Opportunistic microorganisms in patients undergoing antibiotic therapy for pulmonary tuberculosis

Antimicrobial therapy may cause changes in the resident oral microbiota, with the increase of opportunistic pathogens. The aim of this study was to compare the prevalence of Candida, Staphylococcus, Pseudomonas and Enterobacteriaceae in the oral cavity of fifty patients undergoing antibiotic therapy for pulmonary tuberculosis and systemically healthy controls. Oral rinsing and subgingival samples were obtained, plated in Sabouraud dextrose agar with chloramphenicol, mannitol agar and MacConkey agar, and incubated for 48 h at 37ºC. Candida spp. and coagulase-positive staphylococci were identified by phenotypic tests, C. dubliniensis, by multiplex PCR, and coagulase-negative staphylococci, Enterobacteriaceae and Pseudomonas spp., by the API systems. The number of Candida spp. was significantly higher in tuberculosis patients, and C. albicans was the most prevalent specie. No significant differences in the prevalence of other microorganisms were observed. In conclusion, the antimicrobial therapy for pulmonary tuberculosis induced significant increase only in the amounts of Candida spp.

Research on Candida dubliniensis in a Brazilian yeast collection obtained from cardiac transplant, tuberculosis, and HIV-positive patients, and evaluation of phenotypic tests using agar screening methods.

The aim of this study was to research Candida dubliniensis among isolates present in a Brazilian yeast collection and to evaluate the main phenotypic methods for discrimination between C. albicans and C. dubliniensis from oral cavity. A total of 200 isolates, presumptively identified as C. albicans or C. dubliniensis obtained from heart transplant patients under immunosuppressive therapy, tuberculosis patients under antibiotic therapy, HIV-positive patients under antiretroviral therapy, and healthy subjects, were analyzed using the following phenotypic tests: formation and structural arrangement of chlamydospores on corn meal agar, casein agar, tobacco agar, and sunflower seed agar; growth at 45 °C; and germ tube formation. All strains were analyzed by polymerase chain reaction (PCR). In a preliminary screen for C. dubliniensis, 48 of the 200 isolates on corn meal agar, 30 of the 200 on casein agar, 16 of the 200 on tobacco agar, and 15 of the 200 on sunflower seed agar produced chlamydoconidia; 27 of the 200 isolates showed no or poor growth at 45 °C. All isolates were positive for germ tube formation. These isolates were considered suggestive of C. dubliniensis. All of them were subjected to PCR analysis using C. dubliniensis-specific primers. C. dubliniensis isolates were not found. C. dubliniensis isolates were not recovered in this study done with immunocompromised patients. Sunflower seed agar was the medium with the smallest number of isolates of C. albicans suggestive of C. dubliniensis. None of the phenotypic methods was 100% effective for discrimination between C. albicans and C. dubliniensis.

Staphylococcus spp., Enterobacteriaceae and Pseudomonadaceae oral isolates from Brazilian HIV-positive patients. Correlation with CD4 cell counts and viral load.

The aim was to evaluate the presence of Staphylococcus spp., Enterobacteriaceae and Pseudomonadaceae in the oral cavities of HIV-positive patients. Forty-five individuals diagnosed as HIV-positive by ELISA and Western-blot, and under anti-retroviral therapy for at least 1 year, were included in the study. The control group constituted 45 systemically healthy individuals matched to the HIV patients to gender, age and oral conditions. Oral rinses were collected and isolates were identified by API system. Counts of microorganisms from HIV and control groups were compared statistically by a Mann-Whitney test (α=5%). The percentages of individuals positive for staphylococci were similar between the groups (p=0.764), whereas for Gram-negative rods, a higher percentage was observed amongst HIV-positive (p=0.001). There was no difference in Staphylococcus counts between HIV and control groups (p=0.1008). Counts were lower in the oral cavities of patients with low viral load (p=0.021), and no difference was observed in relation to CD4 counts (p=0.929). Staphylococcus aureus was the most frequently isolated species in HIV group, and Staphylococcus epidermidis was the prevalent species in the control group. Significantly higher numbers of enteric bacteria and pseudomonas were detected in the oral cavities of the HIV group than in the control (p=0.0001). Enterobacter cloacae was the most frequently isolated species in both groups. Counts of enteric bacteria and pseudomonas were significantly lower in patients with low CD4 counts (p=0.011); however, there was no difference relating to viral load. It may be concluded that HIV group showed greater species diversity and a higher prevalence of Enterobacteriaceae/Pseudomonadaceae.

