most common opportunistic disease in Aids patients. This study was designed to investigate species distribution, serotype and antifungal susceptibility profile among Candida spp. isolated from the oral cavity of Aids patients recruited from six Brazilian university centers. Oral swabs from 130 Aids patients were plated onto CHROMagar Candida medium and 142 isolates were recovered. Yeast isolates were identified by classical methods and serotyped using the Candida Check« system-Iatron. Antifungal susceptibility testing was performed according to the NCCLS microbroth assay. C. albicans was the most frequently isolated species (91 percent), and 70 percent of the isolates belonged to serotype A. We detected 12 episodes of co-infection (9 percent), including co-infection with both serotypes of C. albicans. Non-albicans species were isolated from 12 episodes, 50 percent of them exhibited DDS or resistance to azoles. Otherwise, only 8 out 130 isolates of C. albicans exhibited DDS or resistance to azoles. Brazilian Aids patients are infected mainly by C. albicans serotype A, most of them susceptible to all antifungal drugs

Avaliaçäo fenotípica e genotípica de leveduras isoladas da cavidade oral de pacientes com ids / Phenotypic and genotypic evaluation of yeasts isolated from oral cavity of aids patients

Objetivos: 1. Caracterizar a distribuiçao de espécies, sorotipos, genotipos e perfil de susceptibilidade de amostras de Candida spp. coletadas da cavidade oral de pacientes com AIDS. 2. Avaliar o impacto da terapêutica ánti-retroviral combinada sobre as características microbiológicas da colonizaçao e infecçao por leveduras na cavidade oral de pacientes com AIDS. Métodos: Conduzimos a coleta de amostras de acordo com dois desenhos: transversal e longitudinal. No estudo transversal foi realizada coleta única de pacientes com AIDS e candidíase orofaríngea, recrutados em 6 centros universitários brasileiros. No estudo longitudinal foram realizadas três coletas nos momentos "zero", 6 e 12 meses, de pacientes com AIDS que estivessem na iminência de iniciar terapêutica anti-retroviral combinada, recrutados de 2 centros universitários brasileiros. A coleta do material de cavidade oral foi realizada com zaragatoa e o conteúdo foi semeado em CHROMagar Candidar. As colônias de cor verde foram submetidas à prova de microcultivo e ao teste de crescimento em 42ºC. Isolados com crescimento reduzido a esta temperatura foram analisados por RAPD, com 2 "primers", para identificaçao de Candida dubliniensis. Aqueles que apresentaram crescimento a 42ºC e clamidoconídios foram identificados como Candida aibicans. Os isolados nao-albicans foram identificados pelo sistema ID32C (bioMérieux). Para identificaçao dos sorotipos A e B de C. albicans foi empregado o sistema Candida Check (latron). Os testes de susceptibilidade aos antifúngicos foram realizados através do método de microdiluiçao em caldo (NCCLS). A cariotipagem eletroforética foi utilizada para a determinaçao do perfil genômico de C. albicans e C. dubliniensis. Resultados: No estudo transversal, um total de 130 cepas de Candida foram isoladas da cavidade oral de 116 pacientes. Candida albicans foi a espécie mais freqüente, contando com 90 por cento das amostras. O sorotipo A (74 por cento) predominou sobre o B e entre as espécies nao-albicans, predominaram C. glabrata e C. tropicalis. Coinfecçao por mais de uma espécie ou sorotipo de levedura na mesma coleta foi observada em 13 pacientes (11 por cento). Em relaçao ao perfil de susceptibilidade, 14/130 (11 por cento) dos isolados foram S-DD ou resistentes aos azóis, com maior resistência das espécies nao-albicans sobre C. albicans. No estudo longitudinal, foram avaliados 38 pacientes em uso de terapêutica anti-retroviral combinada durante 12 meses...(au)