Doutorado em Medicina Tropical e Saúde Pública (IPTSP)
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Item Caracterização de microrganismos isolados em manipuladores e dietas enterais de dois hospitais públicos de Goiânia(Universidade Federal de Goiás, 2010-03-19) BORGES, Liana Jayme; ANDRÉ, Maria Cláudia Dantas Porfirio Borges; http://lattes.cnpq.br/1475834090578722; SERAFINI, álvaro Bisol; http://lattes.cnpq.br/9849440763539845Enteral feeding means the nutrition for special purposes, with controlled intake of nutrients. The advantages of its use often become secondary to complications arising from its contamination, which may be associated with infectious complications. The microbial contamination of enteral feeding may occur during all steps being the handling, particularly critical. Considering the importance of enteral feeding as a therapeutic tool in hospitals and the need to guarantee the microbiological quality of the products offered to critical patients, the present work aimed to evaluate the hygienic and sanitary quality of diets and their ingredients and to identify and characterize phenotypic and genotypically, using the antibiogram and pulsed-field gel electrophoresis, strains of Escherichia coli and Staphylococcus aureus obtained from handlers hands and noses, water, module and enteral nutrition from two public hospital in Goiânia, Brazil in order to investigate the probable source of microbological contamination. A total of 80 samples were collected from enteral nutrition and 140 from hands and noses of handlers involved in the diets manufacturing in hospital 1 (H1), between october/2007 and november/2008 and 80 samples from enteral nutrition and 80 from hands and noses of handlers in hospital 2 (H2), between october/2008 and november/2008. From both hospitals were collected 40 samples from water and module. The samples were submitted to microbiological analysis to verify the presence and numbers of pathogenic and indicator microorganisms. E. coli and S. aureus strains were submitted to antibiogram and PFGE. According to antibiogram, all S.aureus isolates (15) from H1 were susceptible to oxacillin, vancomycin, ciprofloxacin and gentamicin. Resistence profile was observed in 10 (66.7%) isolates for penicillin, four (26.7%) isolates for tetracycline and nine (60.0%) isolates for erythromycin, allowing to classify the strains in six different phenotypes (A-F), but it was not efficient for the determination of the bacterial source for the diets. In the H1, all (08) E. coli strains were susceptible to trimethoprim, ciprofloxacin, cephalothin, gentamicin, ceftazidime and tetracycline. Resistence was observed in six (75.0%) isolates for ampicilin. In H2, all strains isolated (12) were susceptible to trimethoprim, ciprofloxacin, gentamicin and ceftazidime and resistence was observed in 11 isolates (91.7%) for cephalothin and 12 (100.0%) for tetracycline and ampicillin, grouping them into five different phenotypes (A-D). Microorganisms showed the same phenotypic profile from handlers and diet samples (phenotypes A and C), suggesting that in these cases, the source of microorganisms for the final product was the food handler. The genotypic typing of S. aureus strains by PFGE generated seven different DNA banding profiles and the E. coli genotyping generated five profiles. Based on the results, two E. coli strains isolated from diets were identical to one strain isolated from food handler from H2 and two of S. aureus isolated from diets were identical to one strain isolated from food handler from H1. This study shows that the enteral feedings showed unsatisfactory sanitary-hygienic conditions in both hospitals and the hand contact is probably one of the sources of greatest significance for enteral diets contamination in the hospital environment.Item Estudo epidemiológico e molecular da infecção pelo vírus da hepatite B em Afro-descendentes de comunidade isolada no Estado de Goiás (Kalungas)(Universidade Federal de Goiás, 2007-12-18) MATOS, Márcia Alves Dias de; MARTINS, Regina Maria Bringel; http://lattes.cnpq.br/2582896795892370Hepatitis B virus (HBV) infection occurs throughout the world. In Africa, this infection is highly endemic, with the majority of individuals becoming infected during childhood. Although Brazil has been globally considered a country of HBV intermediate endemicity, variable rates have been found in all five Brazilian regions and even inside the same region. This study aimed to investigate the epidemiological and molecular profile of the HBV infection among the Kalunga population in Goiás, Central Brazil, which is considered the largest Afro-Brazilian isolated community. A total