RESUMO
Here, we investigate the transcriptome profiles of two S. Enteritidis and one S. Schwarzengrund isolates that present different persister levels when exposed to ciprofloxacin or ceftazidime. It was possible to note a distinct transcript profile among isolates, time of exposure, and treatment. We could not find a commonly expressed transcript profile that plays a role in persister formation after S. enterica exposure to beta-lactam or fluoroquinolone, as only three DEGs presented the same behavior under the conditions and isolates tested. It appears that the formation of persisters in S. enterica after exposure to ciprofloxacin is linked to the overexpression of genes involved in the SOS response (recA), cell division inhibitor (sulA), iron-sulfur metabolism (hscA and iscS), and type I TA system (tisB). On the other hand, most genes differentially expressed in S. enterica after exposure to ceftazidime appeared to be downregulated and were part of the flagellar assembly apparatus, citrate cycle (TCA cycle), glycolysis/gluconeogenesis, carbon metabolism, bacterial secretion system, quorum sensing, pyruvate metabolism pathway, and biosynthesis of secondary metabolites. The different transcriptome profiles found in S. enterica persisters induced by ciprofloxacin and ceftazidime suggest that these cells modulate their response differently according to each stress.
Assuntos
Anti-Infecciosos , Salmonella enterica , Fluoroquinolonas/farmacologia , Antibacterianos/farmacologia , Salmonella enterica/genética , Ceftazidima/farmacologia , Transcriptoma , Ciprofloxacina/farmacologia , Anti-Infecciosos/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
Bacillus sporothermodurans produces highly heat-resistant spores that can survive ultra-high temperature (UHT) treatment in milk. Therefore, we developed a rapid, specific and sensitive semi-nested touchdown PCR assay combined with propidium monoazide (PMA) treatment for the detection of viable B. sporothermodurans vegetative cells. The semi-nested touchdown PCR alone proved to be specific for B. sporothermodurans, and the achieved detection limit was 4 CFU/mL from bacterial culture and artificially contaminated UHT milk. This method combined with PMA treatment was shown to amplify DNA specifically from viable cells and presented a detection limit of 10(2) CFU/mL in UHT milk. The developed PMA-PCR assay shows applicability for the specific detection of viable cells of B. sporothermodurans from UHT milk. This method is of special significance for applications in the food industry by reducing the time required for the analysis of milk and dairy products for the presence of this microorganism.
Assuntos
Bacillus/crescimento & desenvolvimento , Bacillus/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Coloração e Rotulagem/métodos , Animais , Azidas/química , Bacillus/química , Bacillus/genética , Bovinos , Leite/microbiologia , Propídio/análogos & derivados , Propídio/química , Esporos Bacterianos/química , Esporos Bacterianos/genética , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Bacterianos/isolamento & purificaçãoRESUMO
AIM: To evaluate the pH and antibacterial activity of Endo CPM Sealer and MTA Fillapex by two different methods, using white MTA and Endofill as references for comparison. METHODOLOGY: Antibacterial activity was evaluated against Enterococcus faecalis (ATCC 29212). The agar diffusion test (ADT) was performed to evaluate the effect before setting. The materials were placed in four equidistant wells made in ten agar plates. After incubation at 37 °C for 48 h, the inhibition zones were measured using a digital paquimeter. The direct contact test (DCT) was performed to assess the antibacterial effect after setting. Suspensions of crushed materials were prepared and mixed with E. faecalis. After different periods of time (1, 6, 15 and 60 min), the survival of bacteria was assessed by using 10-fold serial dilution and cultivated on agar plates in triplicate. Colony-forming units (CFU) mL(-1) were calculated after incubation. pH values were also measured in triplicate. Comparison between sealers in the ADT and DCT was performed by the Kruskal-Wallis test. RESULTS: In the ADT, inhibition zones were found with MTA Fillapex and Endofill. They were similar to each other and greater than the other sealers (P < 0.05). None of the tested sealers demonstrated antibacterial activity in the DCT, and thus, all sealers had similar bacterial counts compared with the negative control group (P > 0.05). White MTA and Endo CPM Sealer suspensions had pH values >11, whilst MTA Fillapex and Endofill had lower values. CONCLUSIONS: MTA Fillapex and Endofill had an antibacterial effect against E. faecalis before setting, but none of the sealers maintained antibacterial activity after setting, despite the high pH of the MTA-based materials.
