A bacteriophage cocktail (designated ECP-100) containing three Myoviridae phages lytic for Escherichia coli O157:H7 was examined for its ability to reduce experimental contamination of hard surfaces (glass coverslips and gypsum boards), tomato, spinach, broccoli, and ground beef by three virulent strains of the bacterium. The hard surfaces and foods contaminated by a mixture of three E. coli O157:H7 strains were treated with ECP-100 (test samples) or sterile phosphate-buffered saline buffer (control samples), and the efficacy of phage treatment was evaluated by comparing the number of viable E. coli organisms recovered from the test and control samples. Treatments (5 min) with the ECP-100 preparation containing three different concentrations of phages (10 10 , 10 9 , and 10 8 PFU/ml) resulted in statistically significant reductions (P ‫؍‬ <0.05) of 99.99%, 98%, and 94%, respectively, in the number of E. coli O157:H7 organisms recovered from the glass coverslips. Similar treatments resulted in reductions of 100%, 95%, and 85%, respectively, in the number of E. coli O157:H7 organisms recovered from the gypsum board surfaces; the reductions caused by the two most concentrated phage preparations were statistically significant. Treatment with the least concentrated preparation that elicited significantly less contamination of the hard surfaces (i.e., 10 9 PFU/ml) also significantly reduced the number of viable E. coli O157:H7 organisms on the four food samples. The observed reductions ranged from 94% (at 120 ؎ 4 h posttreatment of tomato samples) to 100% (at 24 ؎ 4 h posttreatment of spinach samples). The data suggest that naturally occurring bacteriophages may be useful for reducing contamination of various hard surfaces, fruits, vegetables, and ground beef by E. coli O157:H7.

Bacteriophages are increasingly being utilized and considered for various practical applications, ranging from decontaminating foods and inanimate surfaces to human therapy; therefore, it is important to determine their concentrations quickly and reliably. Traditional plaque assay (PA) is the current "gold standard" for quantitating phage titers. However, it requires at least 18 h before results are obtained, and they may be significantly influenced by various factors. Therefore, two alternative assays based on the quantitative real-time polymerase chain reaction (QPCR) and NanoSight Limited (NS) technologies were recently proposed for enumerating phage particles. The present study compared the three approaches' abilities to quantitate Listeria monocytogenes-, Escherichia coli O157:H7- and Yersinia pestis-specific lytic phages quickly and reproducibly. The average coefficient of variation (CVS) of the PA method including all three phages was 0.15. The reproducibility of the PA method decreased dramatically when multiple investigators performed the assays, and mean differences of as much as 0.33 log were observed. The QPC R method required costly equipment and the synthesis of phage-specific oligonucleotide primers, but it determined phage concentrations faster (within about 4 h) and more precisely than did PA (CVS = 0.13). NS technology required costly equipment, was less precise (CVS = 0.28) than the PA and QPCR methods, and only worked when the phages were suspended in clear medium. However, it provided results within 5 min. After the overall correlation is established with the PA method, either of the two assays may be useful for quickly and reproducibly determining phage concentrations.

Fabrication of a chemiresistive biosensor for detection of biomolecules is demonstrated on a high surface area, flexible electro‐spun nylon fiber mat. For the first time, the –OH functionalized conducting copolymer of 3,4‐ethylenedioxythiophene (EDOT) and 3‐thiopheneethanol (3‐TE) is synthesized and conformally deposited on the electro‐spun mats by oxidative chemical vapor deposition (oCVD). The free –OH functional groups of the copolymer are available for immobilization of analyte specific biomolecules. Here, avidin and biotin molecules are employed as the analyte‐specific molecule and analyte respectively for their high specificity to each other. The sensitivities of avidin immobilized conducting copolymer on electro‐spun mats are tested against micro‐molar to nano‐molar concentrations of biotin in aqueous solutions. Application of electro‐spun fiber mat in this case enhances the sensor response 6 times when compared to a flat substrate and also significantly lowers the response time. In addition to the experimental studies, current work also includes modeling of the kinetics of the change of response for the biotin‐avidin interactions as a function of time. Most importantly, this fabrication technique promises an extremely sensitive and field deployable method for the detection of other biomolecules, for example, food pathogens.

No abstract

No abstract