Source: UNIV OF MARYLAND submitted to
SYNERGISTIC INTERACTION BETWEEN ULTRAVIOLET LIGHT AND NOVEL PHOTOSENSILIZERS FOR ENHANCED MICROBIAL SAFETY OF FRESH PRODUCE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
EXTENDED
Funding Source
Reporting Frequency
Annual
Accession No.
1008740
Grant No.
2014-67017-24848
Project No.
2015-11585
Proposal No.
2015-11585
Multistate No.
(N/A)
Program Code
A1361
Project Start Date
Jul 15, 2015
Project End Date
Jul 14, 2018
Grant Year
2016
Project Director
Tikekar, R.
Recipient Organization
UNIV OF MARYLAND
(N/A)
COLLEGE PARK,MD 20742
Performing Department
AGNR-Nutrition and Food Scienc
Non Technical Summary
The overall goal of the proposed research is to investigate synergistic combination of novel, food-grade photosensitizers (fructose and gallic acid) and Ultraviolet light (UV) to improve the bacterial and viral inactivation rate in wash water and fresh produce. The underlying hypothesis of the proposed research is that photosensitizers dissolved in wash water will produce free radicals when exposed to UV (254 nm) light. These free radicals will act synergistically with the UV to enhance microbial inactivation rate. This hypothesis is based on literature and our prior studies. UV processing is an attractive technology for bacterial and viral inactivation. However, its limited depth of penetration and non-homogeneous treatment due to rough and contoured shape of produce has impeded its use in produce industry. Conventional chlorine and hydrogen peroxide based sanitizers used for washing produce have significant limitations such as limited microbial inactivation and ineffectiveness in destroying internalized bacteria and viruses. The proposed approach addresses these limitations. The specific objectives are- (1) Investigate the chemical nature of free radicals generated from UV exposure of fructose and gallic acid and determine the factors that impact their rate of generation, (2) Investigate the efficacy of proposed approach in inactivating bacteria and viruses in liquid medium, produce surface and internalized within produce using novel imaging based approaches and (3) identify the impact of proposed technology on quality of fresh produce. Successful development of this novel technology will lead to a cost effective method for improved sanitation of fresh produce with extended shelf-life.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
5017410200033%
5017410110033%
5017410202034%
Goals / Objectives
Non-thermal food processing technologies have demonstrated potential to inactive pathogens in food systems while maintaining the nutritional and quality attributes of the food and achieving significant efficiency in energy consumption. Among various non-thermal technologies, UV based technologies have made a significant impact on decontamination of processing equipment, process waste water and packaging materials with annual sale of UV decontamination equipment nearing 1 billion dollars. Although UV technology has significant potential for reducing microbial load in food systems, particularly beverage products and fresh produce, its application has been limited due to challenges of low penetration depth of UV light in a food matrix (0.1- 1 cm depending upon nature of food matrix). This research proposal aims to address this significant limitation by evaluating the synergistic combination of novel, food-grade photosensitizers (fructose and gallic acid) and UV-C light (254 nm) to improve the microbial (bacterial and viral) inactivation rate in wash water and fresh produce. The underlying hypothesis of the proposed research is that photosensitizers dissolved in aqueous media will produce free radicals when exposed to UV-C (254 nm). These free radicals will act synergistically along with the UV light to enhance the rate of microbial inactivation in wash water and on surface and within fresh produce. This hypothesis is supported by the current literature, our previous studies and preliminary data that demonstrate that exposure of photosensitizers to UV light results in generation of oxidizing free radicals. Extensive prior work shows that microorganisms are