In vitro antifungal susceptibility of Candida spp. oral isolates from HIV-positive patients and control individuals

Oropharyngeal candidiasis is the most common fungal infection among HIV-positive patients. This condition can be treated with either systemic or topical antifungal agents; treatments are usually indicated empirically on the basis of clinical data. The knowledge of in vitro antifungal susceptibility is important to determine correct therapeutic guides for the treatment of fungal infections. Therefore, the objective of this study was to determine the antifungal susceptibility profile of oral Candida isolates from HIV-positive patients and control individuals. Amphotericin B, fluconazole, flucytosine, nystatin and ketoconazole were tested according to the methodology of microdilution proposed by the Clinical and Laboratory Standards Institute (CLSI); results were recorded in values of minimal inhibitory concentration (MIC). A total of 71 Candida isolates from HIV-positive patients were examined with the following species represented: C. albicans (59), C. tropicalis (9), C. glabrata (1), C. guilliermondii (1) and C. krusei (1). A total of 15 Candida isolates were evaluated from control individuals comprised of 11 C. albicans and 4 C. tropicalis samples. Our results demonstrated that the tested antifungal agents showed good activity for most isolates from both groups; however, variability in MIC values among isolates was observed.

In vitro antifungal susceptibility of Candida spp. oral isolates from HIV-positive patients and control individuals.

Oropharyngeal candidiasis is the most common fungal infection among HIV-positive patients. This condition can be treated with either systemic or topical antifungal agents; treatments are usually indicated empirically on the basis of clinical data. The knowledge of in vitro antifungal susceptibility is important to determine correct therapeutic guides for the treatment of fungal infections. Therefore, the objective of this study was to determine the antifungal susceptibility profile of oral Candida isolates from HIV-positive patients and control individuals. Amphotericin B, fluconazole, flucytosine, nystatin and ketoconazole were tested according to the methodology of microdilution proposed by the Clinical and Laboratory Standards Institute (CLSI); results were recorded in values of minimal inhibitory concentration (MIC). A total of 71 Candida isolates from HIV-positive patients were examined with the following species represented: C. albicans (59), C. tropicalis (9), C. glabrata (1), C. guilliermondii (1) and C. krusei (1). A total of 15 Candida isolates were evaluated from control individuals comprised of 11 C. albicans and 4 C. tropicalis samples. Our results demonstrated that the tested antifungal agents showed good activity for most isolates from both groups; however, variability in MIC values among isolates was observed.

Opportunistic microorganisms in patients undergoing antibiotic therapy for pulmonary tuberculosis.

Antimicrobial therapy may cause changes in the resident oral microbiota, with the increase of opportunistic pathogens. The aim of this study was to compare the prevalence of Candida, Staphylococcus, Pseudomonas and Enterobacteriaceae in the oral cavity of fifty patients undergoing antibiotic therapy for pulmonary tuberculosis and systemically healthy controls. Oral rinsing and subgingival samples were obtained, plated in Sabouraud dextrose agar with chloramphenicol, mannitol agar and MacConkey agar, and incubated for 48 h at 37°C. Candida spp. and coagulase-positive staphylococci were identified by phenotypic tests, C. dubliniensis, by multiplex PCR, and coagulase-negative staphylococci, Enterobacteriaceae and Pseudomonas spp., by the API systems. The number of Candida spp. was significantly higher in tuberculosis patients, and C. albicans was the most prevalent specie. No significant differences in the prevalence of other microorganisms were observed. In conclusion, the antimicrobial therapy for pulmonary tuberculosis induced significant increase only in the amounts of Candida spp.

Oral distribution of Candida species and presence of oral lesions in Brazilian leprosy patients under multidrug therapy.

