Source: NANOGUARD TECHNOLOGIES, LLC submitted to
HVACP PROCESSING OF ALMONDS TO REDUCE AFLATOXIN
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1016776
Grant No.
2018-33610-28949
Cumulative Award Amt.
$599,997.00
Proposal No.
2018-03207
Multistate No.
(N/A)
Project Start Date
Sep 1, 2018
Project End Date
Aug 31, 2022
Grant Year
2018
Program Code
[8.5]- Food Science & Nutrition
Recipient Organization
NANOGUARD TECHNOLOGIES, LLC
1100 CORPORATE SQUARE DR STE 229
SAINT LOUIS,MO 63132
Performing Department
(N/A)
Non Technical Summary
High Voltage Atmospheric Cold Plasma (HVACP) treatment of agricultural products is an emerging non-thermal processing (NTP) technology that effectively reduces aflatoxins (AF) and microorganisms on almonds, other tree nuts and peanuts. HVACP-based NTP provides enhanced food safety by reducing incidences of food-borne illness. It is a cost-effective and environmentally friendly processing technology that addresses several NIFA National Challenge Areas. These include: 1) Global food security and hunger alleviation by increased protection of agricultural products from disease and pests and reduced food losses; 2) Low to no impact on climate change by providing an environmentally friendly, low energy and low carbon footprint alternative to chemical and thermal processes; and 3) Food safety by reducing the incidence of food-borne illnesses and death through an improved food processing technology. Industry adoption of this novel NTP technology will provide higher quality almonds, other tree nuts and peanuts to the consumers and increase global demand for U.S. almonds and other tree nuts.Atmospheric cold plasma is a natural phenomenon that has been harnessed and adapted for industrial purposes. The first atmospheric, low temperature plasma was invented by Werner von Siemens in 1857 (Siemens, 1857). Air was passed between two parallel plates containing an electrical spark (corona discharge) to generate ozone. Plasma can be generated in any gas environments (e.g. air, O2, He, N2, Ar, Ne, CO2) with either DC or AC current (Roth, 1995). HVACP capitalizes on the chemical properties of gases and the physics of electric field. When a gas is subjected to a voltage gradient of adequate strength; electrons can be stripped from the gas molecules (ionization). These electrons react with the bulk gas generating light, UV, and unique reactive gas species (ozone, oxides of nitrogen, peroxides, atomic oxygen, etc.), many of which can alter mycotoxins on the surface of almonds, other tree nuts, peanuts as well as on maize, wheat etc. and render mycotoxins non-toxic. In the 2017 USDA-NIFA-SBIR-005943 Phase I grant, NanoGuard demonstrated that a non-thermal processing using HVACP can significantly reduce aflatoxins on almonds without impacting quality. NanoGuard has also demonstrated that the reactive gas species (RGS) in HVACP have excellent bactericidal properties and they could therefore inactivate bacteria, fungi and mold on tree nuts, peanuts, corn, etc. as an added benefit.Mycotoxins are secondary metabolites produced by specific molds and fungi that are toxic to humans and livestock at low part per billion (ppb) levels (Pankaj et al., 2018; Sarangapani et al., 2018; Shi et al.,2017; Bourke et al., 2017;). These mycotoxins are responsible for a range of health issues that have tremendous economic impact in both the human and animal nutrition industries. The Food and Agriculture Organization (FAO) has estimated that 25% of the world's crops are affected by mycotoxins each year, with annual losses of around one billion metric tons of food and food products. Economic losses occur because of: 1) yield loss due to diseases induced by toxigenic fungi; 2) reduced crop value resulting from mycotoxin contamination; 3) losses in animal productivity from mycotoxin-related health problems; and 4) human health costs. Additional costs associated with mycotoxins include the cost of management at all levels - prevention, sampling, analysis, mitigation, litigation, and research costs. Significant economic losses resulting from mycotoxins are well documented along the food and feed supply chains: crop