Progress 06/15/21 to 06/14/24
Outputs
Target Audience:The target audience for this project was three-fold: animal scientists and the poultry industry, my undergraduate research mentees, and students enrolled in the class ANSI 4843 - Application of Biotechnology in Animal Science. Results from this project were presented to animal scientists and the poultry industry at three national conferences and five local meetings. In addition, two first-author publications related to the project were published and a third manuscript will be submitted this year. Throughout the project, I have had the pleasure of mentoring > 15 graduate and undergraduate researchers. Two of my undergraduate research mentees were awarded a Wentz Research Scholarship and a Niblack Research Scholarship, respectively, at Oklahoma State University. Furthermore, I have led both lab and lecture sessions of ANSI 4843 (Application of Biotechnology in Animal Science) for four years. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?In addition to learning classic epigenetic research techniques, this project has provided several opportunities to engage in professional development through the University and at conferences. This spring, I successfully defended my dissertation in addition to presenting my research at the Biological Sciences Symposium and at the Conference of Research Workers in Animal Diseases. In total, my research was shared with the scientific community at three national conferences, two local research symposiums, and in an additional three research presentation competitions. For the past year, I have been the Vice President of the Animal Science Graduate Student Association (ASGSA) and was responsible for coordinating the department's annual Totusek Lectureship event. Previously, I held the President and Secretary positions in the ASGSA for one-year terms. Throughout my graduate program, I have enrolled in numerous professional development courses offered by Oklahoma State University (OSU). The Career Skills and Professionalism course invited guest speakers from diverse career paths to present their experiences from working in academia, industry, government, or leadership. I was given the opportunity to complete a mock application and interview for an academic position. The Teaching in Agriculture course emphasized best practices for effective and engaging teaching. One of the class requirements was to write and receive feedback on my teaching philosophy. In 2021, I completed a Grantsmanship course to enhance my writing skills. Furthermore, I attended the annual 2021-2023 Graduate Teaching Assistant Conferences offered by the Institute for Teaching & Learning Excellence at OSU. There were many other workshops provided by the 360° Professional Development Program that I participated in, for example, how to use EndNote20 with Word. In 2023, I also attended a seminar series offered by the NIH called, "Becoming a Resilient Scientist." My research, professional service, and student mentoring culminated in earning me the outstanding PhD student award from the Department of Animal and Food Sciences at OSU. How have the results been disseminated to communities of interest?The results of this project have been published in Frontiers in Immunology (2022; 13: 874706), the Journal of Animal Science and Biotechnology (2024; 15: 29), and Advances in Nutrition (2024; 15:9), and have been presented on multiple occasions. This spring, I successfully defended my dissertation, "Regulation of Endogenous Host Defense Peptide Synthesis as an Antibiotic-Free Approach for Disease Control in Poultry." In addition, I presented my research during the Biological Sciences Symposium at Oklahoma State University (OSU) in February and at the Conference of Research Workers in Animal Diseases (CRWAD) in January. I also competed in the inaugural Three Minute Thesis competition at the 2024 CRWAD. Since the start of my predoctoral fellowship, I have presented the results from this project at a total of three national conferences, two local symposiums, and in an additional three presentation competitions. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We found several combinations of histone deacetylase inhibitors (HDACi) and histone methyltransferase inhibitors (HMTi) or DNA methyltransferse inhibitors (DNMTi) that synergistically increased the expression of host defense peptides (HDPs). Two chicken macrophage cell lines were treated with structurally distinct HDACi, HMTi, or DNMTi individually or in combinations, followed by HDP gene expression analysis. Each epigenetic compound was found to be capable of inducing HDP expression. To our surprise, a combination of HDACi and HMTi or HDACi and DNMTi showed a strong synergy to induce the expressions of most HDP genes. The HDP-inducing synergy between butyrate, an HDACi, and BIX01294, an HMTi, were further verified in chicken peripheral blood mononuclear cells. Furthermore, tight junction proteins such as claudin 1 were also synergistically induced by