Source: MICHIGAN STATE UNIV submitted to NRP
RELATIONSHIP BETWEEN STRESS AND GASTROINTESTINAL DISORDERS IN SWINE: MECHANISMS AND INTERVENTIONS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1008915
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Mar 1, 2016
Project End Date
Feb 28, 2021
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
MICHIGAN STATE UNIV
(N/A)
EAST LANSING,MI 48824
Performing Department
Large Animal Clinical Sciences
Non Technical Summary
The susceptibility to adult onset of chronic, debilitating gastrointestinal diseases and reduced feed efficiency in agricultural animals is profoundly influenced by environmental influences occurring early in life. We will study the mechanisms by which early life stress triggers in pigscauses long-lasting defects in intestinal epithelial barrier function, a primary causative factor in the development of GI diseases. These studies have the potential to unravel the mechanisms of early life stress-induced GI disorders leading to the identification of novel therapeutic targets for the prevention and treatment of a number of GI diseases of humans and animals.
Animal Health Component
25%
Research Effort Categories
Basic
75%
Applied
25%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31135101020100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3510 - Swine, live animal;

Field Of Science
1020 - Physiology;
Goals / Objectives
Our long-term goal is to understand fundamental mechanisms by which early life stress influences the development and severity of GI disease in later life. The objective of the current proposal is to determine the precise mechanisms by which early life stress alters the development of the enteric cholinergic nervous system leading to chronic intestinal mast cell activation and long-term impairment of intestinal barrier function. Four specific aims (SA1-SA4) were designed to achieve the objective of this research.SA1: Determine the mechanism by which early life stress triggers long-term amplification of enteric cholinergic nervous system activity. We will utilize in vivo approaches, ex vivo electro-physiologic assays, and confocal imaging of the porcine intestine to test our working hypothesis that early intestinal inflammatory pathways alter development of intestinal cholinergic neurons leading to increased activity and long-term intestinal barrier dysfunction. Upon completion of this objective, we expect to have a fundamental understanding of how stress impacts enteric nervous system development and how this modulates key aspects of intestinal epithelial function, including intestinal barrier function and intestinal absorptive and secretory mechanisms. These findings are directly translatable to growth and feed efficiency and disease resistance in agricultural animals such as the pig. SA2: Investigate the mechanisms by which cholinergic signaling triggers persistent mast cell activation. We will utilize innovative porcine animal models and in vitro cell culture systems to test our working hypothesis that cholinergic neurons trigger persistent mast cell activation via M3 muscarinic receptors on mast cells. Upon completion of this objective, we expect we will have gained mechanistic insight into how stress modulates the intestinal immune system via altering mast cell function. These findings will have direct relevance to how agricultural production stressors impairs the ability of the animals immune system to mount an effective immune response to immunological and pathogenic challenges. SA3; Understand the impact of early life stress and cholinergic nerve-mast cell signaling on alterations in epithelial tight junctions We will conduct in-depth imaging analysis in porcine and human intestinal models to test our working hypothesis that early life stress and cholinergic-mast cell signaling induce specific alterations in tight junction proteins that cause impaired barrier function. Upon completion of this objective, we expect to have a fundamental understanding of how alteration in the enteric nervous system and mast cells in stressed pigs is associated with tight junction protein disruption and intestinal permeability. Given the importance of intestinal barrier function in driving nutrient absorption and secretion while protecting the body against the potentially hostile luminal environment, this information will have direct relevance to animal disease and production efficiency.SA4: Determine whether dietary modifications can influence intestinal barrier function in stressed pigs. We will conduct controlled feeding trial experiments and test whether specific feed additive or pharmaceuticals targeted at stress mechanism described above (e.g. mast cell activation, enteric nervous system development, tight junction proteins) are able to modulate gut barrier and immune function in stressed pigs. A few dietary and(or) pharmaceutical strategies under consideration here include CRF receptor antagonists, mast cell stabilizers, long chain polyunsaturated fatty acids, antimicrobials, etc. Upon completion of these studies, we hope to have identified nutritional strategies or feed additives that could modulate gut biology in the stressed animal and therefore promote intestinal defense and improve feed efficiency