susceptible to death by interaction with free radicals such as hydroxyl, acyl, peroxyl, hydroxyalkyl and superoxide radicals through diverse mechanisms such as damage to nucleic acid, proteins and lipids. In our preliminary study, we showed that fructose accelerated quenching of fluorescein, a free radical sensitive fluorescent dye, in presence of UV light (254 nm) indicating generation of free radicals. Prior studies show that these free radicals could be hydroxyalkyl, acyl, or peroxyl radicals. Another study showed that gallic acid exhibited strong antimicrobial activity exclusively in the presence of UV light, possibly through formation of oxidizing free radicals. Therefore, it is expected that generation of oxidizing free radicals coupled with antimicrobial action of UV itself will increase the rate of microbial inactivation compared to UV processing alone. The specific objectives of this proposed research are-Investigate free radicals generation from novel photosensitizers (fructose and gallic acid) upon exposure to UV light in aqueous solutions and a model 3-dimensional solid systemEvaluate efficacy of simultaneous action of photosensitizers and UV exposure on bacterial and viral inactivation in aqueous medium and fresh produce (surface and internalized)Measure the impact of proposed approach on quality of fresh produce
Project Methods
Objective 1: Investigate generation of free radicals from fructose and gallic acid upon exposure to UV light in aqueous solutions and a model 3 dimensional solid systemUnderstanding the effect of various parameters in wash water such as pH, organic load, presence of metal ions and temperature on generation of free radicals by photosensitizers is critical for translation of optimal sanitation process conditions for fresh produce. This objective addresses following gaps in understanding-(1) what is the chemical nature of free radicals generated by fructose and gallic acid? (b) What factors influence the rate of generation of free radicals in these photosensitizers? and (c) What are the diffusion characteristics of these free radicals in a 3-dimensional matrix to accomplish inactivation of internalized microorganisms?This will be accomplished using a combination of fluorescence and EPR based techniques.Objective 2: Evaluate efficacy of simultaneous action of photosensitizers and UV exposure on bacterial and viral inactivation in aqueous medium and fresh produce.We will investigate synergistic effect of photosensitizers and UV exposure on bacterial and viral inactivation in liquid medium representing wash water and fresh produce. Innovations based on bioluminescence imaging and spectroscopy techniques will be used for rapid enumeration of microorganisms. We propose to use lettuce, basil and parsley as model produce systems. Rationale behind choosing these samples are- (a) These green vegetables and herbs are almost always consumed raw, (b) Basil and parsley are added as seasoning and therefore should not contain any residual off flavors or tastes (as might come from use of hypochlorite in washing) and (c) Significant surface area and delicate structure of leafy greens make removal of microorganism difficult with washing. To chieve this goal conventional microbial techniques such as plate counting and additional fluorescence based novel techniques developed in our lab will be used.Objective 3: Measure the impact of proposed approach on quality of fresh produceSince both UV and free radicals generate oxidative stress in food matrix, it is expected that such oxidative stress can affect product quality as well. This approach will evaluate the extent of impact of proposed approach on product quality. Such understanding will enable optimization of process.Produce will be exposed to their respective fructose concentrations and UV doses capable of accomplishing at least 5-log reduction of surface pathogens and or 3 log reductions in internalized microorganisms. After exposure, the change in these quality parameters of produce immediately after processing and during storage will be evaluated using a combination of chemical anlysis, texture analysis and sensory panel testing

