Progress 06/01/24 to 05/31/25
Outputs
Target Audience: The targeted audience of this project include Egg producers, Professionals working with the egg industry, Peer researchers,and Extension educators. Changes/Problems:Tight biosecurity requirement of egg farms caused by HPAI outbreak still creates challenges in recruiting egg producers/farms to collaborate on field data collection at commercial egg farms. Recruitment of a graduate student in agricultural engineering is delayed initially by the vacancy of a graduate coordinator position in the PI's department and then currently visa application restriction for international students. We are expecting a no-cost Extension for the project due to the above changes. What opportunities for training and professional development has the project provided?In addition to the PhD student hired specifically for this project, two other PhD students and a research assistant from Dr. Nazmi's lab have actively contributed to the research. Through this project, they have gained hands-on experience in multiple areas, including layer bird management, assessment of egg quality parameters using the Texture Analyzer (Stable Micro Systems), bacterial culture and plating, and measurement of blood biochemical markers using the i-STAT system. They have also been trained in performing flow cytometry for analyzing intraepithelial lymphocytes isolated from the ileum. Furthermore, in preparation for our upcoming study on low pathogenic avian influenza (LPAI), one of our PhD students has acquired specialized skills in in-ovo inoculation, hemagglutination (HA) and hemagglutination inhibition (HI) tests, as well as EID?? determination for virus propagation, isolation, and detection. He has successfully propagated the LPAI (H7N2) strain in 10-day-old embryonated chicken eggs and titrated the harvested virus using the HA test. He is now well-prepared to lead the upcoming chicken trials. Another Ph.D. student had been trained with monitoring of indoor dust concentration, ammonia concentration, air temperature, relative humidity, and airflow speed. In addition, the student was trained to build computational fluid dynamic model for development a new upward airflow displacement ventilation system for cage-free layer houses. How have the results been disseminated to communities of interest?See above accomplishments in Obj. 4. In addition, the students working on the project also presented the preliminary results from our second study as a poster at the Research Forum and Poster Competition organized by the College of Food, Agricultural, and Environmental Sciences (CFAES) at The Ohio State University in April 2025. Additionally, our students participated Poultry Science Association Annual Meeting in Raleigh, NC from July 14-17, where they delivered three separate presentations showcasing the findings from both studies. What do you plan to do during the next reporting period to accomplish the goals?Objective 1.We will continue participating the annual meetings with the stakeholder advisory board, annual EIC Industry Issue Forum, and Ohio Poultry Association Annual Meeting to exchange experiences and issues with egg producers on emerging cage-free layer housing, management challenges, and producers' needs for research and education in hen health and production efficiency. We will reach out egg producers that showing interest to our study and develop an innovative way to conduct indoor environmental monitoring and microbial sampling to protect the biosecurity of the participating farms. This has been a significant challenge and is preventing us form collecting data at commercial farms. Objective 2.We will continue seeking opportunity to collaborate with egg producers for field data collection on indoor environmental quality, microbiome, and bird performance and health at commercial cage-free houses. We plan to conduct further experiments in controlled environment poultry research center at the Ohio State University with laying hens reared in cage-free housing to investigate the effects of HS, CS, dust levels, and ammonia concentrations on colonization and transmission of Avian Pathogenic E. coli (APEC) and low pathogenic avian influenza (LPAI). As in our previous studies, we will assess egg production and quality, quantify IELs in the ileum, and evaluate the expression of oxidative stress-related, antioxidant and inflammatory genes in the ileum, liver, and shell gland. To examine microbiota diversity across treatment groups, cecal samples will be collected, processed, and submitted for sequencing. The resulting data will be analyzed to assess differences in microbial diversity among the treatments. Objective 3.We will continue to develop the CFD model for the new UADV ventilation system using the ANSYS Fluent 24.0. The compatibility challenges between ANSYS Fluent 14.0 and ANSYS Fluent 24.0 need to be resolved. Several of the user-defined functions (UDFs) need to be developed. The CFD simulations will focus on evaluate and optimized ventilation effects on thermal environment conditions and disease transmission in the new cage-free layer houses. Objective 4.We will continue develop Extension workshops and/or online trainings based on feedback from stakeholders and the participants of the Virtual Forum on Dust and Disease. The project team with continue to collaborate Dr. Brett Ramirez, the interim Director of the Egg Industry Center, Paige Kelly, Sustainability & Environmental Manager of Oho Poultry Association, and Dr. Lilong Chai, the faculty organizer of the Georgia Precision Poultry Farming Conference.
