SYSTEMS-BASED INTEGRATED PROGRAM FOR ENHANCING THE SUSTAINABILITY OF ANTIBIOTIC-RESTRICTED POULTRY PRODUCTION

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: ACTIVE
• Funding Source: AFRI COMPETITIVE GRANT
• Reporting Frequency: Annual
• Accession No.: 1023193
• Grant No.: 2020-69012-31823
• Cumulative Award Amt.: $10,000,000.00
• Proposal No.: 2019-08297
• Project Start Date: Sep 15, 2020
• Project End Date: Sep 14, 2025
• Grant Year: 2020
• Program Code: [A9201]- Sustainable Agricultural Systems

Recipient Organization
UNIV OF CONNECTICUT
438 WHITNEY RD EXTENSION UNIT 1133
STORRS,CT 06269

Performing Department
CAHNR Dean`s Office

Non Technical Summary
The US has the largest broiler industry producing 9 billion birds annually. Antibiotics have been an integral part of poultry production contributing in part to the sustained growth of broiler production, where antibiotic growth promoters (AGP's) are used to enhance poultry performance, feed efficiency and general health. Research indicates that antibiotic-restricted production can adversely affect bird health, production efficiency, and the environment. Therefore, in light of the FDA's directive to phase out AGPs in poultry production, it is critical to ensure sustainability of broiler production for meeting future demand.This proposal's goal is to promote sustainability of broiler production by targeting three major system components in the poultry sector, namely Chicken, Humans and the Environment. To achieve this, we have assembled an interdisciplinary team of researchers, educators and extension professionals with the following objectives: 1) control necrotic enteritis and coccidiosis in broilers; 2) develop novel feed ingredients for improved performance; 3) promote bird welfare; 4) increase broiler meat safety; 5) control antimicrobial resistance dissemination in broiler production; 6) develop strategies for recycling of poultry litter; 7) enhance consumer confidence and acceptability of broiler meat; 8) develop an outreach program targeting poultry producers and other stakeholders; 9) empower the future workforce with skills needed for sustainable broiler production; and 10) assess the socio-economic-environmental impact of proposed interventions.This project will enhance broiler production sustainability by improving bird health, human health and environmental health, and increasing consumer acceptability and economic returns to farmers.

Animal Health Component

50%

Research Effort Categories

Basic

(N/A)

Applied

50%

Developmental

50%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
308	3260	1103	100%

Knowledge Area
308 - Improved Animal Products (Before Harvest);

Subject Of Investigation
3260 - Poultry meat;

Field Of Science
1103 - Other microbiology;

Keywords

antibiotic-restriction

bird health

broiler

environment

human health

performance

sustainability

Goals / Objectives
This proposal's goal is to promote sustainability of broiler production by targeting three major system components in the poultry sector, namely Chicken, Humans and the Environment. The specific objectives include1) Devise novel approaches for controlling necrotic enteritis and coccidiosis in broiler chickens.; 2) Identify novel alternative feed ingredients for enhanced sustainability of broiler production; 3) Develop strategies to promote bird welfare in intensive broiler production; 4) Enhance broiler meat safety by controlling Salmonella and Campylobacter in chickens; 5) control antimicrobial resistance dissemination; 6) develop strategies for recycling of poultry litter; 7) enhance consumer confidence and acceptability of broiler meat; 8) develop an outreach program targeting poultry producers and other stakeholders; 9) empower the future workforce with skills needed for sustainable broiler production; and 10) assess the socio-economic-environmental impact of proposed interventions.

Project Methods
Devise novel approaches for controlling necrotic enteritis and coccidiosis in broiler chickens.Development of recombinant vaccines against avian coccidiosis. Molecular constructs for the Eimeria acervulina EF-1α gene will be cloned into an inducible pET32a expression system and expressed in competent E. coli BL21 (DE3) cells. One-day-old commercial broiler chick will be vaccinated by oral gavage with increasing CFUs (108-109) of E. coli expressing EF-1α/pET32a, or the empty pET32a vector, in mucosal delivery adjuvants. An identical second booster dose will be administered on day 8. On day 15, EF-1α- and empty vector-vaccinated birds will be inoculated by oral gavage with 5x104 E. acervulina oocysts/bird, and coccidiosis disease parameters will be assessed at 6 days post-infection.Development of multi-subunit recombinant vaccines against necrotic enteritis (NE) causing C. perfringens (CP). A comparative transcriptomic analysis to identify NE specific virulence factors with a specific focus on in vivo expressing CP genes. Recombinant CP proteins expressed in E. coli will be used to vaccinate chicks on days 1 and 8, by oral gavage with increasing CFUs (108 -109) of E. coli expressing CP proteins comprising multiple bacterial virulence factors. As controls, chickens will be vaccinated with E. coli expressing the empty pET32a vector.Additionally, the effects of dietary GTE (125 - 500 mg/kg diet) on NE in broiler chickens will be determined.Identify novel alternative feed ingredients and probiotics to promote growth and enhance sustainability of broiler production Three species of insects including the yellow mealworm, the house cricket, and the soldier fly will be produced and will be fed with reference diets optimized for each of the three species. Dry insects will be grounded using a high-speed food processing mixer/grinder. A total of four hundred, day-of-hatch male chicks will be obtained from a local hatchery and randomly assigned to either control or insect-based meal diet with 8 replications/treatment. Each replication will consist of 25 chickens housed in separate floor pen with pine shavings and ad libitum access to feed and water throughout the study period (42 d). Birds will be fed with a starter diet from 0-21 d, and grower diets from 22-42 d. Feed consumption and bodyweight of the birds will be recorded weekly. Additionally, cecal contents will be collected from the birds at each sampling to measure ammonia emissions.For the probiotic trial, birds will be fed with feed supplemented with probiotic cocktail (8 log CFU/Kg feed) for 6 weeks, starting ay hatch. Birds (n=35/group) will be sampled on weeks 1, 3 and 6 for morphometric and body weight measurements. Feed consumption will be recorded weekly to obtain FCR. Muscle (breast muscle weight and total breast muscle cross sectional area) and bone (tibia/femur length, weight, diameter and bone strength) parameters will be measured to assess overall growth. Additionally, cecal contents will sampled to measure ammonia emission and microbiome analysis. To assess meat quality, the pectoralis muscle will be assessed for color, pH, drip loss, cook loss, and shear analysis using standard protocols.Develop strategies to promote bird welfare in intensive broiler production. This objective is aimed at developing approaches to alleviate potential environmental stressors, develop automated and quantitative non-invasive methods to assess stress levels, and to communicate the approaches and methods developed to the poultry industry. These will include improved ventilation to reduce litter moisture and improve broiler welfare, ammonia control products to improve litter and air quality and bird welfare, and phytogenic feed additive supplementation and early life thermal conditioning to mitigate heat stress and improve bird well-being.Enhance broiler meat safety by controlling Salmonella and Campylobacter in chickens.This experiment will be done to ascertain the efficacy and optimize dosages of nanoemulsions of two phytochemicals, namely trans-cinnamaldehyde (TC) and caprylic acid (CA) supplemented through water (0, 0.25% and 0.50% v/w) to decrease Salmonella and C. jejuni in broiler chickens. The nanoemulsions of TC and CA in water will be prepared using emulsifiers (ethanol, lecithin). The size, zeta potential and polydispersity index of nanoemulsion will be determined.Day of hatch broiler chicks will be weighed and randomly allocated to various treatments of the study (n=42 chicks/treatment/trial with at least two replicated trials). The birds will be supplemented with phytochemical nanoemulsions (0, 0.25, 0.5 % v/w) in drinking water for 42 days. On day 7, the birds will be orally challenged with a four-strain cocktail of S. Enteritidis or C. jejuni (~ 6 log CFU/bird). On days 10, 14, 21, 28, 35, and 42, six birds from each treatment will be sacrificed to determine pathogen load in the ceca. In addition, the effect of phytochemical treatments on cecal microbiome will be studied.Delineate the bacterial resistome in poultry production environment for developing strategies for reducing antimicrobial resistance (AMR) spread. Sampling will be done over a period of two years and a total of 10 antibiotic-restricted (ABR) broiler farms will be sampled. We will sample a cohort of birds (cloacal swabs, n=30/farm) and their environment (soil, n=15; litter, n=15) at regular intervals (three times) as they continue along the production line through hatchers and grow out farms. Samples will be processed for Salmonella and Campylobacter and the isolates will be screened for antimicrobial sensitivity. Resistome and plasmidome in the samples will be identified.Conduct consumer outreach on sustainability and poultry meat from ABR broiler production. We will conduct focus groups and develop questionnaires to determine concerns about poultry meat production. We will develop outreach materials that highlight the importance of responsible antibiotic use and the impact on poultry production. We will highlight the methods that producers use to reduce the use of antibiotics. We will also conduct consumer outreach at both national and grassroots levels and partner with stakeholders to integrate our work with existing networks, conferences, and venues.Develop, communicate and evaluate a comprehensive outreach program to educate poultry farmers and other stakeholders, including 4-H youth on sustainable methods of ABR poultry production. This will be accomplished through workshops, train-the-trainer program, on-farm demonstrations, and by conducting symposia in scientific conferences.Educate and equip the future agricultural workforce with the knowledge and skills for sustainable broiler production. This will include developing instructional materials, sequential incorporation of content into existing curricula and certificate programs, developing a new online course on poultry sustainability and developingLearning-abroad programs (India and Mexico) for comparative sustainability experience.Develop strategies for improved poultry litter recycling. We will valorize litter into nitrogen and phosphorus super capacitor (SC) electrodes via pyrolysis; systematically test the performance of the SC electrodes via electrochemical techniques; and, evaluate the environmental feasibility of the litter-to-SC process via a life cycle assessment.Assess the socio-economic-environmental impact of proposed interventions using a comprehensive cost-benefit analysis. We will identify the diverse economic, social, and environmental factors impacted by the adoption of ABR production practices, and interrelationships between these factors. We calculate financial returns and assess other considerations affecting producer willingness to adopt ABR production strategies. In addition, we will identify consumer preferences for ABR poultry products through simulated shopping experiments.

