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Testes de microdiluição em caldo e diluição em ágar para avaliação da suscetibilidade in vitro de dermatófitos
(Universidade Federal de Goiás, 2007-05-31) ARAUJO, Crystiane Rodrigues de; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725
Dermatophytes are keratinophilic fungi that colonize and invade the stratum corneum of the skin, hair and nails causing the dermatophytosis. An increasing number of antifungal agents has become available for the treatment of dermatophytosis, however not all species have the same susceptibility pattern and may occur relative or absolute resistance of some dermatophytes. The document M38-A developed by Clinical and Laboratory Standards Institute (CLSI) for determining the minimal inhibitory concentration (MIC) of different antifungal agents against filamentous fungi, has not included the dermatophytes. The broth microdilution method has been evaluated by various researchers, and some parameters as inoculum size, temperature and duration of incubation and endpoint determination has been investigated. In this study, the in vitro activity of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine against 60 dermatophyte isolates, belong to three species, using the broth microdilution technique, with modifications at temperature and incubation time was used. Additionally, the MIC values obtained by broth microdilution method were compared with those obtained by the agar dilution technique. The results obtained by broth microdilution method showed that all isolates produced clearly detectable growth at 28oC and the MIC values could be determined after 4 days of incubation for the isolates of Trichophyton mentagrophytes and 5 days for T. rubrum and Microsporum canis isolates. Itraconazole, ketoconazole and terbinafine had the lowest MIC values (0.03 μg/ml) for 33.3%, 31.6% and 15% of the isolates, respectively. A good concordance was observed between the agar dilution and broth microdilution methods. The levels of agreement were 91.6% with ketoconazole and griseofulvin, 83.3% with itraconazole, 81.6% with terbinafine and 73.3% with fluconazole for all the tested isolates. In summary, the results of this study suggest that an incubation time of 5 days and temperature at 28oC used in broth microdilution and agar dilution methods can contribute to define and to better interpret the MIC values. Beside, until a reference method for testing the susceptibilities of dermatophytes is standardized, the similar results with broth microdilution method become the agar dilution useful for testing the susceptibility of these fungi.
Surto de infecção após videoscopias causado por Mycobacterium massiliense em Goiânia-GO : análise molecular e determinação da suscetibilidade aos antimicrobianos
(Universidade Federal de Goiás, 2009-12-03) CARDOSO, Alessandra Marques; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; KIPNIS, André; http://lattes.cnpq.br/4434965360286741
In recent years the number of infections caused by microbacteria non-tuberculous mycobacteria (NTM) has increased mainly due to opportunistic infections in individuals imunocompormetidos and improvement of farming techniques and identification of MTN. Mycobacterium massilienese is an emerging body associated with wound infections, abscesses and pneumonia. An outbreak of infection after videoscopy occurred between 2005 and 2007 in seven hospitals in Goiânia-GO, in central Brazil. The objective of this study was to identify NTM isolated from patients with infection after arthroscopy and lararoscopia by PCR followed by analysis of fragment length polymorphism restrção (PRA-hsp65), compared by gel electrophoresis pulsed-field gel (PFGE), sequencing of the partial rpoB gene and determination of antimicrobial susceptibility in vitro. NTM were recovered from samples (exudate abscess subcutâneio) of 18 patients involved in the outbreak. In the period leading up to this study there was no reported case of infection after videoscopy caused by MTN in Goiania. The 18 isolates were identified as M, massiliene and genotyped as a single clone, indicating that they had a common origin, suggesting a common source of infection for the patients involved in the outbreak. The epidemic isolates were susceptible to amikacin (MIC90 4 micrograms / ml) and clarithromycin (MIC90 <1 ug / ml), but resistance to ciprofloxacin (MIC90 <128g/ml), tobramycin (MIC90 32 micrograms / ml) and intermediate susceptibility to cefoxitin (MIC90 64 ug / ml). In conclusion this study demonstrated the clonality of strains of M. massiliense involved in infections after procedures videoscopes and that they are susceptible to drugs indicated for the treatment