HDACi and HMTi. These results have been published in Frontiers in Immunology (2022, 13: 874706). In addition, we revealed that butyrate, a short-chain fatty acid and a well-known histone deacetylase inhibitor (HDACi), synergizes strongly with deoxycholic acid (DCA) in inducing the expression of host defense peptide (HDP) and tight junction protein genes. To investigate the possible synergistic effect between DCA and butyrate in regulating HDP synthesis and barrier function, we treated chicken HD11 macrophage cells and jejunal explants with DCA and sodium butyrate (NaB), either individually or in combination, for 24 h. Subsequently, we performed RNA isolation and reverse transcription-quantitative PCR to analyze HDP genes as well as the major genes associated with barrier function. To further determine the synergy between DCA and NaB in enhancing resistance to necrotic enteritis, we conducted two independent trials with Cobb broiler chicks. In each trial, the diet was supplemented with DCA or NaB on the day of hatch, followed by NE induction through sequential challenges with Eimeria maxima and Clostridium perfringens on days 10 and 14, respectively. We recorded animal mortality after infection and assessed intestinal lesions on day 17. We found that the combination of DCA and NaB synergistically induced multiple HDP genes in both chicken HD11 cells and jejunal explants. Additionally, the gene for claudin-1, a major tight junction protein, also exhibited synergistic induction in response to DCA and NaB. Furthermore, dietary supplementation with a combination of 0.75 g/kg DCA and 1 g/kg NaB led to a significant improvement in animal survival and a reduction in intestinal lesions compared to either compound alone in a chicken model of NE. In conclusion, DCA synergizes with NaB to induce HDP and claudin-1 expression and enhance NE resistance, with potential for further development as cost-effective antibiotic alternatives. This study has been published in the Journal of Animal Science and Biotechnology (2024, 15: 29). A subsequent study employed a 2 × 2 factorial design to investigate the protective mechanisms of DCA in healthy and NE-affected broiler chickens. Animal survival, weight gain, intestinal lesions, and the intestinal microbiome composition were assessed. We confirmed that dietary supplementation of 1.5 g/kg DCA offers robust protection against NE. Animal survival significantly increased, growth inhibition was reversed, and intestinal lesion severity was alleviated. Furthermore, DCA exhibited selective inhibition C. perfringens, Escherichia, and E. cecorum colonization, accompanied by the enrichment of major lactic acid bacteria (LAB) in both the ileum and cecum. Major short chain fatty acid-producing bacteria like Faecalibacterium were also partially restored by the supplementation of DCA to broiler chicks. These combined effects of DCA likely contribute to its protective efficacy against NE. DCA and these DCA-enriched LAB present a promising antibiotic-free alternative for mitigating NE in chickens. Overall, this project successfully demonstrated two antibiotic-free approaches to bolster host defense peptide synthesis and improve poultry health.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Whitmore MA, Tobin I, Burkardt A, Zhang G. (2024) Nutritional modulation of host defense peptide synthesis: a novel host-directed antimicrobial therapeutic strategy? Advances in Nutrition 15: 9.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Yang Q, Liu J, Wang X, Robinson K, Whitmore MA, Stewart SN, Zhao J, Zhang G. (2021) Identification of an intestinal microbiota signature associated with the severity of necrotic enteritis. Frontiers in Microbiology 12: 703693.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Yang Q, Whitmore MA, Robinson K, Lyu W, Zhang G. (2021). Butyrate, forskolin, and lactose synergistically enhance disease resistance by inducing the expression of the genes involved in innate host defense and barrier function. Antibiotics (Basel) 10: 1175.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Yang Q, Liu J, Robinson K, Whitmore MA, Stewart SN, Zhang G. (2021) Perturbations of the ileal mycobiota by necrotic enteritis in broiler chickens. Journal of Animal Science and Biotechnology 12: 107.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Liu J, Stewart SN, Robinson K, Yang Q, Lyu W, Whitmore MA, Zhang G. (2021) Linkage between the intestinal microbiota and residual feed intake in broiler chickens. Journal of Animal Science and Biotechnology 12: 22.
- Type:
Journal Articles
Status:
Other
Year Published:
2024
Citation:
Whitmore MA, Zhang G. Epigenetic regulation of host defense peptide synthesis. Intended for Frontiers in Immunology.
- Type:
Journal Articles
Status:
Other
Year Published:
2024
Citation:
Whitmore MA, Guo J, Kim DM, Liu J, Tobin I, Zhao Z, Zhang G. Deoxycholic acid protects chickens from necrotic enteritis by inhibiting pathobionts while enriching specific lactic acid bacteria. Intended for Pathogens.