Progress 09/15/23 to 09/14/24

Outputs
Target Audience:Target audience include poultry scientific community interested in antimicrobial resistance, organic poultry, educators in sustainable agriculture, largescale organic poultry production, and small-scale organic poultry production. Changes/Problems:Objective 3: Although the addition of TPX with PLT or alum did reduce soluble phosphorus in poultry litter, it did not result in the reduction of soluble phosphorus by orders of magnitude like we had observed in earlier studies in the laboratory.? What opportunities for training and professional development has the project provided?Objective 1: ARS: Two research fellows, Dr. Youngsub Lee and Dr. Samiru S. Wickramasuriya were hired under the Oak Ridge Institute for Science and Education (ORISE) fellowship program. (ORISE is a U.S. Department of Energy (DOE) institute managed by Oak Ridge Associated Universities (ORAU) through a contract between ORAU and DOE. Objective 2: Training for 2 PhD students Objective 3: Two postdocs and one graduate student worked on Objective 3 of the project during the past year. Objective 4: : One graduate student (Trushen Shah) completed his general exam and PhD proposal and is a PhD candidate now. Tentative graduation-Summer 2025. Two more PhD students are working on this objective. This project provided funding and training for two Post-docs (USDA-ARS Fayetteville) working on the project. Obj 5 - none Obj 6 - na Objective 7: University of Minnesota: Grace Dewi, the doctoral student on the grant, defended her thesis. A public seminar was part of the defense. The last chapter of her thesis was on the consumer focus group research done to understand consumers' perspectives that could design how the producers can incorporate sustainability values into their production and processing systems. This chapter was funded entirely through the grant. The grant was acknowledged during the presentation and in the slides. Grace has obtained a full-time job in the poultry vaccine industry. The training in the grant provided significant insights into her understanding of the poultry sector and how to make sustainable progress in her workplace, advancing sustainable approaches such as vaccination in disease control. Two new MS students are being trained on the grant. Undergraduate students are being trained in farm management, FFA activities, and Presidential Turkeys care and management. Obj 8 Obj 9 Obj 10 -none How have the results been disseminated to communities of interest?Objective 2: Project results were presented at the annual meeting of the Poultry Science Association and the symposium on Synergy in Poultry Production, Food Science, and Public Health for Global Well-Being. Objective 3: Four newsletters covering methodology to manage moisture levels in poultry houses were published. Additionally, several posts were made on our social media Facebook page regarding opportunities to better manage poultry house moisture. Objective 8 - Outreach Program accomplishment summary Penn State University: · Participated in the Sustainable Small Scale Poultry Processing Workshop on August 23rd on layer nutrition and management as well as the principles of small-scale broiler processing University of Connecticut: · Efforts have been focused primarily on strengthening the outreach team as well as outreach directly to local producers and processors. · A booth was held at the Connecticut "Ag Day at the Capitol" in March, where outreach was done mostly involving students and those involved in local agricultural industries. · A grant meeting was planned and subsequently hosted during the Poultry Science Association annual meeting in July. · A booth was held at the Tolland County 4-H Fair in August for 3 days, where outreach was done with locals interested in involving themselves with agriculture. Many questions related to poultry production and the grant project were answered. · A 1-day workshop on Small Scale Broiler Processing was hosted in August, where there were multiple speakers including team members from other objectives. · Continued work has been done on the grant website, including minor layout changes in addition to publications being added as they were made available. University of Georgia: Poultry 411 app is available for iOS and Android. In this reporting period app updates were released that included the addition of a litter moisture drying calculator and adding a fourth language option. Language options now include English, Spanish, Portuguese and Vietnamese. · Czarick, M., 2024. Basic circulation fans system design. University of Georgia Cooperative Extension Service Poultry Housing Tips. v35(2). · Czarick, M., C. Mou and B.D. Fairchild, 2023. Using circulation fans to improve foot pad health. UGA Poultry Housing Tips 35:3. · Czarick, M., C. Mou and B.D. Fairchild, 2023. Vertical vs. horizontal circulation fan systems. UGA Poultry Housing Tips 35:4. University of Maryland: Continued outreach efforts to help commercial poultry growers to improve bird welfare and improve (recycle) litter. Five meetings were held at multiple locations covering different topics of litter management and bird welfare. University of Minnesota: Several activities were done during this project period. Several talks were delivered on the updates from the project on regional, national, and international platforms. Students presented at conferences. A graduate student defended her doctoral thesis relevant to the outreach component. Two new MS students are being trained on the grant. Undergraduate students are being trained in farm management, FFA activities, and Presidential Turkeys care and management. Objective 9. Workforce Development. Provided theory-based undergraduate education with sustainable poultry courses. Sequential incorporation of sustainable poultry topics into existing courses: Spring 2024 (January-March): Sarah Bramall taught an 8-week course Sustainable Integrated Small Animal Farming. There were 19 students enrolled. Course evaluations - 4.75/5. (Comments from students regarding information taught: Overall, this was an interesting course that provided information about Small Integrated Animal Farming. It is good to learn the important details of putting up a small farm. This course is very interesting especially for people that do not really live on a farm. You learn a lot of background knowledge.) New courses: A. Fanatico developed and taught a new course in Fall 2023 for the App State Honors College HON2515 (Section 102) Eating Meat in the Anthropocene, with links to many issues in poultry production. The course focused on understanding the ecology of sustainable meat (and alternative protein) production in agriculture. Students learned to think critically about the impact on climate and social justice. We included topics on grazing animals and the role of grassland in carbon sequestration, alternative feeds for monogastrics and their role in nutrient cycling. We discussed the need for biodiversity in agriculture and included topics on swine, poultry, aquaculture, insects, agroforestry. Students had 3 field trips to local farms with the local livestock extension agent. A. Upadhyay developed a syllabus for ANSC 3695/5695 Current Trends in Sustainable Poultry Production. The course will discuss sustainability topics, including the sustainability of broilers and the impact of restricted antibiotic use. It will use an approach of bird health, human health, and planetary health and will highlight outcomes of the research project. A. Fanatico set up an Undergraduate Research Experience (REU) site at App State with another project. In summer 2024 Fanatico hosted three undergraduate students for 10 weeks (Genesis Vargas from University of Puerto Rico, Abby Golembieski from Michigan, and Ameris Chadband from West Virginia State University). Students work in local communitybased food system, nutrient cycling, and developing a production system for black solider fly larvae with a local farmer as a chicken feed. What do you plan to do during the next reporting period to accomplish the goals?Work will continue work as stated in the proposal. Complete data analysis and publish manuscripts under preparation. Objective 3: We plan to finalize our analysis of the solar brood data and publish it. In addition, we are working on two more calculators that we hope to add to the Poutlry411 app pending. The calculators still need to undergo some validation and beta testing before incorporating it into the app. Additional trials on mitigation of heat stress will be performed, and a manuscript based on the effects of early life thermal conditioning on tissue gene expression will be submitted for publication. Objective 8: Penn State team plans for 2025 include running the 2nd round of Brooder School. University of Connecticut plans for the upcoming year include further strengthening communication both within the outreach team as well as between objectives. In addition, we plan to host more workshops and seminars using the information we have received from prior feedback. A 2-day meeting of the entire project team covering project progress, enhancement, and future activities is planned. University of Georgia plans to finalize analysis of the solar brood data and publish it. In addition, they are working on two more calculators that will add to the Poutlry411 app pending. The calculators still need to undergo some validation and beta testing before incorporating it into the app. University of Maryland will continue outreach efforts to growers. Peter Bina, Naveena Maheswarappa, Kumar Venkitanarayanan, Anup Kollanoor Johny are developing an undergraduate experiential learning program in sustainable poultry production - a student learning abroad experience. Objective 9 - Current efforts include scale-up of the electrodes to power a computer fan to demonstrate a proof-of-concept of the idea. Simultaneously, the environmental feasibility of the litter-to-SC process is being performed. Objective 10 - Dr. Collins has set up a model framework to include environmental impacts. We have not made progress in the environmental impacts because we are waiting for inputs from other objectives. Dr. Collins has a new job with SUNY and we are in the process of hiring another Post Doc to take on previous works.