Tipagem molecular da cápsula de Haemophilus influenzae isolados da nasofaringe de crianças de creches de Goiânia
(Universidade Federal de Goiás, 2010-03-19) CARVALHO, Camila Xavier de; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899; KIPNIS, André; http://lattes.cnpq.br/4434965360286741
Haemophilus influenzae (Hi) causes infection in children, and is presented in two ways: with six encapsulated serotypes a-f and non-encapsulated or nontypeable (NTHi). Capsulated strains are responsible for a variety of invasive diseases, with meningitis being the most frequent. Nontypeable strains are responsible for respiratory tract infections and acute otitis media in children under 24 months. Children who attend day care centers have increased risk of developing otitis media when colonized with NTHi. Our goal was to describe the prevalence of colonization by Hi and risk factors associated with carrier status in children attending day care centers. Nasopharyngeal swabs collected from 1192 healthy children under five years of age who attended one of 62 daycare centers in Goiânia - Goiás, Brazil were analyzed. The samples were placed on chocolate agar plates and incubated in an atmosphere containing 5% CO2 at 37 ° C overnight. Hi were identified according with colony morphology in culture, Gram staining, and their requirement for V (hemin) and X (NAD) factors. Capsular typing and the presence of the genes TEM1 and ROB1 for resistance to β-lactams were evaluated by PCR. Differences between proportions and means were tested using Chi-square and Student's t test, respectively. Estimates of relative risk (odds ratio) were evaluated by univariate and multivariate logistic regression, p values less than 5% were considered statistically significant. The prevalence of colonization among the 1192 children was 32.1% and 23.3% for HiNT and 8.8% for encapsulated strains. The prevalence of strains carrying the gene TEM1 was 38.4%. Among HiNT strains the prevalence of TEM1 gene was 43.2%. Previous hospitalization of children in the last 6 months was independently associated with the risk carrier by H. influenzae typeable. The data described in this study will aid investigation on the impact of the 10-valent pneumococcal vaccine (PHiD-CV) introduction.
Caracterização bioquímica da endoxilanase recombinante (HXYN2r) do fungo termofílico Humicola grisea var. thermoidea e sua aplicação na sacarificação de resíduos agrícolas
(Universidade Federal de Goiás, 2008-06-30) CARVALHO, Wagner Rodrigues de; FARIA, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003
Xylanases have been used in the biobleaching of paper pulps, in bioconversion of plant biomass, for food and feed industries, among others. For successful selection of xylanases suitable for specific industrial applications it is important to characterize enzymes isolated from different sources. The thermophilic fungus Humicola grisea var. thermoidea is described as a good producer of extracellular endoxylanases and the Hxyn2 gene from this fungus was isolated and expressed in yeast Pichia pastoris. The aim of this project was focused on the production, purification and biochemical characterization of the recombinant HXYN2 (HXYN2r) enzyme and application on enzymatic hydrolysis of lignocellulosics substrates. The culture conditions of P. pastoris in flasks, using the 12.3 transformant and BMMY-U medium, were optimized and the best xylanolytic activity was 478.2 U/mL after 96 h, with a protein concentration of 100 mg/L, using 2.34% (w/v) of nitrogen sources (yeast extract plus urea 1.34:1.0% - w/v), 1% (v/v) of methanol, and OD600 of 10. The HXYN2r enzyme was purified by gel filtration chromatography with a yield of 6.4% and showed optimum pH and temperature values of 6.5 and 60ºC, respectively, maintaining 100% of initial activity after 4 h of incubation at pH 5.5, 6.5 and 7.5. The half-life time was 18 min at 60ºC and the enzyme was 100% stable after 3 h of incubation at 50ºC. Km and Vmax values were 7.9 mg/mL e 235.4 μmol/(mL.min), respectively. The HXYN2r enzyme were used on the enzymatic hydrolysis of sugarcane bagasse (SCB), corn cob (CC) and foliar sample of sugarcane (FSC) alone or with the enzymes of xylanolytic and cellulolytic system produced by H. grisea fungus. For enzymatic hydrolysis experiments, the substrates were milled and pretreated with 0.25% (w/v) of H2SO4 by 30 min. On the enzymatic hydrolysis the best conversion yield of hemicellulosic fractions were obtained using PpHXYN2r supplemented with EHg: 42.8% to CC, 9.6% to SCB and, a mean of 20% to foliar samples.