of 878 individuals were interviewed about sociodemographic characteristics, risk factors and HBV vaccination. Blood samples were collected from all participants and serum samples were screened by enzyme-linked immunosorbent assay for the presence of HBsAg, anti-HBc and anti-HBs serological markers. HBsAg-positive samples were submitted to HBeAg and anti-HBe detection. HBsAg and anti-HBc positive samples were tested for HBV DNA detection by polymerase chain reaction and genotyping by subsequent restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing of preS/S region. The overall prevalence of HBV infection was 35.4% (95% CI: 32.3-38.7). HBsAg carrier rate was 1.8% (95% CI: 1.1- 3.0). Multivariate analysis of risk factors showed that increased age, male gender, illiteracy and history of multiple sexual partners were associated with this infection. Isolated anti-HBs was found in 301 (34.3%) individuals who were immune for hepatitis B. HBV DNA was detected in 75% (12/16) of the HBsAg positive samples, in 100% (2/2) of the HBeAg and in 83.3% (10/12) of the anti-HBe positive samples. An occult HBV infection rate of 1.7% (5/295) was found among anti-HBc positive individuals. All genotyped isolates belonged to genotype A by RFLP analysis. Nucleotide sequencing of preS/S region confirmed the circulation of genotype A (subgenotype Aa) in this community. The epidemiological findings indicate that preventive measures, such as additional health education and HBV vaccination programs, are needed to control HBV infection in this population. In addition, the molecular results suggest the introduction of genotype A, subgenotype Aa in Brazil from Africa during the slave trade.Item Contribuição ao estudo da doença de Chagas(Universidade Federal de Goiás, 2011-07-18) OSTERMAYER, Alejandro Luquetti; NETTO, Joaquim Caetano de Almeida; http://lattes.cnpq.br/3444498706763045The main papers published on the last five years on the area of human Chagas disease, mainly on parasitological, serological and therapeutic aspects, reflecting areas of recent involvment of the author, were selected. As a baseline, briefly comments on publications on the last 40 years were included. Parasitemia profile measured by hemoculture during the chronic infection was the subject of the first two. The first (published in 2006) included six hemocultures from each of 27 patients, the last three after specific treatment. Results were compared with those of 13 non treated, infected patients. The supressive effect of benznidazole was demonstrated in 89% of the patients and treatment failure was registered in three cases (11%) during the two year follow-up. In the second paper (2011) a single hemoculture was performed in 152 infected women, 101 pregnant. Parasitemic pregnant women doubled the number of non-pregnant, mainly during the first months of pregnancy. A new ELISA test was developed (2010) by employing two recombinant proteins and two synthetic peptides. Sensitivity was 99.3% on 165 positive sera, and specificity of 100% (216 negatives). A multicentric study was done (2009) with participation of laboratories of North America (Mexico), Central America (Honduras) and South America (Brazil) with 98 serum samples from patients of Mexico employing reagents made from Tc1 and Tc2 strains. Results showed good agreement among laboratories demonstrating the feasibility of using reagents prepared from both types of T. cruzi. In another multicentric study (Bolivia, Argentina and Brazil) xenodiagnosis performed in 17 patients after a 60 days course of allopurinol treatment, remained positive, showing the lack of effect of this drug in the chronic phase. A national serological survey in children born after insecticide spraying (below five years old) with filter paper, involved 104,954 samples, tested with ELISA and indirect immuofluorescence. Only 11 samples (0.01%) were identified as by vector transmission (negative mothers) mainly from the northeast of the country, areas without Triatoma infestans. These findings (2011) confirm the effectivity of house spraying for the control of the disease. As a sub-product of this investigation, an unusual number of cases of congenital transmission (n = 12) was found only in the State of Rio Grande do Sul, where TcV-TcVI circulates. When compared with other states (n=8) the proportion was 10 times higher. This is the first report on geographical differences related to congenital transmission in Brazil. We considered that all these findings contributed significantly to a better knowledge on different aspects of Chagas disease.Item Avaliação dos monócitos na Leishmaniose Tegumentar Americana(Universidade Federal de Goiás, 