Human protothecosis: A case report in northeastern Brazil.

We describe a case of extensive human cutaneous protothecosis in the northeastern of Brazil. The diagnosis was made through histopathological examination of skin samples and culturing on Sabouraud dextrose agar, with identification through mass spectrometry. Treatment with itraconazole failed and was switched to liposomal amphotericin followed by voriconazole with clinical cure. Prothotecosis should be a diagnostic hypothesis when there are skin lesions after trauma, specially in situations of atypical evolution.

Molecular Detection of Drug-Resistance Genes of blaOXA-23-blaOXA-51 and mcr-1 in Clinical Isolates of Pseudomonas aeruginosa.

Pseudomonas aeruginosa has caused high rates of mortality due to the appearance of strains with multidrug resistance (MDR) profiles. This study aimed to characterize the molecular profile of virulence and resistance genes in 99 isolates of P. aeruginosa recovered from different clinical specimens. The isolates were identified by the automated method Vitek2, and the antibiotic susceptibility profile was determined using different classes of antimicrobials. The genomic DNA was extracted and amplified by multiplex polymerase chain reaction (mPCR) to detect different virulence and antimicrobial resistance genes. Molecular typing was performed using the enterobacterial repetitive intergenic consensus (ERIC-PCR) technique to determine the clonal relationship among P. aeruginosa isolates. The drug susceptibility profiles of P. aeruginosa for all strains showed high levels of drug resistance, particularly, 27 (27.3%) isolates that exhibited extensively drug-resistant (XDR) profiles, and the other isolates showed MDR profiles. We detected the polymyxin E (mcr-1) gene in one strain that showed resistance against colistin. The genes that confer resistance to oxacillin (blaOXA-23 and blaOXA-51) were present in three isolates. One of these isolates carried both genes. As far as we know from the literature, this is the first report of the presence of blaOXA-23 and blaOXA-51 genes in P. aeruginosa.

Chromoblastomycosis in an Endemic Area of Brazil: A Clinical-Epidemiological Analysis and a Worldwide Haplotype Network.

Chromoblastomycosis (CBM) is a neglected implantation mycosis prevalent in tropical climate zones, considered an occupational disease that affects impoverished rural populations. This retrospective study described clinical aspects of CBM in a hyperendemic area in Brazil and constructed a worldwide haplotype network of Fonsecaea spp. strains. The variables were collected from medical records using a standard report form, reporting 191 patients with CBM from Maranhão, Brazil. The mean age was 56.1 years, 168 (88%) patients were male and predominantly farmers (85.8%). The mean time of evolution of the disease until diagnosis was 9.4 years. Lower limbs (81.2%) and upper limbs (14.2%) were the main sites affected. Most patients exhibited verrucous (55%) and infiltrative plaque (48.2%). Fonsecaea spp. were identified in 136 cases and a haplotype network constructed with ITS sequences of 185 global strains revealed a total of 59 haplotypes exhibiting high haplotypic and low nucleotide diversities. No correlation was observed between the different haplotypes of Fonsecaea species and dermatological patterns, severity of disease or geographic distribution inside Maranhão. Data from this area contributed to better understanding the epidemiology of CBM. For the first time, a robust haplotype network with Fonsecaea strains reveals an evolutionary history with a recent population expansion.

Evaluation of the interaction between polymyxin B and Pseudomonas aeruginosa biofilm and planktonic cells: reactive oxygen species induction and zeta potential.

BACKGROUND: Although the most widely accepted mechanism of action for polymyxins is related to bacterial lysis via disruption, we hypothesized that this antimicrobial drug class could have other effects on Pseudomonas aeruginosa planktonic and sessile cells. Little is known regarding oxidative burst and zeta potential (ZP) data associated with the interaction between polymyxin B and P. aeruginosa cells. The present study evaluated endogenous reactive oxygen species (ROS) production and changes in the net charges of biofilm and planktonic cells in response to polymyxin B. RESULTS: Polymyxin B induced concentration-dependent killing at all concentrations tested in planktonic and sessile cells from P. aeruginosa strains. Sublethal concentrations of polymyxin B induced oxidative burst. ROS production was higher in resistant planktonic cells than in biofilm cells but this was not observed for susceptible cells. Moreover, no net surface charge alterations were observed in planktonic cells from a susceptible strain treated with polymyxin B, but a significant increase of ZP was noted in planktonic cells from a resistant strain. CONCLUSION: Oxidative burst generated by planktonic and sessile cells from P. aeruginosa strains against polymyxin B indicates that ROS may have an important role in the mechanism of action of this drug. ZP data revealed that electrostatic interactions of the cationic peptide with the anionic surface of the cells are strain-dependent. Therefore, we suggested that the intracellular effects of polymyxin B should be further investigated to understand polymyxin B-induced stress in P. aeruginosa.