Impacts
What was accomplished under these goals? Objective 1: 1) Identified, cloned and characterized chicken Interleukin-23 (IL-23), a recently identified member of the IL-12 family of cytokines playing a critical role in regulating T helper cell function. IL-12 and IL-23 share a common p40 subunit, but differ in their p35 and p19 subunits, respectively. Here, we report the functional characterization and immunomodulatory properties of chicken IL-12 and IL-23 using the panels of newly developed mouse anti-IL-12p40, IL-12p35-α and IL-23p19 monoclonal antibodies (mAbs). The levels of chicken IL-12 and IL-23 in the circulation of Eimeria maxima (E. maxima) and Eimeria tenella (E. tenella)-infected chickens were determined. Collectively, results validate the specificity and significance of 2 newly developed antigen-capture immunoassays for chIL-12 and chIL-23 which will expand our understanding of the functional characteristics of IL-12 and IL-23 and their role in normal and diseased chickens. 2) This study investigated the best oral delivery strategy (gavage or feed) for the B. subtilis expressing chicken anti-microbial peptide cNK-2 (B. subtilis-cNK-2) compared to monensin in chickens challenged with E. acervulina. The oral administration of B. subtilis-cNK-2 improved growth performance, enhanced local protective immunity, and reduced fecal oocyst shedding in chickens infected with E. acervulina, demonstrating potential use of B. subtilis-cNK-2 as an alternative to antibiotics to protect chickens against coccidiosis. Objective 2: Feeding trials to test the impact of 2 different insect meal-based diets on production performance and meat quality parameters were completed. Also, we determined the feasibility of in ovo probiotic application in modulating microbiome acquisition in hatchlings. These studies are completed, and three manuscripts are under preparation. Objective 3: An effective system for tracking air quality with potential for expansion and improvements has been developed. The system includes sensors connected to Raspberry Pi GPIO, I2C and UART. It provides real-time air quality metrics, VOC, PM levels, CO2, and CO. We completed the evaluation of the solar brood system. The data are currently being analyzed and summarized. During year 4, a small plot rainfall simulation study was conducted using the poultry litter obtained from pen trials conducted in year 3 to determine the effect of various litter amendments on phosphorus runoff and yields of tall fescue. The objective was to determine how land application of litter treated with new amendments, such as alum mud litter amendment (AMLA), made from the biproduct of alum; or TPX, a calcium-based nanoparticle compared to amendments that are already in use by the poultry industry. The treatments were: (1) unfertilized control, (2) normal poultry litter, (3) AMLAtreated litter, (4) alum-treated litter, (5) PLT-treated litter, (6) alum plus TPX-treated litter, and (7) PLT plus TPX-treated litter. Soluble phosphorus runoff from litter treated with AMLA, alum, and alum plus TPX-treated was reduced by 50, 60, and 38%, respectively, compared to normal litter, which was similar to that observed for total phosphorus loss in runoff. Forage yields were significantly higher for all plots fertilized with litter compared to the unfertilized control plots. Nitrogen uptake was significantly higher for litter treated with alum, PLT, or alum plus TPX than normal litter. Effects of early life thermal conditioning and heat stress on mRNA levels for candidate genes expressed in the hypothalamus, pituitary, adrenal, liver, adipose, heart, and breast and thigh muscle were analyzed, and a manuscript has been drafted. Objective 4. 4a: Completed bird study to validate the efficacy of in-feed phytochemical treatments, Trans-cinnamaldehyde (TC) and Caprylic acid (CA), in reducing Salmonella and Campylobacter in chickens under field conditions in a commercial farm setting. We published one manuscript in the journal of Poultry Science, a second manuscript evaluating the effect of infeed inclusion of these phytochemicals on production performance and cecal microbiome is in preparation. Objective 4b: In-water supplementation of CA nanoemulsion reduces colonization of S. Enteritidis (SE) in broiler chickens. CA is a GRAS-status phytochemical with significant anti-Salmonella efficacy in chickens. However, the challenge working with CA oil is its low solubility in water. To overcome this and enhance dispersion of CA oil in water, CA nanoemulsions (CANE) were prepared with food grade Gum Arabic and lecithin emulsifiers. Thereafter, the efficacy of in-water supplementation of CANE in reducing SE colonization in 28-day-old broiler chickens was studied. In two separate trials, 96, day of hatch broiler chickens (24 birds/treatment/trial) were randomly allocated to 4 groups: Control, emulsifier control, CA 0.1% and CANE 0.1%, TC 0.06, TCNE 0.03, TCNE 0.06 for challenge (to be infected with SE) and non-challenge (no infection), respectively. Inwater supplementation of CANE at 0.1% reduced SE colonization by ~ 1 log CFU/g of cecal content (P<0.05) on days 21 and 28. No reduction in feed consumption, water intake or body weight gain was observed in any treatment groups compared to controls (P>0.05). CA nanoemulsion could be used to control SE colonization in chickens. Follow up analysis on the effect of treatments on gut histopathology and cecal microbiome is currently underway. Objective 5: No updates since last report. Objective 6: Work completed on a process to convert poultry litter into supercapacitors (SC) for energy storage. Methods: Representative samples of poultry litter were obtained from a commercial farm near Raleigh, NC. The litter was pyrolyzed at 400°C, 500°C, 600°C, and 700°C (3 hours) under a nitrogen environment to synthesize raw biochar. Results: Based on the data analyses, pyrolysis at 600°C was found to provide the highest specific capacitance. Objective 7: The Consumer and Stakeholder Engagement objective team works as part of the overall SAS team. Appalachian State University (Fanatico) and University of Minnesota (Johny) worked together in Year 4. We focused on responsible antibiotic use and other aspects of sustainability. We explored how messages can make a difference in beliefs and feelings for stakeholders and impact their behavior and action. Intervention: Use Public Interest Communication strategies for science-based strategic communication and engagement. Messages focus on shared human values: Companies: Be transparent, engage with your stakeholders, build trust. Consumers: Explore and be part about your supply chain. Fanatico summarized our draft messages on the Sustainable Development Department website Poultry in Sustainable Food Systems. "Reduce routine antibiotic use; treatments with nonmedically important antibiotics may be needed; change the world with your chicken." These messages can be used in different ways and with different messengers. Objective 8 - Outreach program updates reported in the "How have results been disseminated to communities of interest" section. Objective 9: Work force development updated reported in the "How have Results been disseminated to communities of interest" section. Objective 10: Completed two Agent-based models in different structures. NetLogo was used as a simple template to design and create a logical flow of agents, interactions, and networks. Repast Suite was used to expand the complexity of the ABM framework by introducing multiple buyers and producers, with flexibility of market prices and production costs. We have a space holder for incorporating the environmental impacts and will use input from other objectives to simulate consequences.