Assuntos
Compostos de Alumínio/farmacologia , Antibacterianos/farmacologia , Compostos de Cálcio/farmacologia , Óxidos/farmacologia , Materiais Restauradores do Canal Radicular/farmacologia , Silicatos/farmacologia , Compostos de Alumínio/química , Antibacterianos/química , Carga Bacteriana/efeitos dos fármacos , Compostos de Cálcio/química , Combinação de Medicamentos , Enterococcus faecalis/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio , Imunodifusão , Teste de Materiais , Viabilidade Microbiana/efeitos dos fármacos , Óxidos/química , Transição de Fase , Materiais Restauradores do Canal Radicular/química , Silicatos/química , Temperatura , Fatores de TempoRESUMO
Cloacal swabs were collected from 280 captive psittacine birds belonging to 13 species. Samples of dna were tested by PCR using a pair of primers that amplify a 284 base pair fragment of the Salmonella genus invA gene, and the PCR-positive samples were tested by standard microbiological techniques. Thirteen per cent of the samples were positive by PCR, but negative by microbiological techniques. The infection rates were significantly different among the 13 species, the most commonly infected being Amazona amazonica (28 per cent) and Amazona pretrei (20 per cent). Specific tests for Salmonella Typhimurium Salmonella Enteritidis, Salmonella Pullorum and Salmonella Gallinarum did not produce positive results.
Assuntos
Doenças das Aves/diagnóstico , Psittaciformes/microbiologia , Salmonelose Animal/diagnóstico , Salmonella/isolamento & purificação , Animais , Cloaca/microbiologia , DNA Bacteriano/química , DNA Bacteriano/genética , Feminino , Masculino , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Especificidade da EspécieRESUMO
ABSTRACT Salmonella Enteritidis (SE) is an important pathogen, causing both food poisoning outbreaks in humans and economic losses to the poultry industry, being also widely spread in the environment. This work aimed to identify SE phage types and to standardize the Random Amplified Polymorphic DNA (RAPD) for evaluating SE isolates obtained from different origins. To do so, 238 SE strains were selected, of which 104 were isolated from broiler carcasses, 106 from food samples and human biological materials involved in food poisoning outbreaks and 28 from different poultry materials. Among these 238 SE isolates, 111 were phage typed, and 57.7% (64/ 111) corresponded to phage type (PT) 4, 32.4% (36/111) to PT 4a, 3.6% (4/111) to PT 6a and 0.9% (1/111) to PT 7, whereas 5.4% .6/111) of the strains were not typeable (RDNC, reacts but does not conform). After the standardization of amplification conditions, all 238 SE isolates were submitted to RAPD/PCR. Among these, 91.8% (217/238) were classified as pattern A. Twenty-one isolates were differentiated into four patterns and into seven subtypes with the use of primer 1254, and into four patterns and ten subtypes using primer OPB 17. The combination of phage typing and RAPD/PCR proved to be a useful tool in epidemiological investigations. RAPD/PCR can be easily used as a routine laboratory method, thus helping with the monitoring of SE isolates and contributing to the establishment of effective Salmonella Enteritidis control and preventive programs.