Impacts
What was accomplished under these goals?
Objective 1. In the past year, we continued our efforts to communicate with Ohio Poultry Associations, the Egg Industry Center, and egg producers, and professionals of the egg industry to recruit participating egg producers and seek their feedbackon challenges and workforce training needs for cage-free layer operations. See Objective 4 for detailed activities. Objective 2. We completed two independent layer chicken trials focusing on the effects of heat and cold stress on laying chickens raised on cage free housing. Our first trial was focused on the detrimental effects of heat-stress (HS) in laying birds. A total of 240 Hy-Line W-36 layers (26 weeks old) were randomly assigned to two temperature treatments: thermoneutral (TN, 22°C) and cyclic heat stress (HS, 35°C for 12 h followed by 22°C for 12 h) for 8 weeks. Each treatment had six pens (20 birds/pen) at a stocking density of 0.139 m2/bird. Egg production rate and weight were recorded daily, while body weight, feed intake, and cloacal temperature were measured weekly. Egg quality along with the blood gases and electrolyte levels were assessed at the end of study. Results showed no significant difference in the daily egg production rate between groups, however, the HS group produced significantly (p < 0.05) lower egg weight. Moreover, chronic HS significantly (p < 0.05) reduced eggshell breaking strength, while shell thickness, Haugh unit, yolk color, and yolk index remained similar between groups. Despite the HS chickens consumed significantly (p < 0.05) less feed, their feed conversion ratio (based on egg weight) improved (p < 0.05) compared with TN birds. However, body weight gain did not differ significantly (p > 0.05) between groups. Cloacal temperature was significantly higher (p < 0.05) under HS conditions. Additionally, chronic HS significantly (p < 0.05) decreased blood pH, while increasing PCO2 (mmHg), and ionized calcium (iCa, mmol/L) levels. In our second trial, we assessed the negative effects of both the HS and cold stress (CS) in chickens' performance, blood biochemistry, disease susceptibility to Salmonella Enteritidis and immune responses. A total of 216 Hy-Line W-36 layers (48 weeks old) were randomly assigned to six groups housed in separated rooms: thermoneutral control (TNC, 22°C), HS control (HSC, 35°C), CS control (CSC, 10°C), thermoneutral + infected (TNI, 22°C), HS + infected (HSI, 35°C), and CS + infected (CSI, 10°C) groups, with 36 birds in each group (density of 0.139 m2/bird). After a week, nine birds in the TNI, HSI and CSI groups were inoculated orally with Salmonella Enteritidis (108 CFU/mL). Egg production and quality, body weight, feed intake, and cloacal temperature were recorded as in the first trial. At 2 days post-inoculation (dpi), 7 dpi and 14 dpi, birds from each of the treatments were euthanized and tested for the presence of Salmonella in cecal contents, liver, shell gland, and eggs (albumin and yolk). Moreover, the intraepithelial lymphocyte (IEL) populations were isolated from ileum, stained with anti-chicken antibodies. Blood biochemical parameters were measured at pre- and post-infection. Birds under HS condition consumed less feed and produced smaller eggs yet had a better feed conversion ratio (based on egg weight) than the TN birds. Similarly, chickens under CS condition produced lighter eggs but consumed more feed compared to the TNC birds. Eggs from HS birds exhibited significantly (p < 0.05) lower shell thickness and vitelline membrane strength, while shell breaking strength and Haugh unit remained comparable between groups. HS also significantly (p < 0.05) reduced blood gases and electrolyte levels. Notably, CS birds had significantly increased shell thickness before and after infection, with no changes in egg breaking strength. Blood parameter analysis showed that HCO? levels were significantly higher in CSC and TNC than CSI and TNI (p < 0.01). Neither HS or the CS impact Salmonella colonization and transmission at 2 dpi, 7 dpi and 14 dpi. Eggs from all the groups were tested negative for Salmonella. Moreover, there was no difference between groups in the number of IEL population. We