No abstract

Highly pathogenic avian influenza virus H5N1 is a continuous threat to public health and poultry industry. The recurrence of the H5N1 led us to develop a robust, specific, and rapid detection method for the virus. In this study, an impedance aptasensor was developed for the virus detection using specific H5N1 aptamer and a gold interdigitated microelectrode. Streptavidin was immobilized on the microelectrode surface and biotin labeled H5N1 aptamer was bound to the immobilized streptavidin. The microelectrode was blocked with the polyethylene glycol and the bound aptamer captured the virus. The impedance change caused by the captured virus was measured using an impedance analyzer. To enhance impedance signal, a nanoparticle-based amplifier was designed and implemented by forming a network-like gold nanoparticles/H5N1-aptamer/thiocyanuric acid. The detection limit of the impedance aptasensor was 0.25 HAU for the pure virus and 1 HAU for the tracheal chicken swab samples spiked with the H5N1 virus. The detection time of aptasensor without employing the amplifier was less than an hour. The amplifier increased impedance by a 57-fold for the 1 HAU samples. Only negligible impedance change was observed for non-target viruses such as H5N2, H5N3, H7N2, H1N1, and H2N2. This aptasensor provides a foundation for the development of a portable aptasensor instrument.

The potential of food-borne pathogens to survive and grow during refrigerated and frozen storage has raised serious concerns over the safety of stored poultry products. In this study, the effect of refrigeration and freezing temperatures (-20, -12, 0, 4, and 8°C) on growth and survival of Listeria innocua and Salmonella enterica serovar Typhimurium in raw chicken breasts for storage times of 3, 7, 10, 14, and 21 d were investigated. A modified Weibull model was also developed to analyze the microbial behavior of both microorganisms in raw chicken breasts under different refrigerated storage conditions over time. The results showed that the bacterial loads of L. innocua at 4 and 8°C and Salmonella Typhimurium at 8°C were significantly different (P < 0.05) from those at other refrigerated and frozen storage temperatures over storage times. The loads of both bacteria at frozen storage temperatures did not change significantly over time. At a storage time of 7 d, the increase in bacterial loads of L. innocua at 4 and 8°C was 2.1 log cfu/g and 3.7 log cfu/g, respectively, and that of Salmonella Typhimurium at 8°C was 1.2 log cfu/g. The root mean square errors, median relative error, mean absolute relative error, and the plot of predicted versus observed bacterial loads showed a good performance of the model. The results from this study provided useful information regarding the behavior of Listeria and Salmonella in raw chicken breast meat during refrigerated and frozen storage, which would be helpful in giving insight over the safety of poultry products storage.

E. coli O157:H7, one of the major foodborne pathogens, can cause a significant threat to the safety of foods. The aim of this research is to develop an activated biochar-based immunosensor that can rapidly detect E. coli O157:H7 cells without incubation in pure culture. Biochar was developed from corn stalks using proprietary reactors and then activated using steam-activation treatment. The developed activated biochar presented an enhanced surface area of 830.78 m2/g. To develop the biosensor, the gold electrode of the sensor was first coated with activated biochar and then functionalized with streptavidin as a linker and further immobilized with biotin-labeled anti-E. coli polyclonal antibodies (pAbs). The optimum concentration of activated biochar for sensor development was determined to be 20 mg/mL. Binding of anti-E. coli pAbs with E. coli O157:H7 resulted in a significant increase in impedance amplitude from 3.5 to 8.5 kΩ when compared to an only activated biochar-coated electrode. The developed immunosensor was able to detect E. coli O157:H7 cells with a limit of detection of 4 log CFU/mL without incubation. Successful binding of E. coli O157:H7 onto an activated biochar-based immunosensor was observed on the microelectrode surface in scanning electron microscopy (SEM) images.