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IncQ-type plasmids have become important vectors in the dissemination of blaGES among different bacterial genera and species from different environments around the world, and studies estimating the occurrence of Guiana extended-spectrum (GES)-type ß-lactamases are gaining prominence. We analyzed the genetic aspects of two IncQ1 plasmids harboring different blaGES variants from human and environmental sources. The blaGES variants were identified using polymerase chain reaction (PCR) in Aeromonas veronii isolated from hospital effluent and Klebsiella variicola isolated from a rectal swab of a patient admitted to the cardiovascular intensive care unit in a different hospital. Antimicrobial-susceptibility testing and transformation experiments were performed for phenotypic analysis. Whole-genome sequencing was performed using Illumina and Oxford Nanopore platforms. The comparative analysis of plasmids was performed using BLASTn, and the IncQ1 plasmids showed a high identity and similar size. A. veronii harbored blaGES-7 in a class 1 integron (In2061), recently described by our group, and K. variicola carried blaGES-5 in the known class 1 integron. Both integrons showed a fused gene cassette that encodes resistance to aminoglycosides and fluoroquinolones, with an IS6100 truncating the 3'-conserved segment. The fused genes are transcribed together, although the attC site is disrupted. These gene cassettes can no longer be mobilized. This study revealed a mobilome that may contribute to the dissemination of GES-type ß-lactamases in Brazil. Class 1 integrons are hot spots for bacterial evolution, and their insertion into small IncQ-like plasmids displayed successful recombination, allowing the spread of blaGES variants in various environments. Therefore, they can become prevalent across clinically relevant pathogens.
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Extended-spectrum beta-lactamase producing and ciprofloxacin-non-susceptible Escherichia coli are clinical and environmental issues. We evaluated the susceptibility profile of fosfomycin in non-susceptible E. coli isolated from urine and the environment. We measured the activity of fosfomycin against 319 and 36 E. coli strains from urine and environmental isolates, respectively, collected from rivers. Fosfomycin resistance profiles were investigated using the minimal inhibitory concentration (MIC), according to the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST) guidelines. Antibiotic susceptibility testing revealed that 5% and 6.6% of urine samples were non-susceptible to fosfomycin according to CLSI and EUCAST guidelines, respectively. The fosfomycin MIC50/90 was 0.5/4 mg/L. Of the 36 E. coli isolates from river water, 11.1% and 13,8% were non-susceptible to fosfomycin according to CLSI and EUCAST, respectively (range ≤0.25 ≥512 mg/L). All the isolates with MIC ≥512 mg/L for fosfomycin showed the fosA3 gene. Fosfomycin resistance was more frequent in the environment than in clinical samples.
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Infecções por Escherichia coli , Fosfomicina , Humanos , Fosfomicina/farmacologia , Ciprofloxacina/farmacologia , Escherichia coli/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Infecções por Escherichia coli/tratamento farmacológico , beta-Lactamases/genética , Testes de Sensibilidade MicrobianaRESUMO
ABSTRACT Extended-spectrum beta-lactamase producing and ciprofloxacin-non-susceptible Escherichia coli are clinical and environmental issues. We evaluated the susceptibility profile of fosfomycin in non-susceptible E. coli isolated from urine and the environment. We measured the activity of fosfomycin against 319 and 36 E. coli strains from urine and environmental isolates, respectively, collected from rivers. Fosfomycin resistance profiles were investigated using the minimal inhibitory concentration (MIC), according to the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST) guidelines. Antibiotic susceptibility testing revealed that 5% and 6.6% of urine samples were non-susceptible to fosfomycin according to CLSI and EUCAST guidelines, respectively. The fosfomycin MIC50/90 was 0.5/4 mg/L. Of the 36 E. coli isolates from river water, 11.1% and 13,8% were non-susceptible to fosfomycin according to CLSI and EUCAST, respectively (range ≤0.25 ≥512 mg/L). All the isolates with MIC ≥512 mg/L for fosfomycin showed the fosA3 gene. Fosfomycin resistance was more frequent in the environment than in clinical samples.
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Abstract INTRODUCTION. This study aimed to evaluate different methods for differentiation of species of coagulase-negative staphylococci (CoNS) that caused infections in hospitalized immunocompromised patients. METHODS. A total of 134 CoNS strains were characterized using four different methods. RESULTS. The results of matrix assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) analysis were in complete agreement with those of tuf gene sequencing (kappa index = 1.00). The kappa index of Vitek 2® Compact analysis was 0.85 (very good) and that of the conventional method was 0.63 (moderate). CONCLUSIONS . MALDI-TOF MS provided rapid and accurate results for the identification of CoNS (134; 100%).
