Predictive Modeling of Oxygen Transmission Through Micro-Perforations for Packaging Applications

Methods for creating precise perforations in respiring produce packaging are being increasingly adopted. Knowledge of oxygen transfer through perforated packaging and oxygen distribution in packages is necessary for successful packaging design of fresh produce. An approach to modeling perforated packaging performance was developed using a cylindrical chamber with precision perforations using Fick’s second law. The model was simulated using two techniques including Finite Element Method (FEM) using commercially available software and Finite Volume Method (FVM) through programming. Perforations were approximated as a source term in the second method. Both simulation techniques showed trends similar to experimental data

Actuation of chitosan-aptamer nanobrush borders for pathogen sensing

We demonstrate a sensing mechanism for rapid detection of Listeria monocytogenes in food samples using the actuation of chitosan-aptamer nanobrush borders. The bio-inspired soft material and sensing strategy mimic natural symbiotic systems, where low levels of bacteria are selectively captured from complex matrices. To engineer this biomimetic system, we first develop reduced graphene oxide/nanoplatinum (rGO-nPt) electrodes, and characterize the fundamental electrochemical behavior in the presence and absence of chitosan nanobrushes during actuation (pH-stimulated osmotic swelling). We then characterize the electrochemical behavior of the nanobrush when receptors (antibodies or DNA aptamers) are conjugated to the surface. Finally, we test various techniques to determine the most efficient capture strategy based on nanobrush actuation, and then apply the biosensors in a food product. Maximum cell capture occurs when aptamers conjugated to the nanobrush bind cells in the extended conformation (pH \u3c 6), followed by impedance measurement in the collapsed nanobrush conformation (pH \u3e 6). The aptamer-nanobrush hybrid material was more efficient than the antibody-nanobrush material, which was likely due to the relatively high adsorption capacity for aptamers. The biomimetic material was used to develop a rapid test (17 min) for selectively detecting L. monocytogenes at concentrations ranging from 9 to 107 CFU-mL-1 with no pre-concentration, and in the presence of other gram-positive cells (Listeria innocua and Staphylococcus aureus). Use of this bio-inspired material is among the most efficient for L. monocytogenes sensing to date, and does not require sample pretreatment, making nanobrush borders a promising new material for rapid pathogen detection in food

Review of Challenges and Advances in Modification of Food Package Headspace Gases

Modified Atmosphere Packaging (MAP) has been widely used as an effective way to preserve foods. Fresh produce, meat and meat products, seafood, and dairy products can benefit from modified gaseous atmospheres, which are usually achieved by reducing oxygen and increasing carbon dioxide concentrations, within limits, defined by product tolerances. MAP of fresh produce is particularly challenging because products are living and respiring. Respiration rates depend on several factors including temperature, oxygen, and carbon dioxide concentrations. Balancing package permeation with respiration is challenging, often due to limited selection of practical packaging materials. Failing to remain within tolerance limits of products leads to rapid quality loss. Gas barrier properties of packages determined rate of gas exchange with the external environment and is a critical factor for achieving tolerable levels. Availability of packaging materials that meet requirement of specific produce is essential. Relative permeability of common films to carbon dioxide is about 3 to 6 times of that to oxygen, often leading to package collapse for package atmospheres that benefit from carbon dioxide. Films often fail to provide desired oxygen transmission rates, high carbon dioxide to oxygen selectivity and desired mechanical properties simultaneously. Despite advances, minimal availability and high cost of selective barrier films limit applications of MAP for fresh produce packaging. Therefore, active packaging components and films are being developed and designed to overcome these limitations. Inserts or films that contain active mixtures as gas emitters and/or scavengers are now commercially available. “Clean label” trends are motivating alternative approaches using active packaging components

Effect of Red Cabbage Extract on Minced Nile Perch Fish Patties Vacuum Packaged in High and Low Oxygen Barrier Films

Oxidation of polyunsaturated fatty acids (PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in fish causes loss of product quality. Oxidative rancidity causes loss of nutritional value and undesirable color changes. Therefore, powerful antioxidant extracts may provide a relatively low cost and natural means to reduce oxidation, resulting in longer, higher quality and higher value shelf life of foods. In this study, we measured synergistic effects of red cabbage antioxidant and vacuum packaging on lipid oxidation in fresh tilapia patties using thiobarbituric acid reactive substances (TBARS) assay, peroxide value (PV), pH and color analysis. Concentrated red cabbage extract was obtained using an efficient freeze/thawed method developed in our laboratory (citation). Fresh tilapia patties were prepared with solutions containing 68 ppm of extract concentrate for each 50 gr of fish patties. Samples were stored for 15 days at refrigeration conditions (4±1°C) and analyzed interval between two days for pH, color analysis, and lipid oxidation assessments. Results show that treated and vacuum packaged samples had lower oxidation levels than controls. Lipid peroxide values on treated samples showed benefits through day 12. This work shows that synergistic effect of red cabbage antioxidant extracts and vacuum packaging may represent an inexpensive and natural method for retarding oxidative spoilage of fresh fish