also monitored air quality parameters throughout both experiments across all treatment groups. We are currently analyzing the data to identify any potential differences among the treatments. Additionally, we collected cecal samples for microbiome analysis from both studies. DNA will be extracted from these samples and submitted for microbiome sequencing analyses. Objective 3. The CFD model developed using ANSYS Fluent 14.0 for conventional caged layer housing has been successfully exported to the currently supported version, ANSYS Fluent 24.0. Key parameters from the prior model simulation were incorporated into the updated new model. The sensible and latent loads of the facility operating in a cage-free environment were calculated. Relevant user-defined functions (UDFs) were identified, but not working properly. Several of these UDFs were built, loaded and hooked into the new model. Objective 4. Dr. Zhao presented "Measurement, Modeling, and Mitigation of Indoor Environment, Air Quality, and Air Emissions at Poultry Facilities" at the Ohio Poultry Sustainability Summit organized by Ohio Poultry Association on Oc. 10, 2024. The new IEQ interventions and technologies were reviewed. Many large Ohio poultry producers participated in the Summit. The PI also presented "Engineering Technologies for Air Quality Control in Poultry Houses" at the 2025 Georgia Precision Poultry Farming Conference -Virtual, held on May 6, 2025. A Virtual Forum "Dust & Disease in Egg Production" was organized on June 12. 2025 through collaborative efforts among the team, industry partners (Ohio Poultry Association and Egg Industry Center), and peers (Brett Ramirez, Egg Industry Center, Associate Prof. Iowa State University; and Lilong Chai, Associate Professor & Engineering Specialist, Department of Poultry Science, The University of Georgia). Presentations about dust, poultry diseases, animal and human health protections, best available dust control technologies and management practices, and egg producers' experiences and perspectives were shared by academic researchers, USDA government agency researchers, private consulting companies, and egg producers. Research and extension education needs are shared by producer panel members. The Forum reached peak attendance of more than 170 audiences. Effectiveness of the Forum presentations, format, and future educational needs were also assessed through survey after the forum. The extension presentations were recorded and will be used to form online multidisciplinary seminars for college students on advanced management strategies and indoor environmental control technologies to sustain poultry health and performance in the new emerging cage-free housing.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Shuja Majeed, Bikash Aryal, Bikas R. Shah, Nimra Khalid, Lisa Bielke, Qiuhong Wang, Lingying Zhao, Ali Nazmi. The impact of cold stress on performance, blood parameters, mucosal immune response and Salmonella susceptibility in cage-free layers. Poster. CFAES Research Forum and OARDC Annual Research Conference. April 10, 2025. The Ohio State University, Columbus, OH.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Bikash Aryal, Shuja Majeed, Bikas R. Shah, Nimra Khalid, Lisa Bielke, Qiuhong Wang, Lingying Zhao, Ali Nazmi. Assessing the detrimental effects of chronic heat stress on commercial layers in cage-free system. Oral. Poultry Science Association (PSA) Annual Meeting. July 1417, 2025. Raleigh, NC.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Bikash Aryal, Shuja Majeed, Bikas R. Shah, Nimra Khalid, Lisa Bielke, Qiuhong Wang, Lingying Zhao, Ali Nazmi. Effect of heat stress on egg production, blood biochemistry, immune response, and disease susceptibility in cage-free laying hens infected with Salmonella Enteritidis. Oral. PSA Annual Meeting. July 1417, 2025. Raleigh, NC.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Shuja Majeed, Bikash Aryal, Bikas R. Shah, Nimra Khalid, Lisa Bielke, Qiuhong Wang, Lingying Zhao, Ali Nazmi. Cold stress in cage-free laying hens: effect on performance, mucosal immunity, blood biomarkers, and Salmonella susceptibility. Oral. PSA Annual Meeting. July 1417, 2025. Raleigh, NC.