Phenotypic and molecular detection of the bla KPC gene in clinical isolates from inpatients at hospitals in São Luis, MA, Brazil.

BACKGROUND: Bacteria that produce Klebsiella pneumoniae carbapenemases (KPCs) are resistant to broad-spectrum ß-lactam antibiotics. The objective of this study was to phenotypically and genotypically characterize the antibiotic susceptibility to carbapenems of 297 isolates recovered from clinical samples obtained from inpatients at 16 hospitals in São Luis (Maranhão, Brazil). METHODS: The study was conducted using phenotypic tests and molecular methods, including polymerase chain reaction (PCR), sequencing and enterobacterial repetitive intergenic consensus (ERIC)-PCR. The nonparametric chi-square test of independence was used to evaluate the associations between the bacterial bla KPC gene and the modified Hodge test, and the chi-square adherence test was used to assess the frequency of carbapenemases and their association with the bla KPC gene. RESULTS: The most frequently isolated species were Acinetobacter baumannii (n = 128; 43.0%), K. pneumoniae (n = 75; 25.2%), and Pseudomonas aeruginosa (n = 42; 14.1%). Susceptibility assays showed that polymixin B was active against 89.3% of the bacterial isolates. The Acinetobacter spp. and K. pneumoniae strains were susceptible to amikacin and tigecycline, and Pseudomonas spp. were sensitive to gentamicin and amikacin. Among the 297 isolates, 100 (33.7%) were positive for the bla KPC gene, including non-fermentative bacteria (A. baumannii) and Enterobacteriaceae species. Among the isolates positive for the bla KPC gene, K. pneumoniae isolates had the highest positivity rate of 60.0%. The bla KPC gene variants detected included KPC-2, which was found in all isolates belonging to species of the Enterobacteriaceae family. KPC-2 and KPC-3 were observed in A. baumannii isolates. Importantly, the bla KPC gene was also detected in three Raoultella isolates and one isolate of the Pantoea genus. ERIC-PCR patterns showed a high level of genetic diversity among the bacterial isolates; it was capable of distinguishing 34 clones among 100 strains that were positive for bla KPC and were circulating in 11 of the surveyed hospitals. CONCLUSIONS: The high frequency of the bla KPC gene and the high degree of clonal diversity among microorganisms isolated from patients from different hospitals in São Luis suggest the need to improve the quality of health care to reduce the incidence of infections and the emergence of carbapenem resistance in these bacteria as well as other Gram-negative pathogens.

Isolation and identification of Candida species in patients with orogastric cancer: susceptibility to antifungal drugs, attributes of virulence in vitro and immune response phenotype.

BACKGROUND: Because of the inherent immunosuppression of cancer patients opportunistic infections by Candida spp, occur frequently. This study aimed to identify Candida species in the oral mucosa of 59 patients with orogastric cancer (OGC) and to analyze the immunological phenotype of these patients. METHODS: The yeasts were identified by MALDI-TOF mass spectrometry (MS). For all isolates, we performed phospholipases and proteinases assays, in vitro adherence to buccal epithelial cells (BEC), minimum inhibitory concentration of antifungal drugs and determined the cytokine profile by Cytometric Bead Array flow citometry assay. RESULTS: C. albicans was the most prevalent species in OGC patients (51.6 %) and control group (66.7 %). Candida spp. strains isolated from OGC patients exhibited better adherence to BEC (p = 0.05) than did the control group. Phospholipases production by Candida strains from OGC patients was lower (51.6 %) than in the control group (61.9 %). Proteinases were detected in 41.9 % and 4.8 % of the yeasts from OGC patients and control group, respectively. Significant differences were found in the serum of OGC patients compared to the control group for IL-2, IL-10, TNF-α, IFN-γ and IL-17. CONCLUSIONS: The results of this work suggest increased virulence of yeasts isolated from OGC patients and, that this may interfere with the immune phenotype.

Extended-spectrum ß-lactamase-producing enterobacteriaceae in community-acquired urinary tract infections in São Luís, Brazil.

The number of ESBL-producing Enterobacteriaceae in community-acquired urinary tract infections worldwide is probably underestimated because of the technical difficulties encountered with their detection. In this study, out of 5,672 urine samples analyzed, 916 were positive for uropathogens, 472 of them being enterobacteria of which 7.6% produced ß-lactamases. Analysis of the isolated from 36 patients showed a high level of antibiotic resistance, with 52.7% and 80.5% of isolates expressing bla TEM and bla CTX-M, respectively.

