Doutorado em Biologia da Relação Parasito-Hospedeiro (IPTSP)

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    Análise da proliferação de amastigotas de Leishmania (Viannia) braziliensis em macrófagos murinos
    (Universidade Federal de Goiás, 2021-09-30) Teixeira, Mirian Vieira; Oliveira, Milton Adriano Pelli de;; Oliveira, Milton Adriano Pelli de; Vinaud, Marina Clare; Afonso, Luís Carlos Crocco; Gomes, Clayson Moura; Baeza, Lilian Cristiane
    Leishmania (Viannia) braziliensis is the main species responsible for American tegumentary leishmaniasis in Brazil. However, the use of this parasite species to study Leishmania infection in a murine model has been less conducted when compared to other Leishmania species. Control of murine Leishmania infection has been associated with nitric oxide (NO) produced by inducible NO synthase (iNOS) from M1 macrophage, while arginase expressed by M2 macrophages is related to Leishmania proliferation. The aim of this study was to analyze the ability of L. (V.) braziliensis to proliferate within murine macrophages in vitro for a period of 9 days. Macropha-ges were derived from bone marrow precursors (BMDM) of wild-type mice and were cultured with IFN-γ and LPS, or IL-4, or BMDM iNOS knockout (iNOS KO), and nitric oxide production, arginase activity, and infection with L. (V.) braziliensis. The number of infected macrophages and parasite load were determined by light microscopy. Promastigotes of L. (V.) braziliensis parasites were inoculated (106) into the paw of wild-type and iNOS-deficient mice and lesion progression was measured weekly. Wild-type BMDM were observed to not support proliferation of amastigotes of L. (V.) braziliensis strains after day 3 infection, even within IL-4-treated ma-crophages or iNOS KO macrophages. Arginase activity was higher in iNOS KO macrophages than in IL-4 treated macrophages, showing that the absence of proliferation is arginase inde-pendent. L. (V.) braziliensis was able to cause uncontrolled disease in iNOS KO mice in vivo. The data obtained suggest that murine macrophages do not support proliferation of L. (V.) braziliensis amastigotes, even in the absence of nitric oxide and presence of high arginase ac-tivity. Therefore, further studies related to the requirements of amastigotes internalized in host cells are needed, for the search of better methods to interfere in the diversity of leishmaniasis forms caused by different Leishmania spp.
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    Análise dos genes de virulência cagA, vacA e dupA de Helicobacter pylori e associação com doenças gástrica
    (Universidade Federal de Goiás, 2021-06-07) Silva, Lucas Luiz de Lima; Carneiro, Lilian Carla;; Carneiro, Lilian Carla; Silva, Antonio Márcio Teodoro Cordeiro; Gama, Aline Rodrigues; Moraes Filho, Aroldo Vieira de; Santos, Mônica de Oliveira
    Helicobacter pylori (H. pylori) is a Gram-negative bacterium associated with the development of different gastropathies. The different clinical outcomes of infection by H. pylori are the result of the parasite-host relationship, and the virulence factors produced by the different strains of H. pylori are determinant in the pathogenesis of the infection. Based on the importance of the topic, the aim of this study was to assess the prevalence of H. pylori infection; compare the Polymerase Chain Reaction (PCR) technique with the histopathological examination for the detection of the microorganism; associate the genotypes of cagA, vacA and dupA virulence with the gastropathies presented by the patients and evaluate the origin of the local circulating strains. For this, gastric tissue biopsies from 117 dyspeptic patients were collected and used for DNA extraction. The histopathological examination report of the study patients was also obtained. The screening for H. pylori infection was performed using the 16S rRNA gene (hpx) and the positive H. pylori samples were subjected to the detection and sequencing of the virulence genes vacA, cagA and dupA. The dupA gene sequences obtained from the PCR were used for phylogenetic analysis. The prevalence of H. pylori infection found in this study was 64.1%, when considering only the PCR technique and 72.3% when associated with histopathological examination. A high frequency of positive cagA strains was found (80.0%). The cagA gene was detected in all strains present in patients with severe pathologies, while the isolated vacA gene was not detected in this group. Seriously ill patients had fewer virulence genes in the infecting strain. The presence of the dupA gene in the infecting strain was not associated as a risk factor or a protective factor for gastropathies, however, in women the infection by H. pylori dupA positive, increased the chance of developing gastritis by two. In addition, molecular phylogeny demonstrated that the dupA gene isolated in this study showed homology with genes from H. pylori strains isolated from western countries. The circulating strains of H. pylori in central-western Brazil show high heterogeneity in the frequency of cagA, vacA and dupA virulence genes and the combination of several virulence factors can influence the clinical outcomes of patients.