Project Methods
Define the role of intestinal inflammation in the long-term amplification of intestinal cholinergic nervous system in pigs subjected to early life stress. Nursing piglet litters (Yorkshire x Large White x Hampshire cross) will be randomly assigned to one of three experimental treatments. A subset of piglets from each litter will be weaned from the sow at 16 d of age (early weaning stress) while the remaining piglets will remain with the sow until they are weaned at 28 days of age (Control). Early weaned pigs will be transferred to the Swine Metabolism Unit or J-Building veterinary research farm (MSU). Sow parity will be evenly distributed across experimental groups to control for potential immunological and behavioral differences between different age sows. Early weaned pigs will be randomly assigned to receive either daily IM injections with saline vehicle (Treatment 2) or hydrocortisone (anti-inflammatory treatment, Treatment 3) until to 28 d of age. To control for nutritional differences between early weaned pigs and controls, all early weaned pigs will be placed on a liquid milk replacer diet (with similar nutritional content to sow's milk) until 28 days of age. From 28 days of age onward, all pigs will be maintained in the same research facility under identical environmental and nutritional conditions (see animal care procedures for detailed description of housing and diets). To assess the impact of experimental treatments on the development of long-term enteric cholinergic function in the pig, we will perform functional electrophysiologic analyses and imaging analyses of enteric cholinergic neuron activity on post-weaning days 0 (weaning), 2, 28, 60, 120 and 210 (n=8 pigs/treatment/time point) as described below.Electrophysiologic Assessment of intestinal cholinergic nervous system activity in porcine intestine: At each post-weaning time point, pigs from each experimental group will be sacrificed. Intestinal tissues will be immediately harvested and prepared as flat sheets of ileal and colonic mucosa and mounted on Ussing chambers(13). To investigate cholinergic secretomotor neuronal function, we will use an electrical field stimulation (EFS) technique, where an electrical current will be passed across the intestinal tissues on Ussing chambers resulting in the firing of all enteric nerves that will be measured by recording elevations in transepithelial short-circuit current (Isc) (14, 15). To determine the contribution of cholinergic neurons to the EFS induced Isc response, EFS will be conducted in the presence of various inhibitors including tetrodotoxin (a neurotoxin that will confirm neural-mediated Isc responses), atropine (an acetylcholine muscarinic receptor antagonist, to confirm cholinergic-mediated Isc responses mediated via muscarinic receptors, 5 µM), hexamethonium (a nicotinic, ganglionic blocker to confirm cholinergic-mediated Isc responses via nicotinic receptors, 100 µM) and VPAC1 (a VIP receptor antagonist to determine VIPergic contribution, 1µM). In addition we will measure tissue acetylcholine release by measuring acetylcholine levels in Ussing chamber supernatants via ELISA (EMD Millipore). All reagents proposed here have been established previously in porcine intestinal tissues(14, 16, 17).Data analysis. The numbers of pigs proposed in these experiments is based from a power calculation and our previous experience with these techniques with the goal of achieving appropriate statistical power (P=0.8) and being able to detect significance between treatments at the p<0.05 level. For EFS experiments, Isc data will be recorded at 1 sec intervals will be expressed either as real time data tracings or as the relative change in Isc from baseline in response to EFS. Statistical significance will be determined by ANOVA with or without repeated measures analysis (for time course data) followed by a Tukey's post hoc analysis. Differences will be considered significant at the p<0.05 level. For Western blot results, densitometry analysis of protein bands for each marker will be performed and expressed relative to the pan-neuronal marker PGP 9.5 (EMD Millipore). Determine the influence of cholinergic neural activity on mast cell activation Established DRG-mast cell (BMMC or HMC-1 cells) co-cultures will be treated with select pharmacologic modulators of the cholinergic system (Table 2). To stimulate cholinergic signaling we will treat DRG-mast cell co-cultures with the acetycholinesterase inhibitor physostigmine which will elevate endogenous Ach.(22) In addition we will conduct studies in normal (non-cocultured) mast cell cultures where we will stimulate BMMCs and HMC-1 cells with the muscarinic receptor agonists Ach and carbachol at dose dependent concentrations. Treatments will also be tested on non-cocultured DRGs and mast cell cultures as controls In addition, all treatments will be tested in the absence or presence of atropine and a selective M3 receptor inhibitor to confirm that the effects are via muscarinic receptor-dependent pathways. Mast cell activation will be determined at 0, 1, 4, 6, 12, and 24 hrs after treatments by histological and biochemical assays. Histological evidence of mast cell degranulation will be determined by toluidine blue stainingData analysis: For mast cell activation studies in Experiments SA2a-c, β-hexosaminidase will be presented as the % release of β-hexosaminidase into culture supernatants at each time point according to routine calculations.