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2024
Citation:
Whitmore MA. Regulation of endogenous host defense peptide synthesis as an antibiotic-free approach for disease control in poultry. PhD Defense, Department of Animal and Food Sciences, Oklahoma State University
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Wu Y, Zhang X, Liu X, Li Y, Han D, Pi Y, Whitmore MA, Xingmiao L, Zhang G, Zheng J, Wang J. (2023) Strain specificity of lactobacilli with promoted colonization by galactooligosaccharides administration in protecting intestinal barriers during Salmonella infection. Journal of Advanced Research 56.
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Progress 06/15/22 to 06/14/23
Outputs
Target Audience:The target audience included animal scientists and the poultry industry, my undergraduate research mentees, and students enrolled in the class ANSI 4843 - Application of Biotechnology in Animal Science. The results of my research project were presented for the Three Minute Thesis competition at Oklahoma State University and at the Poultry Science Association Annual Meeting. Over the past year, I mentored two undergraduate researchers in experiment design, project execution, analysis, and in scientific communication. One of my mentees received a Wentz Research Grant for the 2022-2023 academic year. During the spring 2023 semester, I led the majority of the lectures for the Biotechnology class to further polish my teaching and mentoring skills. Changes/Problems:No major changes or problems have been encountered for this project. What opportunities for training and professional development has the project provided?The University presented me with the opportunity to enroll in two professional development classes during the Fall 2022 and Spring 2023 semesters. The Career Skills and Professionalism course invited guest speakers from many career types to share their experiences with the class. I was given the opportunity to complete a mock application to an academic position, including submitting my CV and cover letter and participating in a practice interview. The course instructor provided feedback on the application materials and the quality of the interview. In the College Teaching of Agriculture and Natural Resources, the course instructor as well as the Department Head of Agricultural Education shared best practices for effective and engaging teaching. Two major projects in the class prompted me to design a course syllabus and create the first draft of my teaching philosophy. How have the results been disseminated to communities of interest?The results of this project have been published in Frontiers in Immunology (2022, 13: 874706) and have been presented on multiple occasions. My research findings were summarized for the Three Minute Thesis competition at Oklahoma State University, and a second presentation was given at the Poultry Science Association Annual Meeting in July of 2022. A second manuscript has also been submitted to the Journal of Animal Science and Biotechnology. What do you plan to do during the next reporting period to accomplish the goals?Not only do HDACi and HMTi synergize for the gene induction of chicken HDPs, but we have also found a prominent synergy between HDACi and a third class of epigenetic compounds named DNA methyltransferase inhibitors (DNMTi). During the next reporting period, I will test for synergy between DNMTi and HMTi to ensure we have found the most effective combination of epigenetic compounds to enhance the production of chicken HDPs. Concurrently, I will evaluate the mechanism of action for synergy between HDACi and HMTi through chromatin accessibility assays, western blot, and ChIP-qPCR. Additionally, I will evaluate the impact of these epigenetic modifiers and their combinations on the host physiology, gut microbiome, and disease resistance.
Impacts
What was accomplished under these goals?
We revealed that butyrate, a short-chain fatty acid and a well-known histone deacetylase inhibitor (HDACi), synergizes strongly with deoxycholic acid (DCA) in inducing the expression of host defense peptide (HDP) and tight junction protein genes. To investigate the possible synergistic effect between DCA and butyrate in regulating HDP synthesis and barrier function, we treated chicken HD11 macrophage cells and jejunal explants with DCA and sodium butyrate (NaB), either individually or in combination, for 24 h. Subsequently, we performed RNA isolation and reverse transcription-quantitative PCR to analyze HDP genes as well as the major genes associated with barrier function. In conclusion, DCA synergizes with NaB to induce HDP and claudin-1 expression, with the potential for further development as a cost-effective antibiotic alternative.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2023
Citation:
Kim DM, Liu J, Whitmore MA, Tobin I, Zhao Z, Zhang G. (2023) Two intestinal microbiota-derived metabolites, deoxycholic acid and butyrate, synergize to enhance host defense peptide synthesis to enhance host defense peptide synthesis and alleviate necrotic enteritis. Journal of Animal Science and Biotechnology.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Robinson K, Yang Q, Stewart S, Whitmore MA, Zhang G. (2022) Biogeography, succession, and origin of the chicken intestinal mycobiome. Microbiome 10: 55.