Identificação e caracterização de moléculas envolvidas na interação de Paracoccidioides brasiliensis com o hospedeiro
(Universidade Federal de Goiás, 2009-03-31) DANTAS, Sabrina Fonseca Ingênito Moreira; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637
Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM), a systemic mycosis presenting clinical manifestations ranging from mild to severe forms. A P. brasiliensis cDNA expression library was produced and screened with pooled sera from PCM patients adsorbed against antigens derived from in vitro-grown P. brasiliensis yeast cells. Sequencing DNA inserts from clones reactive with PCM patients sera indicated 35 open reading frames presenting homology to genes involved in metabolic pathways, transport, among other predicted functions. The complete cDNAs encoding aromatic L-amino acid decarboxylase (Pbddc), lumazine synthase (Pbls) and a homologue of the high affinity copper transporter (Pbctr3) were obtained. Recombinant proteins PbDDC and PbLS were obtained; a peptide was synthesized for PbCTR3. The proteins and the synthetic peptide were recognized by sera of patients with confirmed PCM and not by sera of healthy patients. Using the vivo-induced antigen technology (IVIAT) we identified immunogenic proteins expressed at high levels during infection. Quantitative real - time RT-PCR demonstrated high transcript levels of Pbddc, Pbls and Pbctr3 in yeast cells infecting macrophages. Transcripts in yeast cells derived from spleen and liver of infected mice were also measured by qRT-PCR. Our results suggest a putative role for the immunogenic proteins in the infectious process of P. brasiliensis.
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2009-02-27) FERNANDES, Maria Regilda de Araújo; PEREIRA, Maristela; http://lattes.cnpq.br/1345781867765758
The pathogenic fungus Paracoccidioides brasiliensis is the etiologic agent of Paracoccidioidomycosis (PCM). The main route of contamination is the inhalation of fungal propagules that convert to yeast in the host tissues. The urease is among the factors of virulence described for dimorphic fungus. It hydrolyzes the urea producing molecules of ammonia and carbamate. Pbure has 3,012 bp, corresponding to a predicted protein of 837 amino acids, predicted molecular mass of 90 kDa and pI 6.0. PbURE has signature to nickel-dependent enzyme for its activity. The phylogenetic relationship between PbURE and urease from other fungi was evaluated. The cDNA that encodes PbURE was inserted into the expression vector pET-32a and recombinant protein of 103 kDa was expressed. Polyclonal antibody were produced against PbUREr and used on Western blot, Far-Western blot and ELISA. The results showed that PbUREr interferes on interaction between P. brasiliensis and pulmonary epithelial cells A549. PbUREr was able to bind to proteins fibronectin and type IV collagen, components of the extracellular matrix (ECM). The interaction between PbURE and calnexin protein was identified by the technique of two-hybrid.
Suscetibilidade in vitro e alterações morfologicas em células melanizadas e não melanizadas na presença de voriconazol, anfotericina B e extrato bruto da folha de P. pseudocaryophyllus (Gomes) L. R. Landrum
(Universidade Federal de Goiás, 2010-10-25) FERNANDES, Orionalda Fatima Lisboa; LINO JÚNIOR, Ruy de Souza; http://lattes.cnpq.br/0372118837748010; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725
Aspectos epidemiológicos e genótipos do vírus da Hepatite C em caminhoneiros de rota longa do Brasil
(Universidade Federal de Goiás, 2008-02-22) FREITAS, Nara Rubia de; MARTINS, Regina Maria Bringel; http://lattes.cnpq.br/2582896795892370
Globally, the prevalence of hepatitis C virus (HCV) infection is about 3%, with a significant variation according to geographic areas and population groups. However, studies concerning this infection in truck drivers population are rare. This study aimed to determine the HCV infection prevalence, to analyse associated risk factors and also to identify this virus genotypes in a population of long distance truck drivers in Brazil. In 2005-2006, 641 truck drivers who were circulating on BR-153 Federal road were interviewed and blood samples collected. Blood samples (sera) were tested for anti-HCV antibodies detection by enzyme linked immunosorbent assay. Anti-HCV reactive sera were retested for confirmation by immunoblot and also for HCV RNA detection by polymerase chain reaction. HCV RNA positive samples were genotyped by line probe assay (LiPA). Nine samples were anti-HCV positive, resulting in a prevalence of 1.4% (95% CI: 0.7-2.7). By multivariate analysis, use of illicit drug and hepatitis B virus seropositivity (anti-HBc marker) were risk factors for this infection. Genotyping of HCV RNA positive samples revealed the presence of genotypes 1 (37.5%), 2 (25.0%) and 3 (37.5%). The findings of this study indicate an intermediate endemicity for HCV infection in truck drivers in Brazil, the relevance of drug use in the transmission of this agent and the circulation of genotypes 1, 2 and 3 in the studied population.