(23) Tryptase will be presented at the mg tryptase/mL in the supernatant. Other mast cell mediators measured via ELISA will be presented as pg/mL. For barrier function analyses in SA2.d., data will be analyzed at each time point using a standard 2-way ANOVA with treatment and time as the main factors. A post-hoc Tukey's test will be used to determine differences between experimental treatments. Based on our experience we expect that an n=6-8 mice/timepoint/treatment will be needed to detect statistically significant differences between treatments at the P<0.05 level and with a Power of 0.8.Determine the impact of early life stress on the development and profile of TJ proteins. Intestinal tissues will be harvested from early weaned pigs (early weaning stress) and control pigs (late weaned) at Day 0, 2, 28, 60, and 120 post-weaning. We will utilize tissue from experiments outlined in Specific Aim 1 to minimize animal use. For IF studies, intestinal tissues will be embedded in OCT and sectioned at 5 micrometer and processed followed by TJ protein profiling focused on occludin, ZO-1, (which tend to remain in place during developmental and injured states) and claudins 1-5, 7, 8, 10.12. 15, 17 (which differ in presence and expression according to development and injury) will be analyzed using sucrose density gradients and qPCR of cell membranes using sucrose density gradients, followed by western blotting, and qPCR (26, 27),(28). Data Analysis: Barrier function data (TER and FD4 flux) will be expressed as either real-time data over time or as the % change after experimental treatments. Specifically, FD4 flux data will be expressed as the rate (mg FD4/min) of mucosal-to-serosal FD4 flux across the intestine or in vitro cell culture system. Data that is expressed as the % change after treatments will be analyzed using a standard 2-way ANOVA with pig and treatment as the main effects. Data measured from the same tissue/culture over time will be analyzed with a ANOVA on repeated measures. P<0.05 will be considered as a significant difference and P values between .051 and .10 will be considered trends. For TJ protein analysis by Western blotting, protein bands for each TJ protein within each sucrose gradient fraction will be expressed relative to beta actin using densitometry pixel analysis (Adobe Photoshop).

Progress 03/01/16 to 02/28/21

Outputs
Target Audience:The target audiences include the swine industry (integrators, producers and pharma) and academic (scientists, extension, students) groups. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Students partcicipating with this research have presented at national conferences including CRWAD, Annual Conference of the American Association of Immunilogists and annual meeting of the Organization for the Study of Sex Differences. Students participated in manuscript and abstract writing and felloship applictions. How have the results been disseminated to communities of interest?Research has been presented at university, national and internation venues. These include basic science meetings with universities, production animal integrators, nurtitionists and industry. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? SA1: Determine the mechanism by which early life stress triggers long-term amplification of enteric cholinergic nervous system activity. 1) Major activities completed / experiments conducted: Characterized the impact of early weaning and biological sex on development of enteric cholinergic nervous system expression and function in pigs. 2) Data collected; expression of cholinergic proteins by qPCR, Western blot and immunohistochemistry. Ussing chamber analysis of cholinergic mediates secretory function in the small intestine. Fecal analysis. 3) Summary statistics and discussion of results. These studies revealed that early weaned pigs exhibit alterations in the development of the enteric nervous system, specifically cholinergic neurons compared with later weaned pigs. Early weaned pigs had greater numbers of cholinergic neurons and increased excitability resulting in enhanced secretory function compared with later weaned pigs. Further, early weaned female gilts exhibited greater cholinergic response compared with male castrates. Enhanced cholinergic function and heightened secretory function corresponded with increased incidence of diarrhea in early weaned pigs. 4) Key outcomes or other accomplishments realized. The intestinal cholinergic system is a major regulator or gut functions including motility, secretion and immune activity. Here we showed that common stressful production practices in early life, such as early weaning, can lead to long-lasting upregulation of the intestinal cholinergic nervous system in pigs. Given the role of the cholinergic system in normal GI functions in health and disease, these findings have significant implications to pig health and suggest that the cholinergic system may be a novel target for health interventions. SA2: Investigate the mechanisms by which cholinergic signaling triggers persistent mast cell activation. 