Publications

• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Wickramasuriya S.S., Park I., Lee, Y., Richer L.M., Przybyszewski C., Gay C.G., van Oosterwijk J.G., Lillehoj H.S. 2024. Effect of orally administered B. subtilis-cNK-2 on growth performance, immunity, gut health, and gut microbiome in chickens infected with Eimeria acervulina and its potential as an alternative to antibiotics1. Poultry Science 103:104156.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Park I., Nam H., Lee, Y., Smith A.H., Rehberger T.G., Lillehoj H.S. 2024. Effect of ?-Alanine Metabolite on Gut Integrity and Immunity in Commercial Broiler Chickens Infected with Eimeria maxima. Animals 14:2558.2024
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Lee Y., and Lillehoj, H.S., 2024. Differential detection of chicken heterodimeric cytokines, interleukin 12 and 23 using their subunit-specific mouse monoclonal antibodies. PSA Annual meeting, Louisville, KY, USA.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Wickramasuriya S.S., Park I., Lee Y., Lillehoj, H.S. 2024. Functional analysis of gut microbiome changes in broiler chickens treated with B. subtilis-cNK-2 to mitigate avian coccidiosis. PSA Annual meeting, Louisville, KY, USA.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Gao M, Ren Y, Lu S, Reddyvari R, Venkitanarayanan K, Amalaradjou MA. 2024. In ovo probiotic supplementation supports hatchability and improves hatchling quality in broilers. Poult. Sci. 103(6), 103624.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Gao M, Ren Y, Lu S, Reddyvari R, Amalaradjou MA. 2024. In ovo probiotic application modulates microbiota development and acquisition in broiler embryos and hatchlings. 2024 PSA Annual Meeting, July 15-18, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: At the crossroads: Sustainable poultry production through a One Health lens. Symposium on Synergy in Poultry Production, Food Science, and Public Health for Global Well-Being, Thessaloniki, GR, June 19-20, 2024.
• Type: Theses/Dissertations Status: Other Year Published: 2024 Citation: Gao, M. 2024. Promoting Hatchability and Modulating Microbiome Acquisition in Broilers using in ovo Probiotic Supplementation. Doctoral Dissertation.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Greene ES, Ardakani MA, Dridi S. Effects of an herbal adaptogen feed-additive on feeding-related hypothalamic neuropeptides in chronic cyclic heat-stressed chickens. Neuropeptides. 2024 May 21:106:102439. doi: 10.1016/j.npep.2024.102439
• Type: Other Status: Published Year Published: 2024 Citation: Greene ES, Tabler TW, Orlowski SK, Dridi S. Effect of heat stress on the hypothalamic expression of water channel- and noncoding RNA biogenesis-related genes in modern broilers and their ancestor red jungle fowl. Brain Res. 2024 May 1;1830:148810. doi: 10.1016/j.brainres.2024.148810. Epub 2024 Feb 15
• Type: Other Journal Articles Status: Accepted Year Published: 2024 Citation: Mou, C. T., M. Czarick, and B. D. Fairchild, 2024. Evaluation of the effects of maintaining a moderate humidity (50-60%) and increased air movement on litter moisture and footpad health in a commercial broiler house. Journal of Applied Poultry Research Accepted-InPress.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Lyte, J. M., Arsi, K., Caputi, V., Liyanage, R., Assumpcao, A. L. F. V., Jesudhasan, P. R., and Donoghue, A. M. 2024. Inclusion of trans-cinnamaldehyde and caprylic acid in feed results in detectable concentrations in the chicken gut and reduces foodborne pathogen carriage. Poultry Science, 103(2), p.103368.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Arsi, K., Lyte, J. M., Caputi, V., Liyanage, R., Assumpcao, A. L. F. V., Jesudhasan, P. R. R., and Donoghue, A. M. Inclusion of trans-cinnamaldehyde and caprylic acid in feed results in detectable concentrations in the chicken gut and reduces foodborne pathogen carriage. Poultry Science Association Annual Meeting, Louisville, KY, July 15-18, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Shah, T., Shah C., Allen, J., Zhu, C., Walunj, A., Balasubramanian, B., Leticia, A., Rankin, K., Mishra, N., Upadhyaya, I., Venkitanarayanan, K., Upadhyay, A. Advancing sustainable broiler production: Harnessing phytochemical nanoemulsions to mitigate Salmonella colonization in chickens and enhance human health and safety. IFT-First Annual Meeting and Expo, Chicago, IL, July 14-17, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Shah, T., Shah, C., Allen, J., Zhu, C., Walunj, A., Rodriguez, A. L., Mishra, N., Upadhyaya, I., Venkitanarayanan, K., Upadhyay, A. In-water supplementation of caprylic acid nanoemulsion reduces colonization of Salmonella Enteritidis in broiler chickens. Poultry Science Association Annual Meeting, Louisville, KY, July 15-18, 2024.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Liang, C. and Collins, B. (2024), Applying Agent-Based Modeling to Examine Business Strategies  Tools and Examples for Researchers and Practitioners, revised and re-submitted to the Small Business Institute Journal, published by the National Small Business Institute (top journal in Small Business and Entrepreneurship)
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Collins, B. And Liang, C. (2024, forthcoming), Using NetLogo to Build an Agent-Based Model for Teaching Purposes at the Graduate Student Level, Applied Economics Teaching Resources Journal, Published by the Agricultural and Applied Economics Association.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Presentation: Liang, C. (2024). Best practices for furthering innovation and capturing the benefits of the bioeconomy. Panelist, Agricultural and Applied Economics Annual Meeting, July 30, New Orleans, LA.
• Type: Other Journal Articles Status: Accepted Year Published: 2024 Citation: Mou, C. T., M. Czarick, and B. D. Fairchild, 2024. Evaluation of the effects of maintaining a moderate humidity (50-60%) and increased air movement on litter moisture and footpad health in a commercial broiler house. Journal of Applied Poultry Research Accepted-InPress.
• Type: Websites Status: Other Year Published: 2024 Citation: Newsletter (available at www.poultryventilation.com)
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Upadhyaya, I.(2024) Co-hosted and presented at the International Symposium "Synergy in Poultry Production, Food Science, and Public Health for Global Well-Being", American Farm School of Thessaloniki, Greece, June, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Upadhyaya I., Upadhyay A. (2024). Ensuring Sustainable Meat and Poultry Safety: HACCP standards and regulatory guidelines. IMSACON XII, Mathura, India. Oral Presentation, Policy issues, regulatory frameworks and extension strategies for bringing paradigm shift in meat sector, IMSACON annual meeting, Mathura, India, September, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Colbert Elementary. Raising chickens for eggs and keeping hens healthy. Colbert, GA February 2024.  presentation done 3 times for (3rd, 4th and 5th grades).
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Perdue Farms Grower Meetings. Using water meters and circulation fans to manage poultry house moisture. September 18-19, 2023. Forsyth, GA, Barnesville, GA, Perry, GA, Reynolds, GA
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Georgia Broiler Conference. Watkinsville, GA. September 27, 2023. o UGA Poultry Housing research update o Introduction and review of new features of the Poultry411 app
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Carroll County Young Farmers Meeting. Broiler Light Management: Duration and Intensity. Carroll County Ag Center, Carrollton, GA October 31, 2023.
• Type: Other Status: Other Year Published: 2024 Citation: Delaware AG Week: Poultry Program. January 8, 2024. o Importance of bird distribution o Brooding fundamentals
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J. (2024) Poultry the Shore and More. Rooted in Our Community: Climate-Smart Farming, Grow Cecil.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Madison County Young Farmers Meeting. UGA Poultry Housing research and activities update. Danielsville, GA, April 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Livestock Poultry Learning Community Webinar. Poultry house tightness and fuel usage. December 15, 2023.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Queensland Alliance for Agriculture and Food Innovation Seminar. UGA poultry housing research update. University of Queensland- St Lucia Campus, Brisbane QLD Australia, February 15, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Poultry house environmental management and impact on broiler body temperature. Brisbane, Australia. February 16, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Australian Poultry Science Symposium. Moisture control in poultry houses during cold weather. Sydney, Australia. February 21, 2024.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Merck Poultry Management Seminar. Aibonito, Puerto Rico. March 5-7, 2024. o Broiler chick brooding fundamentals o Broiler hot weather management o Broiler house moisture control
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J. (2024) Key indicators of Poultry welfare and management. Sustainable Small Scale Poultry Processing, University of Connecticut.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J. (2024) Raising Birds on a Small Scale: Backyard and Pasture Poultry. Sustainable Small Scale Poultry Processing, University of Connecticut.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J. (2024) Mortality Management. Delaware Ag Week Small Flock Session, University of Delaware.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J. (2024) Strategies to Improve Paw Quality. Delaware Ag Week Commercial Poultry (evening session), University of Delaware.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Dridi S. Heat stress and poultry production sustainability: challenges and potential mechanism-based strategies. PSA Meeting, July 12-18, Louisville KY, USA.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Moyle, J., Thilmany, E., Lansing, S., Dill, S., Rhodes, J., Semler, J., Franceschi, R., MacDonald, J., Nunn, N., Everts, K., Alves, P., Delp, D. (2024) Maryland Animal Waste Technology Fund Assessment. 24th Congress of the International Farm Management Association (litter recycling)
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: A.C. Fanatico. 2024. Sustainable Poultry Education. Symposium: Defining sustainability within the poultry industry, Poultry Science Association Annual Meeting July 15-18, 2024. Louisville, KY.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Fanatico, A. and R. Udale. The Rich Heritage of Chicken Breeds. Episode 354. ATTRA Voices from the Field. https://attra.ncat.org/episode-354-the-rich-heritage-of-chicken-breeds/. May 22, 2024. National Center for Appropriate Technology, Fayetteville, AR.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Selecting the right breed for integrated and pastured poultry production panel. USDA BFRDP Integrated Crop-Poultry Production Seminar. October 19, 2023. Webinar hosted by Pitowski Lab at the University of California at Davis.
• Type: Websites Status: Other Year Published: 2024 Citation: Fanatico maintains a website on App States Sustainable Development page to provide a 2-prong approach to sustainable poultry production. One approach is efficiency (large-scale) and the other approach is robust (small-scale). Fanatico modified this to include a website: The Importance of Poultry Production in Sustainable Agriculture. Sustainable poultry production is based on processes in nature, such as nutrient cycling and biodiversity.
• Type: Other Journal Articles Status: Published Year Published: 2024 Citation: Syed Fahad Bin Haque, Sanjay Shah, Joe Sagues, Ran Ding, John Ferraris, Praveen Kolar. 2024. Poultry litter-derived biochar for supercapacitor applications, Next Energy, 5, 100171.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Maynard C, Greene ES, Dridi S. Effect of heat stress on hypothalamic autophagy pathway in modern broilers and their ancestor jungle fowl. 10th Proteasome & Autophagy Congress, Clermont-Ferrand, France, November 13-15, 2024.
• Type: Theses/Dissertations Status: Other Year Published: 2023 Citation: Thesis titled Valorization of poultry litter into engineered biochar submitted to NC State University graduate school (Fall 2023), Student: Nur-Al-Sarah Rafsan
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Perdue Farms Grower Meetings. Using water meters and circulation fans to manage poultry house moisture. September18-19, 2023.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Dridi S. Heat stress and poultry production sustainability: challenges and potential solutions. International Symposium on Sustainable Animal Industry and Biotechnology Applications- NCHU, Taiwan, November 8-10, 2024
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Dridi S. Fine Effects of High Environmental Temperature on Poultry Production Efficiency. AVEM, March 2024, Mexico city, Mexico
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Perretti A, Pennicott C, Oyeniran V, Graham D, Forga A, Hargis BM, Porter TE, Weimer S (2023) Effects of early-life thermal conditioning on the behavior of broilers during heat stress later in life. Program 2023 Annual Meeting of the Poultry Science Association
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Leya Susan Viju, Divya Joseph, Poonam Gopika Vinayamohan, Kendra Maas, Annie M. Donoghue, and Kumar Venkitanarayanan. 2024. In-feed supplementation of linalool reduces Campylobacter jejuni colonization in broiler chickens. Oral Presentation at the annual meeting of the Poultry Science Association, July 15-18, Louisville, KY.
• Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Leya Susan Viju, Divya Joseph, Poonam Gopika Vinayamohan, Sydney Puchol, Yi Wang, Arya Niraula, Yangchao Luo, Chaoyu Zhai, Abhinav Upadhyay, Annie M. Donoghue, and Kumar Venkitanarayanan. 2024. Efficacy of linalool in reducing Salmonella Enteritidis on shelled eggs. Poster Presentation at the annual meeting of the Poultry Science Association, July 15-18, Louisville, KY.