Detecção de metalo beta lactamase em Pseudomonas aeruginosa isoladas de pacientes hospitalizados
(Universidade Federal de Goiás, 2009-02-18) GONÇALVES, Diana Christina Pereira Santos; PIMENTA, Fabiana Cristina; http://lattes.cnpq.br/2230554075502158
P. aeruginosa is frequently isolated in hospitals and the clinical importance has been increased due to gravity of infections. The metallo-beta-lactamase (MBL) production is an emergent mechanism of resistance in P. aeruginosa. The study aimed to determine the antimicrobial susceptibility profile of P. aeruginosa isolated of patients admitted in a hospital in Goiânia, to verify the MBL production by diffusion test and detect MBL genes by PCR technique. A total of 75 samples were evaluated, isolated of various clinical samples, in the period of January/2005 to January/2007. The biochemical identification was performed by automation technique system (API 20E ®) and antimicrobial susceptibility profile by Kirby- Bauer method. The 75 P. aeruginosa presented multi-drug resistance and, the resistance profile was: 90.7% to ceftazidime: 30.7% to aztreonam, 97.3% to ciprofloxacin; 48.0% of resistance to piperacilin/tazobactam, 88.0% to cefepime; amicacin, gentamicin and tobramicina whit resistance profile of 78.7%, 84.0% and 77.4%, respectively. The MBL production by difusion disc method was 46.7% (35/75). The gene blaSPM-1 was detected in 39 (52.0%) and gene blaIMP-1 in three (4.0%) isolates. The high frequency of P. aeruginosa resistant and MBL production alert to necessity of control the dissemination of bacteria multi-drug resistant in hospital, as well as the adoption of preventive actions and explanation of the health workers about rational use of antibiotics.
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas
(Universidade Federal de Goiás, 2010-03-11) NOGUEIRA, Sarah Veloso; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637
Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis, prevalent in Latin America. Extracellular matrix (ECM) is a complex net where collagens, laminin and fibronectin can be found and, when exposed, is the first site for the fungus adhesion. Our aim was to study genes involved in the adhesion process using Representational Difference Analysis (RDA). RDA is a PCR-coupled subtractive method that allows the isolation of genes differentially expressed in two different cDNA populations. Hence, cDNAs were synthesized from RNAs extracted from P. brasiliensis yeast cells adhered to collagen and fibronectin to identify overexpressed genes. Genes involved in a wide range of cellular process were found and PbCtr3 (cooper transporter) and enolase (PbEno) were chosen to further studies. A synthetic peptide (PbCTR3) and the recombinant enolase (rPbEno) were utilized together with the anti-rPbEno polyclonal antibody in functional analysis with ECM components and plasminogen. The studies suggest that P. brasiliensis enolase, in the surface, is able to generate plasmin from plasminogen by plasminogen activator. Therefore, it was also demonstrated that this protein is secreted and able to promote fungus adhesion and invasion to cells. These findings clearly establish the role of enolase in the patogenicity of P. brasiliensis.