RESUMO Salmonella Enteritidis (SE) é um patógeno de importância destacada como causa de toxinfecções alimentares em humanos, prejuízos ao setor produtivo e ampla disseminação no ambiente. Este trabalho objetivou identificar fagotipos e padronizar a RAPD/PCR (DNA polimórfico amplificado ao acaso) para a avaliação de isolados de SE. Foram selecionadas 238 amostras, sendo 104 oriundas de carcaças de frango, 106 de alimentos e material biológico de humanos isolados em episódios de toxinfecções alimentares e 28 de materiais de origem avícola. Foram fagotipadas 111 amostras sendo 57,7% (64/111) do fagotipo 4, 32,4% (36/111) fagotipo 4a, 3,6% (4/111) fagotipo 6a e 0,9% (1/111) fagotipo 7, enquanto 5,4% (6/111) não foram fagotipáveis (RDNC, reagent do not conform). Para a RAPD/PCR foram utilizados 238 isolados de SE. Destes, 91,8% (217/238) foram enquadrados no padrão A e 21 isolados (8,8%) foram diferenciados em quatro padrões e sete subtipos com o primer 1254 e em quatro padrões e dez subtipos com o primer OPB 17. A facilidade de execução da RAPD/PCR, após padronizada, habilita a sua implantação em uma rotina laboratorial, auxiliando na monitoria dos isolados de SE e, conseqüentemente, contribuindo para a elaboração de programas efetivos de controle e prevenção de S. Enteritidis.
RESUMO
Ornithobacterium rhinotracheale (ORT) is a recently described species of bacterium associated with respiratory disease, growth retardation, mortality, and decreased egg production in chickens and turkeys. Pneumonia, pleuritis, and airsacculitis characterise the infection. ORT has been isolated in many countries but it is still considered exotic in Brazil. Up to date it is prohibited to import and produce reagents for diagnostic and vaccination control. The aim of this study was to isolate and identify the bacteria in chickens. Four isolates were obtained from tracheal swabs of broilers. They were isolated in blood agar with gentamicin and showed biochemical, morphological, antigenic and genetic characteristics of ORT. The results confirm that ORT is present in Brazil.
Assuntos
Galinhas/microbiologia , Ornithobacterium/classificação , Ornithobacterium/isolamento & purificação , Animais , BrasilRESUMO
AIMS: To compare PCR combined with enrichment media with the standard microbiological techniques (SMT) and to determine the most sensitive method for the detection of Salmonella and the identification of Salm. typhimurium (ST), Salm. enteritidis (SE), Salm. gallinarum (SG) and Salm. pullorum (SP). METHODS AND RESULTS: We analysed 87 samples from poultry using PCR and SMT, PCR being performed from non-selective (NS) and Rappaport-Vassiliadis (RV) media. PCR-NS was less sensitive than PCR-RV and SMT for the detection and identification of Salmonella. PCR-RV detected more positive samples of Salmonella sp. than SMT but both these methods showed similar sensitivity regarding the identification of Salmonella serovars. CONCLUSIONS: PCR-RV was more sensitive and decreased the time necessary to detect and identify Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-RV is a powerful tool for the rapid and accurate detection and identification of Salmonella and can be implemented in diagnostic and food analysis laboratories.
Assuntos
Técnicas Bacteriológicas , Microbiologia de Alimentos , Reação em Cadeia da Polimerase/métodos , Aves Domésticas/microbiologia , Salmonella/genética , Salmonella/isolamento & purificação , Animais , Técnicas Bacteriológicas/estatística & dados numéricos , Sequência de Bases , Primers do DNA/genética , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase/estatística & dados numéricos , Intoxicação Alimentar por Salmonella/diagnóstico , Intoxicação Alimentar por Salmonella/microbiologia , Salmonelose Animal/diagnóstico , Salmonelose Animal/microbiologia , Salmonella enteritidis/genética , Salmonella enteritidis/isolamento & purificação , Salmonella typhimurium/genética , Salmonella typhimurium/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
A polymerase chain reaction (PCR) assay was developed for the generic detection of Salmonella sp. and the identification of S. Enteritidis (SE), S. Gallinarum (SG), S. Pullorum (SP) and S. Typhimurium (ST) in material collected in the field from poultry. The specificity and sensitivity of the assay combined with Rappaport-Vassiliadis selective enrichment broth (PCR-RV) were determined, and field samples were analyzed to verify the validity of the method application. Specificity of the assay was tested using 29 SE, 11 SG, 10 ST and 10 SP strains, along with 75 strains of 28 other Salmonella serovars and 21 strains of other bacterial genera. The assay was 100% specific for Salmonella detection and ST identification. The primer pair for SE, SG and SP also detected S. Berta. PCR detection limits for Salmonella at the genus level were 2 ST, 8 SE, 1.1x10(3) SG and 1.8x10(5) SP cells. At the serovar level, detection limits were 7 ST, 1.2x10(3) SE, 4.4x10(7) SG and 1.8x10(6) SP cells. At the genus level, PCR-RV detected approximately 128% more positive field samples than the standard microbiological techniques and results were ready in 48h instead of 7 days. PCR-RV method is diagnostic of Salmonella at the genus level and ST at the serovar level, although other tests are needed to identify SE, SG and SP to serovar level.