Ants in a hospital environment and their potential as mechanical bacterial vectors.

INTRODUCTION: We studied the richness and abundance of ant species, their bacteria and the bacteria isolated from patient clinical samples. METHODS: Ants were collected with baited traps at 64 sites in a public hospital in São Luis, State of Maranhão, Brazil. RESULTS: In total, 1,659 ants from 14 species were captured. The most frequent species were Crematogaster victima, Solenopsis saevissima, Tapinoma melanocephalum, Camponotus vittatus and Paratrechina fulva. Forty-one species of bacteria were isolated from the ants and 18 from patients. CONCLUSIONS: Ants are potential vehicles for pathogenic and opportunistic bacteria, and they can represent a risk factor in nosocomial infections.

Ants in a hospital environment and their potential as mechanical bacterial vectors

Introduction We studied the richness and abundance of ant species, their bacteria and the bacteria isolated from patient clinical samples. Methods Ants were collected with baited traps at 64 sites in a public hospital in São Luis, State of Maranhão, Brazil. Results In total, 1,659 ants from 14 species were captured. The most frequent species were Crematogaster victima, Solenopsis saevissima, Tapinoma melanocephalum, Camponotus vittatus and Paratrechina fulva. Forty-one species of bacteria were isolated from the ants and 18 from patients. Conclusions Ants are potential vehicles for pathogenic and opportunistic bacteria, and they can represent a risk factor in nosocomial infections. .

Phenotypic detection of metallo-ß-lactamases in Pseudomonas aeruginosa and Acinetobacter baumannii isolated from hospitalized patients in São Luis, State of Maranhão, Brazil.

INTRODUCTION: Acquired metallo-ß-lactamases (MßL) are emerging determinants of resistance in Pseudomonas aeruginosa and Acinetobacter baumannii. The objectives of this study were to phenotypically detect MßL in imipenem-resistant P. aeruginosa and A. baumannii, to investigate the association between MßL-positive strains and hospitals, and to compare the resistance profiles of MßL-producing and non-MßL-producing strains. METHODS: The approximation disk and combined disk assay methods were used in this study. RESULTS: A total of 18 (38.3%) P. aeruginosa isolates and 1 (5.6%) A. baumannii isolate tested positive for the presence of MßL. CONCLUSIONS: These results demonstrate the need for strict surveillance and for the adoption of preventive measures to reduce the spread of infection and potential outbreaks of disease caused by MßL-producing microorganisms.

Extended-spectrum β-lactamase-producing enterobacteriaceae in community-acquired urinary tract infections in São Luís, Brazil

The number of ESBL-producing Enterobacteriaceae in community-acquired urinary tract infections worldwide is probably underestimated because of the technical difficulties encountered with their detection. In this study, out of 5,672 urine samples analyzed, 916 were positive for uropathogens, 472 of them being enterobacteria of which 7.6% produced β-lactamases. Analysis of the isolated from 36 patients showed a high level of antibiotic resistance, with 52.7% and 80.5% of isolates expressing blaTEM and blaCTX-M, respectively.

Nosocomial infection and characterization of extended-spectrum ß-lactamases-producing Enterobacteriaceae in Northeast Brazil.

INTRODUCTION: Extended spectrum ß-lactamases (ESBLs) are enzymes that degrade ß-lactam antibiotics and have been reported to be an important cause of nosocomial infection in worldwide. METHODS: During 2009, 659 enterobacteria strains were isolated from different clinical specimens and tested for ESBL production. The disk approximation test, combined disk method and addition of clavulanic acid were used for phenotypic detection of the ESBL-producing strains and PCR for detection of the bla(TEM) and bla(CTX-M) genes. RESULTS: Among the isolates, 125 were ESBL producers. The bla(CTX-M) and bla(TEM) genes were detected in 90.4% and 75% of the strains, respectively. Most strains were isolated from urine. Klebsiella pneumoniae was the most prevalent organism. Microorganisms presented high resistance to the antibiotics. CONCLUSIONS: These results support the need for extending ESBL detection methods to different pathogens of the Enterobacteriaceae family because these methods are only currently standardized by the CLSI for Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis. Carbapenems were the antibiotic class of choice for the treatment of infections caused by ESBL-producing Enterobacteriaceae.

