Programa de Pós-graduação em Biologia da Relação Parasito-Hospedeiro
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Navegando Programa de Pós-graduação em Biologia da Relação Parasito-Hospedeiro por Por Orientador "Carneiro, Lilian Carla"
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Item Detecção, tipificação e filogenia molecular de Papilomavírus bovino em bovinos leiteiros(Universidade Federal de Goiás, 2017-03-31) Albuquerque, Winnie Castro Amorim e; Braga, Carla Afonso Da Silva Bitencourt; http://lattes.cnpq.br/7673897995590123; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Barbosa, Mônica Santiago; Santos, Mônica Oliveira; Moraes Filho, Aroldo Vieira de; Souza, Guilherme Rocha Lino deBovine papillomavirus is the etiological agent of bovine papillomatosis, a disease that triggers warts throughout the skin, udder, roofs, genitalia and in more severe cases can develop extensive papillomas, cause neoplasia in the digestive tract and bladder, weaken the animal's health and cause losses in the Productivity and losses for livestock. The present study aims to detect and typify bovine Papillomavirus present in bovine tissue and blood samples with papillomatosis, to sequence the isolated viral types, to analyze the nucleotide sequences and the phylogeny of the detected viral types. As a result, amplification was obtained in five tissue samples (papilloma) from different bovines, not being successful in the amplification of blood samples. PCR reactions revealed the presence of BPV-1 in 60%, BPV-5 in 40%, BPV-9, BPV-10, BPV-13 and BPV-14 in 20% and BPV-12 in 40% of the analyzed samples. The presence of coinfection was verified in 60% of the lesions analyzed, with up to four viral types infecting the same sample. Alignments of viral type sequences 1, 5 and 14 were validated with identity ranging from 74% to 95%. The phylogenetic diagram showed a genetic approximation between viral types 1 and 14, both belonging to the genus Deltapapillomavirus, and distancing between nucleotide sequences of viral types 5, 9 and 14. Papillomaviruses of types 5 and 9 belong to different genera, Epsilonpapillomavirus and Xipapillomavirus, Respectively, the phylogenetic distance between these viral types, verified in the diagram, is justified.Item Indicadores de contaminação viral em amostras de água consumida em regiões rurais de Goiás(Universidade Federal de Goiás, 2022-11-28) Bordoni, Graziela Picciola; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Albuquerque, Antônio João Carvalho de; Oliveira, Bruno Francesco Rodrigues deRural communities are located far from urban centers, where the adequate supply of potable water is hampered. In the lack of a public distribution network, water is obtained from surface and groundwater sources, where the microbial quality is often unknown. Goiás has 9.7% of the population in rural areas and previous studies carried out in some of these rural and traditional communities have already demonstrated the vulnerability of water for the consumption of these residents by waterborne pathogens, such as enteric viruses. Therefore, the aim of the work was to continue the evaluation of fecal contamination of water samples from individual and collective sources, from 24 communities in rural areas of the State of Goiás. 160 samples were collected, whose water sources were shallow tubular well, deep tubular well, dug well and spring, surface water, rainwater stored in cisterns and water truck. The enteric viruses Human Adenovirus (HAdV), Rotavirus (RV) and Enterovirus (EV) were used as indicators of fecal contamination, analyzed by qPCR. In all, 30% of the samples showed signs of fecal contamination. Detection rates were 20% for RV, 9.4% for HAdV and 4.4% for EV. The geometric means of concentration of these indicators were 1.5x106 GC/L, 1.9x106 GC/L and 7.1x105 GC/L, respectively. RV was the most prevalent viral indicator in underground sources, being statistically associated with shallow tubular wells. In spring samples, HAdV stands out with 11.8%. In the cistern samples, both HAdV and RV were found in the same prevalence. These results reveals that the analyzed population is vulnerable to waterborne diseases caused by enteric pathogens, serving to guide future decisions and improve daily water treatment practices in these communities.Item 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; http://lattes.cnpq.br/5637230378599476; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Vieira, José Daniel Gonçalves; Santos, Mônica de Oliveira; Galvão Filho, Arlindo Rodrigues; Moreira, Ellen FláviaWater 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.Item Produção e caracterização de celulases e xilanases produzidas por Streptomyces thermocerradoensis I3 em fermentação semi-sólida(Universidade Federal de Goiás, 2016-09-14) Gama, Aline Rodrigues; Bataus, Luiz Artur Mendes; http://lattes.cnpq.br/5637230378599476; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Jesuíno, Rosália Santos Amorim; Kipnis, André; Lino, Guilherme; Paula, Fabrícia deThe use of carbohydrates derived from agricultural waste is important in the industry of food, textiles, paper, detergents, animal feed and in the production of bioethanol. Some filamentous bacteria are used in the enzymatic degradation processes, such as Streptomyces thermocerradoensis I3, which was isolated from soil and this work was selected for the production of cellulases and xylanases by growing in semi-solid fermentation (SSF) in medium supplemented with wheat bran (WB) as carbon source. S. thermocerradoensis I3 it was maintained for 4 days at 37 ° C and showed a higher production of Endoglucanase (2,92 U/mL) and Xylanase (12,34 U/mL) after 72 hours of cultivation and β-glucosidase (0,023 U/mL) and FPase (2,82 U/mL) after 96 hours of cultivation. The 96-hour extract was concentrated. The enzyme present in the concentrated extract presented molecular mass of 17 kDa. It showed activity for cellulase and xylanases confirmed by zimograms and enzymatic activities. The crude extract (EB) and the concentrated extract (EC) were analyzed for pH and temperature