Sponsoring Institution
National Institute of Food and Agriculture
Project Status
Funding Source
Reporting Frequency
Accession No.
Grant No.
Project No.
Proposal No.
Multistate No.
Program Code
Project Start Date
Sep 1, 2017
Project End Date
May 31, 2018
Grant Year
Project Director
George, C.
Recipient Organization
Performing Department
Non Technical Summary
Non-thermal pasteurization using High Voltage Atmospheric Cold Plasma (HVACP) is expected to reduce mycotoxins and microorganisms on tree nuts resulting in reduced incidences of food-borne illness. Non?thermal pasteurization of food products using HVACP is a novel platform technology which addresses several NIFA National Challenge Areas. These include: 1) Global food security and hunger by increased protection of agricultural products from disease and pests and reduced food waste; 2) Climate change by providing an environmentally friendly and low carbon footprint alternative to chemical and thermal treatments; and 3) Food safety by reducing the incidence of food-borne illnesses and death through an improved food processing technology. Industry adoption of this disruptive platform technology will deliver higher quality food products at reduced cost to consumers, while improving overall food safety and security.
Animal Health Component
Research Effort Categories

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
Goals / Objectives
Goal 1. Demonstrate the feasibility of HVACP technology to reduce mycotoxins on tree nuts.Objective 1: Identify voltage level, gas composition and electrode gap that can achieve 50% reductions in aflatoxin B1 on shelled almonds.The voltage level, gas composition and electrode gap will affect the plasma field intensity resulting in varying reactive gas species and their concentrations. The composition of the inducer gas will directly impact the reactive oxygen species (ROS) and reactive nitrogen species (RNS) generated. Voltage level, gas composition and electrode gap will be investigated to identify 50% aflatoxin B1 reduction on shelled almonds.Objective 2: Demonstrate product stability after HVACP treatment through total fatty acid analysis and peroxide value determination of oil extracted from HVACP-treated almonds.Fatty acid analysis and peroxide value determination will be conducted using published methodsObjective 3: Demonstrate product quality by examining the texture and color of shelled almonds before and after HVACP treatment.Visually analyze the pre- and post- treated almonds to assess if any physical changes have occurred due to HVACP treatment.Goal 2. Conduct in vitro model studies to demonstrate that HVACP deactivates microorganisms.Objective 1: Identify voltage level, gas composition and electrode gap that can achieve 2log10 deactivation of gram positive and gram negative bacteria on filter paper soaked in bacteria culture.Goal 3. Evaluate the commercial viability of HVACP in tree nut treatment by estimating the approximate treatment cost per pound of nuts. Objective 1: Establish critical control parameters to assure consistent treatment results and to confirm that the operating envelope for the treatment is large enough to obtain consistent HVACP treatment results. Objective2: Determine the equipment and capital requirements for installing a commercial scale HVACP system for shelled almond treatment.Goal 4. Develop additional knowledge to strengthen NanoGuard's intellectual property position on HVACP in food safety.Objective 1: Discern patentable information from Phase I work and have consultation with a patent lawyer to see how the outcome of the project can strengthen NanoGuard's patent estate. ?
Project Methods
The focus of this study will be on aflatoxin B1 reduction on shelled almonds. We will also demonstrate that HVACP can reduce microorganisms such as bacteria by conducting in vitro studies using representative gram positive and gram negative bacteria.A lab scale HVACP equipment will be designed and fabricated for the proposed work. Voltage level, gas composition and electrode gap will be adjusted to achieve the desired outcome of 50% reductions in aflatoxin B1 on shelled almonds and 2log10 in vitro deactivation of representative gram positive and gram negative bacteria. Voltage, amperage and wattage measurements will be made in each HVACP treatment experiment to understand the relationship between electrical measurements and the desired outcome of 50% reduction in aflatoxin B1 on shelled almond and in vitro 2 log10 deactivation of bacteria.Aflatoxin B1 analysis of shelled almond before and after HVACP treatment will be conducted using validated ELISA method. Aerobic plate counting will be used to measure the extent of deactivation of bacteria by HVACP.A preliminary evaluation of any impact to product stability due to HVACP treatment of shelled almonds will be conducted by comparing fatty acid composition and peroxide value of oil extracted from shelled almonds before and after treatment.Effect on product quality, if any, due to HVACP treatment of shelled almonds will be assessed based on color and texture changes by visual inspection or even microscopic analysis of whole almonds and nutmeat before and after treatment.

