Effects of conservation buffer systems on adsorption of fluorescent-labeled Escherichia coli.

The magnitude of bacterial transport through runoff into surface water or infiltration into groundwater is influenced by the adsorption processes in soil. The objective of this study was to evaluate fluorescent-labeled Escherichia coli (E. coli) adsorption by soil under agroforestry buffer (AB), grass buffer (GB), and row crop (RC) management. Adsorption experiments were conducted by inoculating three masses (0.5, 1, and 10 g) of each treatment (AB, GB, and RC) with E. coli O157:H7-GFP with concentration ranges of 105 -108 colony-forming units (cfu) ml-1 . Adsorption data were evaluated using Langmuir, Freundlich, and Temkin adsorption isotherm models. The Freundlich isotherm model described the observed data well for all treatments using the 10-g soil mass, with the R2 values closer to unity in all treatments. The Freundlich Kf parameter, an indicator of adsorption capacity, was higher for the AB treatment (9.93 cfu ml-1 ) compared with the GB and RC treatments (2.32 and 1.27 cfu ml-1 , respectively). The multiple pairwise comparisons test (Tukey test) of the Freundlich 1/nf parameter demonstrated a significant difference (p < .05) between the AB treatment and the RC and GB treatments. Similarly, the Kf values were significantly (p = .05) higher for the 10-g mass under the same test conditions, but no significant differences were observed in the 0.5- and 1-g masses. This study demonstrated that AB has a higher E. coli adsorption capacity and the potential for mitigating the effects of E. coli O157:H7 transport to surface or groundwater through the soil ecosystem.

A Review of SARS-CoV-2 Disease (COVID-19): Pandemic in Our Time.

Development and deployment of biosensors for the rapid detection of the 2019 novel severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) are of utmost importance and urgency during this recent outbreak of coronavirus pneumonia (COVID-19) caused by SARS-CoV-2 infection, which spread rapidly around the world. Cases now confirmed in February 2022 indicate that more than 170 countries worldwide are affected. Recent evidence indicates over 430 million confirmed cases with over 5.92 million deaths scattered across the globe, with the United States having more than 78 million confirmed cases and over 920,000 deaths. The US now has many more cases than in China where coronavirus cases were first reported in late December 2019. During the initial outbreak in China, many leaders did not anticipate it could reach the whole world, spreading to many countries and posing severe threats to global health. The objective of this review is to summarize the origin of COVID-19, its biological nature, comparison with other coronaviruses, symptoms, prevention, treatment, potential, available methods for SARS-CoV-2 detection, and post-COVID-19 symptoms.

A microfluidic biosensor for rapid simultaneous detection of waterborne pathogens.

A microfluidic based biosensor was investigated for rapid and simultaneous detection of Salmonella, Legionella, and Escherichia coli O157:H7 in tap water and wastewater. The biosensor consisted of two sets of focusing electrodes connected in parallel and three sets of interdigitated electrodes (IDE) arrays. The electrodes enabled the biosensor to concentrate and detect bacteria at both low and high concentrations. The focusing region was designed with vertical metal sidewall pairs and multiple tilted thin-film finger pairs to generate positive dielectrophoresis (p-DEP) to force the bacteria moving toward the microchannel centerline. As a result, the bacterial pathogens were highly concentrated when they reached the detection electrode arrays. The detection IDE arrays were coated with three different antibodies against the target bacterial pathogens and a cross-linker to enhance the binding of antibodies to the detection electrode. As the binding of bacterial pathogen to its specific antibodies took place, the impedance value changed. The results demonstrated that the biosensors were capable of detecting Salmonella, Legionella, and E. coli 0157:H7 simultaneously with a detection limit of 3 bacterial cells/ml in 30 - 40 min.

Nanoparticle immuno-fluorescent probes as a method for detection of viable E. coli O157:H7.

Development of revolutionary sensitive biosensors for detecting the presence of harmful biological species in the environment is a necessity for countering disease outbreaks. This work examined the interaction of fluorescence-labeled antibody on amine functionalized gold nanoparticles (GNP) as a model system. The synthesized tetramethylrhodamine isothiocyanate (TRITC) labeled antibody-amine functionalized GNP interaction was characterized using UV-Vis spectroscopy and Fluorescent Microscopy imaging. Transmission Electron Microscopy (TEM) was also used to observe the morphology of the GNP. In contrast to TEM, the fluorescence microscopy imaging revealed the coating of the TRITC labeled antibody on the surface of the GNP. The signals were measured using a Photon Technology Inc. fluorometer at excitation of 541 nm and emission at 555 nm to 650 nm. Tests were conducted at near real-time with results obtained using the biosensor assay within 5 min. Results indicated that there was a shift of the wavelength from lower to higher wavelength (blue to red shift) when conjugated GNP (anti-E. coliO157:H7; IgY-TRITC-GNP) are compared to free GNP, a difference of about 28 nm. The GNP demonstrated a quenching capability when compared to the TRITC labeled antibody (degree of labeling of 15.41 mol dye per mole of IgY) using fluorometer. The lower and upper detection range of this method was found to be 103-105 CFU/mL with observed fluorescence of about 42,000 counts per seconds as against 24,000 counts per seconds that was observed when the specificity of the sensor was tested using Salmonella enterica.