producers, animal producers, distributors, processors, consumers, and society as a whole (due to health care impacts and productivity losses). Estimates of the costs of mycotoxins in the United States vary: one report estimated $0.5 to $1.5 billion/year (Council for Agricultural Science and Technology, 2003) and another estimated $5 billion/year for the U.S. and Canada (WHO/FAO, 2002).The highest valued tree nut crop in the United States is almonds, generating $6.4 billion in 2014 with a total production of approximately 2 billion pounds (USDA ERS, 2015). The initial subjective forecast for the 2017 California almond production is 2.2 billion pounds (USDA NASS, 2017). Mycotoxin contamination in almonds, primarily aflatoxins, is a constant concern. Aflatoxins are produced by the fungi Aspergillus flavus. Good agricultural practices in tree nuts are recommended for prevention of contamination and growth of Aspergillus flavus, such as keeping product dry during storage. However, aflatoxin production can occur seasonally to varying degrees and is most problematic under wet harvesting conditions or periods of heavy rain when air humidity is high, leading to favorable growth conditions. The tree nut as well as the peanut industries are looking for a simple, cost effective and environmentally friendly solution for reducing mycotoxins on nuts without compromising product integrity. NanoGuard's HVACP-based non-thermal processing technology is a perfect solution to mitigate the post-harvest mycotoxin problem facing the almond industry, in particular, and tree nut as well as peanut industry in general.There is an increasing global demand for food that is fresh, free of pathogens and toxins, and has longer shelf-life. Consumer attention to nutrition, especially awareness and concerns of the potential hazards of preservatives and additives during processing, is also fueling demand for minimally processed food that is safe but retains high nutritional value, texture, and flavor. The recent reports on microbial contaminated food has significantly elevated consumer awareness on food safety. Because non-thermal processing through HVACP reduces mycotoxins and microorganisms of agricultural products, it must be of public interest to bring this technology to commercialization. HVACP-based non-thermal processing assures global food security and reduce hunger by increased protection of agricultural products from disease and pests and reduced food losses. This post-harvest processing technology is cost-effective and environmentally friendly, and it will promote the export of U.S. agricultural products, bringing prosperity to farmers and growers. With the help of this state-of-the-art technology, U.S. tree nut growers and exporters can stay competitive in the global market place by reducing the losses from aflatoxin contamination.
Animal Health Component
90%
Research Effort Categories
Basic
5%
Applied
90%
Developmental
5%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31412122020100%
Goals / Objectives
Technical Objectives1. Build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how.2. Perform 60-minutes treatment of almonds in the newly built 450-kg HVACP system. a. Examine the quality of the above HVACP-treated almonds. b. Examine almond oil quality of HVACP-treated almonds. c. Conduct sensory evaluation of HVACP-treated almonds.3. Examine toxicity of HVACP-treated aflatoxin on almonds.4. Implement intellectual property strategy.5. Develop regulatory approval pathway.6. Implement market and business development plan (commercialization plan for almonds).7. Expand the business opportunities for HVACP technology beyond almonds.
Project Methods