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Progress 06/01/23 to 05/31/24
Outputs
Target Audience:The targeted audience of this project include Peer researchers, Stakeholders of the egg industry, Egg producers, Professionals working with the egg industry, Extension educators, and College students majored in animal production, health, facility and environmental control, and sustainable animal production. In the past year, we have focused on communications with egg producers, stakeholders and professionals of the egg industry, and the general public to introduce the goals and objectives of new project and seek their feedbacks on challenges and workforce training needs for cage-free layer operations. We participated the 2023 Egg Industry Issue Forum, interacted with egg producers, egg industry professionals, and peer researchers. We published a popular press article "Control Indoor Environment to Abate Heat Stress and Diseases in Poultry Production" at Ohio Country Journal to introduce this study in June 2023. In addition, we have communicated with peer researchers about the study to seek collaboration in the research and education objectives of the study. Changes/Problems:In the first project year, two co-PIs of the project had changed their jobs. Dr. Thaina, who was in charge of Obj. 4 had left OSU in fall of 2023. Dr. Bielke, who was in charge of Obj. 2 had moved to North Carolina State University (NCSU) in fall of 2023. Dr. Ali Nazmi was recruited to join the project to replace Dr. Bielke's roles and Dr. Bielke had taken the responsibilities of Dr. Thaina to work on Obj. 4. A subcontract was established with NCSU and a re-budgeting process has been completed. With the newly assembled project team, we will be able to conduct the project objectives/activities as planned in the proposal. HPAI outbreak still creates challenges in recruiting egg producers/farms to collaborate on field data collection at commercial egg farms. Recruitment of a graduate student in agricultural engineering is delayed by the vacancy of a graduate coordinator position in the PI's department. We are expecting a no-cost Extension for the project due to the above changes. What opportunities for training and professional development has the project provided?We recruited a PhD student with DVM and MS in Poultry Science in Autumn 2023. The student has completed 11 of 15 credits for course work required by our PhD program. In the lab, he has mastered general virological techniques, such as cell culture, virus isolation, microwell virus infectivity assay, plaque assay, immunofluorescent assay, and inoculation of embryonated chicken eggs (ECE) for influenza virus propagation. Also, he has learned and practiced basic sequence analyses, such as BLAST searching, building phylogenetic trees, and performing recombination analysis. He is ready to lead the experiments proposed for this project. We also recruited a post-doctoral research associate working on the project for the past year. He was trained on equipment and measurement methods and experiment design for indoor air quality monitoring at commercial poultry layer houses and research farms. The post-doctoral research associate also participated the 2024 CFAES Research Forum of the Ohio State University and the annual international conference of American Society of Agricultural and Biological Engineers to present some preliminary environmental measurement methods. How have the results been disseminated to communities of interest?We participated the 2023 Egg Industry Issue Forum, discussed the study objectives with egg producers, egg industry professionals, and peer researchers. We published a popular press article "Control Indoor Environment to Abate Heat Stress and Diseases in Poultry Production" at Ohio Country Journal to introducer this study in June 2023. We developed a poster presentation on ammonia measurement methods and presented at the 2024 CFAES Research Forum of the Ohio State University and the annual international conference of American Society of Agricultural and Biological Engineers. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. Reach out to CF egg producers to learn about their management challenges and needs for research and education programs We will continue participating the annual meetings with the stakeholder advisory board, annual EIC Industry Issue Forum, the annual UEP meeting, and Ohio Poultry Association Annual Celebration Banquet to exchange experiences and issues with egg producers on emerging cage-free layer housing, management challenges, and producers' needs for research and education in hen health and production efficiency. Objective 2. Assess the impacts of cage-free housing environments on microbiota, immune development, disease susceptibility and transmission, and bird performance We plan to continue working with the Egg Industry Center (EIC), Ohio Poultry Associate (OPA) and their advisors or stakeholders to identify collaborating layer farms and develop innovative data collection methods in consideration of limited access to commercial egg farms due to HPAI outbreak. Trainings will be developed and conducted for staff helpers of commercial collaborating farms. We will start simulation of heat stress tests in controlled environment poultry houses of the OSU Poultry Research Center. At 25 weeks of age, the birds will be divided into three groups and exposed to different temperature conditions: high (35-38°C), thermoneutral (22-25°C), or low (10-15°C). During the experiment, we will monitor relative humidity, ammonia levels, and airborne particulate matter (PM) concentrations. We will also collect weekly samples from the room environment (dust, water, floor, and air) and the birds for microbiome analysis. Blood samples will be collected for the analysis of serum cytokines, immune suppression markers, and quantification of peripheral blood mononuclear cells (PBMCs). Additionally, we will observe all performance traits, including body weight, feed consumption, and egg production and quality. Data analysis will be developed and the protocols for environmental monitoring, microbial sampling and analysis, and bird performance and health tests will be refined and finalized. Objective. 3 Development of an Upward Airflow Displacement Ventilation (UADV) System for Cage-Free Layer Houses We will start to design and develop a CFD model for the new UADV ventilation system. Ventilation effects on thermal environment conditions and disease transmission will be evaluated and optimized by the CFD simulations. Objective 4. Disseminate the advanced IEQ intervention strategies and technologies to the egg industry's current and future workforce. We will develop Extension workshops and/or online trainings on the following topics in collaborations with Dr. Brett Ramirez, the interim Director of the Egg Industry Center and Dr. Lilong Chai, the faculty organizer of the Georgia Precision Poultry Farming Conference Fundamental poultry health, immunity, and performance Poultry infectious diseases (HPAI, colibacillosis, salmonellosis) and current treatment and prevention of the disease Current cage-free layer housing and ventilation and environmental control systems Heat stress and indoor air quality in layer houses
Impacts
What was accomplished under these goals?