An impedance biosensor for simultaneous detection of low concentration of Salmonella serogroups in poultry and fresh produce samples.

This paper reports the design, fabrication and testing of a microfluidic based impedance biosensor for rapid and simultaneous detection of three Salmonella serogroups. The microfluidic device consists of three microchannels, each one includes a region for focusing the Salmonella cells into the centerline of the microchannel and direct them toward the sensing region to obtain highly concentrated samples using positive dielectrophoresis force. A region for bacteria sensing consists of interdigitated electrode (IDE) array with 10 pairs of fingers. Three types of Salmonella antibodies (type B, D and E) were mixed separately with the cross-linker (Sulfo-LC-SPDP) to enhance the immobalization of the antibodies to the detection electrodes. The electrode surfaces was then functionalized with the three mixtures, one for each channel. As target antigen binds to the antibody, it results in impedance change. The Salmonella samples were spiked with Salmonella type B, introduced into the biosensor via the sample inlet into the focusing region, and then toward the sensing region where they bind to the immobilized antibody, causing a change in the impedance. The performance of the devices was tested using single Salmonella serotype B and two Salmonella serotypes B, and D, with a limit of detection of 7 cells/ml. The biosensor was also able to differentiate live from dead bacteria eliminating the false positive results. Finally, the device was also able to detect Salmonella selectively when other type of pathogen was present.

An integrated impedance biosensor platform for detection of pathogens in poultry products.

This paper presents an impedance-based biosensor for rapid and simultaneous detection of Salmonella serotypes B, D, and E with very low concentration. The biosensor consists of a focusing region, and three detection regions. The cells focusing was achieved using a ramp down electroplated vertical electrode pair along with tilted thin film finger pairs that generate p-DEP forces to focus and concentrate the bacterial cells into the center of the microchannel, and direct them toward the detection region. The detection regions consist of three interdigitated electrode arrays (IDEA), each with 20 pairs of finger coated with a mixture of anti-Salmonella antibody and crosslinker to enhance the adhesion to IDEA. The impedance changes as the target Salmonella binds to the antibody. The biosensor has showed excellent performance as proven by the detection of a single Salmonella serotype B, and simultaneous detection of two Salmonella serotypes B and D with a limit of detection (LOD) of 8 Cells/ml in ready-to-eat turkey samples, the addition of focusing capability improved the measured signal by a factor of between 4-4.5, the total detection time of 45 minutes, selectivity of the sensor on different types of bacterial cells, and the ability to distinguish between dead and live cells.

A rapid and highly specific immunofluorescence method to detect Escherichia coli O157:H7 in infected meat samples.

Developing rapid and sensitive methods for the detection of pathogenic Escherichia coli O157:H7 remains a major challenge in food safety. The present study attempts to develop an immunofluorescence technique that uses Protein-A-coated, magnetic beads as the platform. The immunofluorescence technique described here is a direct detection method in which E. coli O157:H7 cells are labeled with tetramethylrhodamine (TRITC) fluorescent dye. TRITC-labeled bacteria are captured by the desired antibody (Ab), which is immobilized on the Protein-A magnetic beads. Fluorescence of the captured cells is recorded in a fluorescence spectrophotometer, where the fluorescence values are shown to be directly proportional to the number of bacteria captured on the immunobead. The formation of an immunocomplex is evidenced by the fluorescence of the beads under microscopy. The Ab immobilization procedure is also evidenced by microscopy using fluorescein isothiocyanate (FITC)-labeled Ab. The total experimental time, including preparation of the sample, is just 1h. The minimum bacterial concentration detected by this method is 1.2±0.06×10(3)CFUml(-1). The high specificity of this method was proved by using the specific monoclonal Ab (MAb) in the test. The proposed protocol was successfully validated with E. coli O157:H7-infected meat samples. This approach also opens the door for the detection of other bacterial pathogens using Protein-A magnetic beads as a detection platform.