82 research outputs found

    Antibiotic Resistance Genes in the Bacteriophage DNA Fraction of Environmental Samples

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    Antibiotic resistance is an increasing global problem resulting from the pressure of antibiotic usage, greater mobility of the population, and industrialization. Many antibiotic resistance genes are believed to have originated in microorganisms in the environment, and to have been transferred to other bacteria through mobile genetic elements. Among others, β-lactam antibiotics show clinical efficacy and low toxicity, and they are thus widely used as antimicrobials. Resistance to β-lactam antibiotics is conferred by β-lactamase genes and penicillin-binding proteins, which are chromosomal- or plasmid-encoded, although there is little information available on the contribution of other mobile genetic elements, such as phages. This study is focused on three genes that confer resistance to β-lactam antibiotics, namely two β-lactamase genes (blaTEM and blaCTX-M9) and one encoding a penicillin-binding protein (mecA) in bacteriophage DNA isolated from environmental water samples. The three genes were quantified in the DNA isolated from bacteriophages collected from 30 urban sewage and river water samples, using quantitative PCR amplification. All three genes were detected in the DNA of phages from all the samples tested, in some cases reaching 104 gene copies (GC) of blaTEM or 102 GC of blaCTX-M and mecA. These values are consistent with the amount of fecal pollution in the sample, except for mecA, which showed a higher number of copies in river water samples than in urban sewage. The bla genes from phage DNA were transferred by electroporation to sensitive host bacteria, which became resistant to ampicillin. blaTEM and blaCTX were detected in the DNA of the resistant clones after transfection. This study indicates that phages are reservoirs of resistance genes in the environment

    Generation and characterization of standardized forms of trehalose dihydrate and their associated solid-state behavior

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    Trehalose dihydrate is a nonreducing disaccharide which has generated great interest in the food and pharmaceutical industries. However, it is well recognized that considerable batch to batch variation exists for supposedly identical samples, particularly in terms of the thermal response. In this investigation, two standardized forms of trehalose dihydrate were generated using two distinct crystallization pathways. The two batches were characterized using scanning electron microscopy, X-ray powder diffraction, and FTIR. The thermal responses of the two forms were then studied using modulated temperature differential scanning calorimetry (MTDSC) and thermogravimetric analysis (TGA). In particular, we describe the technique of quasi-isothermal MTDSC as a means of studying the change in equilibrium heat capacity as a function of temperature. Finally, variable temperature FTIR was utilized to assess the change in bonding configuration as a function of temperature. SEM revealed significant differences in the continuity and grain structure of the two batches. The TGA, MTDSC, and quasi-isothermal MTDSC studies all indicated significant differences in the thermal response and water loss profile. This was confirmed using variable temperature FTIR which indicated differences in bond reconfiguration as a function of temperature. We ascribe these differences to variations in the route by which water may leave the structure, possibly associated with grain size. The study has therefore demonstrated that chemically identical dihydrate forms may show significant differences in thermal response. We believe that this may assist in interpreting and hence controlling interbatch variation for this material

    Optimal enrichment time for isolation of Vibrio parahaemolyticus from seafood.

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    The growth of Vibrio parahaemolyticus in a liquid medium was compared with that of human fecal flora and estuarine flora. No marked differences were noted between growth at 25 and 37 degrees C for V. parahaemolyticus. However, the marine organisms were strongly inhibited when incubated at 37 degrees C. Incubation for 8 h in an enrichment broth yielded V. parahaemolyticus growth, even with a small inoculum, whereas the marine and fecal floras were inhibited. Therefore, enrichment for 8 h at 37 degrees C appears to be optimal for isolation of V. parahaemolyticus, permitting more rapid results in seafood analysis

    Effect of high energy milling on molecular phases

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