Nosocomial infection and characterization of extended-spectrum β-lactamases-producing Enterobacteriaceae in Northeast Brazil / Infecção hospitalar e caracterização de enterobactérias produtoras de β-lactamases de espectro ampliado no nordeste do Brasil

INTRODUCTION: Extended spectrum β-lactamases (ESBLs) are enzymes that degrade β-lactam antibiotics and have been reported to be an important cause of nosocomial infection in worldwide. METHODS: During 2009, 659 enterobacteria strains were isolated from different clinical specimens and tested for ESBL production. The disk approximation test, combined disk method and addition of clavulanic acid were used for phenotypic detection of the ESBL-producing strains and PCR for detection of the blaTEM and blaCTX-M genes. RESULTS: Among the isolates, 125 were ESBL producers. The blaCTX-M and blaTEM genes were detected in 90.4% and 75% of the strains, respectively. Most strains were isolated from urine. Klebsiella pneumoniae was the most prevalent organism. Microorganisms presented high resistance to the antibiotics. CONCLUSIONS: These results support the need for extending ESBL detection methods to different pathogens of the Enterobacteriaceae family because these methods are only currently standardized by the CLSI for Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis. Carbapenems were the antibiotic class of choice for the treatment of infections caused by ESBL-producing Enterobacteriaceae.

INTRODUÇÃO: As β-lactamases de espectro ampliado (ESBLs) são enzimas que degradam os antibióticos β-lactâmicos e têm sido reportadas como uma importante causa de infecções hospitalares em todo o mundo. MÉTODOS: Em 2009, 659 enterobactérias foram isoladas de diferentes espécimes clínicos e testadas quanto à produção de ESBL. Os testes de aproximação do disco, disco combinado e adição do ácido clavulânico foram utilizados na detecção fenotípica das amostras produtoras de ESBL e PCR para a detecção dos genes blaTEM e blaCTX-M. RESULTADOS: Entre os isolados, 125 foram produtores de ESBL. Os genes blaCTX-M e blaTEM foram detectados em 90,4% e 75% das amostras, respectivamente. Com relação ao espécime clínico, a maioria das amostras foi isolada de urina. Klebsiella pneumoniae foi a espécie mais prevalente e o teste de susceptibilidade antimicrobiana mostrou uma elevada resistência dos microorganismos aos antibióticos testados. CONCLUSÕES: Estes resultados suportam a necessidade de se ampliar os métodos de detecção das ESBLs para os diferentes patógenos da família Enterobacteriaceae, uma vez que esses métodos estão padronizados pelo CLSI apenas para Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca e Proteus mirabilis. Os carbapenens foram os antibióticos de escolha para o tratamento de infecções causadas por enterobactérias produtoras de ESBL.

Effect of chlorhexidine gel containing saccharin or aspartame in deaf children highly infected with mutans streptococci

Aim: Since deaf children are unable to comprehend or cooperate with dental treatment due to lack of communication, preventive measures may be an important way to control the high prevalence of dental caries in these patients. The aim of the this study was to evaluate the effect of an intensive treatment with chlorhexidine (CHX) gel, containing either saccharin or aspartame, in deaf children highly infected with mutans streptococci (MS). Methods: Eighteen children were randomly divided into two groups, according to the sweetener used to improve the CHX gel bitter taste: saccharin or aspartame. Before CHX treatment, saliva samples were collected to establish baseline microbial data for MS. CHX gel was applied on two consecutive days, four times the first day and three times the second day. Saliva samples were then taken after 7, 30, 60, 90 and 120 days to evaluate MS oral recolonization. Results: CHX gel containing saccharin was not effective on the reduction of MS levels, while the gel containing aspartame decreased significantly MS levels after treatment (P<.05). Conclusions: Although a new CHX application may be necessary after 60 days to control caries risk and MS levels, CHX treatment should be individually controlled because of variations in the response of subjects.

Fonsecaea pedrosoi infection induces differential modulation of costimulatory molecules and cytokines in monocytes from patients with severe and mild forms of chromoblastomycosis.

The host defense mechanism in chromoblastomycosis has not been thoroughly investigated. It has been suggested that cell-mediated immunity in patients with long-standing chromoblastomycosis is somehow impaired. As a result, these individuals became unable to develop an efficient immune reaction. Many studies have shown that monocyte-derived macrophages exhibit critical activities in immunity to microorganisms. Moreover, the ability of cells from the monocytic lineage to process and present antigens, to produce cytokines, and to provide costimulatory signals confirms their pivotal role in the initiation of specific immune responses. In the present study, it was observed that monocytes from patients with a severe form of disease had a higher production of IL-10 and a lower expression of HLA-DR and costimulatory molecules when stimulated with specific antigen or LPS. Immune modulation with recombinant IL-12 or anti-IL-10 can restore the antigen-specific Th1-type immune response in chromoblastomycosis patients by up-regulating HLA-DR and costimulatory molecules in monocytes. Therefore, our data show that monocytes from patients with different clinical forms of chromoblastomycosis present distinct phenotypic and functional profiles. This observation suggests possible mechanisms that control the T cell response and influence their role in the development of pathology.