2012-08-31) PEREIRA, Ledice Inacia de Araujo; DIAS, Fátima Ribeiro; http://lattes.cnpq.br/5741031258926403American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan that infects mononuclear phagocytic cells leading to cutaneous or mucosal lesions. The mechanisms responsible for parasite control or persistence are not completely known. Human monocytes are blood cells subdivided into three subsets (classical, nonclassical and intermediate) according to the CD14 and CD16 expression that is associated with different phenotypical and functional characteristics. The aim of this study was to evaluate the profile of monocytes in ATL. First, it was analyzed the nonclassical monocytes (CD14loCD16+) and CD56+CD16+ NK cell frequencies in peripheral blood (by using flow cytometry) of a patient with diffuse cutaneous leishmaniasis (DCL) before, during and after immunochemotherapy (chemotherapy treatment together with L. (L.) amazonensis and L. (V.) braziliensis monovalent vaccines plus BCG). Then, in localized cutaneous leishmaniasis (LCL) patients (n = 32) and healthy donors, the three monocyte subsets (CD14hiCD16-, CD14hiCD16+, CD14loCD16+) were evaluated by flow cytometry; in the whole blood cultures we evaluated the expression of cytokines in CD14+ monocytes activated with lipopolysaccharide (LPS, by flow cytometry), and the secretion of proinflammatory (tumor necrosis factor, TNF) and antiinflammatory (interleukin 10, IL-10) after activation with Toll-like receptor agonists (TLR2 (Pam3Cys), TLR4 (LPS)) or L. (V.) braziliensis antigen (Ag; ELISA was used). In DCL patient it was detected an increase in non classical monocytes and NK cell frequencies probably associated to BCG stimulation, contributing to the clinical cure of several lesions caused by L. (L.) amazonensis. In LCL patients, the CD16+ monocyte subsets were increased before treatment. A similar TNF and IL-10 expression in CD14+ monocytes activated with LPS was found in patients and controls. It was not possible to identify which monocyte subpopulation was the responsible for the TNF or IL-10 production due to alterations of the percentages of each subset after LPS stimulation. Activation through TLR2, TLR4 or with Ag leads to a higher production of TNF but not IL-10 in whole blood cultures of patients than of controls. Whereas in healthy controls there was a positive correlation between TNF and IL-10 production after different stimulations (LPS, Pam3Cys and Ag), this was not observed in LCL patients, except for TLR2 stimulation. A positive correlation was detected between amount of TNF in serum or in activated-whole blood cultures and the number of cutaneous lesions. These results suggested that nonclassical monocytes can contribute to the control of infection in DCL patient, a clinical form in which patients do not present acquired cellular immune response. In this case, activation of innate cells as monocytes can cause lesion regression decreasing costs with medicines and improving the life quality of the patients. On the other hand, in LCL patients monocyte subsets and cytokine imbalance, especially with increase of nonclassical and intermediate monocytes (CD14loCD16+, CD14hiCD16+) and TNF, can contribute to leishmaniasis immunopathogenesis. To understand the role of monocyte subsets in ATL can open new avenues to the identification of biological markers of disease severity as well as new therapeutic targets to ATL treatment.Item Estudo sobre contatos de pacientes com tuberculose pulmonar na região metropolitana de Goiânia(Universidade Federal de Goiás, 2011-07-07) REIS, Michelle Cristina Guerreiro dos; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987Tuberculosis (TB) affects millions of people every year and it is estimated that one third of world‟s population is infected with Mycobacterium tuberculosis (Mtb). The identification of infection and monitoring of close contacts of TB patients would contribute to disease dissemination control. In the present study the epidemiological characteristics that may facilitate contact‟s infection were evaluated. Additionally, the antibody profile against Hspx from Mtb, among individuals that were in frequent contact of pulmonary tuberculosis (PTB) patients, was investigated as a marker for risk of active disease development. All participants were submitted to tuberculin skin test (TST) and their blood were harvested to determine the antibody profile (IgM/IgG - ELISA). Forty-five PTB patients and 177 contacts were followed for one year. During the follow-up period, only daily contact with patient was associated with positive TST at the moment of diagnosis. Two TST positive contacts developed active PTB, eight