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    Identificação de bactérias ambientais obtidas em amostras de água em comunidades quilombolas de Goiás
    (Universidade Federal de Goiás, 2020-03-17) Gama, Aline Rodrigues; Bataus, Luiz Artur Mendes;; Carneiro, Lilian Carla;; Vieira, José Daniel Gonçalves; Santos, Mônica de Oliveira; Galvão Filho, Arlindo Rodrigues; Moreira, Ellen Flávia
    Water quality has already been related to only aesthetic and sensory aspects, such as color, odor and taste. Today, in response to advances in understanding the relationship between contaminated water and disease, a series of regulations are proposed by the World Health Organization and the Ministry of Health that regulate the potability of water for human consumption. Based on studies and surveys on the way of life of two quilombola communities located in Aparecida de Goiânia and Silvânia, in the State of Goiás, the present work proposed the verification of the microbiological, physicochemical and toxicological quality of the water consumed by the residents of these communities. For the analyzes, samples of water from artesian wells and cisterns that supply the residences were collected and used for consumption in the Quilombola communities mentioned. A physical-chemical analysis was carried out by means of pH verification, alkalinity, hardness, physicality, turbidity, nitrate concentration and potentially toxic metals. The process of demonstration for a microbiological analysis began with the detection of coliforms followed by iso-regulation and phenotypic identification of isolated colonies, with the final recognition of two genera / species, which were subsequently submerged in the analysis of antibiotic resistance profile. The DNA of two bacterial isolates has been extracted and used for amplification and sequencing of the 16S rRNA gene. A comparison between the results obtained from the phenotypic identification and the ones based on the genomic sequence was carried out. In addition, at hyperspectral image (HSI) two isolates were obtained in camara SWIR, for classification, based on spectral signature. The results show that, as we have obtained, the Quilombola community of Aparecida de Goiânia has 16 bacterial isolates, considering that Klebsiella pneumoniae was more frequent (four isolates) with the collected points; followed by Escherichia coli (three isolated). As we have collected from the community of Silvânia, we show that there are no significant values for physicochemical analyzes and potentially toxic metals. The microbiological data showed 32 isolates, weaning, Acinetobacter lwoffii was a species more frequent (nine isolates), followed by Staphylococcus auricularis (seven isolates), with a large number of bacteria showing resistance to multiple antibiotics. Expressive were the differences generated between a phenotypic identification and genomic sequences; less than 20% two isolates achieved gender level agreement. It was possible, by means of HSI analyzes, to separate the bacterial isolates in different groups, and is not a group of isolates, in genus and species. Based on the technique used, there were no significant values for the physical-chemical and potentially toxic metal analyzes. Based on two observed results, it becomes important to perform actions, by the public power, that aim to improve water quality and in consequence of the life quality of people that live in studied communities. In addition, the use of appropriate methodologies for identifying environmentallybased bacteria, not being confidential or using phenotypic characteristics, is highlighted. Through the need for bacterial identification methodologies, it is considered that or use of HSI for this purpose, it will briefly be a common practice, for the construction of more robust data banks.
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    Situação atual de mães cronicamente infectadas pelo tripanosoma cruzi no estado de Goiás e triagem sorológica para infecção congênita em recém-nascidos pelo teste do pezinho na região metropolitana de Goiânia
    (Universidade Federal de Goiás, 2016-07-28) Gomes, Taynara Cristina; Castro, Ana Maria de;; Castro, Ana Maria de; Soares, Joanna D’arc Aparecida Herzog; Alves, Daniela de Sousa Mendes Moreira
    The congenital transmission is considered one of the main pathways of Trypanosoma cruzi infection. Retrospective studies of pregnant women population with Chagas disease (CD) allow the study of the epidemiological profile of this population which contributes to the knowledge of the congenital CD prevalence in Brazil. This study evaluated the triage of T. cruzi congenital infection in new-borns through the newborn screening aiming the precocious detection of congenital transmission as well as its retrospective analysis of the seroepidemiological profile of the infected pregnant women dwelling in Goias State in a period of three years (2013-2015). The retrospective study used the data base of the Association of Parents and Friends of Special Needs Individuals (APAE) of Goiania. The analysis showed that the prevalence of CD infected pregnant women from 0.24% to 0.17%, with a decrease throughout the years. Most of these pregnant women had ≥31 years old, which is in accordance with the success of Triatoma infestans vectorial transmission control and transfusional control programmes, improvement in dwellings and better efficacy in diagnostics and therapeutics. The prospective study used the IFI and ELISA techniques for serology and filter paper detection. A total of 967 samples of dry blood in filter paper were collected in three public health units in Goiania and Aparecida de Goiania. From these samples 19 (1.96%) which were collected in filter paper from newborns were from mothers who declared to have CD. Results showed that 8 (42%) from the 19 samples were reagent to IgG anti-T. cruzi in both technique used. Peripheral blood was collected in seven mothers and their respective children. The serum samples were analyzed and the results confirmed IgG positive serology anti-T. cruzi with rates and titulation similar between mother and their respective newborn. Also there was accordance between the serology and the filter paper results which validates this strategy in the newborn screening triage from children from chronically infected mother. This study highlights the importance of the prenatal and neonatal care that precociously identifies the T. cruzi infection and also of programmes of health education about CD to the pregnant women population.