ABSTRACT
Abstract Bacteriocins are peptides produced by various species of bacteria, especially lactic acid bacteria, which exhibit a large spectrum of action against spoilage bacteria and foodborne pathogens. Successful application of techniques for quantitative or qualitative bacteriocin determination relies not only on the sensitivity of the test-microorganisms, but also on the agar-medium employed. Cell free supernatants are routinely used to preliminary screen for antimicrobial activity of bacteria by means of the agar well diffusion method, but the supernatant may also include other molecules (such as medium components and/or intracellular compounds) accidentally released during cell free supernatant preparation, which may interfere with the assay. Reproducibility of bacteriocin activity against the same test-microorganisms is an important factor to be considered. Unfortunately, no specific information about bioassays standardization to determine bacteriocin activity is available in the literature. In this work, growth inhibition by means of the agar well diffusion assays were carried out on different agar-media showing a strong dependence on the agar-medium used, indicating that the inhibitory effects could also depend on the diffusion of exudates that are included in the cell-free supernatant. The results presented in this communication show that selection of the agar-medium is crucial for the bioassay response.
Subject(s)
Bacteriocins/analysis , Agar/analysis , Agar/pharmacokineticsABSTRACT
La tuberculosis es una enfermedad causada por bacterias pertenecientes al Complejo Mycobacterium tuberculosis. El diagnóstico se dificulta por su sintomatología inespecífica y los métodos bacteriológicos que ofrecen resultados tardíos. El objetivo del presente estudio fue comparar el desempeño del sistema automatizado BacT/ALERT® 3D con los métodos convencionales de cultivo en medio Lowenstein-Jensen (L-J) y Ogawa-Kudoh (O-K) para el aislamiento de micobacterias. Se procesaron 266 muestras provenientes de pacientes con sospecha de tuberculosis entre mayo y octubre de 2013. Se aislaron 63 bacilos acido-resistentes: 46 M. tuberculosis y 17 micobacterias no tuberculosas (MNT). Al comparar los tres métodos, se observó que todos presentaron desempeño similar en el aislamiento de M. tuberculosis. Las tasas de recuperación obtenidas no mostraron diferencia estadísticamente significativa (p>0,05) sin embargo, con BacT/ALERT® 3D se aislaron mayor número de MNT, con diferencia significativa respecto al L-J (p<0,05). El tiempo de detección promedio de M. tuberculosis fue de 11 días por BacT/ALERT® 3D, 20 días en L-J y 23 días en O-K. La contaminación cruzada, fue de 0,38%. Se concluyó que BacT/ALERT® 3D es una excelente herramienta para el aislamiento de M. tuberculosis, mejora la recuperación de las MNT y reduce significativamente el tiempo de diagnóstico, lo que permitiría un tratamiento oportuno, con mayor probabilidad de sobrevida del paciente.
Tuberculosis is a disease caused by species of the Mycobacterium tuberculosis complex. The diagnosis is difficult due to nonspecific symptoms and late results of bacteriological culture methods. The aim of this study was to compare the efficiency of the BacT/ALERT® 3D automated system with conventional methods of culture on Lowenstein-Jensen (L-J) and Ogawa-Kudoh (O-K). A total of 266 specimens from patients with clinical suspected tuberculosis were studied from May to October 2013. We recovered, 63 acid fast bacilli isolates: 46 identified as M. tuberculosis and 17 as nontuberculous mycobacteria (NTM). The three methods had a similar performance for isolation of M. tuberculosis; recovery rates obtained showed no statistically significant difference (p> 0.05). However, with the BacT/ALERT® 3D system, a larger number of MNT were isolated, with significant statistic difference for L-J (p <0.05). The average detection time for M. tuberculosis was 11 days with the BacT/ALERT® 3D system, with significant statistic difference for LJ (20.4 days) and O-K (23.2 days). Additionally, cross-contamination was determined as 0.38%. The study results showed that the BacT/ALERT® 3D system is an excellent tool for isolation of M. tuberculosis and it improves the recovery of NTM. Also, the time of diagnosis is significantly reduced, leading to earlier treatment that could improve patient recovery.
ABSTRACT
Poucas são as informações referentes ao perfil enzimático de bactérias rizobiais. Em meio de cultura solidificado, foi conduzido um experimento em fatorial 7 x 3 x 3 para avaliar o efeito do pH (5,0; 6,5 e 8,0) e da temperatura (25, 35 e 42 ºC) sobre a atividade amilolítica de sete isolados de rizóbio. As maiores atividades amilolíticas foram observadas em ambientes ácidos, com algumas bactérias também produzindo níveis significativos em pH 8,0. Entre as interações significativas (P<0,01), os isolados INPA R-110 e R-822 apresentaram máximas atividades em pH 5,0 e 25 ºC, com o R-822 também sendo um bom produtor de amilase nas temperaturas de 35 e 42 ºC. Em termos gerais, os isolados INPA R-110 e R-822 foram os melhores produtores de amilases, com atividades enzimáticas maiores do que 2,0.
Information is scarce regarding the enzymatic profiles of rhizobia bacteria. On solid medium, a 7 x 3 x 3 factorial experiment was conducted to evaluate the effect of pH (5.0, 6.5 and 8.0) and temperature (25, 35 and 42 ºC) on amylase activities of seven rhizobia strains. The highest enzymatic activities were found in acid environments, with some bacteria also producing significant levels at pH 8.0. Among the significant interactions (P<0.01), INPA strains R-110 and R-822 exhibited maximum activities at pH 5.0 and 25 ºC, with the R-822 being also a good amylase producer at 35 and 42 ºC. In general terms, INPA strains R-110 and R-822 were the best enzyme producers, with amylolytic enzymatic activities higher than 2.0.