contacts converted their TST to positive, 6 individuals presented the booster effect, 44 remained TST negative and 20 dropped out of the study. The IgM-Hspx levels were 0,841±0,40 for TST positive contacts, 0,807±0,32 for TST negative, 0,732±0,218 for TST converted and 0,961±0,48 to contacts that boosted their TST. The IgG-Hspx levels were 0,242±0,10 for TST positive contacts, 0,237±0,10 for TST negative, 0,140±0,02 for TST converted and 0,255±0,22 for contacts that boosted their TST, with significant statistical difference (p=0,019). Our data show that daily contact is associated with TST positivity and the risk of disease development and antibodies against Hspx (both, IgM and IgG) were not associated among recent latently infected contacts of PTB patients.Item Análise de resistência a antimicrobianos de cepas de Mycobacterium tuberculosis isoladas no Estado de Goiás(Universidade Federal de Goiás, 2010-12-07) SANTOS, Lorena Cristina; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; KIPNIS, André; http://lattes.cnpq.br/4434965360286741Tuberculosis (TB) is a serious global public health. In Brazil for the confirmed TB cases is recommended a multi-drug therapy regimen which combines different drugs during at least 6 month. However, because of treatment inconsistency, the emergency and spread of drug resistant M. tuberculosis become a serious threat. Actually, strains resistant to at least one drug used in the TB treatment have been one of the main factor that avoid the effective TB control. According to WHO M. tuberculosis strains that are resistant to at least INH and RMP, the key drugs used in the TB treatment, are considered multidrug resistant (MDRTB). The main mutations responsible for INH and RMP resistance occur at some specific regions in the katG, inhA and rpoB genes. We analyzed by phenotypic and genotypic methods the susceptibility profile of M. tuberculosis isolated from 132 patients treated at a reference hospital in Goiânia-Goiás, between January of 2006 and July of 2007 and then performed the resistant strains genotypic identifications by RFLP-IS6110. Additionally, clinical and epidemiological informations from the patients was collected. A high frequency of drug resistance was observed in previously untreated patients (13.6% to at least one antibiotic and 6.1% MDR-TB), and a high DNA polymorphism was observed among these strains. Our results suggest that the prevalence of resistant TB in Goiás is underestimated and that resistance in new TB cases was not associated with an outbreak in this region.Item Caracterização dos genes de NSP4 e VP6 de amostras de rotavírus do grupo A provenientes de crianças da região Centro-Oeste do Brasil(Universidade Federal de Goiás, 2008-04-28) TAVARES, Talissa de Moraes; BRITO, Wilia Marta Elsner Diederichsen de; http://lattes.cnpq.br/7605775995731168; CARDOSO, Divina das Dôres de Paula; http://lattes.cnpq.br/9770835116155857Group A rotaviruses are the major cause of gastroenteritis in children throughout the world. Epidemiological surveys and molecular analysis of rotavirus strains are required for gastroenteritis control and prevention. Studies using VP6, an important immunogenic structural protein, and NSP4, a transmembrane nonstructural glycoprotein which is critical to rotavirus morphogenesis and pathogenesis, have been performed. In this study, 330 rotavirus-positive fecal samples previously obtained from children with or without diarrhea, between 1987 and 2003, in three cities of Central West Region of Brazil (Goiânia, Brasília and Campo Grande), were characterized for VP6- and NSP4-encoding genes. The VP6 and NSP4 genes were amplified by reverse transcription- polymerase chain reaction followed by sequencing and phylogenetic analysis. Detection rates of 84.8% and 78.5% were observed for VP6 and NSP4 genes, respectively. Two distinct genotypes could be recognized for NSP4 (A and B). It was observed that the G9P[6] samples were associated with genotype A, whereas the G1P[6], G1P[8], G2P[8], G3P[8], G4P[8] and G9P[8] samples were associated with genotype B. The analysis of VP6 gene allowed genogrouping of samples in two clusters, genogroups I and II. The G2P[4], G3P[4] and G9P[6] samples were identified as genogroup I, whereas the G1P[6], G1P[8], G2P[8], G4P[6], G4P[8] and G9P[8] samples were identified as genogroup II. In addition, it was showed that samples identified as VP6 genogroup I were associated with NSP4 genotype A and samples identified as VP6 genogroup II were associated with NSP4 genotype B. This investigation described different genetic groups representing diversity of group A rotavirus samples circulating in the Central West Region of Brazil.