OBJECTIVE: The aim of this study was to evaluate the prevalence of Candida spp. and presence of oral lesions in Brazilian leprosy patients under multidrug therapy (MDT). METHODS: Thirty-eight individuals (18 males and 20 females, median age 53 years) clinically and microbiologically diagnosed as leprosy (lepromatous variant), and under MDT for at least 45 days were studied. The control group constituted by 38 healthy individuals (median age 53.5), matched to the test group in relation to age, gender and oral conditions. Oral rinses were collected and the Candida identification was performed by phenotypic tests. The existence of Candida dubliniensis among the isolates was analyzed using a validated multiplex PCR assay. Twenty-nine leprosy patients were examined intra-orally for the presence of lesions. Data were analyzed by z- and Mann-Whitney tests (alpha = 5%). RESULTS: Yeast carriage rate between leprosy patients (65.8%) and controls (47.4%) was similar (P = 0.099), and no significant difference between yeast counts was observed (P = 0.1004). Candida albicans was the most frequently isolated species in both groups. In the leprosy group, Candida tropicalis and Candida parapsilosis were also identified. In the control group, we additionally identified Candida tropicalis, Candida glabrata and Candida kefyr. Candida dubliniensis was not detected. No leprosy-related oral lesion was registered. CONCLUSION: Within the limits of the study, we concluded that Brazilian leprosy patients under MDT showed similar levels of carriage and Candida species distribution in relation to the controls.

Efetividade de um novo produto a base de gel de triclosan e Pimpinella anisum na anti-sepsia de mãos em Odontologia / Effectiveness of a new product with triclosan and Pimpinella anisum for hand antisepsis in Odontology

O objetivo deste estudo foi avaliar a efetividade antimicrobiana de um novo anti-séptico para mãos a base de triclosan e Pimpinella anisum. Foi realizado estudo in vitro incluindo cepas de Staphylococcus aureus (ATCC 6.538), Escherichia coli (ATCC 25.922), Proteus mirabilis, Citrobacter freundii (CCUT 22.001), Enterobacter aerogenes (IAL 2.482) e Candida albicans (20 isolados clínicos). A metodologia utilizada foi de diluição em ágar (Müeller Hinton ou RPMI/MOPS) e os resultados foram expressos em valores de concentração inibitória mínima (CIM). A CIM para Escherichia coli, Citrobacter freundii e Entterobacter aerogenes foi de 0,03%; para Staphylococcus aureus 6,25%; Proteus mirabilis 12,5% e Candida albicans 12,5%. Para avaliação in vivo foram incluídos 60 voluntários dividos em três grupos: Produto A (triclosan, álcool etílico e Pimpinella anisum - Erva doce); Produto B (irgasan e álcool etílico a 70%) e controle (clorexidina 2% gel). Para a realização da coleta, as mãos foram divididas em oito partes iguais sendo que a coleta inicial e a final foram realizadas em quatro partes alternadas diferentes das mãos. As amostras foram coletadas com auxílio de swab esterilizado, umedecido em solução salina esterilizada e semeadas em duplicata em ágar sangue. Após a incubação, por 48 horas/37ºC, foram obtidos valores de unidades formadoras de colônias. O estudo in vivo demonstrou redução significativa das contagens em todos os grupos estudados (p < 0,001), sendo de 92,5% para o produto A, 91,1% para o produto B e 98,4% para o controle. Não houve diferença estatisticamente significativa entre as reduções de contagens dos grupos testados (p = 0,30). Concluiu-se que o novo produto apresentou alta efetividade antimicrobiana similar a dos demais grupos testados.