Progress 09/15/22 to 09/14/23

Outputs
Target Audience:The target audience reached this reporting period included researcher and students interested in sutatinable poultry production, as well as the community of poultry farmers, broiler industry, veterinarians, and food microbiologists. Changes/Problems:Team member, Shanjumar Mooyottu, started a new job at Auburn University. The subcontact to his form institution, Iowa State University, was terminated 9/30/2023. Dr. Mooyottu's project was delayed in project year 3 while the new subcontact to Auburn was finalized. What opportunities for training and professional development has the project provided?Accomplishments continued. Consumer Education and Workforce Development: Education on sustainable poultry production is needed for undergraduate students who will make up the future workforce. Innovative education will help students understand concepts and apply them. The team is currently in the final phase of developing a sustainable poultry production course, which will be offered to students next academic year. Outreach: Focus has been primarily on direct outreach efforts, and strengthening communication within the grant teams. Stakeholder meetings were hosted at the International Production & Processing Expo and at the Poultry Science Association annual meeting. There has been continued work done on the grant website (https://sas.poultryproject.uconn.edu/), The University of Maryland group continued on-farm demonstrations and other outreach regarding the use of high capacity circulation fans to improve bird welfare. The University of Minnesota has been working more with outreach to consumers, processors and producers by hosting webinars, workshops, and conference talks. NC State University Completed 265 screening surveys to gather information from consumers about their recognition, awareness, and preferences in choosing types of chicken breast products from stores. NCSU developed an Agent Based Model to simulate grower-buyer interactions under different conditions, characteristics, factors, and decisions. The NetLogo Program was applied to set up the first phase of the modeling procedure. NCSU organized educational forums when we conducted the screening surveys - such as July Small Farm Unit Field Day (150+ participants), September Bioeconomy conference (62 participants). University of PA conducted a Brooder School consisting of 6 consecutive weeks of programming. Poultry Litter- Poultry litter collected from a commercial broiler facility was converted into biochar via pyrolysis at 600°C for 3 h followed by activating with potassium hydroxide and re-pyrolyzed at to obtain activated biochar. Results suggested that activation protocol increased the specific capacitance (electric charge) from 12.7 ± 0.5 and 74.5 ± 1.6 F/g. Environment Assessment: Hired a Post Doc to work on the Agent Based Model Simulations. Completed basic model construction using NetLogo, which is a basic ABM model for multiple agents (multiple buyers and sellers with different preferences and characteristics). The results of NetLogo will be published in a journal article under preparation. This project provided training and professional development for graduate students and undergraduate students. Two research fellows, Dr. Youngsub Lee and Dr. Samiru S. Wickramasuriya were hired under the Oak Ridge Institute for Science and Education (ORISE) fellowship program. (ORISE is a U.S. Department of Energy (DOE) institute managed by Oak Ridge Associated Universities (ORAU) through a contract between ORAU and DOE. • Postdoctoral Fellows (Dr. Khan and Dr. Adetunji), and a Ph.D. student (Tunde Ogundare) are currently working on the SAS project at NCAT. ISU: One graduate (PhD) student (Akhil Vinitakumari) at ISU is on SAS project. • During this reporting period, graduate students were engaged in conducting the poultry trials, necropsy, sample collection and tissue processing. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data as part of team 3. • Mentoring and training graduate students is on going as part of project. How have the results been disseminated to communities of interest?Results have been presented at the annual meeting of the Poultry Science Association. The annual TAMUS Pathways Student Research Symposium, and journal articles, presentations and through the extension newsletters and videos as detailed in the Publications section of this report. PD Venkitanarayanan conducted an Indo-US workshop entitled "Systems-based approaches for enhancing the sustainability of the poultry sector in India and the US at the National Meat Research Institute, India on September 5, 2023. The workshop was attendedby over 60 stakeholders, including scientists, students, farmers, poultry industry personnel, and people from regulatory agency. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Broiler Health: Accomplishment 1: A study was conducted to develop an antigen-capture ELISA that detects an immunodominant antigen of Eimeria. Results show this new immunoassay is a sensitive diagnostic tool to monitor coccidiosis in a large field population in the commercial poultry farms before clinical symptoms develop using serum, feces, and gut samples during the entire period of infection cycle starting from 1 d after infection. Accomplishment 2: A study was conducted to investigate the synergistic effects of orally delivered B. subtilis-cNK-2 on vaccination with rEF-1α against E. maxima infection in broiler chickens. Results found that collectively, rEF-1α vaccination induced significant protection against E. maxima infection in the broiler chickens, and the efficacy of rEF-1α vaccination was further enhanced by co-administration with orally delivered B. subtilis spores expressing cNK-2. Accomplishment 3: The synergistic effects of orally-delivered chicken NK-lysin peptide 2 (cNK-2) or recombinant chicken IL-7 (rchIL-7) on vaccination with recombinant Eimeria elongation factor-1α (rEF-1α) against Eimeria maxima (E. maxima) infection was investigated in broiler chickens. Collectively, these results showed that the efficacy of rEF-1α vaccination was significantly enhanced when rEF-1α vaccine co-immunized with chIL-7 or cNK-2. Accomplishment 4: Studies conducted found dietary mixture containing antibodies to NetB and EFTu provides protection against experimental NE in chickens through passive immunization. Accomplishment 5: The objective of this study was to first compare detectable CNA and NetB levels in biological samples from NE-afflicted chickens, and then evaluate jejunal digesta samples from various commercial farms in the United States using our newly developed Mouse monoclonal antibodies (mAbs) based CNA- and NetB-specific sandwich ELISAs. Study results indicate these new C. perfringens antigen-specific sandwich ELISAs offer a sensitive and specific means to detect C. perfringens CNA and NetB proteins as biomarkers of early NE occurrence in field samples from commercial broiler chickens. Accomplishment 6: A study was conducted to further investigate the effects of a higher dose of an oral B. subtilis-cNK-2 treatment on coccidiosis, intestinal health, and gut microbiota composition. Results show NK treatment restored dysbiosis incurred by E. acervulina infection and showed the general protective effects of orally delivered B. subtilis-cNK-2 on coccidiosis infection. This includes reduction of fecal oocyst shedding, enhancement of local protective immunity, and maintenance of gut microbiota homeostasis in broiler chickens. Accomplishment 7: Both in vitro and in vivo studies were conducted to evaluate the beneficial effects of green tea extract (GT), cinnamon oil (CO), and pomegranate extract (PO) on avian coccidiosis. Study findings provide scientific support for the development of a novel phytogenic feed additive formula that enhances the growth and intestinal health of broiler chickens infected with coccidiosis. Broiler Production: Novel Feed Ingredients. Conducted a feeding trial to test the impact of 2 different insect meal-based diets on production performance and meat quality parameters in comparison to control diet in broiler chickens. These studies are completed, and three manuscripts are under preparation. Studies are underway to evaluate the role of probiotics in promoting the energy status of the late-term embryo and hatchling by analyzing the glycogen content in tissue samples and elucidating the underlying modulation in target gene expression. Broiler Welfare:The circulation fan study evaluating the use of high volume circulation fans to create more air movement at floor level in poultry houses was completed with all of the data collected. A combination of maintaining relative humidity below 60% and increased air movement of 150 ft per minute resulted in drier floors, less foot pad dermatitis, better chick distribution throughout the house without negatively impacting performance. Poultry 411 app was developed and available for iOS and Android devices as a free download from the AppStore or GooglePlay. In 2023, three addition calculators and additional language option were added to the app (Litter drying time, Pad cooling, Tunnel ventilation design). Plasma hormone analysis continued on the heat stress trial performed in year 2. Thermal conditioning on day 4 reduced stress levels and gut permeability in response to heat stress on day 28. The litter amendment pen trial has been completed and two rainfall simulations were conducted using the litter. We're currently analyzing the data. Meat Safety: Objective 4a: Completed bird studies to validate the efficacy of in-feed phytochemical treatments, Trans-cinnamaldehyde (TC) and Caprylic acid (CA), in reducingSalmonellaandCampylobactercolonization in chickens under field conditions in a commercial farm setting. Conducted a study to investigate how the in-feed provision of these phytochemicals may protect against environmental acquisition ofCampylobacter jejuniandSalmonellaspp. Data from our study showed that in-feed addition of trans-cinnamaldehyde and caprylic acid, results in detectable concentrations in the broiler chicken gastrointestinal tract. Results suggest that TC nanoemulsion could potentially be used to control SE colonization in broiler chickens. Follow up analysis on the effect of treatments on gut histopathology and cecal microbiome is currently underway. ?Poultry Production: AMR Surveillance. Successfully finished sampling thebackyard farms to reach the goal of sampling 10 backyard and 10 commercial farms for antimicrobial surveillance.Since completion of farm sampling in late Spring 2023, focus has shifted entirely to in-lab experiments, including completion of antimicrobial susceptibility testing and whole genome sequencing analysis. Antimicrobial susceptibility testing was completed for all isolates from all three pathogens collected. Accomplishment continued in the section below (Opportunities for training)