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Humanos , Staphylococcus/genética , Técnicas Bacteriológicas/métodos , Coagulase/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fenótipo , Staphylococcus/efeitos dos fármacos , Staphylococcus/enzimologia , Reprodutibilidade dos Testes , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Antibacterianos/farmacologiaRESUMO
Segundo o Ministério da Saúde, há uma carência de informações epidemiológicas relativas a fatores de risco relacionados às doenças crônicas não transmissíveis e que podem ser evitáveis, em sua maioria, com a modificação de estilo de vida, detecção precoce e o controle oportuno, Dentre estas, destacam-se as doenças cardiovasculares, as quais, segundo dados da Secretaria de Saúde do Paraná, apresentam maior impacto sobre a mortalidade, O objetivo deste trabalho foi avaliar a saúde dos servidores da Universidade Estadual de Ponta Grossa por meio do risco total presumido em desenvolver doenças do aparelho circulatório, de acordo com o Escore de Framingham e ocorrência de fatores de risco modificáveis, Para tanto, aplicou-se inquérito epidemiológico e determinaram-se parâmetros antropométricos, parâmetros laboratoriais e pressão arterial, As análises laboratoriais foram realizadas no Laboratório Universitário de Análises Clínicas em rotina automatizada, Os resultados revelaram o risco absoluto de infarto (calculado usando o Escore de Framingham) e morte em 10 anos, Segundo este critério, 78 dos 86 voluntários enquadraram-se em baixo risco no desenvolvimento de doenças cardiovasculares, Entretanto, a ocorrência de fatores de risco modificáveis, como sobrepeso e obesidade, tabagismo e hipertensão, aponta para a necessidade de medidas educativas e preventivas em relação aos fatores de risco observados nos servidores da Universidade, os quais devem ser alvos de um programa de saúde que contemple esses aspectos...
According to the Ministry of Health there is a lack of epidemiological information regarding risk factors related to chronic diseases that may be preventable, mostly, with the modification of lifestyle, early detection and timely control. Among these, there are the cardiovascular diseases, which, according to the Department of Health of Paraná, have greater impact on mortality. The aim of this study was to evaluate the health of servers from the Paraná State University of Ponta Grossa through the assumed total risk in developing cardiovascular diseases, according to the Framingham Score and the occurrence of modifiable risk factors. Therefore, it was applied an epidemiological survey and anthropometric parameters, laboratory parameters and blood pressure were determined. Laboratory analyzes were performed at the University Laboratory of Clinical Analyses in automated routine. The results revealed the absolute risk (calculated using the Framingham Score) of myocardial infarction and death in 10 years. According to this criterion, 78 of the 86 volunteers were classified at low risk of developing cardiovascular disease. However, the occurrence of modifiable risk factors, such as overweight and obesity, smoking and hypertension, points to the need for educational and preventive measures in relation to the risk factors noted in the Universitys servers, which should be targeted for a health program that addresses these issues...
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Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Doenças Cardiovasculares/prevenção & controle , Valor Preditivo dos Testes , Prevenção Primária , Prevenção Secundária , Fatores de RiscoRESUMO
BACKGROUND: Animal models are useful to evaluate the efficacy of antimicrobials in experimental sepsis. AIM: To elucidate the steps of producing an experimental model for the treatment of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae sepsis METHODS: Several ESBL inoculums ranging from 1.5x109 colony-forming units per milliliter (CFU/mL) to 2.0x1010 CFU/mL were administered by peritoneal injection in adults Wistar rats. Outcomes and microbiological data of quantitative peritoneal and blood cultures were observed in untreated animals. Animals which received 2.0x1010 CFU/mL inoculums were treated with single meropenem dose (30mg/kg) after one hour and those which received 1.0x1010 CFU/mL inoculums were treated immediately with three doses of meropenem 50 mg/kg. Outcomes were observed for 24 hours after inoculation. RESULTS: Solutions with 1.5 x109 and 6.0x109 CFU/mL were not lethal within 24 hours. Inoculums of 1.0x1010 CFU/mL were lethal in 80% and solutions with 2.0x1010 CFU/mL were lethal in 100% of animals. ESBL lethal sepsis (1.0x1010CFU/mL) was treated immediately with 50 mg/kg of meropenem every eight hours for 24 hours and presented 40% mortality compared with 80% mortality of the control group (p=0.033). Quantitative cultures of peritoneal fluid presented 104 CFU/mL or less for treated animals compared to more than 105 for untreated animals (p=0.001). CONCLUSION: Inoculums of 1.0x1010CFU/mL achieved the best results to study a model of lethal sepsis and this model of treatment of carbapenem-susceptible Enterobacteriaceae can serve as control to further evaluation of treatment of carbapenemase-producing Enterobacteriaceae models. .