NanoGuard's HVACP technology can deliver significant aflatoxin reduction on almonds and the technology can be scaled to larger treatment volumes without restriction. In Phase II, further development of the technology will occur in constructing a treatment system to treat 450 kg of almonds for sixty minutes. NanoGuard will work with several vendors to design and build a commercial grade HVACP system that can treat 450 kg (1000 × scale-up of the Phase I HVACP system) of almonds per hour for aflatoxin reduction. To build this system, NanoGuard will use its proprietary knowledge and expertise along with its partners to scale-up the electrodes, plasma generator, and contactor developed during the Phase I research. The system will be tested for efficacy and robustness. Operational data from the scaled-up system will be used to develop a techno-economic model (TEM) for the treatment of almonds for aflatoxin reduction. The purpose of the TEM is to determine an operating point or a set of operating points that fall inside an operating envelope of the HVACP process that provides a cost to treat that makes it commercially viable.Approximately 450 kg of almonds will be treated for 60 minutes in this scaled-up commercial grade HVACP system for aflatoxin reduction. The treated almonds and almond oil derived from the treated product will be examined for quality and sensory attributes to be consistent with commercial standards. Analysis of the air above almonds in storage bins (head-space analysis) is commonly used to assess the level of rancidity of almonds during storage. Rancidity causes nuts to have an unpleasant smell or taste. It is due to hydrolysis and/or autoxidation of fats into short-chain aldehydes and ketones which are objectionable in taste and odor. When these processes occur in almonds, undesirable odors and flavors can result. Head-space analysis is an analytical method used to evaluate the rancidity of food oil. The same method will be conducted on HVACP-treated and untreated almonds soon after treatment and during storage to assess the effect (if any) of HVACP treatment on rancidification of almonds. The current plan is to conduct the headspace analysis in an outside lab who does this routinely for the edible oils with guidance from scientists at the Almond Board of California (ABC). Almonds after treatment will be extracted using hexane and the oil will be analyzed for peroxide value (PV), anisidine value (AV), totox value, acid value, thiobarbituric acid (TBA) value, iodine value (IV) and fatty acid composition for oil quality. Peroxide, anisidine, totox and thiobarbituric acid values are indicators of fatty acid oxidation.For conducting the sensory testing, NanoGuard will identify a set of key sensory attributes that are relevant to almond industry as part of the testing protocol. NanoGuard will seek guidance from ABC in identifying the sensory attributes. Appropriate experimental design and statistical method will be used to generate and interpret the sensory data, and to assess the effect of HVACP treatment, if any, on the quality and integrity of almonds. Once satisfactory analytical measures are found from HVACP treatment of almonds, a taste testing of the HVACP-treated almonds by a trained sensory panel will be initiated. This sensory testing will be overseen by Dr. Keener and will be used to evaluate the taste, aroma, appearance, flavor and texture characteristics of both HVACP-treated and untreated almonds immediately after treatment and after 28 days of storage.The analytical methods (HPLC and ELISA) that NanoGuard have used for the efficacy of HVACP treatment showed only the level of aflatoxin reduction on almonds due to treatment. These methods measured the quantity of aflatoxin left on almonds after treatment; but did not confirm that the treatment rendered aflatoxin non-toxic or whether the degradants of aflatoxin resulted from HVACP treatment are non-toxic. To confirm that the aflatoxin degradants from HVACP treatment are non-toxic, NanoGuard will develop an invivo toxicology model system using brine shrimp (Artemia). Brine shrimp have been extensively used to evaluate the toxicity in pollution research as a test organism and it is an acceptable alternative to the toxicity testing of mammals in the laboratory. The fact that millions of brine shrimp are so easily reared has been an important help in assessing the effects of a large number of samples on the shrimps under well controlled experimental conditions. Regulatory agencies around the world consider the brine shrimp toxicology results acceptable in environmental safety assessment of new products and/or processes.