Isolation of Fonsecaea pedrosoi from the shell of the babassu coconut (Orbignya phalerata Martius) in the Amazon region of Maranhão Brazil.

Fonsecaea pedrosoi, a dematiaceous fungus and the main causative agent of chromoblastomycosis, has been isolated in worldwide from different natural sources in regions where the disease is endemic. In the Amazon region of Maranhão, Brazil, where the disease is prevalent, the breaking of the babassu coconut (Orbignya phalerata Martius) represents an important agricultural activity. In order to determine the presence of this fungus on this plant and on other natural substrates, material was collected in the Fortaleza Village Municipality of Pinheiro, Maranhão, in April and September 2002. A total of 68 samples, including 18 (26.5%) obtained from the shell of the babassu coconut, were analyzed. Samples were cultured using a standard method. Isolates were identified based on macromorphological aspects of the colonies on Sabouraud dextrose agar and based on the micromorphology of the conidia after growth on potato dextrose agar. Exophiala sp. was the most prevalent fungus isolated from the different natural substrates analyzed, while Cladophialophora sp. was only isolated from decomposing wood. Fonsecaea pedrosoi was isolated from one sample of babassu coconut shell suggesting that this coconut represents an important source of infection of chromoblastomycosis during extraction of the plant product in this region.

Identificación de especies del género cándida en la cavidad oral de pacientes con SIDA / Identification of candida species in oral cavity in AIDS patients

Fueron aisladas especies del género candida y otras levaduras de la cavidad oral de pacientes con SIDA y se las relacionó con las manifestaciones clínicas. Fueron estudiadas 58 pacientes internados en el Hospital Estatal Getúlio Vargas en Sao Luis Maranhao, entre julio y diciembre de 2000. Se aislaron levaduras de la cavidad oral de 45 pacientes con la siguiente distribución: C. albicans en 58,4 por ciento; C. krusei y C. parapsilosis, se verificó en 8,3 por ciento, C. tropicalis en 6,7 por ciento, C. glabrata y S. cerevisiae en 5,0 por ciento, C. guilliermondii y Trichosporon spp2 en 3,3 por ciento, y menos frecuentemente C. lusitaniae en 1,7 por ciento. Se encontró en el 26,7 por ciento de los pacientes, conjuntamente con C. albicans, otras especies. En el 73,3 por ciento de los pacientes se encontró sólo un tipo de levadura. La forma clínica de candidiasis oral predominante fue la forma eritematosa en el 33,3 por ciento de los enfermos, seguida por la forma pseudomembranosa asociada a la eritematosa en 22,2 por ciento. Se comprobó que en los pacientes con SIDA portadores de C. albicans se presentaba con mayor frecuencia manifestaciones clínicas (66,7 por ciento) que los portadores de otra especie de levaduras (22,2 por ciento). Se observó que, el 75,5 por ciento de los pacientes presentaron uno o más episodios de infección. El aislamiento de levaduras fue mayor en pacientes que presentaron recuentos de linfocitos T CD4 entre <50 y 200 céls/mm3 (64,5 por ciento)

Cromoblastomicose tratada com itraconazol sistêmico associado a anfotericina B intralesional / Chromoblastomycosis treated with systemic itraconazole associated with intralesional amphotericin B

A cromoblastomicose constitui infecçäo fúngica granulomatosa com relativa freqüência em alguns estados do Brasil, acreditando-se que 95 porcento dos casos ocorram devido a fungos pertencentes ao gênero Fonsecaea. No Estado do Maranhäo a maioria dos casos ocorre em lavradores. Em funçäo das dificuldades financeiras dos pacientes e de a rede de saúde pública näo dispor de medicamentos atuantes na doença, de modo geral os pacientes ficam anos sem tratamento ou o fazem irregularmente, tornando-se refratários às medicaçöes instituídas. Em virtude de tais fatos, säo sempre buscadas alternativas terapêuticas e, entre elas, a associaçäo medicamentosa. Neste presente estudo, relata-se o tratamento de três pacientes portadores de cromoblastomicose causada por fungos da espécie Fonsecaea pedrosi usando o medicamento itraconazol sistêmico associado a anfotericina B intralesional, discutindo-se as perspectivas de utilizaçäo dessa associaçäo e a evoluçäo terapêutica