RACIONAL: Modelos animais são importantes para avaliar a eficácia de antimicrobianos e a validação do sítio de infecção e a carga bacteriana. OBJETIVO: Definir a concentração do inóculo bacteriano, a dose e o tempo de administração de antimicrobianos a fim de validar um modelo experimental para o tratamento de Klebsiella pneumoniae produtora de betalactamase de amplo espectro em sepse letal. MÉTODO: Inóculos de Klebsiella pneumoniae produtora de betalactamase de espectro estendido de 1,5x109 unidades formadoras de colônias por mililitro (UFC/ml) a 2,0x1010 UFC/ml foram administrados via injeção peritoneal em ratos Wistar adultos. Sobrevida e dados microbiológicos de hemoculturas e culturas quantitativas de fluido peritoneal foram avaliados inicialmente em animais não tratados. Animais inoculados com 2,0x1010 UFC/ml foram tratados dose única de meropenem (30mg/kg) e animais inoculados com 1,0x1010 UFC/ml foram tratados imediatamente com meropenem (50 mg/kg) por 24 horas e os desfechos foram avaliados após 24 horas da inoculação. RESULTADOS: Soluções com 1,5 x109 e 6,0x109 UFC/ml não foram letais. Inóculos de 1,0x1010 UFC/ml e de 2,0x1010UFC/ml foram letais em 80% e 100% dos animais respectivamente. Sepse letal (1.0x1010CFU/mL) com tratamento imediato e por 24 horas apresentou 40% de mortalidade, comparada com 80% nos controles (p=0.033). Culturas quantitativas de fluido peritoneal apresentaram ≤104 UFC/ml enquanto que controles sem tratamento apresentaram >105 UFC/ml (p=0.001). CONCLUSÃO: Modelo experimental com inóculo de 1,0x1010UFC/ml submetido ao tratamento imediato e por 24 horas foi capaz de avaliar resposta microbiológica e de sobrevida podendo ser modelo de embasamento e de controle para tratamento de sepse letal por Klebsiella pneumoniae produtora de carbapenemase. .
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Animais , Feminino , Masculino , Ratos , Anti-Infecciosos/uso terapêutico , Modelos Animais de Doenças , Infecções por Klebsiella/tratamento farmacológico , Klebsiella pneumoniae/enzimologia , Sepse/tratamento farmacológico , beta-Lactamases/biossíntese , Ratos WistarRESUMO
INTRODUÇÃO: A presença de hemácias dismórficas na urina é um forte indicativo da origem glomerular do sangramento, sendo uma ferramenta importante no diagnóstico de glomerulonefrites. Os cilindros hemáticos geralmente acompanham as hemácias dismórficas, sendo também fortes indicadores de hematúria glomerular, embora não sejam encontrados com frequência no exame parcial de urina. OBJETIVO: Comparar duas técnicas de concentração de amostras em uma série de exames de urina com hematúria dismórfica. MATERIAL E MÉTODOS: Foram selecionadas 249 amostras com hematúria dismórfica a partir de 4.277 amostras de urina de rotina. As amostras foram processadas utilizando-se duas técnicas: a convencional e a de concentração. O percentual de identificação dos cilindros hemáticos foi comparado de acordo com a metodologia utilizada. RESULTADOS: A presença de cilindros hemáticos pela técnica de concentração foi estatisticamente maior (52,6 por cento) em comparação com a positividade pela metodologia convencional (8,4 por cento) (p < 0,001). DISCUSSÃO E CONCLUSÃO: Sugere-se que a técnica convencional não concentrou suficientemente a amostra de urina e os cilindros hemáticos ficaram no sobrenadante, sendo descartados. A utilização da técnica de concentração aumentou a sensibilidade técnica para a pesquisa dos cilindros hemáticos. Portanto, a técnica de concentração, associada à presença de hemácias dismórficas, mostrou-se útil para aumentar a concordância dos dois parâmetros laboratoriais para a detecção da hematúria de origem glomerular como auxílio diagnóstico das glomerulopatias, importante causa de doença renal crônica.