1) Major activities completed / experiments conducted; determined how Salmonella typhimurium challenge altered the expression of cholinergic proteins in the pig intestine. Measured inflammatory cytokine and histological analysis of intestinal mucosa and expression and localization of cholinergic proteins. 2) Data collected; expression of marker of the cholinergic system intestinal mucosa, lymph nodes and blood from challenged and control pigs. histological analysis of intestinal injury. 3) Summary statistics and discussion of results; Upon challenge of pigs with Salmonella typhimurium, the enteric cholinergic system is profoundly upregulated, suggesting an important role of this system in enteric inflammatory disease pathogenesis. Moreover, we discovered that upregulation of the cholinergic system was not restricted to the enteric nervous system as a significant increase in expression was observed in intestinal epithelial cells. the level of cholinergic marker expression correlated with histological inflammatory scores and cytokines 4) Key outcomes or other accomplishments realized. These experiments suggest that the cholinergic system in the intestine is acutely responsive to enteric pathogenic challenges. These studies provide the rationale for future investigations on the role of cholinergic activation in the gut in mediating inflammatory and defense mechanisms, which in turn could be a target for manipulation in enteric disease challenges. SA3; Understand the impact of early life stress and cholinergic nerve-mast cell signaling on alterations in epithelial tight junctions 1) Major activities completed / experiments conducted. Characterized the impact of early weaning and biological sex on intestinal mast cells and intestinal barrier function. Conducted studies evaluating the effects of histamine receptor antagonist drugs on weaning induced immune responses. 2) Data collected; intestinal mast cells and barrier function analyzed. qPCR and protein analysis of expression of cytokines and proinflammatory mediators , and histological assessment of histamine receptor location and expression in the intestine of weaned pigs. 3) Summary statistics and discussion of results. Early weaning increased in early and lasting increased mast cell activation. Upregulation of histamine receptors in the intestine prompted studies investigating the role ofof HR antagonists in modulating intestinal inflammation associated with weaning stress. 4) Key outcomes or other accomplishments realized. The studies demonstrated that intestinal mast cells increase in number and activity in response to early weaning and that mediate intestinal permeability changes. Histamine, a major product of mast cells is elevated along with its receptors (H1, H2, and H4) in the intestine. SA4: Determine whether dietary modifications can influence intestinal barrier function in stressed pigs. 1) Major activities completed / experiments conducted; Measurement of nutrient transporter expression and function in early weaned and late weaned pigs. conducted metabolic and inflammatory analyses. 2) Data collected; Glucose transporter expression by qPCR and Western blot. Glucose transporter function in Ussing chambers. Plasma levels of glucose, inflammatory cytokines, and fatty acids. 3) Summary statistics and discussion of results: Early weaning induces lasting developmental alterations in glucose transporter expression and function. early weaning increased adiposity of visceral adipose stores and gene expression. 4) Key outcomes or other accomplishments realized. These studies demonstrated that the development of long-term nutrient transport in the pig intestine is significantly influenced by early weaning. This coincides with inflammation. These studies have laid the foundation for future investigations focusing on the role of nutrient transporters in the modulation of chronic inflammation.

Publications

  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Sun M, Shankar R, Ko M, Chang CD, Yeh SJ, Li S, et al. Sex differences in viral entry protein expression, host responses to SARS-CoV-2, and in vitro responses to sex steroid hormone treatment in COVID-19. Res Sq. 2020.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Williams ES, Manning CE, Eagle AL, Swift-Gallant A, Duque-Wilckens N, Chinnusamy S, et al. Androgen-dependent excitability of mouse ventral hippocampal afferents to nucleus accumbens underlies sex-specific susceptibility to stress. Biological psychiatry. 2020;87(6):492-501.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Thelen K, Wilson N, Berrios-Vazquez G, Moeser AJ. Mast cell histamine mediates intestinal immune response to early life stress in piglets via histamine 1 receptor (H1R). American Association of Immunologists; 2020.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Bali V, Mackey E, Moeser A. Perinatal androgen exposure impacts mast cell development and mediator storage. American Association of Immunologists; 2020.