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: T. Shah, C. Shah, C. Zhu, A, Walunj, J. Allen, B. Balasubramanian, A. Leticia, K. Rankin, N. Mishra, I. Upadhyaya, K. Venkitanarayanan, A. Upadhyay. 2023. In-water supplementation of Trans-cinnamaldehyde nanoemulsion reduces colonization of Salmonella Enteritidis in broiler chickens. PSA Annual Meeting, Philadelphia, PA, July 10-13, 2023
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Dridi S. Molecular signatures for heat stress response and metabolic disorders in broilers. 3�me Congr�s de l�cole Nationale de M�decine V�t�rinaire de Sidi Thabet. October 11-13, 2023, Tunis, Tunisia.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Muyyarikkandy, M. S., Parzygnat, J., & Thakur, S. (2023). Uncovering changes in microbiome profiles across commercial and backyard poultry farming systems. Microbiology Spectrum, e01682-23.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Lee Y, Lillehoj HS. 2023. Development of a new immunodiagnostic tool for poultry coccidiosis using an antigen-capture sandwich assay based on monoclonal antibodies detecting an immunodominant antigen of Eimeria. Poultry Science 102:102790. https://doi.org/10.1016/j.psj.2023.102790
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Lee Y, Park I., Wickramasuriya SS, Lillehoj HS. 2023. Bacillus subtilis Expressing Chicken NK-2 Peptide Enhances the Efficacy of EF-1? Vaccination in Eimeria maxima-Challenged Broiler Chickens. Animals 13:1383. https://doi.org/10.3390/ani13081383
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Lee Y, Park I, Lillehoj HS. 2023. Oral administration of chicken NK-lysin or recombinant chicken IL-7 improves vaccine efficacy of Eimeria tenella elongation factor-1? (EF-1?) against coccidiosis in commercial broiler chickens. Poultry Science 102:102611. https://doi.org/10.1016/j.psj.2023.102611
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Goo D, Gadde UD, Kim WK, Gay CG, Porta EW, Jones SW, Walker S., Lillehoj HS. 2023. Hyperimmune egg yolk antibodies developed against Clostridium perfringens antigens protect against necrotic enteritis. Poultry Science 102:102841. https://doi.org/10.1016/j.psj.2023.102841
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Goo D, Park I, Nam H, Lee Y, Sawall J, Smith AH, Rehberger TG, Li C, Lillehoj HS. 2023. Collagen adhesin protein and necrotic enteritis B-like toxin as biomarkers for early diagnosis of necrotic enteritis in commercial broiler chickens. Poultry Science 102:102647. https://doi.org/10.1016/j.psj.2023.102647
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Wickramasuriya SS, Park I, Lee Y, Richer LM, Przybyszewski C, Gay CG, van Oosterwijk JG Lillehoj HS. 2023. Orally delivered Bacillus subtilis expressing chicken NK-2 peptide stabilizes gut microbiota and enhances intestinal health and local immunity in coccidiosis-infected broiler chickens. Poultry Science 102:102590. https://doi.org/10.1016/j.psj.2023.102590
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Park I, Nam H, Wickramasuriya SS, Lee Y, Wall EH, Ravichandran S, Lillehoj HS. 2023. Host-mediated beneficial effects of phytochemicals for prevention of avian coccidiosis. Frontiers in Immunology 14:1145367. https://doi.org/10.3389/fimmu.2023.1145367
• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Komala Arsi, Mary Anne Amalaradjou, Mairui Gao, Yuying Ren, Ragini Reddyvari, Si Lu, John W. Boney, Juan Morales Ramos, Maria G. Rojas, Casey M. Owens, Palmy R. Jesudhasan and Annie M. Donoghue. 2023. Improving broiler production efficiency using alternate ingredients: Probiotics and Insects. Symposium on Multi-disciplinary Approaches for Enhancing the Sustainability of Broiler Production at the 2023 PSA Annual meeting, July 10-13, 2023, Philadelphia, PA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Mayra Herrera, Komala Arsi, Palmy R. Jesudhasan, Juan Morales Ramos, Maria G. Rojas, John W. Boney, Annie M. Donoghue, and Casey M. Owens. 2023. The impact of insect-based protein Inclusion in broiler diets on meat quality. Poster presentation at the 2023 PSA Annual meeting, July 10-13, 2023, Philadelphia, PA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Mairui Gao, Yuying Ren, Si Lu, Praveen Kosuri, Ragini Reddyvari, Mary Anne Amalaradjou. 2023. In ovo probiotic application improves embryonic development and modulates microbiome acquisition in broiler chicks. Oral presentation at the 2023 PSA Annual meeting, July 10-13, 2023, Philadelphia, PA.
• Type: Journal Articles Status: Under Review Year Published: 2023 Citation: J. M. Lyte, K. Arsi, V. Caputi, R. Liyanage, A. L. F. V. Assumpcao, P. R. R. Jesudhasan and A. M. Donoghue. 2023. Inclusion of trans-cinnamaldehyde and caprylic acid in feed results in detectable concentrations in the chicken gut and reduces foodborne pathogen carriage. Poultry Science (under Review)
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Dridi S. Identification of molecular signatures to improve poultry production sustainability: heat stress, meat quality, and muscle myopathy. The American Meat Science Associations (AMSA) 76th Reciprocal Meat Conference (RMC) on June 25-28, 2023, MN.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Loujain ALOUI, Sara ORLOWSKI, Raouia BEN RHOUMA, Walter BOTTJE, Sami DRIDI. EFFECT OF HEAT STRESS ON HYPOTHALAMIC GENE EXPRESSION PROFILE IN DIVERGENTLY SELECTED BROILERS FOR LOW- OR HIGH-WATER EFFICIENCY. XIV International Agriculture Symposium"AGROSYM 2023", 5-8 October 2023, Bosnia and Herzegovina.

Progress 09/15/21 to 09/14/22

Outputs
Target Audience:The target audience reached this reporting period included researcher and students interested in sutatinable poultry production, as well as the community of poultry farmers, broiler industry, veterinarians, and food microbiologists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Two research fellows, Dr. Youngsub Lee and Dr. Samiru S. Wickramasuriya were hired under the Oak Ridge Institute for Science and Education (ORISE) fellowship program. (ORISE is a U.S. Department of Energy (DOE) institute managed by Oak Ridge Associated Universities (ORAU) through a contract between ORAU and DOE. Postdoctoral Fellows (Dr. Khan and Dr. Adetunji), and a Ph.D. student (Tunde Ogundare) are currently working on the SAS project at NCAT. ISU: One graduate (PhD) student (Akhil Vinitakumari) at ISU is on SAS project. During this reporting period, graduate students were engaged in conducting the poultry trials, necropsy, sample collection and tissue processing. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data as part of team 3. Mentoring and training graduate students is on going as part of team 2 activities. How have the results been disseminated to communities of interest?The results have been disseminated via publications, conferences, and communications with scientific, students, industry, and stakeholders. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Control coccidiosis and necrotic enteritis in broilers In efforts to develop novel coccidiosis control measures, Eimeria coli-expressed E. tenella elongation factor-1α (ETEF-1α) protein was evaluated as an oral vaccine along with chicken IL-7 to show that this vaccine formulation elicited protective immunity against E. maxima and E. tenella challenge infections. In vivo vaccination studies using a cross-protective antigen of Eimeria injected with chicken IL-7 and antimicrobial peptides together improved the efficacy of EF-1α recombinant vaccine. The efficacies of these vaccines were evaluated using various host immunity and gut integrity biomarkers in commercial broiler chickens following E. maxima challenge infection. Chickens in the EF-1α/IL-7 group showed significantly (P < 0.01) higher body weight gains post-E. maxima challenge infection (dpi) with reduced gut lesions in the jejunum at 6 dpi. The EF-1α/IL-7/NK2 group showed reduced fecal oocyst shedding compared to the non-immunized and infected chickens (NC). Recombinant Bacillus subtilis spores with different protein expression levels were studied for sustainable delivery systems to evaluate NK-2 peptide delivery to the gut in chickens as a prevention strategy against coccidiosis. Chicken NK-lysin peptide 2 (cNK-2) is a natural lytic peptide with cytotoxicity against Eimeria. We developed a stable strain of B. subtilis that carries cNK-2 to the gut to determine its effectiveness in ameliorating the negative impacts of coccidiosis and to replace the use of antibiotics in controlling coccidiosis in broilers. The results showed that oral treatment of young chickens with B. subtilis-cNK-2 improved growth performance, enhanced gut integrity, and reduced fecal oocyst shedding. The effect of green tea extracts (GTE), Caffeinated (GTEC) and Decaffeinated (GTED) on Clostridium perfringens toxin genes (netB) was studied. The potency of GTE in reducing the expression of netB was compared to BMD (Bacitracin Methylene Disalicylate). Results indicated that GTEC and GTED reduced C.perfringens proliferation albeit at a higher concentration compared to BMD. In addition, both extracts reduced the level of expression of toxin encoding genes associated with NE in chickens. Novel feed ingredients for improved broiler growth, performance and sustainability A feeding trial was conducted to test the impact of two different insect meal-based diets on production performance and meat quality parameters in broiler chickens. The treatment groups include a control diet and two insect-based diets, one diet with addition of adult cricket meal powder and one diet supplemented with mealworm larvae powder. Body weights, feed intake, and feed conversion ratio (FCR) for starter (0-8 d), grower (9-18 d), finisher 1 (19-28 d), and finisher 2 (29-35d) were recorded. The birds were also processed at the end of the study period (n=10 birds/replicate pen) to determine the impact of feeding insect meals on chicken meat quality and organoleptic parameters. Additionally, ceca samples (n=10 birds/replicate pen) were collected aseptically for microbial and microbiome analysis. Currently, these studies are in progress and data being collected. In another study to investigate the effect of in-ovo probiotic supplementation on hatchability and hatchling quality in broilers, two trials containing a total of 1080 Ross 308 eggs were conducted. In each trial, eggs were sprayed with 200 μl of phosphate buffered saline (PBS; control) or probiotics [~9 log CFU/egg of Lactobacillus rhamnosus (LR) or Lactobacillus paracasei (LP)] on D0, D3, D7, D10, D14 and D18 of incubation. Hatchability was calculated at D21.5. Hatched chicks were scored for their quality using the Tona and Pasgar score. Additionally, morphometric measurements including hatchling weight and length were recorded at necropsy. Overall, in-ovo probiotic supplementation improved hatchability and hatchling quality. Specifically, the application of LP improved hatchability by ~5% when compared to the control. Further, both LR and LP significantly(p<0.05) improved hatchling quality. Moreover, probiotic treated groups tended to have a significantly higher body weight at hatch when compared to the control group(p<0.05). Promote bird welfare: A study to determine the beneficial role of herbal adaptogens in ameliorating growth performance of broilers under heat stress challenges was initiated. Currently, the mode of action of adaptogens is studied. Experiments to study the effect of heat stress on mechanical properties of isolated red and white blood cells from four modern commercial broiler lines and their ancestor jungle fowl are underway. Increase broiler meat safety: The the efficacy of in-feed phytochemical treatments, Trans-cinnamaldehyde (TC) and Caprylic acid (CA), in reducing Salmonella and Campylobacter colonization in chickens was determined. The treatment groups included in this study are control, CA (0.5 or 0.75%) or TC (0.5 or 0.75%) supplemented in feed for 6 weeks,market age, data being analyzed. Completed a study evaluating the application of Trans-cinnamaldehyde nanoemulsion (TCNE) as a natural sanitizer for inactivating Salmonella Enteritidis (SE) in poultry drinking water system. Poultry drinking water was inoculated with SE (~ 6 log CFU/ml) followed by addition of TCNE at 0, 0.01, 0.02, 0.03, 0.04, 0.05 or 0.06 % concentration. All TCNE treatments (except 0.01%), reduced SE in poultry drinking water by at least 3 log CFU/ml in 24 h. By day 5 of incubation, all TCNE treatments reduced SE in poultry drinking water by ~ 5 log CFU/ml (P<0.05). Control Antimicrobial Resistance Dissemination in poultry A longitudinal systems-based approach in sampling backyard and commercial broiler farms was followed, where each farm is visited three times across production. Backyard farms were sampled at days 10, 31, and 52 days of production; commercial farms were sampled at days 10, 24, and 38 days. At each visit, feces (n=10), litter/compost (n=5), soil (n=5), and swabs of feeders and waterers (n=6) were collected for in-lab processing for Salmonella, Campylobacter, and extended-spectrum beta-lactamase (ESBL) E.coli. The team was able to complete 5 backyard farms in year one of the project. In year two, completed sampling for 10 commercial farms during winter months. Of all 468 samples collected from 6 backyard farms, 15.81% tested positive for Salmonella, 26.07% tested positive for Campylobacter, and 14.10% tested positive for ESBL E. coli. Of all 780 samples collected from 10 commercial farms, 52.18% tested positive for Salmonella, 13.08% tested positive for Campylobacter, and 2.31% tested positive for ESBL E. coli. Consumer Education: We are currently working on a collaborative education-extension program around poultry production in India, Mexico, and the US. Extension/Outreach: A website for the grant is established for project dissemination and continued communication with stakeholders. The team at the University of Georgia and Maryland continue to demonstrate how circulation fans can reduce heat stress and improve bird welfare. The team at Penn State University interpreted survey data and drafted a "Brooder School" agenda for a Spring 2023 kickoff. Data from a national broiler producer survey was also interpreted. Workforce development: Currently developing instructional materials for incorporation of sustainable poultry production practices content into existing curricula and certificate programs. Develop strategies for recycling of poultry litter: Performed chemical and physical characterization of representative litter samples sourced from a commercial broiler facility, analyzed data. Assess the socio-economic-environmental impact: We created a list of market prices for poultry products in different categories. Will incorporate these prices into the cost-benefit analysis after more information is collected from other objectives.