Assuntos
Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia , Salmonella/isolamento & purificação , Animais , DNA Bacteriano/química , DNA Bacteriano/genética , Eletroforese em Gel de Ágar/veterinária , Reação em Cadeia da Polimerase/métodos , Aves Domésticas , Doenças das Aves Domésticas/diagnóstico , Salmonella/classificação , Salmonella/genética , Salmonelose Animal/diagnósticoRESUMO
The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10(-7), 10(-8) or 10(-9) CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10(-9) CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.
Assuntos
Galinhas/microbiologia , DNA Bacteriano/isolamento & purificação , Produtos Avícolas/microbiologia , Salmonella/isolamento & purificação , Animais , DNA Bacteriano/análise , Reação em Cadeia da Polimerase , Fatores de TempoRESUMO
INTRODUCTION: Knowledge accumulation is overfilling the thematic content of medical graduation. Medical Schools must be alert to define a minimal content related with the most frequent disturbs. We intent to suggest topics for the minimal content, to the program of Neurology in medical graduation. METHOD: 1. To identify the places where young doctors are working outside the School Hospital (SH): we analysed the answers of the letters sent to 6415 resident - doctors (RD) in São Paulo's State and we made personal interview with 201 RD. 2. To verify the most frequent neuropsychiatric disturbs (ND) in the population: we made the analysis of the patient's diagnostic records in emergency room of three institutions: Municipal of Taubaté-SP, Municipal of São José dos Campos-SP and Faculty of Medicine of São Paulo Santa Casa de Misericórdia. RESULTS: 1. The RD are young and, outside the SH, they work in other emergency services. 2. The most frequent diagnosis in institutions were listed: alcoholism, cerebrovascular disease, coma, cranial trauma, dementia, dizziness, epilepsy, facial paralysis, faint, headache, hemiplegia or paraplegia, meningitis, others paralysis, periferical neuropathy and psychiatric disorders. CONCLUSIONS: Those diagnostics most frequent are relevant topics of the programmatic content to the program of Neurology in medical graduation.
Assuntos
Currículo , Educação de Graduação em Medicina , Doenças do Sistema Nervoso/epidemiologia , Neurologia/educação , Brasil/epidemiologia , Feminino , Humanos , Masculino , Doenças do Sistema Nervoso/diagnósticoRESUMO
The authors performed a case study using Dorothea Orem's self-care model as a framework in the treatment by llizarov's method, administering care and specific orientations for recovery and prevention of further disorders. The operational system of the study consisted of procedures related to demands of universal and therapeutic care during hospitalization. Demands were detected in the pre-operative and care developed in the postoperative phase, providing the patient with the knowledge and achievement of the necessary procedures to be followed at home.
Assuntos
Técnica de Ilizarov/reabilitação , Educação de Pacientes como Assunto/métodos , Cuidados Pós-Operatórios/métodos , Autocuidado/métodos , Adulto , Terapia por Exercício/métodos , Humanos , Masculino , Modelos de Enfermagem , Alta do PacienteRESUMO
The study aims to explore the utilization of the Formative Evaluation by the microcomputer in the teaching of primary attention to the health. The experiment is composed of eight modules, programmed in Basic software "Formative Evaluation". For each module, a "Question Bank" with three sorts of feedbacks was organized. The sample includes 16 subjects. The results show that the Formative Evaluation by Microcomputer proportioned to the students a cognitive domain significantly higher than the traditional instruction. However, it didn't provide a significantly superior performance to the students who experienced this system.