Análise Morfométrica de Staphylococcus aureus Meticilina Resistentes Cultivados em Diferentes Concentrações de Cloreto de Sódio e Oxacilina
(Universidade Federal de Goiás, 2010-04-15) OLIVEIRA, Ana Cláudia Alves de; LINO JÚNIOR, Ruy de Souza; http://lattes.cnpq.br/0372118837748010
Methicillin-resistant Staphylococcus aureus (MRSA) has been one of the most prevalent microorganisms that cause hospitals infections worldwide. Several studies show that this microorganism is often found colonizing health professionals. MRSA can cause skin infections from the severe pneumonia, with high resistance to different antimicrobial agents. This study aimed to evaluate morphometric changes in MRSA isolated from the saliva of health professionals, through the use of different concentrations of oxacillin and sodium chloride. The identification of morphological changes was assessed by growing the isolates in the following concentrations of Sodium Chloride and oxacillin: 2μg, 4μg and 6μg and 2%, 4%, 6% and 7.5% and using the means of computerized morphometry. This technique was tested by microscopic computed by employing the software Image J 1.38 (HIH USA). Statistical analysis was performed using the Sigma Stat, version 2.03, and the differences between the groups were compared using the nonparametric Kruskal-Wallis (p <0.05). The 20 MRSA analyzed showed no alterations. The present study showed that Sodium Chloride and oxacilin at concentrations did not alter the development and morphology of MRSA.
Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
(Universidade Federal de Goiás, 2009-03-16) OLIVEIRA, Natalia Carvalhaes de; VIEIRA, José Daniel Gonçalves; http://lattes.cnpq.br/1742731776579730
Endophytic microorganisms live inside plants showing no apparently damage for the host, often assisting in survival of plants, helping its growth with production of phytohormones, phosphates solubilization, nitrogen fixation and enzymes production, or they can metabolize organic contaminants, like petroleum and derivates. This work aimed to isolated and identified endophytic microorganisms of plants present in impacted areas, as well as test their ability in petroleum and its derivatives degradation, identify bacteriocin production, to test their nitrogen fixation capability, phosphate solubilization, indol-acetic acid (IAA) and enzymes production. Plant samples were collected, in an area impacted with asphaltic and mud, were superficially disinfected using 70% ethanol, sodium hypochlorite and sterile distilled water. After macerated and fragmented, the samples were incubated at 30°C for about 72 hours, when growth of microorganisms was observed in culture media: Nutrient Agar, TSA (Tryptone Soya Agar) and King medium. The verification of petroleum and derivatives degradation capacity was performed in ELISA plates, exposing the bacteria to a solution of Minimal Medium, the dye DCPIP solution (2,6-dicloroindofenol sodium salt) and petroleum or derivative tested (burning oil, lubricating oil, diesel oil and gasoline). A positive reading for degradation was observed by discoloration of DCPIP. Among nine bacteria tested, three showed degradative activity in different fractions of petroleum, diesel oil and gasoline, and others showed different profiles. This result shows the potential of these microorganisms for application in bioremediation processes. The production of IAA was tested on TSA medium supplemented with tryptophan and only one sample was positive, while none was considered phosphate solubilizing. Testing the nitrogen fixing, samples were inoculated for a free nitrogen culture media, showing growth after 24 hours of incubation (positive results), however it s isolated did not show positive results in the acetylene reduction test, which confirms activity of the enzyme nitrogenase. The samples showed different profiles for the production and activity of bacteriocins against the clinical isolates tested. There were no detected activity of endoglucanase, cellulolytic and pectinolytic enzymes, but esterases, ß-glucosidases, proteases and amylases were positive. Some isolates were identified from the sequencing of 16S DNA, whose sequence was compared to database (GenBank), and then diagnosed as Bacillus cereus, Staphylococcus pasteuri and Pseudomonas sp. The endophytic bacteria isolated from Cerrado plants confirmed their potential for use in bioremediation, plant growth promotion and diverse enzyme production, with potential for use in pharmaceutical, food, textiles, among others.