Laser Scribed Graphene Biosensor for Detection of Biogenic Amines in Food Samples Using Locally Sourced Materials

In foods, high levels of biogenic amines (BA) are the result of microbial metabolism that could be affected by temperatures and storage conditions. Thus, the level of BA is commonly used as an indicator of food safety and quality. This manuscript outlines the development of laser scribed graphene electrodes, with locally sourced materials, for reagent-free food safety biosensing. To fabricate the biosensors, the graphene surface was functionalized with copper microparticles and diamine oxidase, purchased from a local supermarket; and then compared to biosensors fabricated with analytical grade materials. The amperometric biosensor exhibits good electrochemical performance, with an average histamine sensitivity of 23.3 µA/mM, a lower detection limit of 11.6 µM, and a response time of 7.3 s, showing similar performance to biosensors constructed from analytical grade materials. We demonstrated the application of the biosensor by testing total BA concentration in fish paste samples subjected to fermentation with lactic acid bacteria. Biogenic amines concentrations prior to lactic acid fermentation were below the detection limit of the biosensor, while concentration after fermentation was 19.24 ± 8.21 mg histamine/kg, confirming that the sensor was selective in a complex food matrix. The low-cost, rapid, and accurate device is a promising tool for biogenic amine estimation in food samples, particularly in situations where standard laboratory techniques are unavailable, or are cost prohibitive. This biosensor can be used for screening food samples, potentially limiting food waste, while reducing chances of foodborne outbreaks

Flexible Laser-Induced Graphene for Nitrogen Sensing in Soil

Flexible graphene electronics are rapidly gaining interest, but their widespread implementation has been impeded by challenges with ink preparation, ink printing, and post-print annealing processes. Laser-induced graphene (LIG) promises a facile alternative by creating flexible graphene electronics on polyimide substrates through a one-step laser writing fabrication method. Herein we demonstrate the use of LIG, created through a low-cost UV laser, for electrochemical ion selective sensing of plant-available nitrogen (i.e., both ammonium and nitrate ions: NH4+ and NO3-) in soil samples. The laser used to create the LIG was operated at distinct pulse rates (10, 20, 30, 40, and 50 ms) in order to maximize the LIG electrochemical reactivity. Results illustrated that a laser pulse rate of 20 ms led to a high percentage of sp2 carbon (77%) and optimal peak oxidation current of 120 uA during ferricyanide cyclic voltammetry. Therefore, LIG electrodes created with a 20 ms pulse rate were consequently functionalized with distinct ionophores specific to NH4+(nonactin) or NO3- (tridodecylmethylammonium nitrate) within polyvinyl chloride (PVC)-based membranes to create distinct solid contact ion selective electrodes (SC-ISEs) for NH4+ and NO3-ion sensing respectively. The LIG SC-ISEs displayed near Nernstian sensitivities of 51.7 ± 7.8 mV/decade (NH4+) and -54.8 ± 2.5 mV/decade (NO3-), detection limits of 28.2 ± 25.0 uM (NH4+) and 20.6 ± 14.8 uM (NO3-), low long-term drift of 0.93 mV/hr (NH4+) sensors and -5.3 μV/hr (NO3-) sensors and linear sensing ranges within 10-5-10-1 M for both sensors. Moreover, soil slurry sensing was performed and recovery percentages of 96% and 95% were obtained for added NH4+and NO3-, respectively. These results, combined with a facile fabrication that does not require metallic nanoparticle decoration, make these LIG electrochemical sensors appealing for a wide range of in field or point-of-service applications for soil health management

Post hoc support vector machine learning for impedimetric biosensors based on weak protein-ligand interactions

Impedimetric biosensors for measuring small molecules based on weak/transient interactions between bioreceptor and target analyte are a challenge for detection electronics, particularly in field studies or in analysis of complex matrices. Protein-ligand binding sensors have enormous potential for biosensing, but accuracy in complex solutions is a major challenge. There is a need for simple post hoc analytical tools that are not computationally expensive, yet provide near real time feedback on data derived from impedance spectra. Here, we show use of a simple, open source support vector machine learning algorithm for analyzing impedimetric data in lieu of using equivalent circuit analysis. We demonstrte two different protein-based biosensors to show that the tool can be used for various applications. We conclude with a mobile phone-based demonstration focused on measurement of acetone, an important biomarker related to onset of diabetic ketoacidosis. In all conditions tested, the open source classifier was capable of performing as well, or better, than equivalent circuit analysis for characterizing weak/transient interactions between a model ligand (acetone) and a small chemosensory protein derived from tsetse fly. In addition, the tool has a low computational requirement, facilitating use for mobile acquisition systems such as mobile phone. The protocol is deployed through Jupyter notebook (an open source computing environment available for mobile phone, tablet, or computer use) and the code was written in Python. For each of the applications we provide step-by-step instructions in English, Spanish, Mandarin, and Portuguese to facilitate widespread use. All codes were based on scikit-learn, an open source software machine learning library in the Python language, and were processed in Jupyter notebook, an open-source web application for Python. The tool can easily be integrated with mobile biosensor equipment for rapid detection, facilitating use by a broad range of impedimetric biosensor users. This post hoc analysis tool can serve as a launchpad for convergence of nanobiosensors in planetary health monitoring applications based on mobile phone hardware