Publications

• Type: Journal Articles Status: Published Year Published: 2022 Citation: Lee Y, Park I, Wickramasuriya S, Ben Arous J, Kozoil, M-E, Lillehoj, HS. 2022. Co-administration of chicken IL-7 or NK-lysin peptide 2 enhances the efficacy of Eimeria elongation factor-1? vaccination against Eimeria maxima infection. Poultry Science 101:102013. https://doi.org/10.1016/j.psj.2022.102013
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Lee Y, Lu M, Lillehoj HS. 2022. Coccidiosis: Recent Progress in Host Immunity and Alternatives to Antibiotic Strategies. Vaccines 2022, 10(2), 215, https://doi.org/10.3390/vaccines10020215.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Wickramasuriya SS, Park I, Lee K, Lee Y, Kim WH, Nam H, Lillehoj HS. 2022. Role of Physiology, Immunity, Microbiota, and Infectious Diseases in the Gut Health of Poultry. Vaccines (Basel). 10(2):172. doi: 10.3390/vaccines10020172.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Wickramasuriya S, Park I, van Oosterwijk J, Gay CG, Przybysewki C, Lillehoj HS. 2021. Bacillus subtilis expressing chicken NK-2 peptide protects against Eimeria acervulina challenge infection in broiler chickens. Front. Vet, Sci. 8:684818. doi: 10.3389/fvets.2021.684818.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Elizabeth S Greene, Clay Maynard, Casey M Owens, Jean-Fran�ois Meullenet, Sami Dridi. Effects of Herbal Adaptogen Feed-Additive on Growth Performance, Carcass Parameters, and Muscle Amino Acid Profile in Heat-Stressed Modern Broilers. Front Physiol. 2021 Nov 24;12:784952.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Elizabeth S. Greene, Elizabeth Adeogun, Sara K. Orlowski, Karthik Nayani, Sami Dridi. Effects of heat stress on cyto(chemo)kine and inflammasome gene expression and mechanical properties in isolated red and white blood cells from 4 commercial broiler lines and their ancestor jungle fowl. Poultry Science Volume 101, Issue 6, June 2022, 101827.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Trushenkumar Shah, Chen Zhu, Brindhalakshmi Balasubramanian, and Abhinav Upadhyay. 2022. Application of Trans-cinnamaldehyde nanoemulsion as a natural sanitizer for inactivating Salmonella Enteritidis in poultry drinking water system. IFT 2022, Chicago.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Youngsub Lee, In-kyung Park, Samiru S. Wickramasuriya and Hyun S Lillehoj. J Immunol., Co-administration of recombinant chicken IL-7 or NK-lysin peptide 2 increases efficacy of Eimeria tenella elongation factor-1? in Eimeria maxima-infected chickens. May 15, 2022, 208 (1 Supplement) 102.01; American Association of Immunologist annual meeting, Portland, OR., May 6-10, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Kelsy Robinson, Komala Arsi, Kristin Duffield, Tawni L. Crippen, Kim Cook, and Annie M. Donoghue. 2022. Debugging a Poultry Sustainability: Developing Insects as a Sustainable Alternative Protein Source through a USDA Grand Challenge Research Approach. Poultry Science Annual meeting (PSA), San Antonio, July 11-14, 2022.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Gao M, Ren Y, Reddyvari R, Lu S, Amalaradjou MA. 2022. Early probiotic application improves hatchability and hatchling quality of broiler chicks. 2022 Poultry Science Annual meeting (PSA), San Antonio, July 11-15, 2022.