Progress 09/01/17 to 05/31/18

Target Audience:The target audience for NanoGuard's USDA-NIFA funded project "Non-Thermal Pasteurization of Tree Nuts to Reduce Microbes and Mycotoxins" (USDA-NIFA-SBIR-005943) are the U.S. almond producers, The Almond Board of California (ABC), almond exporters, almond distributors and almond consumers. The presence of mycotoxins on almonds, particularly aflatoxin, is a serious concern of almond growers and consumers. Mycotoxins are present on all tree nuts as well as peanuts and they cause detrimental health problems to humans and farm animals even at parts per billion (ppb) levels. Currently, there are no good technologies available to mitigate mycotoxin on tree nuts. Good agricultural practice is the primary intervention available to manage mycotoxins, primarily aflatoxin, contamination. The estimated value of the U.S. tree nut market is approximately 9 billion dollars. It has been estimated that the industry spends approximately $100 million annually for routine analysis and disposal of rejected lots of mycotoxin-contaminated tree nuts. The estimated public health cost for mycotoxin exposure exceeds 2 billion dollars annually. The NIFA-funded almond project (USDA-NIFA-SBIR-005943) demonstrated that NanoGuard's non?thermal pasteurization (NTP) using High Voltage Atmospheric Cold Plasma (HVACP) generated from air deactivates aflatoxin and microorganisms (bacteria, mold and fungi) on almonds. NanoGuard's Non-Thermal Pasteurization using cold plasma from air is safe, effective, cheap and environmentally friendly for reducing aflatoxin on tree nuts and peanuts, which increase their market value, reduce aflatoxin testing and management costs, and provide safer as well as higher quality products to consumers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The USDA-NIFA almond project (USDA-NIFA-SBIR-005943) provided significant professional development opportunities for NanoGuard scientists and several summer interns from Washington University, St. Louis, MO, the University of Notre Dame, Notre Dame, IN and Missouri University of Science and Technology, Rolla, MO. The summer interns were engineering students who got significant exposure to HVACP technology during their internship at NanoGuard. They learned how to fabricate HVACP components, troubleshoot the HVACP system and conduct spectroscopic measurements of reactive gas species. The interns also learned techniques such as Enzyme Linked Immunosorbent Assay (ELISA), almond oil extraction, peroxide value determination of almond oil etc., which engineering students are not generally familiar with. They also had the opportunity to do project-related presentations at team meetings and develop problem solving skills. The summer interns were empowered to solve difficult HVACP-related problems on a day-to-day basis as part of their leadership development skills. Some of these interns were eager to come back and work for NanoGuard in subsequent years to pick up where they left off. Overall the almond project provided unique professional development opportunities to several summer interns as well as veteran NanoGuard scientists, which wouldn't have been possible without the NIFA grant. How have the results been disseminated to communities of interest?NanoGuard shared the NIFA almond project results with several commercial organizations who have vested interest in mitigating aflatoxin on tree nuts and peanuts. The Almond Board of California (ABC) was one such organization who was overly enthusiastic about the outcome of the USDA-NIFA funded almond project. The ABC has been on the lookout for good processes for reducing aflatoxin on almonds without compromising almond quality and integrity. Other commercial organizations such as 'The J. M. Smucker Company, Orrville, OH', 'ADM, Chicago, IL', 'Mars Inc., McLean, VA' etc. were also highly appreciative of the almond results and showed sincere interest in working with NanoGuard to streamline the technology for their respective uses in mitigating aflatoxin and microorganisms on tree nuts and peanuts. We continue to discuss the almond results with U.S. tree nut and peanut companies who are interested in Non-Thermal Pasteurization using High Voltage Atmospheric Plasma for mitigating aflatoxin contamination. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

What was accomplished under these goals? The areas of focus of USDA-NIFA-SBIR-005943 Phase I were: 1) establish the feasibility of NanoGuard's Non-Thermal Pasteurization using High Voltage Atmospheric Cold Plasma (HVACP) to reduce aflatoxin on almonds, 2) confirm that HVACP treatment does not compromise the quality and integrity of almonds, 3) identify critical control parameters in establishing an operating envelop for consistent aflatoxin reduction on almonds, and 4) assess the economic viability of HVACP treatment at a meaningful scale to the almond industry.NanoGuard successfully investigated all four PhaseI focus areas.All the proposed Phase I technical objectives were successfully met within the specified duration of the project. Phase I results showed that NanoGuard's HVACP technology can deliver significant aflatoxin reduction on almonds and the technology can be scaled to larger treatment volumes without restrictions.Phase I results also demonstrated that there are no limitations for treating almonds up to sixty minutes in ambient air with HVACP.In Phase I, NanoGuard identified a working HVACP operating condition for effective treatment of almonds for aflatoxin reduction.Under this operating condition HVACP treatment was found to significantly reduce aflatoxin (60% - 81%) on several 450-gram almond samples.HVACP?treated almonds showed similar oxidative stability as untreated almonds as determined using peroxide values.The fatty acid profile of almond oil extracted from HVACP-treated almonds (60-minute treatment) showed no change compared to oil extracted from untreated almonds.These results demonstrate the potential for building a larger HVACP almond treatment system that could even be portable allowing for in-plant trials at almond producers' storage facilities.Additionally, preliminary work has shown that cold plasma from air is quite effective in reducing microorganisms (>2 log10 reduction) such as bacteria, fungi, and mold.