Planar Interdigitated Aptasensor for Flow-Through Detection of Listeria spp. in Hydroponic Lettuce Growth Media

Irrigation water is a primary source of fresh produce contamination by bacteria during the preharvest, particularly in hydroponic systems where the control of pests and pathogens is a major challenge. In this work, we demonstrate the development of a Listeria biosensor using platinum interdigitated microelectrodes (Pt-IME). The sensor is incorporated into a particle/sediment trap for the real-time analysis of irrigation water in a hydroponic lettuce system. We demonstrate the application of this system using a smartphone-based potentiostat for rapid on-site analysis of water quality. A detailed characterization of the electrochemical behavior was conducted in the presence/absence of DNA and Listeria spp., which was followed by calibration in various solutions with and without flow. In flow conditions (100 mL samples), the aptasensor had a sensitivity of 3.37 ± 0.21 kΩ log-CFU−1 mL, and the LOD was 48 ± 12 CFU mL−1 with a linear range of 102 to 104 CFU mL−1. In stagnant solution with no flow, the aptasensor performance was significantly improved in buffer, vegetable broth, and hydroponic media. Sensor hysteresis ranged from 2 to 16% after rinsing in a strong basic solution (direct reuse) and was insignificant after removing the aptamer via washing in Piranha solution (reuse after adsorption with fresh aptamer). This is the first demonstration of an aptasensor used to monitor microbial water quality for hydroponic lettuce in real time using a smartphone-based acquisition system for volumes that conform with the regulatory standards. The aptasensor demonstrated a recovery of 90% and may be reused a limited number of times with minor washing step

Rapid and Label-Free Detection of Interferon Gamma via an Electrochemical Aptasensor Comprising a Ternary Surface Monolayer on a Gold Interdigitated Electrode Array

A label-free electrochemical impedance spectroscopy (EIS) aptasensor for rapid detection (\u3c35 \u3emin) of interferon-gamma (IFN-γ) was fabricated by immobilizing a RNA aptamer capture probe (ACP), selective to IFN-γ, on a gold interdigitated electrode array (Au IDE). The ACP was modified with a thiol group at the 5′ terminal end and subsequently co-immobilized with 1,6-hexanedithiol (HDT) and 6-mercapto-1-hexanolphosphate (MCH) to the gold surface through thiol–gold interactions. This ACP/HDT-MCH ternary surface monolayer facilitates efficient hybridization with IFN-γ and displays high resistance to nonspecific adsorption of nontarget proteins [i.e., fetal bovine serum (FBS) and bovine serum albumin (BSA)]. The Au IDE functionalized with ACP/HDT-MCH was able to measure IFN-γ in actual FBS solution with a linear sensing range from 22.22 pM to 0.11 nM (1–5 ng/mL) and a detection limit of 11.56 pM. The ability to rapidly sense IFN-γ within this sensing range makes the developed electrochemical platform conducive toward in-field disease detection of a variety of diseases including paratuberculosis (i.e., Johne’s Disease). Furthermore, experimental results were numerically validated with an equivalent circuit model that elucidated the effects of the sensing process and the influence of the immobilized ternary monolayer on signal output. This is the first time that ternary surface monolayers have been used to selectively capture/detect IFN-γ on Au IDEs

Sense–Analyze–Respond–Actuate (SARA) Paradigm: Proof of Concept System Spanning Nanoscale and Macroscale Actuation for Detection of Escherichia coli in Aqueous Media

Foodborne pathogens are a major concern for public health. We demonstrate for the first time a partially automated sensing system for rapid (~17 min), label-free impedimetric detection of Escherichia coli spp. in food samples (vegetable broth) and hydroponic media (aeroponic lettuce system) based on temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) nanobrushes. This proof of concept (PoC) for the Sense-Analyze-Respond-Actuate (SARA) paradigm uses a biomimetic nanostructure that is analyzed and actuated with a smartphone. The bio-inspired soft material and sensing mechanism is inspired by binary symbiotic systems found in nature, where low concentrations of bacteria are captured from complex matrices by brush actuation driven by concentration gradients at the tissue surface. To mimic this natural actuation system, carbon-metal nanohybrid sensors were fabricated as the transducer layer, and coated with PNIPAAm nanobrushes. The most effective coating and actuation protocol for E. coli detection at various temperatures above/below the critical solution temperature of PNIPAAm was determined using a series of electrochemical experiments. After analyzing nanobrush actuation in stagnant media, we developed a flow through system using a series of pumps that are triggered by electrochemical events at the surface of the biosensor. SARA PoC may be viewed as a cyber-physical system that actuates nanomaterials using smartphone-based electroanalytical testing of samples. This study demonstrates thermal actuation of polymer nanobrushes to detect (sense) bacteria using a cyber-physical systems (CPS) approach. This PoC may catalyze the development of smart sensors capable of actuation at the nanoscale (stimulus-response polymer) and macroscale (non-microfluidic pumping)