Progress 09/01/21 to 08/31/22

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? One of the main objectives of the project is to build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how. The RGS generator, a key component of the HVACP system, was delivered in early 2020 and was later installed at NanoGuard Technologies Inc. in St. Louis. The new system was tested extensively to confirm the scaleup parameters, which included the treatment of artificially contaminated almonds. The treatment of aflatoxincontaminated almonds similar to the Phase I work confirmed that the new system was scaling up as expected.

Publications


    Progress 09/01/18 to 08/31/22

    Outputs
    Target Audience:NanoGuard is working with a major US grain and oil seed processors for commercialization of HVACP for microorganism and mycotoxin reduction. We have NDAs in place with several companies to develop the technology for reducing mycotoxin on almonds, peanuts, and hazelnuts. HVACP technology can be tailored for many applications in the food and feed industry for reducing mycotoxin and microorganisms. To date more than 50 food and feed companies including multinationals have approached NanoGuard to explore the possible use of HVACP in their respective businesses. Several of these companies have initiated feasibility studies with NanoGuard to see if HVACP technology solves microorganism and mycotoxin issues in their products and processes. Changes/Problems:Agricultural commodities such as corn, peanuts, almonds etc. show a high degree of non-uniformity in aflatoxin contamination due to the uneven presence of the aflatoxin on them. Because of high aflatoxin non-uniformity, obtaining reliable control and test samples that have the same relative initial aflatoxin contamination is near impossible. Several attempts at homogenizing the almonds for aflatoxin occurred during the early stages of testing. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? One of the main objectives of the project was to build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how. The plasma or Reactive Gas Species (RGS) generator, a key component of the HVACP system was fabricated at Phenix Technologies (Accident, MD) and delivered to NanoGuard in early 2020. The plasma generator was subsequently installed at NanoGuard Technologies Inc. in St. Louis and tested for performance and other operational parameters. The other part of the HVACP system is the contactor where RGS exiting out of the HVACP generator is immediately allowed to contact the almonds. A fluid bed type of contactor was connected to the plasma generator and was used for treating almonds. The plasma generator consists of a series of epoxy encased metal electrodes that are placed in an electrode holder so that they are parallel to one another at a specified gap. The top and bottom surfaces of the electrodes are covered with glass to provide a durable non oxidative, inert surface for plasma generation. The industrially hardened small commercial unit has a gap of 1.5" between the electrodes, and the gap determines the operating potential necessary to breakdown or ionize the air passing between the electrodes in the gap. For this gap size, the operating potential is approximately 83kV. The electrodes slide into the holder and alternate in the stack, with one being at a positive potential and the adjacent electrode being at a negative potential. The holder is fabricated from a special dielectric material. The walls of the holder and the positive and negative electrode form a plenum through which gas is transported at a certain velocity and is ionized when the high voltage is on (Figure 1). At the top and the bottom of the high voltage electrode stack are ground electrodes. These are identical in construction to the high voltage electrodes but are connected to earth ground to stabilize the electrical field in the plasma generator. NanoGuard completed all the proposed USDA-NIFA-SBIR-006824 grant work. The 450-kg capacity HVACP system needed to treat almonds for aflatoxin reduction was constructed with the help of Phenix Technologies and the device was installed at NanoGuard Technologies Inc. in St. Louis., MO. Approximately 1600 lbs. of aflatoxin-contaminated almonds were procured for HVACP treatment through Summation Lab (California).

    Publications


      Progress 09/01/20 to 08/31/21

      Outputs
      Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

      Impacts
      What was accomplished under these goals? One of the main objectives of the project is to build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how. The RGS generator, a key component of the HVACP system, was delivered in early 2020 and was later installed at NanoGuard Technologies Inc. in St. Louis. The new system was tested extensively to confirm the scaleup parameters, which included the treatment of artificially contaminated almonds. The treatment of aflatoxin-contaminated almonds similar to the Phase I work confirmed that the new system was scaling up as expected.

      Publications


        Progress 09/01/19 to 08/31/20

        Outputs
        Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

        Impacts
        What was accomplished under these goals? One of the main objectives of the project is to build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how. The RGS generator, a key component of the HVACP system, was delivered in early 2020 and was later installed at NanoGuard Technologies Inc. in St. Louis. The new system was tested extensively to confirm the scale-upparameters, which included the treatment of artificially contaminated almonds. The treatment of aflatoxin-contaminated almonds similar to the Phase I work confirmed that the new system was scaling up as expected.

        Publications


          Progress 09/01/18 to 08/31/19

          Outputs
          Target Audience:NanoGuard continues to have discussions with companies in the following industries: Pet Care, Spices, Coffee, Fresh and Dry Fruits, Fresh Vegetable, Supply Chain Management, Global Corn Wheat and Soy, Flour, Tree Nuts, Ground Nuts, Seed Markets, and Ethanol. Without exception, each of these companies continue to show significant interest in pursuing our technology for mycotoxin and microorganism reduction and some of them have already initiated proof-of-concept work with NanoGuard for their specific applications. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

          Impacts
          What was accomplished under these goals? One of the main objectives of the project is to build and test a 450-kg capacity HVACP system for treating almonds based on Phase I results and know-how. The RGS generator, a key component of the HVACP system, was delivered in early 2020 and was later installed at NanoGuard Technologies Inc. in St. Louis. The new system was tested extensively to confirm the scale-up 2 parameters, which included the treatment of artificially contaminated almonds. The treatment of aflatoxin-contaminated almonds similar to the Phase I work confirmed that the new system was scaling up as expected.

          Publications