Aspectos clínicos, radiográficos e microbianos de uma família com expressiva prevalência de doença periodontal / Clinical, radiographics and microbiological aspects of a family with expressive prevalence of periodontal disease

A periodontite agressiva é caracterizada por uma rápida perda de inserção conjuntiva e destruição óssea, podendo ocorrer na forma localizada ou generalizada. Na periodontite agressiva localizada ocorre o comprometimento, principalmente, dos dentes incisivos e primeiros molares permanentes. É a forma de doença periodontal mais associada com Actinobacillus actinomycetemcomitans. De modo distinto, a forma generalizada da periodontite agressiva afeta vários dentes em um ou ambos os arcos dentários. O objetivo do presente estudo foi estabelecer através de exames clínicos, radiográficos e microbianos o diagnóstico de uma família com expressiva prevalência de doença periodontal. Foram incluídos 5 indivíduos da mesma família, os quais foram submetidos à exame clínico periodontal e exame radiográfico de todos os dentes. Para a análise microbiológica foram selecionados no mínimo dois dentes para cada indivíduo, e a presença de Actinobacillus actinomycetemcomitans foi determinada pelo método de cultura bacteriana e reação em cadeia da polimerase. Dos indivíduos examinados, dois receberam o diagnóstico de periodontite incipiente, dois de periodontite agressiva localizada e um de periodontite agressiva generalizada. O Actinobacillus actinomycetemcomitans de máxima leucotoxicidade foi detectado em todos os indivíduos. Com base nestes resultados observou-se que os indivíduos apresentavam a mesma característica microbiana e diferenças na expressão e severidade da doença periodontal, sugerindo que outros fatores poderiam estar presentes modificando a manifestação clínica da doença.

Longitudinal clinical evaluation of adjunct minocycline in the treatment of chronic periodontitis.

BACKGROUND: The clinical benefits of minocycline in combination with thorough scaling and root planing (SRP) have been examined in multicenter studies. The aim of this longitudinal investigation was to evaluate the clinical response to scaling and root planing combined with the use of locally delivered minocycline microspheres for 720 days in individuals with advanced chronic periodontitis. METHODS: A total of 26 individuals aged 26 to 69 years (mean: 46.8+/-12.1 years) were included in this double-blind randomized clinical trial. After randomization, 13 individuals were selected for the test group (TG) and treated with SRP plus subgingival minocycline at baseline and 90, 180, and 270 days, and 13 individuals were selected for the control group (CG) and received SRP plus vehicle at the same timepoints. Two homologous sites with probing depth (PD)>or=6 mm were chosen in each subject. To evaluate the clinical response after treatment, PD, plaque index (PI), and gingival index (GI) were assessed at baseline and 90, 180, 270, 360, and 720 days. RESULTS: No statistical differences were found between test and control groups in relation to PD at the different timepoints. The mean values of PD demonstrated a higher reduction in the test group at 270 and 360 days. No statistical differences were observed at 90, 180, and 720 days between TG and CG (P<0.05; Wilcoxon test). There were no statistically significant differences between TG and CG concerning PI and GI (P<0.05; analysis of variance and t test) at all evaluated timepoints. CONCLUSION: Our findings demonstrated that both therapies reduced mean PD from 90 to 360 days; however, SRP combined with the use of subgingival minocycline showed a higher reduction at 270 and 360 days following therapy.

Microrganismos superinfectantes na cavidade bucal de indivíduos submetidos a antibioticoterapia para tratamento de tuberculose pulmonar / Superinfecting microorganisms in the oral cavity of individuals undergoing antibiotic therapy for pulmonary tuberculosis treatment