Objective 1. Utilize the established university and egg industry partnerships to reach out to cage-free egg producers to learn their management challenges and needs for research and education to improve hen health and production efficiency In the past year, we have been focused on communications and interactions with egg producers, stakeholders and professionals of the egg industry to introduce the goals and objectives the project and seek their feedbacks on challenges and workforce training needs for cage-free layer operations. We participated the 2023 Egg Industry Issue Forum, interacted with egg producers, egg industry professionals, and peer researchers. The PI also participated the 2023 Ohio Poultry Association Annual Celebration Banquet. A summary of the study for egg producers has been developed to communicate our needs for producer collaborations on the environmental monitoring and collection of microbiome, production, and bird health data at commercial cage-free layer houses. The summary has been used to recruit potential egg producer collaborators. We published a popular press article "Control Indoor Environment to Abate Heat Stress and Diseases in Poultry Production" at Ohio Country Journal to introduce this study to Ohio farmers in June 2023. Objective 2. Assess the impacts of cage-free housing environments on microbiota, immune development, disease susceptibility and transmission, and bird performance. We developed new IACUC (Institutional Animal Care and Use Committee) and IBC (the Institutional Biosafety Committee) protocols to encompass all animal trials and downstream lab experiments associated with this proposal. These protocols were submitted for review to The Ohio State University (OSU) IACUC and the IBC. The IACUC protocol 2024A00000066 and the IBC protocol 2015R00000086-R1 were both approved on July 24th, 2024. Following approval, we acquired 235-layer pullets at 16 weeks of age, which have been housed in cage-free conditions at the Poultry Research Center under a controlled environment. The birds are currently laying eggs, and we are ready to perform the first animal trial. We have acquired the equipment for thermal environment and air quality monitoring at commercial farms and the OSU Poultry Research Center. The equipment have been calibrated and students are trained to use the equipment. We have developed methods for environmental monitoring and sampling and analysis of microbiome in commercial cage-free layer houses. We have developed experimental plan for tests in controlled environment at OSU Poultry Research Center to evaluate potential impacts of key IEQ parameters on susceptibility to infectious diseases and pathogen transmission in research farms with controlled experiment. Dr. Nazmi's graduate student and lab technician are fully trained to carry out and analyze all proposed experiments, particularly microbiome analysis and immunological parameters.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Li, P. M. Herkins, R. Knight, L.Y. Zhao, S. Akter, L. Wang-Li, J. Q. Ni, and A. Heber. 2024. Comparison of three on-field measurement methods for low-level ammonia concentrations at ambient locations of a poultry layer production facility. Poster presentation at the 2024 CFAES Research Forum, the Ohio State University, April 9, 2024 at the Nationwide & Ohio Farm Bureau 4H Center in Columbus, OH.
- Type:
Other
Status:
Published
Year Published:
2023
Citation:
Zhao. L.Y. 2023. Control Indoor Environment to Abate Heat Stress and Diseases in Poultry Production. Ohios Country Journal (OCJ). June 2023. https://ocj.com/2023/06/control-indoor-environment.
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