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Progress 06/15/21 to 06/14/22
Outputs
Target Audience:The target audience of my project over the past year included animal scientists and the poultry industry, my undergraduate research mentees, and students enrolled in the class ANSI 4843 - Application of Biotechnology in Animal Science. Results of my research project were presented twice at Oklahoma State University, for the Whiteman Research Presentation Competition and at the Biological Symposium Presentation Competition, and for a third time at the Poultry Science Association Annual Meeting. During the 2021-2022 academic year, I mentored two undergraduate researchers in experiment design, statistical analysis, and scientific communication. I also took a more active role in the Biotechnology class this spring and had the opportunity to lead the majority of the lectures, to better develop both my presentation and mentorship skills. Changes/Problems:No major changes or problems have been encountered for this project. What opportunities for training and professional development has the project provided?In addition to learning classic epigenetic research techniques, this project has provided several opportunities to engage in professional development through the University and at conferences. Last year I completed a Grantsmanship course to enhance my writing skills and attended the 2021 Graduate Teaching Assistant Conference offered by the Institute for Teaching & Learning Excellence (ITLE) at Oklahoma State University. There are also many workshops provided by the 360o Professional Development Program that I participated in, for example, how to use EndNote20 with Word. I am now enrolled in a seminar series from the NIH called, "Becoming a Resilient Scientist." This summer, my abstract was accepted by the Poultry Science Association Annual Meeting, where I presented my research and had the chance to network with industry professionals and poultry scientists from across the world. How have the results been disseminated to communities of interest?The results of this project have been published in Frontiers in Immunology (2022, 13: 874706) and have been presented on multiple occasions. My research findings were summarized for two different presentation competitions at Oklahoma State University, including the Whiteman Competition in the Department of Animal and Food Sciences and the campus-wide Biological Symposium. A third presentation was given at the Poultry Science Association Annual Meeting during the summer of 2022. What do you plan to do during the next reporting period to accomplish the goals?We have found that not only do HDACi and HMTi synergize for the gene induction of chicken HDPs, but there is also prominent synergy between HDACi and a third class of epigenetic compounds named DNA methyltransferase inhibitors (DNMTi). During the next reporting period, I will test for synergy between DNMTi and HMTi to ensure we have found the most effective combination of epigenetic compounds to enhance the production of chicken HDPs. Concurrently, I will evaluate the mechanism of action for synergy between HDACi and HMTi through chromatin accessibility assays, western blot, and ChIP-qPCR. The first of these mechanistic studies will identify the optimal time point at which to measure HDP promoter accessibility and changes in histone acetylation or methylation in response to treatment with HDACi and HMTi.
Impacts
What was accomplished under these goals?
Objective 1 has been completed and the results published in Frontiers in Immunology (2022, 13: 874706). We found several combinations of histone deacetylase inhibitors (HDACi) and histone methyltransferase inhibitors (HMTi) or DNA methyltransferse inhibitors (DNMTi) that synergistically increased the expression of host defense peptides (HDPs). Two chicken macrophage cell lines were treated with structurally distinct HDACi, HMTi, or DNMTi individually or in combinations, followed by HDP gene expression analysis. Each epigenetic compound was found to be capable of inducing HDP expression. To our surprise, a combination of HDACi and HMTi or HDACi and DNMTi showed a strong synergy to induce the expressions of most HDP genes. The HDP-inducing synergy between butyrate, an HDACi, and BIX01294, an HMTi, was further verified in chicken peripheral blood mononuclear cells. Furthermore, tight junction proteins such as claudin 1 were also synergistically induced by HDACi and HMTi. For objective two, primers have been designed and confirmed, and the assay kits received for chromatin immunoprecipitation qPCR (ChIP-qPCR) and chromatin accessibility.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Whitmore MA, Li H, Lyu W, Khanam S, Zhang G. Epigenetic Regulation of Host Defense Peptide Synthesis: Synergy Between Histone Deacetylase Inhibitors and DNA/Histone Methyltransferase Inhibitors. Front Immunol. 2022 Apr 22;13:874706. doi: 10.3389/fimmu.2022.874706. PMID: 35529861; PMCID: PMC9074817.
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