Superinfecção resulta no desenvolvimento de patógenos residentes prejudiciais ou oportunistas em conseqüência de alteração da microbiota benéfica e pode representar uma complicação na terapia antimicrobiana. O presente estudo avaliou a presença de leveduras, estafilococos, enterobactérias e Pseudomonas na cavidade bucal e sulco gengival de indivíduos submetidos à terapia antibiótica para tratamento de tuberculose pulmonar e indivíduos controle. Foram incluídos cinqüenta indivíduos diagnosticados com tuberculose pulmonar e que estivessem sob antibioticoterapia por um período mínimo de 45 dias e máximo de seis meses. Para controle foram incluídos indivíduos sistemicamente saudáveis. A coleta de material da cavidade bucal foi realizada pela técnica de enxágüe bucal e do sulco gengival utilizando-se cones de papel absorvente esterilizados. As amostras coletadas foram semeadas em duplicata em placas de Petri contendo ágar Sabouraud dextrose com cloranfenicol, ágar Manitol e ágar MacConkey. As amostras de Candida spp. e de estafilococos coagulase-positivas foram identificadas por provas bioquímicas específicas e as amostras de estafilococos coagulase-negativas, enterobactérias e Pseudomonas foram identificadas utilizando-se o sistema API. Os resultados foram analisados pelo teste estatístico de Mann-Whitney (p<0,05). Candida spp. foram detectadas em número significativamente maior na cavidade bucal de indivíduos com tuberculose pulmonar em relação aos controles, sendo C. albicans mais prevalente em ambos os grupos. Não foram observadas diferenças estatisticamente significativas entre os grupos para Staphylococcus spp., enterocatérias e Pseudomonas. Concluiu-se que os indivíduos sob antibioticoterapia para tratamento de tuberculose pulmonar apresentaram maior prevalência de Candida spp. na cavidade bucal em relação aos controles.

Clinical and microbial evaluation of dental scaling associated with subgingival minocycline in chronic periodontitis subjects.

The aims of this double-blind randomized clinical trial were to evaluate the presence of periodontal pathogens and the clinical response of periodontal pockets treatment to scaling and root planing (SRP) associated with subgingival minocycline (SM). A total of 36 subjects, 26 to 60 years old (40.7 +/- 9.1), who had been previously diagnosed with chronic periodontitis, were included in the present study. Eighteen subjects were selected for the test group (TG), who were treated with SRP plus SM (new treatment), and 18 subjects for the control group (CG) who received SRP plus vehicle (current treatment). Two homologous sites in each subject with a probing depth (PD) > or = 6 mm were chosen. To evaluate the clinical response after treatment, PD was measured at baseline and at 90 days. Microbiological evaluation was performed to detect 7 periodontal pathogens using polymerase chain reaction at baseline, 30, and 120 days. A mean reduction in PD of 2.8 and 2.1 mm was observed in the TG and CG, respectively. At baseline, P. gingivalis was the most prevalent organism in both test (65.8%) and control (48.6%) groups. After 120 days it fell to 30.8% in TG and to 23.1% in CG. There were no statistically significant differences between the test and control groups concerning PD (p > 0.05 by Wilcoxon test) or presence of periodontal pathogens (p > 0.05 by Wilcoxon and chi-square; p > 0.01 by Signal test). The results observed showed that the new treatment was as effective as the current treatment in reducing periodontal pathogens and PD among chronic periodontitis subjects.

Clinical and microbial evaluation of dental scaling associated with subgingival minocycline in chronic periodontitis subjects

O objetivo deste estudo randomizado duplo-cego foi avaliar a presença de periodontopatógenos e o comportamento clínico de bolsas periodontais tratadas com raspagem e aplainamento radicular (RAR) associado à minociclina subgengival (MS). Foram incluídos no presente estudo 36 indivíduos de 26 a 60 anos de idade (40,7 ± 9,1), previamente diagnosticados com periodontite crônica. Dezoito indivíduos foram selecionados para o grupo teste (GT), tratado com RAR e MS (novo tratamento), e 18 indivíduos, para o grupo controle (GC), que recebeu RAR e veículo (tratamento convencional). Foram selecionados dois sítios homólogos em cada indivíduo com profundidade de sondagem (PS) > 6 mm para testar a hipótese proposta. Para avaliar o comportamento clínico após o tratamento, a mensuração da PS foi realizada inicialmente e após 90 dias. A avaliação microbiana foi realizada para detecção de 7 periodontopatógenos através de reação em cadeia da polimerase, inicialmente e após 30 e 120 dias. Observou-se redução média de 2,8 e 2,1 mm na PS nos grupos teste e controle, respectivamente. Inicialmente, P. gingivalis foi o microrganismo mais prevalente tanto no GT (65,8%) quanto no GC (48,6%). Após 120 dias, houve redução para 30,8% no GT e 23,1% no GC. Não houve diferenças estatisticamente significativas entre os grupos em relação à PS (teste Wilcoxon - p > 0,05) e presença dos periodontopatógenos (teste Wilcoxon e qui-quadrado - p > 0,05 e teste do sinal - p > 0,01). Os resultados observados demonstraram que o novo tratamento foi tão efetivo quanto o tratamento convencional na redução de periodontopatógenos e de PS em indivíduos com periodontite crônica.