optima for activity of cellulases and xylanases. The results showed that the highest activity of the total cellulase (FPase) was at pH 6.0 at 60 °C (EB) and pH in the range 3.0 to 6.0 at 80 °C (EC); the endoglucanase has higher activity at pH 6.0 at 55 °C (EB and EC); xylanase showed higher activity at pH 8.0 at 70 °C (EB and EC). The xylanase activity of EB and EC showed thermostability 60% after 2 hours of incubation at 60 °C and the endoglucanase activity of EB and EC remained above 50% after 4 hours of incubation at 50 °C, 60 °C and 70 °C. Qualitative analysis observed by TLC (Thin layer chromatography) showed the production of oligosaccharides and xilooligômeros from the hydrolysis of different substrates. S. thermocerradoensis I3 successfully used the WB in SSF producing enzymes that have the characteristics necessary for their industrial application.Item Escherichia coli, adenovírus e enterovírus em amostras de água consumida em áreas rurais de Goiás(Universidade Federal de Goiás, 2020-06-23) Lima, Fernando Santos; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Scalize, Paulo Sérgio; Santos, Mônica de Oliveira; Gabriel, Ellen Flávia Moreira; Gama, Aline RodriguesThe rural environment lacks basic sanitation services. Thus, the water supply and sewage disposal facilities are often under the initiative of each resident, who may not have the financial resources and technical knowledge to build and keep them functioning properly. Thus, water for human consumption is subject to fecal contamination and, consequently, to the presence of waterborne pathogens, such as enteric viruses. The objective of this work was to evaluate the fecal contamination of water samples from individual sources used for household supply in rural areas of the State of Goiás. The samples were collected from 86 homes, distributed in 15 communities, whose water sources were tubular wells, dug wells, springs, surface waters and rainwater. Escherichia coli (EC) bacteria, analyzed by the defined chromogenic substrate method, and the enteric virus human adenovirus (HAdV) and enterovirus (EV), analyzed by qPCR, were used as fecal contamination indicators. It was observed that 90.7% of the samples showed indications of fecal contamination. Detection rates were 74.4% for EC, 57% for HAdV and 9.3% for EV. The concentration averages of these indicators were, respectively, 8.34 x 101 NMP / 100mL, 8.6 x 105 CG / L and 9.75 x 105 CG / L. The EC indicator was the most prevalent in ground and surface water samples. The HAdV indicator was significantly more detected in groundwater samples than in surface water and was more efficient in indicating contamination in tubular wells. In cistern samples, viral indicators were the most prevalent. There was no association of frequencies or correlation of concentrations between EC and HAdV. HAdV indicated human fecal contamination and performed well as a complementary indicator. These results reveal that the analyzed population is vulnerable to waterborne diseases caused by enteric pathogens.Item Perfil fenotípico e genotípico de enterobactérias resistentes aos beta-lactâmicos(Universidade Federal de Goiás, 2018-06-28) Santos, Andressa Liberal; Braga, Carla Afonso da Silva Bitencourt; http://lattes.cnpq.br/7673897995590123; Carneiro, Lilian Carla; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Bataus, Luiz Artur Mendes; Santos, Mônica de OliveiraEnterobacteria are microorganisms involved with bacteria and the health of women with care. Treatment of bacterial infections is most often done with the use of antibiotics and one of the major classes of antimicrobials is one of the β-lactams. Among the main mechanisms of resistance to the observational β-lactam antibiotics: alteration of the antimicrobial target; alteration of β-lactam permeability; Flow pumps and the entry of enzymatic signals that destroy the β-lactate totally or with the development of an alternative metabolic pathway. These enzymes are known as beta-lactamases and are encoded by specific genes. Thus, the objective of this study was to correlate the resistance profile of enterobacteria, using phenotype methodologies and identifying 14 genes that encode as beta-lactamase enzymes: blaOXA genes; blaIMP; blaNDM; blaSME; blaDHA; blaCMY, blaTEM, blaKPC, blaSPM, blaCTX-M, blaVIM, blaSIM, blaGim and blaSHV. The phenotypic methodologies used were the Antimicrobial Sensitivity Test for Disk-Diffusion (antibiogram), and complementary tests for the detection of resistance mechanisms of beta-lactamases (ESBL, MBL, AmpC and Carbapenemase). The molecular methodology used was Real Time PCR using the Sybr Green system. Among the results found in the tests it was observed that 74.28% were resistant to ampicillin, 34.28% were resistant to aztreonam, 62.85% were resistant to amoxicillin associated with clavalunate, 51.42% were resistant to ceftazidime, 41 , 42% were resistant to cefoxitin, 54.28% were resistant to cefazolin, 44.28% were resistant to cefepime, 41.42% were resistant to cefuroxime, 8.57% were resistant to cefuroxime, 35.71% were resistant an imipenem and 41.42% were resistant to piperacillin associated with tazobactam. Among the total samples, the mechanism of resistance that presented the highest expression was ESBL (17.14%). The genes studied that were detected in a greater number of genera were blaGIM and blaSIM (66.66% of the samples). The gene that was amplified in a smaller number of samples was blaVIM (16.66%). It is concluded that although there is a low correlation between the methodologies analyzed, the levels of antimicrobial resistance in enterobacteria are high and worrying, and a way to minimize the accelerated emergence of resistance includes the development or improvement of techniques that generate diagnoses with high efficiency and speed.Item 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; http://lattes.cnpq.br/6506744224041777; Carneiro, Lilian Carla; Silva, Antonio Márcio Teodoro Cordeiro; Gama, Aline Rodrigues; Moraes Filho, Aroldo Vieira de; Santos, Mônica de OliveiraHelicobacter 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.