Progress 07/15/16 to 07/14/17

Outputs
Target Audience:Food processing industry professionals working in the area of food safety with an emphasis on fresh produce safety through extension, outreach and workshops Academicians through scientific conferences Graduate students and undergraduate students through research opportunities Changes/Problems:PI has requested and been approved a 1 year no-cost extension due to the delay in hiring process. In the subsequent period we will accomplish the not yet completed tasks. What opportunities for training and professional development has the project provided?There are currently 4 graduate students working on several aspects of theis project (3 PhD and 2 MS) Two postdoctoral research associates were also involved in the project Two short-term undergraduate summer internships were offered to students based on this research How have the results been disseminated to communities of interest?PI also has an extension appointment and has an extension program in food safety. He presented some of his work to the stakeholders (industry) through industry advisory board meeting that was held in connection with another project. PI presented his work to USDA NIFA through PD meeting PI presented his work to scientific community through a conference presentation at IFT. What do you plan to do during the next reporting period to accomplish the goals?-Evaluate the role of technology on wider produce categories -Investigate other phenolic compounds discovered during project for their synergistic interaction with UV-A light -Explore scale-up and commercialization opportunities

Impacts
What was accomplished under these goals? .Demonstrated that gallic acid and its derivaties are effective antimicrobial agents in presence of UV-A light that can improve fresh produce safety without damaging the quality Mechanisms behind the underlying synergistic antimicrobial mechanism was investigated. Fruotse was also shows to be an effective photooxidzer in presence of UV light, although it is not a strong antimicrobial agent in presence of UV-C light. In our research, we found several phenolic compounds similar to Gallic acid that have potential to show synergistic antimicrobial activity not only in presence of UV-A light but also in presence of blue light, heat and high-pressure opening a whole new area of investigation that is attrative and not yet explored.

Publications

  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: On mechanism behind UV-A light enhanced antibacterial activity of gallic acid and propyl gallate against Escherichia coli O157:H7. Scientific Reports, in press
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: De Oliveira, E.; Cossu A.; Tikekar R.; Nitin N. (2017). Enhanced Antimicrobial Activity Based on a Synergistic Combination of Sub-Lethal Levels of Stresses Induced by UV-A Light and Organic Acids. Applied and Environmental Microbiology. 83(11), e00383-17.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Cossu A.; Ercan D.; Wang Q.; Peer W.; Nitin N.; Tikekar R. (2016). Antimicrobial effect of synergistic interaction between UV-A light and Gallic Acid against Escherichia coli O157:H7 in fresh produce wash water and biofilm. Innovative Food Science and Innovative Technologies. 37 (part A), 44-52.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Tikekar R. V., Synergistic interactions between UV light and food-grade compounds to improve microbial inactivation and degrade xenobiotic compounds IFT Annual Meeting and Food Expo, Las Vegas, NV, June 27, 2017.


Progress 07/15/15 to 07/14/16

Outputs
Target Audience:Research scientists from industry and academia working in the area of food safety with an emphasis on produce safety undergraduate, graduate and post-doctoral students Stakeholders such as farmers andfood processors Changes/Problems:-The rate of expenditure was lower than expected because although the reporting period started in July, thefunding arrived in Dec-Jan due to several steps in the administrative process. What opportunities for training and professional development has the project provided?- 4 graduate students ( 2MS and 2PhD) students are working on the several aspects of this project -2 undergraduate students were exposed to the research performed in this project -2 post-doctoral research associates were diretcly involved in this project How have the results been disseminated to communities of interest?-Research publications -Research presentations at national and international venues such as IFT and IUFoSt conferences, -Research presentations at the extension events What do you plan to do during the next reporting period to accomplish the goals?-Evaluate more compounds that can enhance UV induced inactivation of bacteria and viral particles. We are investigating gallic acid, benzoic acid and caffeic acid. -Explore the use of lactic acid and EDTA as treatment enhancers -Evaluate the effect of proposed tehcnologies on produce quality -Explore commercialization potential -disseminate the knowledge through publications, conferences and extension events

Impacts
What was accomplished under these goals? -Demonstrated that UV light and certain food grade chemicals can efficiently interact to improve the inactivation of bacteria and viruses in water used for wahing fresh produce. Inactivation as high as 99.999% was observed. -Developed fundamental understanding of the intactions responsible for this enhanced microbial inactivation -Fresh produce such as spinach exposed to this treatment did not show any adverse effect on its color, tetxure and overall quality

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Fructose Accelerates UV-C Induced Photochemical Degradation of Pentachlorophenol in Low and High Salinity Water. Journal of Agricultural and Food Chemistry. 64(21), 4215.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Antimicrobial Effect of Photosensitized Rose Bengal on Bacteria and Viruses in Model Wash Water. Food and Bioprocess Technology. 9, 441.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Synergistic interaction of ultraviolet light and zinc oxide photosensitizer for enhanced microbial inactivation in simulated wash-water. Innovative Food Science and Emerging Technologies. 33, 240.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Synergistic interaction of ultraviolet light and zinc oxide photosensitizer for enhanced microbial inactivation in simulated wash-water. Chemosphere. 144, 1690.