Avaliação Clínica e microbiana da terapia periodontal mecânica em indivíduos com periodontite crônica / Clinical and microbiological evaluation of periodontal mechanical therapy in chronic periodontitis subjects

O objetivo deste estudo foi avaliar o comportamento clínico de bolsas periodontais ò6mm de profundidade em indivíduos com periodontite crônica após tratamento com raspagem e aplainamento radicular (RAR) e a presença de periodontopatógenos. Foram incluídos 35 individuos, de ambos os gêneros, com idade entre 26 e 60 anos (40,7 ± 9,1) previamente diagnosticados com periodontite crônica. Foram selecionados quatro sítios com profundidade de sondagem (PS) ò 6mm em cada indivíduo, totalizando 140 sítios avaliados. Os sítios selecionados foram submetidos aos procedimentos de RAR realizados por um único operador e por um período de tempo padronizado. A mensuração da PS foi realizada no momento inicial e após 90 dias. Através de PCR avaliou-se a presença de A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus, C. rectus, E. corrodens, F.nucleatum subsp. nucleatum e F. nucleatum subsp. vicentii no momento inicial após 120 dias. O produto amplificado por PCR (rDNA 165) foi analisado por ensaio Dot-Blot. Os resultados foram submetidos a tratamento estatistico através dos testes Wilcoxon para PS e Kruskal-Wallis e T-Student para presença de periodontopatógenos. Observou-se redução (p<0,05) quando se comparou PS inicial e após 90 dias. Em relação aos periodontopatógenos examinados, observou-se redução (p<0,05) entre os dois tempos analisados, exceto para C. rectus, E. corrodens e F. nucleatum subsp. nucleatum independentemente do gênero. Assim, concluiu-se que as bolsas periodontais analisadas apresentaram uma resposta clínica e microbiana compatível com a proposta da terapia mecânica periodontal, podendo ser indicada nos individuos com periodontite crônica.

PCR e cultura na detecção subgengival de Actiobacillus actinomycetemcomitans: estudo comparativo / Actinobacillus actinomycetemcomitans detection by PCR and culture technique: comparative study

O objetivo do presente estudo foi comparar a eficácia de duas técnicas, PCR e cultura, frequentemente utilizadas para a detecção de Actinobacillus actinomycetemcomitans em amostras de placa bacteriana subgengival. Foram avaliados 136 indivíduos de ambos os sexos, acima de 14 anos de idade, diagnosticados com gengivite (n = 47), periodontite crônica (n = 70) ou agressiva (n = 19) segundo os critérios propostos pela AAP3 (1999). Para cada indivíduo foram obtidas amostras de placa bacteriana subgengival de 5 bolsas periodontais, localizadas na face mesial de dentes distintos, equivalentes aos maiores valores de profundidade de sondagem (PS). Diante de valores edênticos de Ps o maior grau de inflamação dos tecidos periodontais foi auxiliar na seleção dos sítios. A. actinomycetemcomitans foi cultivado em placas de ágar TSBV e identificado de acordo com a morfologia de colônia, coloração de Gram e provas bioquímicas. A PCR foi realizada utilizando-se primers para amplificação do gene da leucotoxina. Os resultados foram analisados empregando-se teste Qui-Quadrado e MacNemar. Dezenove indivíduos apresentaram resultados positivos para cultura bacteriana e 35 indivíduos resultados positivos para PCR. Foram encontradas sensibilidade de 0,74 e especificidade de 0,82 considerando-se a cultura como teste padrão. PCR esteve associada a sangramento a sondagem (c2 = 7,11) e detectou mais amostras positivas de A. actinomycetemcomitans (p < 0,05). Houve superioridade da PCR em relação à cultura na detecção subgengival de A actinomycetemcomitans.