Recipient Organization
TENNESSEE STATE UNIVERSITY
3500 JOHN A. MERRITT BLVD
NASHVILLE,TN 37209
Performing Department
(N/A)
Non Technical Summary
Antibiotic resistance is becoming a major health issue in the United States. Countries in the worldare striving for ways to eliminate the misuse of antibiotics in feed as growth promoters and illnessprevention intervene in livestock production. Essential oils and polyphenols are natural compoundsthat can be used as antibiotic alternatives, but their use is limited due to their sensitivity to heat andlight, and harsh flavor and taste. Microencapsulation is a promising approach to protect thesecomponents and to deliver them to the target site in the intestinal tract. This proposal fillsknowledge gaps critical to the development of antibiotic alternatives that will enhance food safety,improve poultry performance through modified intestinal microflora, and eliminate antibioticabuse. The project is designed in a series of logical, sequential objectives supported by an industrialcollaborator. The specific objectives are: 1). Optimize microencapsulation formula and methods byusing novel ingredients as wall materials; 2). Investigate the delivery efficiency and releasingprofile using in vitro and in vivo methods; 3). Evaluate poultry intestinal microflora and growthperformance after feeding micro particles through diet. This project fits into the institutional planand confronts a local and national priority, conceiving, designing and evaluating new bioprocessingtechniques in Food Safety. This project addresses the CBG Program Priority Areas, USDAPriorities and NIFA Challenge Areas in Food Safety. Outcomes yielded from the grant willstrengthen the research and teaching capacity in 1890 universities.
Animal Health Component
50%
Research Effort Categories
Basic
(N/A)
Applied
50%
Developmental
50%
Goals / Objectives
Each year in the United States, at least 2 million people become infected with bacteria that are resistant to antibiotics, and at least 23,000 people die each year as a direct result of these infections (Center for Disease Control and Prevention, 2018). The widespread use of antibiotics on both animals and humans has contributed to the microbial resistance to antibiotics. Since January 2017, the FDA has restricted the use of some antibiotics as growth promoters, a practice that leads to antibiotic-resistant bacteria that pose threats to human health (FDA 2018). In awareness of this crisis, the consumers and markets are demanding antibiotic-free products. The world's biggest burger chain, McDonald's, announced that they will implement a new program of only sourcing chickens raised without antibiotics that are important to human medicine (USA Today, March 4, 2015). KFC announced in April 2017 that it will rid its chicken of antibioticsby the end of 2018. Tyson Foods announced that it will go antibiotic free chicken as well (USA Today, May 2, 2017). As a result, alternatives to antibiotics are in urgent need to improve digestive health and growth performance of livestock.Many natural compounds are proven with antibiotic activities and can be used as alternatives to antibiotics. In combination of encapsulation technology, these natural compounds could be delivered at target location in digestive tract, thus improving the gut health and growth performance of livestock. Therefore, this project will develop an antibiotic alternative to sustain poultry production. This project will enhance the research capacity of 1890 institutions with novel ideas and research capacity to resolve emerging issues, and will equip our underrepresented students with advanced research skills and knowledge to become highly competitive in the fast growing technology market. The proposal has obtained support from international food industry, Archer Daniel Midland (ADM), for providing research materials, and technical support regarding new ingredients in feed industry. This Food Safety research is among the top priorities in the College of Agriculture at Tennessee State University (TSU) (Letter of Support from College Dean). Recently, the Food Sciences Laboratory at TSU has received support from the college, and has acquired a number of cutting edge instruments. This project will bring together multidisciplinary expertise to enhance the collaboration among industrial stakeholders and universities.In this proposal, five major goals will be achieved: 1. Develop an antibiotic alternative product by microencapsulating essential oil and polyphenol mixtures; 2. Evaluate the delivery efficiency, releasing profile and antimicrobial effects by in vivo methods; 3. Explore a novel application of a natural ingredient in encapsulation filed; 4. Promote the application of antibiotic alternatives in poultry production; 5. Build the research capacity in Food and Animal Science Degree program at TSU by collaborating with scientists from nationally and internationally recognized institutions and industrial stakeholders.Specific Objectives are:1) Formulate and optimize microencapsulation for antibiotic alternatives; 2) Investigate the delivery efficiency, releasing profile and effects on bacteria of the microparticle using in vitro methods; 3) Evaluate poultry intestinal microflora and growth performance after feeding encapsulates through poultry diet.Objective 1 will formula of the encapsulation and methods will be optimized base on the releasing profile of essential compounds, and their effects on microbial. Microparticles will be produced using three types of technology, spray-drying, freeze-drying, and encapsulator. Objective 1 includes the following tasks:Task 1.1. Prepare water-soluble yellow mustard mucilage (WSM)Task 1.2. Prepare micro particles using Spray-DryingTask 1.3. Prepare microparticles using EncapsulatorTask 1.4. Prepare microparticles using Freeze-DryerTask 1.5. Evaluate encapsulation efficiency