Prevalência de tipos específicos de Papilomavírus humano (HPV) e relação com a severidade da lesão cervical em mulheres com exame citopatológico anormal
(Universidade Federal de Goiás, 2009-12-15) RIBEIRO, Andrea Alves; SANTOS, Sílvia Helena Rabelo dos; http://lattes.cnpq.br/4994826511439492
Human papillomavirus (HPV) is considered the central etiological agent involved in the genesis of cervical cancer. The HPV viruses are classified according to their biological niche, oncogenic potential and phylogenetic position. According to the criteria established by the International Committee on Taxonomy of Viruses (ICTV), the various groups of human papillomaviruses that infect the female genital tract are classified phylogenetically in the Alphapapillomavirus genus, including species classified among phylogenetic species 1 and species 15. The main high risk HPV are classified in species 9 (HPV 16, 31, 33, 35, 52, 58, 67), and in species 7 (18, 39, 45, 59, 56, 66, 68 and 70). HPV 16 is the most prevalent type irrespective of diagnosis, principally in more severe lesions. Coinfection with multiple-types HPV is a common finding of many molecular studies. Some HPV types might interact or act synergistically to induce progression. Few studies have investigated the interactions of viral genotypes or species in multiple-type HPV infections. Therefore, the objective of this study was to evaluate the effect of single or multiple-types HPV infections considering also the phylogenetic groups on the prevalence and severity of cervical intraepithelial neoplasia (CIN) among women undergoing colposcopy following a abnormal cervical smear. Methodology: In this analysis, 198 women attending at the colposcopic clinic, because of an abnormal cervical smear were included. Colposcopy was carried out in all cases and biopsies were done in 193 of 198 women included. All specimens were tested for 27 HPV genotypes by Roche s polymerase chain reaction reverse line blot assay. Results: The overall prevalence of HPV in women with an abnormal cervical smear was 86% (171/198). Of the total of HPV-positive women, 45% (77/171) were infected with HPV 16 as a single or multiple-type infections. HPV 31 and 35 were, respectively, the second and third most prevalent types. The prevalence of HPV 16 in high grade cervical intraepithelial neoplasia (CIN2/3) was 52% (40/76) and it was detected in 88.8% (8/9) in cases of invasive carcinoma. The prevalence of type 31 and 35 in high grade CIN was respectively 10.5% (8/76) and 6.6% (5/76). Single HPV infection for any type was significantly associated with neoplastic diagnosis. High grade neoplastic diagnosis (≥ CIN2) was significantly associated with HPV 16 in single or multiple infections. Also, there was significantly association between HPV 16 and others types of specie 9 and high grade neoplastic diagnosis, but no association was observed considering the HPV 16 and other of groups of species 7 or others types. Conclusion: These results indicated that the type 16 is the most important predictor of high grade cervical neoplasia. Multiple-type infections are predictors of high grade cervical neoplasia when the type 16 is present.
Tipagem molecular de Streptococcus pneumoniae isolados da nasofaringe de crianças no contexto da vacinação pneumocócica
(Universidade Federal de Goiás, 2010-02-18) ROCHA, Cristyane Gonçalves Benicio Bastos; ANDRADE, Ana Lúcia Sampaio Sgambatti de; http://lattes.cnpq.br/7770363683068899; PIMENTA, Fabiana Cristina; http://lattes.cnpq.br/2230554075502158
Objectives (i) Present review article focusing on pneumococcal vaccines and carriage; (ii) to validate sequential multiplex PCR for identifying pneumococcal capsular serotypes from children attending day-care centers; (iii) determine the multilocus sequence typing; (iv) to identify the capsular types of multiple colonies of S. pneumoniae isolates from a single sample of nasopharyngeal secretions of children attending day-care centers in Goiânia. Materials and Methods S. pneumoniae was obtained from health children less than 5 years old attending 62 day care centers of Goiânia. The laboratory procedures were performed according to WHO recommendations. Were selected 217 isolates (penicillin resistant and sensitive) for capsular typing by multiplex PCR technique. MLST was performed for 55 isolates representing the serotypes detected and the different susceptibility patterns for penicillin. Quellung reaction was used for typing isolates serotypes 6A, 6B, 18C and the isolates not typed by multiplex PCR. For 28 presumptive pneumococcal positive NP swabs, 3 colonies were picked to acess possible serotype diversity. Eighty four pneumococci were identified by conventionally procedure and multiplex PCR was performed. Results Serotypes were deduced for 177/217 (81.6%) of the pneumococcal. The most frequent serogroups/serotypes were 14, 6, 23F, 19F and 18. Multiple serotypes were detected in 13 specimens. Were found 19 MLST types and two new ST. Forty (18.4%) were not serotyped by the multiplex PCR and quellung reaction. The analysis of three colonies from the same NP permitted the detection of differente serotypes in 7/28 (25%) NP samples. Conclusion (i) The multiplex PCR is simple and cost-effective method for detecting multiple serotypes in nasopharyngeal isolates; (ii) and thus might be useful for the monitoring of pneumococcal colonization over time; (iii) the use of multiplex PCR can further broaden our understanding of the dynamics of pneumococcal carriage, including multiple serotypes, the effect of vaccination on carriage, and transmission, as well as surveillance of IPD and co-colonization.