Progress 09/15/20 to 09/14/21

Outputs
Target Audience:The target audience reached this reporting period included researcher and students interested in sutatinable poultry production, as well as the community of poultry farmers, broiler industry, veterinarians, and food microbiologists. Changes/Problems:Our team leader for the outreach program targeting poultry producers and other stakeholders, Michael Darre, tragically passed away October 9, 2021. Indu Upadhyaya has taken over the outreach program team leader responsibilities. The impact of COVID and research ramp down on University campus' and field sites led to a significant delay in the study. Specifically, inability of research personnel to come to campus delayed project initiation and limited research progress. What opportunities for training and professional development has the project provided?Objective #1. ARS: Two research fellows, Dr. Youngsub Lee and Dr. Samiru S. Wickramasuriya were hired under the Oak Ridge Institute for Science and Education (ORISE) fellowship program. (ORISE is a U.S. Department of Energy (DOE) institute managed by Oak Ridge Associated Universities (ORAU) through a contract between ORAU and DOE. ISU: One graduate (PhD) student (Akhil Vinitakumari) at ISU. Akhil is being trained on bacteriologic and genomic techniques in collaboration with ISU veterinary Diagnostic Laboratory (ISU-VDL) and Pantheravax Ltd. Objective # 2. Identify novel alternative feed ingredients for enhanced sustainability for broiler production. During this reporting period, graduate students were engaged in conducting the poultry trials, necropsy, sample collection and tissue processing. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data. Objective #3. Develop strategies to promote bird welfare in intensive broiler production. Students were trained in conducting field research. Students set up data loggers, audio and video equipment, assessed animal welfare, took digital and thermal images, and collected litter samples in 4 houses on a commercial broiler farm. Students were also trained in the computer laboratory. They measured footpad lesion severity from digital images and were trained in performing early life thermal conditioning and experimental heat stress. In the Department of Animal Science, University of Connecticut: one graduate student is being trained in Poultry microbiology and safety. The project is also a source of training to several other graduate and undergraduate students who are assisting in the project through independent study. Objective #5. Control antimicrobial resistance dissemination. The project has helped in the training and professional development of undergraduate students, graduate students, and a postdoctoral scholar at North Caroline State University. How have the results been disseminated to communities of interest?Results have been disseminated through peer reviewed publications, conference presentations and new apps. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective #1. Devise novel approaches for controlling necrotic enteritis and coccidiosis in broiler chickens. · A chicken vaccine trial was conducted to develop a recombinant Eimeria tenella elongation factor-1a (EF-1a)­mediated vaccination strategy against avian coccidiosis. Results of in vivo vaccination trial demonstrated that chickens of all groups immunized with recombinant EF-1a developed high serum antibody levels to EF-1a, and co-administration of recombinant chll-7 with the EF-1a vaccine enhanced host protective immune response against coccidiosis. E. maxima infection significantly (P < 0.05) up-regulated the expression level of jejunal proinflammatory cytokines (IL-13, IL-17F) and type I cytokines (IFN-y and IL-10), and those expression levels were decreased in EF-1a-vaccinated groups. These results demonstrated a potential protective effect of EF-1a recombinant vaccine with IL-7. In another study, 20 field strains of C. perfringens isolates from different geographic regions (Iowa, Nebraska, Missouri, Illinois, Wisconsin, Indiana, Ohio, and Arizona) were isolated and cultured for comparative genomic analysis. Whole genome sequencing is underway (ISU-VDL) using comparative genomic analysis to identify NE specific and geographically-associated virulence genes (vaccine candidates) of C. perfringens. Comparative genomic analysis is being done in collaboration with Mr. Chandra Tangudu at the Pantheravaxinc. Objective #2. Identify novel alternative feed ingredients for enhanced sustainability of broiler production. Towards developing novel alternative feed ingredients, insect meal (mealworm larvae powder and adult cricket meal) was prepared and their nutrient composition was analyzed. Initial nutrient analysis revealed that adult cricket meal had a TME (true metabolizable energy) (Kcal/kg) - 3,723; with crude protein (%) - 71; Fat (%) - 11.91 while mealworm larvae powder had TME (Kcal/kg) of 5,471, with crude protein (%) - 47.56; Fat (%) - 36.89. Additionally, preliminary chicken trials were conducted to evaluate the ability of in-ovo probiotic application to promote broiler growth and performance. Results revealed that early probiotic supplementation was associated with an increase in hatchability and improved hatchling quality, including an increase in hatchline weight, yolk-free body mass and hatchling length. Follow up experiments are underway. Objective #3.Develop strategies to promote bird welfare in intensive broiler production. 1) The additional air movement provided by circulation fans can potentially improve air stratification, reduce litter moisture, and improve broiler welfare in commercial houses. We have collected environmental and welfare data from two flocks of organic broilers in 4 commercial houses on the eastern shore of Maryland. 2) The 24" 1/3Hp circulation fans have been installed in one house on two farms in Georgia, and data are being collected. 3) Video data was captured at NC State on 10 pens (4 broilers/pen) for 48 hours using high definition video cameras mounted and angled above the pen. Behavior for each broiler will be scored continuously for 48 hours and then compared to the automatic tracking software program Ethovision XT. 4) Two trials were conducted during this period at Arkansas to evaluate the beneficial effects of herbal adaptogen and tannin on heat stress (HS) responses in broilers. As expected, HS increased core body temperature by approximately 1°C compared to the thermoneutral condition. Supplementation of the adaptogen did not affect the core body temperature nor the skin temperature of birds. However, it improved feed intake, which resulted in dose-dependent higher body weight and better FCR compared to the heat-stressed group. The supplementation of tannin did not improve growth performance in heat-stressed birds. 5) Investigators from Maryland and Arkansas conducted one trial on the effect of four durations of early life thermal conditioning (0, 6, 12, and 24 hours at 35 °C on day 5) on HS responses (8 hours at 35 °C) on days 28 and 35. All data are being analyzed. 6) Our USDA/ARS unit has been basically closed due to covid for the past year. Hence, we just started the first pen trial. 7) A literature investigation of the effects of air quality in a confined poultry house on bird welfare was conducted by investigators at Prairie View A&M. Objective #4. Enhance broiler meat safety by controlling Salmonella and Campylobacter in chickens. We investigated the effect of sub-inhibitory concentrations of trans-cinnamaldehyde on the proteome profile of Salmonella Enteritidis, especially the expression of proteins critical for colonization in chickens, using LC-MS/MS analysis. Approximately 1440 proteins were identified. The expression of proteins involved in stress response or cellular transport (GrolL, TolC, AcrA) was upregulated by trans-cinnamaldehyde (P<0.05). Objective #5. Control antimicrobial resistance dissemination. Sampled five antibiotic-restricted backyard poultry farms during April to October of 2021. Due to the scarcity of backyard broiler farms in the winter months, the remaining five backyard farms will be completed in the Spring of 2022. Each farm was sampled at three-time points: 10, 31, and 52 days of production. On the first visit, the team walked through a questionnaire with the farm owner in order to gather information to help understand potential risk factors that could contribute to the persistence of antimicrobial-resistant pathogens on backyard broiler farms. The questionnaire entails several subjects, including farm management, previous outbreaks, antibiotic use, and weather. Objective #6. Develop strategies for recycling of poultry litter. Objective #7. Enhance consumer confidence and acceptability of broiler meat. Objective #8. Develop an outreach program targeting poultry producers and other stakeholders. Objective #9. Empower the future workforce with skills needed for sustainable broiler production Objective #10. Access the socio-economic-environmental impact of proposed interventions. The project team conducted monthly research meetings to identify literature regarding concerns, analysis, and policies in antibiotic applications in poultry industry. More than 100 published articles and reports were gathered, and summarized based on key information related to our study. Two manuscripts (metareview) are in progress - one for the consumers' aspects and the other for producers' aspects. The articles will help inform our team to construct interviews and survey questions for Year 2.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Anup Kollanoor Johny. 2021. COE Faculty Perspective  Teaching the Future. Midwest Poultry Consortium Annual Meeting, March 29.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Anup Kollanoor Johny. 2021. Outliving the pandemic: Veterinarians to build stronger COVID19/Post-COVID19 local and global communities. Virtual International seminar on the Veterinarian response to the COVID19 crisis organized by the Indian Veterinary Association, Kerala Veterinary and Animal Sciences University, World Veterinary Association, and the College of Veterinary and Animal Sciences, Mannuthy, Kerala, India. May 8.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Anup Kollanoor Johny. 2020. Developing sustainable alternatives for poultry food safety. Virtual Minnesota Nutrition Conference. October 14.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Zoie N. McMillian, Jonathan Moyle, Shawna Weimer. Increasing welfare by placing circulation fans in commercial broiler houses. Poultry Science Association, oral presentation, July 2021, Virtual conference.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Jonathan Moyle. Circulation Fans for Destratification on House Temperature. Penn State University Poultry Sales and Service Conference, September 2021, State College, PA.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Anup Kollanoor Johny. 2021. Exploring sustainable alternatives for poultry food safety. South Dakota State University  Des Moines District Food Safety Summit, August 4.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Fairchild, B.D., 2021. Cold weather moisture control in broiler housing tips. Texas Broiler Symposium: Broiler Session, Virtual Presentation September 15
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Fairchild, B.D., 2021. Breeder house cold weather ventilation basics. Texas Broiler Symposium: Breeder/Hatchery Session, Virtual Presentation September 15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Czarick, M., 2021. Poultry house humidity management and control. University of Georgia Cold Weather Management Online Training Series, October 20.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Fairchild, B.D., 2021. Using the Poultry 411 app for better environmental control. Wilkes Country Young Farmers Meeting, Washington, GA, October 26.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Czarick, M., 2021. Minimizing Litter Caking During Brooding. Koch Foods Live Production Webinar, November 12.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Shah, T., Balasubramanian, B., Upadhyaya, I., Venkitanarayanan, K., Upadhyay, A. (2021). Effect of Trans-cinnamaldehyde, Eugenol and Carvacrol on Salmonella Enteritidis proteome critical for colonization in chickens. Poultry Science Association Annual Meeting (Virtual), 2021.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Liang, C. (2021). Using multifunctional agriculture to improve profitability and sustainability, North Carolina Farm Stewardship Association conference, online, Nov 5 to Nov 14, (79 participants).
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Fanatico, A. and Liang, C. (2021). Sustainable poultry production and social justice, North Carolina Farm Stewardship Association conference, online, Nov 5 to Nov 14, (16 participants).
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Center of Excellence for Small Farmers, Ranchers, and Landowners, Thrust 5 report, Professional Agricultural Workers Conference, online panel, Dec 6, 2021. (62 participants)
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: McMillian, Z., J. Moyle, and S. Weimer. Increasing welfare by placing circulation fans in commercial broiler houses. Poultry Science Association virtual meeting. July 19-22, 2021. (oral)
• Type: Other Status: Published Year Published: 2021 Citation: McMillian, Z. and S. Weimer. 2021. Relationships between chicken vocalizations and health, behavior, and welfare. University of Maryland Extension. FS-1177.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Kyung-Woo Lee and Hyun S. Lillehoj. 2022. Role of Clostridium perfringens Necrotic Enteritis B-like Toxin in Disease Pathogenesis, Vaccines 2022, 10(1), 61; https://doi.org/10.3390/vaccines10010061
• Type: Other Status: Published Year Published: 2021 Citation: McMillian, Z. and S. Weimer. 2021. Chicken vocalizations. Poultry Extension Collaborative. 14: May 2021 Newsletter.