of essential oils and polyphenolsTask 1.6. Particle size measurementObjective 2 will evaluate the effect of micropaticles using in vitro methods in terms of releasing profile, delivery efficiency and effect on microbes. The tasks are listed below:Task 2.1. Digestion of the encapsulated compounds in vitroTask 2.2. Releasing profile Delivery efficiencyTask 2.3. Minimum inhibitory concentration (MIC) for microparticles by in vitro testObjective 3 will Evaluate poultry intestinal microflora and performance after feeding encapsulates through poultry diet. The encapsulated particles will be evaluated in chicken through diet. The study contains three major tasks: 1) microflora evaluation, 2) poultry performance measurement, and 3) delivery efficiency of the microparticles. The experimental plan is listed in Table 3. An antibiotic, clistin, will be used to compare with the encapsulated antibiotic alternatives at various concentrations. The taskes are listed below:Task 3.1 Set up the in vivo experiment: Task 3.2: Retention rate: Task 3.3. Effect of micro particles on intestinal microflora in chickenTask 3.4. Delivery efficiency of encapsulates in intestineTask 3.5. Poultry performance
Project Methods
Methods for objective 1: Formula of the encapsulation and methods will be optimized base on the releasing profile of essential compounds, and their effects on microbial. Microparticles will be produced using three types of technology, spray-drying, freeze-drying, and encapsulator.Statistical Analysis: Each experiment will be repeated three times. Data will be presented as Mean ± SD. Statistical computations will be performed using the GLM procedure of the Statistical Analysis System (SAS Release 9.3, SAS Institute Inc., Cary, NC, USA). The data for encapsulation efficiency and particle size prepared by the 3 different techniques will be examined using ANOVA. Least square means of each of the property factors will be calculated using the option of LSMEANS and statistical differences among the 3 techniques will be identified at p < 0.05 using the option of PDIFF.Methods for objective 2: This objective will evaluate the effect of micropaticles using in vitro methods in terms of releasing profile, delivery efficiency and effect on microbes.Statistical Analysis: Each experiment will be repeated three times. All values will be expressed as Mean ± SD. Mean values will be compared among treatment groups by ANOVA and subsequent Duncan's multiple range test employing SAS software (SAS Version 9.4, SAS Institute Inc., Cary, NC). Differences among means will be considered statistically significant at p < 0.05.Methods for objective 3: The encapsulated particles will be evaluated in chicken through diet. The study contains three major tasks: 1) microflora evaluation, 2) poultry performance measurement, and 3) delivery efficiency of the microparticles. Statistical analysis: All values will be expressed as Mean ± SD. Mean values will be compared among treatment groups by ANOVA and subsequent Duncan's multiple range test employing SAS software (SAS Version 9.4, SAS Institute Inc., Cary, NC). Differences among means will be considered statistically significant at p < 0.05.Ileum and cecal bacterial burden recovered from individual chickens between comparison groups will be evaluated using unpaired t-test by the GraphPad Prism version 6.00 (GraphPad Software, La Jolla, CA, USA, www.graphpad.com). This method can identify over 100 bacteria stains, which will help us to analyze the potential microflora change among treatment groups.Evaluations:1) Each experimental objective is achieved within the planned time frame; 2) Research findings will be disseminated in real time to the general public; 3) The technologies for producing microparticles of antimicrobial alternatives will be shared with other research scientists and poultry growers, feed producers through publications, presentations, educations and collaborations; 4) Up to 7 articles will be published in scientific journals by faculty and students;5) The quality and number of students trained on this project. About 8 students (2 M.S., and 6 B.S.) will complete their dissertation, thesis or senior projects using results generated from this project. Those students must maintain above 3.0 GPA and they will present their research at seminars or conferences.Evaluation Framework: Our project evaluation plan consists of six core assessments and will be monitored by Dr. Reddy, the Dean of the College as indicated by his Letter of Support.1) Self-evaluation within the research team: A quarterly progress report prepared by each PI will be shared with team members. This will allow the team to determine if the objectives are accomplished within the timeframe, and make adjustments in case of pitfalls;2) Institutional evaluation: Every TSU participant will present his (her) research at least once a year to the TSU family, and receive feedback from the faculty and students;3) Evaluation from the industrial partners: We will post project activities on the project website to receive comments from the 'Stake holders';4) Contributions from student research participants: All the junior scientists will submit weekly work reports to their supervisors, and their work will be evaluated by the major professor;5) Input/Project Oversight from ADM and UC Davis: On the second year of the project, Dr. Li from ADM will visit TSU campus to evaluate the project and make recommendations for improvements; Dr. Zhou from UC Davis will visit the TSU campus to give recommendations and technical guidance when the poultry experiment will start (as shown in Letter of Support from Dr. Li and Dr. Zhou),6) Social impact: Developing a tracking system of employment for all students receiving training on this project; citations of the articles and protocols (the top 10 most-read articles) and the visitor times of project websites.