DETECÇÃO DO GENE mecA EM ESTAFILOCOCOS COAGULASE NEGATIVA RESISTENTES A OXACILINA ISOLADOS DA SALIVA DE PROFISSIONAIS DA SAÚDE DE UM HOSPITAL UNIVERSITÁRIO
(Universidade Federal de Goiás, 2008-04-17) ROSA, Juliana de Oliveira; REIS, Cleomenes; http://lattes.cnpq.br/5103786277121402; PIMENTA, Fabiana Cristina; http://lattes.cnpq.br/2230554075502158
Coagulase negative staphylococci (CNS) is in human and animal microbiota, but may cause with infections with significant morbidity and mortality rates. Healthy care workers may be carriers of many microorganisms and spread resistant ECN in the hospital. This study aimed to identify the CNS species isolated from healthy care workers saliva, establishe the oxacillin resistance pattern and detect the mecA gene in resistant isolates. We evaluated 100 ECN, isolated from the saliva of an institution of professional health of large Riberão Preto in Sao Paulo state. The ECN identification was based on biochemical tests, and 41 were identified as S. epidermidis, 25 S. saprophyticus, 18 S. haemolyticus, 8 S. cohnii, 4 S. lugdunenses, 3 S. capitis, and 1 S. simulans. Thirty-two percent were nonsusseptible to oxacillin, 84.4% to mupirocin, 43.7% to cefoxitin, but all were vancomycin susceptible. The oxacillin nonsusseptible ECN, detected by disk diffusion test were grown in agar screening oxacillin (6 μ g) supplemented with sodium chloride (4.0%) and submited to mecA detection by the PCR. Of the 32 nonsusseptible oxacillin CNS,, 93.7% developed in the oxacillin agar and the mecA gene was detected in 75.0%. This is the first report of mecA gene presence in CNS isolated from the saliva of healthy care workers. Attention must be given to CNS species identification, as well as the characterization of the nonsusceptible microorganisms, since healthy care workers may represent a reservoir of CNS
Caracterização molecular de Mycobacterium tuberculosis isolados de pacientes atendidos na cidade de Goiânia-GO, pela técnica de RFLP-IS6110
(Universidade Federal de Goiás, 2008-02-29) SANTOS, Lorena Cristina; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; KIPNIS, André; http://lattes.cnpq.br/4434965360286741
Tuberculosis is a serious public health problem all over the world. It has been demonstrated that different M. tuberculosis strains, characterized by the RFLP-IS6110 standard technique, have different virulence properties and antibiotic resistance. The aim of this study was to characterize M. tuberculosis strains isolated from patients attending two state reference hospitals of Goiânia-Goiás, using the RFLP-IS6110 technique. Positive cultures of M. tuberculosis sampled and isolated from January 2006 to June 2007 had their DNA extracted. A total of 142 viable DNA samples were analyzed, of which 126 samples presented an RFLP-IS6110 profile. Similarities comparisons between samples, as well as with literature reported profiles, were done with Bionumerics software (version 4.0). Forty three percent of the samples could be grouped in 24 clusters, when analyzed by the RFLP method, suggesting recent transmission among individuals belonging to the same cluster, however those patients did not present any epidemiological relationship, suggesting possible casual transmission between members of the same cluster. When compared with strain profile of the MDR-TB, three samples had grouped in the clade formed by families virulent. This study strengthens the importance of determining transmission routes not only within the state of Goiás but in the entire country, since there is the possibility of resistant strains transmissions.
