Background: The Double-Layer Agar (DLA) technique is extensively used in phage research to enumerate and identify phages and to isolate mutants and new phages. Many phages form large and well-defined plaques that are easily observed so that they can be enumerated when plated by the DLA technique. However, some give rise to small and turbid plaques that are very difficult to detect and count. To overcome these problems, some authors have suggested the use of dyes to improve the contrast between the plaques and the turbid host lawns. It has been reported that some antibiotics stimulate bacteria to produce phages, resulting in an increase in final titer. Thus, antibiotics might contribute to increasing plaque size in solid media.Results: Antibiotics with different mechanisms of action were tested for their ability to enhance plaque morphology without suppressing phage development. Some antibiotics increased the phage plaque surface by up to 50-fold.Conclusion: This work presents a modification of the DLA technique that can be used routinely in the laboratory, leading to a more accurate enumeration of phages that would be difficult or even impossible otherwise.
1 December 2013 in Journal of Microbiological Methods
The evidence that biologically active food components are key environmental factors affecting the incidence of many chronic diseases is overwhelming. However, the full extent of such components in our diet is unknown, as is our understanding of their mechanisms of action. Beyond the interaction of these food components with the gut and intestinal immune functions, whey proteins such as lactoferrin are being tested as anticancer agents. Lactoferrin is an iron-binding protein that has been reported to inhibit several types of cancer. In the present work, the effects of bovine milk lactoferrin on human breast cancer HS578T and T47D cells were studied. The cells were either untreated or treated with lactoferrin concentrations ranging from 0.125 to 125 μM. Lactoferrin decreased the cell viability of HS578T and T47D by 47 and 54%, respectively, and increased apoptosis about 2-fold for both cell lines. Proliferation rates decreased by 40.3 and 63.9% for HS578T and T47D, respectively. For the T47D line, cell migration decreased in the presence of the protein. Although the mechanisms of action are not fully known, the results gathered in this work suggest that lactoferrin interferes with some of the most important steps involved in cancer development.
In this study, single and dual species biofilms of Pseudomonas aeruginosa and Escherichia coli, two common bacteria associated with urinary tract infections, were formed in silicon coupons immersed in artificial urine medium. In single species experiments, E. coli appeared to form biofilms more easily than P. aeruginosa. In mixed biofilms, both species apparently benefited from the presence of the other, as the average Log total cells cm−2 of mixed biofilms (7.29 cells cm−2) was higher than obtained for single cultures (6.99 cells cm−2). However, the use of selective media seemed to indicate that P. aeruginosa was the only microorganism to benefit in mixed biofilms (Log 7 CFU of P. aeruginosa cm−2, compared to Log 6 CFU cm−2 obtained in pure cultures). Peptide nucleic acid-fluorescence in situ hybridization combined with confocal laser scanning microscopy confirmed that E. coli was indeed being outnumbered by P. aeruginosa at 48 h. Whereas E. coli is the main causative agent of catheter-associated urinary tract infections, the results from this study indicate that the reason for the higher prevalence of this microorganism is not related to an enhanced ability to form biofilm and outcompete other species that may also be present, but rather to a better ability to form single-species biofilms possibly due to a more frequent access to the catheter surface.