INTRODUCTION: Dysmorphic red blood cells (RBCs) in the urine are a strong indicator of a glomerular bleeding source. RBC casts, which while generally following RBC dysmorphism are not frequently seen on routine urinalysis, are also important indicators of glomerular hematuria. OBJECTIVE: This study tested the superiority of a urine concentration technique (CT) over the standard method (SM) for RBC cast identification in a group of patients suspected of glomerular hematuria. MATERIAL AND METHODS: Of a total of 4,227 routine urinary samples, 249 with dysmorphic hematuria were selected. The samples were processed according to two techniques: standard method (SM) and concentration technique (CT). The percentages of RBC cast identification according to each method were compared. RESULTS: The CT showed a higher rate of RBC casts (52.6 percent) compared to the SM (8.4 percent) (p < 0.001). DISCUSSION AND CONCLUSION: We suggest that the SM did not sufficiently concentrate the urine sample, the RBC casts remaining in the supernatant and being discarded. The CT increased the sensitivity of the RBC cast yield. The CT, associated with the presence of RBC dysmorphism, was useful to increase the agreement of the two parameters used for identification of glomerular-based bleeding and the diagnosis of glomerular diseases, important causes of chronic kidney disease.
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Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Eritrócitos Anormais , Hematúria/patologia , Urinálise/métodos , Urina/citologiaRESUMO
A infecção do trato urinário (ITU) é uma das doenças infecciosas mais comuns, perdendo em freqüência somente para as infecções respiratórias. No presente trabalho foram estudadas 106 uroculturas positivas de pacientes hospitalizados na Santa Casa de Misericórdia de Ponta Grossa, Paraná, Brasil. As bactérias isoladas foram classificadas, segundo sua origem: 68 (64,25%) em comunitária e 38 (35,8%) em hospitalar. Enterobactérias predominaram entre os isolados (82,1%). A susceptibilidade geral foi de 89,8% nitrofurantoína, 84% norfloxacina, 82% gentamicina, 75,3% cefalotina, 63,4% sulfazotrim e 45,7% ampicilina. Dos 94 prontuários estudados em 80,8% (76/94) constavam antibióticos prescritos. Em 62,8% (59/94) deles iniciou-se o tratamento empiricamente e em 18% (17/94) aguardou-se o resultado do antibiograma para a tomada de decisão. Na comparação entre o perfil de susceptibilidade, observado no antibiograma, com as prescrições empíricas, observou-se que em 81,4% (48/59) dos prontuários os antimicrobianos prescritos apresentaram sensibilidade no antibiograma e em 18,6% (11/59) estavam resistentes. O antibiótico mais prescrito empiricamente foi a norfloxacina, onde as Escherichia coli isoladas de infecções comunitárias e hospitalares apresentaram taxa de resistência de 11,6% e 15,8%, respectivamente. Outras Enterobactérias apresentaram taxas de 5,9% e 12,5%. As taxas de resistência inferiores a 20%, entre os principais agentes, mostram que a norfloxacina é adequada para ser utilizada como terapia empírica.
Urinary tract infection (UTI) is one of the most common infectious diseases, second in frequency only to respiratory infections. In the present work, 106 positive urine cultures were studied from patients hospitalized at Santa Casa de Misericórdia de Ponta Grossa, Paraná, Brazil. The isolated bacteria were classified, according to their source: 68 (64.2%) into community and 38 (35.8%) hospital bacteria. Enterobacteria were the most common among the isolates (82.1%). General susceptibility was 89.8% to nitrofurantoin, 84% to norfloxacin, 82% to gentamicin, 75.3% to cephalotin, 63.4% to sulfamethoxazole/trimethoprim, and 45.7% to ampicillin. Among the 94 patient records studied, 80.8% (76/94) indicated that antibiotics had been prescribed. In 62.8% (59/94) of them, treatment was started empirically, and in 18% (17/94) a decision was made only upon receiving the antibiogram result. In the comparison between the susceptibility profile revealed by the antibiogram and the empirical prescriptions, it was verified that the antibiogram showed sensitivity to the prescribed antimicrobials in 81.4% (48/59) of the patient records, while 18.6% (11/59) indicated resistance. The most frequently prescribed antibiotic was norfloxacin wherein Escherichia coli isolated from community and hospital infections showed resistance rates of 11.6% and 15.8%, respectively. Other Enterobacteria showed rates of 5.9% and 12.5%. Resistance rates lower than 20% among the most important agents indicated that norfloxacin is suitable to be used as empirical therapy.