Caracterização e identificação de leveduras do gênero Candida em pacientes transplantados de medula óssea
(Universidade Federal de Goiás, 2011-08-31) SILVA, Hildene Meneses e; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725
Fungal infections in immunocompromised patients, especially in hospitalized individuals, which are submitted to differente types of treatment have increased in recent decades and are cause of death of these patients. The diagnoses of infection as well as the correct identification of the fungus, in addition to the in vitro susceptibility tests are important to control the disease and thus avoid death. This work was conducted to identify the species that causes candidemia in patients with hematologic malignancies and in those submitted to bone marrow transplantation from Araujo Jorge Hospital in the city of Goiânia. The main predisposing factors for the acquisition of candidiasis, the virulence factors inherent to the microorganism, as well as the in vitro susceptibility to different antifungal agents of Candida isolates were verified in this work. Clinical samples from patients were collected and cultived in brain heart infusion and Sabouraud agar dextrose and incubated at room temperature and at 36ºC. The identification of yeasts was performed by culture in cornmeal agar to check the chlamydospore production, by germ tube production in fetal bovine serum plus 1% tween 80, by inoculation onto chromogenic agar, CHROMagar and methods of assimilation of carbohydrates. The isolates were subjected to hemolytic activity and proteinase testing as well as to in vitro susceptibility testing using broth microdilution and Etest methods. Of the 410 clinical samples originated from 144 transplant patients we identified eleven yeasts, included three (3) Candida albicans, five (5) C. parapsilosis, one (1) C. tropicalis and two (2) Candida spp. All isolates of C. parapsilosis were susceptible to all antifungal agents, while two (2) isolates of C. albicans and one (1) C. tropicalis were resistant to three azole derivatives, fluconazole, itraconazole and voriconazole. Candida spp isolates did not show hemolytic activity, they were not able to secrete the enzyme proteinase either, they were resistant to fluconazole, itraconazole and amphotericin B and susceptible to caspofungin and voriconazole. In this study we can conclude that C. parapsilosis was prevalent, showing that the emergence of Candida non albicans. Besides we can conclude that in vitro susceptibility tests are of great importance for the adequate therapy to avoid the death of patients.
CARACTERIZAÇÃO DA ISOCITRATO LIASE E METILISOCITRATO LIASE DO FUNGO PATOGÊNICO HUMANO Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2009-02-26) TROIAN, Rogério Fraga; PEREIRA, Maristela; http://lattes.cnpq.br/1345781867765758
Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of paracoccidioidomycosis (PCM), a prevalent systemic mycosis in Latin America. Pathogenicity appears to be intimately related to the dimorphic transition from the hyphal to the yeast form, which is induced by a shift from environmental temperature to the temperature of the mammalian host. To cope with nutrient deprivation during the infection process, a number of pathogens employ the glyoxylate cycle (GC) to utilize fatty acids as carbon sources. The genes which constitute this pathway have been implicated in pathogenesis. An important aspect in the interaction between P.brasiliensis and your host is the ability to adhere to matrix extracelular components. In this work has shown that the isocitrate lyase of P. brasiliensis (PbICL) is located in the cell wall and also in the cytoplasm. PbICL recombinant and polyclonal antibody were able to inhibit the interaction of P. brasiliensis to epithelial cell cultures in vitro. Was also evaluated the ability of PbICL recombinant to connect the components of the extracellular matrix such as laminin, fibronectin, collagen type I and IV. These results suggest that PbICL is necessary to interaction between molecules of the extracellular matrix and P.brasiliensis, and that this interaction is crucial in adhesion and invasion of the fungus to the host cells. The kinetic parameters of PbICLr were determined. The sequence coding for methylisocitrate lyase of P. brasiliensis (PbmeICL) and the recombinant protein PbmeICLr were obtained.

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