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Lewton JR, Woodward AD, Moser RL, Thelen KM, Moeser AJ, Trottier NL, et al. Effects of a multi-strain Bacillus subtilis-based direct-fed microbial on weanling pig growth performance and nutrient digestibility. Translational Animal Science. 2021;5(3):txab058.
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Gao Y, Duque-Wilckens N, Aljazi MB, Wu Y, Moeser AJ, Mias GI, et al. Loss of histone methyltransferase ASH1L in the developing mouse brain causes autistic-like behaviors. Communications Biology. 2021;4(1):1-10.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Duque-Wilckens N, Maradiaga NC, Szu-ying Y, Nestler EJ, Robison AJ, Moeser AJ. Mast cell-specific inactivation of Fosb exacerbates release of pro-inflammatory mediators in models of systemic anaphylaxis and lipopolysaccharide-induced sepsis. American Association of Immunologists; 2021.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Piliponsky, A.M., et al., Evidence for sexual dimorphism in mast cell-mediated innate immune response against Group B Streptococcus. 2020, American Association of Immunologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Duque-Wilckens, N., et al., Mast cells as Immune modulators of behavior and brain cellular composition. 2020, American Association of Immunologists.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Mackey, E., et al., Perinatal androgens organize sex differences in mast cells and attenuate anaphylaxis severity into adulthood. Proceedings of the National Academy of Sciences, 2020. 117(38): p. 23751-23761.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: McClain, J.L., et al., Histamine-dependent interactions between mast cells, glia, and neurons are altered following early-life adversity in mice and humans. American Journal of Physiology-Gastrointestinal and Liver Physiology, 2020. 319(6): p. G655-G668.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lewton, J.R., et al., 129 Effects of a multi-strain Bacillus subtilis-based direct-fed microbial on growth performance, nutrient digestibility, and colonic pH in diets fed to weanling pigs. Journal of Animal Science, 2020. 98(Supplement_4): p. 101-102
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moeser, A.J., 357 Early life factors shaping gut development and lifelong disease risk. Journal of Animal Science, 2020. 98(Supplement_4): p. 92-92.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pohl, C.S., et al., S. Typhimurium challenge in juvenile pigs modulates the expression and localization of enteric cholinergic proteins and correlates with mucosal injury and inflammation. Autonomic Neuroscience, 2018. 213: p. 51-59.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lewton, J.R., et al., 130 Effects of a multi-strain Bacillus subtilis-based direct-fed microbial on immunity markers, intestinal morphology, and microbial communities in diets fed to weanling pigs. Journal of Animal Science, 2020. 98(Supplement_4): p. 105-106.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Boyd, R.D., et al., Review: innovation through research in the North American pork industry. Animal, 2019. 13(12): p. 2951-2966.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: D'Costa, S., et al., Mast cell corticotropin-releasing factor subtype 2 suppresses mast cell degranulation and limits the severity of anaphylaxis and stress-induced intestinal permeability. J Allergy Clin Immunol, 2019. 143(5): p. 1865-1877 e4.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mackey, E. and A.J. Moeser, Perinatal androgens drive sex differences in mast cell phenotype and severity of mast cell disease. 2019, American Association of Immunologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Maradiaga, N.C., et al., Early Life Adversity Programs Mast Cells Toward a Hyperactive Phenotype into Adulthood. 2019, American Association of Immunologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moeser, A.J., et al., 115 Negative impacts of early weaning on lifelong gut health in the pig. Journal of Animal Science, 2019. 97(Supplement_2): p. 63-64.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Lennon, E.M., et al., Mast Cells Exert Anti-Inflammatory Effects in an IL10(-/-) Model of Spontaneous Colitis. Mediators Inflamm, 2018. 2018: p. 7817360.
  • Type: Other Status: Published Year Published: 2018 Citation: Moeser, A., Gender and stress matter in pig gut health. National Hog Farmer Daily, 2018. 17.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Lennon, E.M., et al., Mast Cells Play a Protective Role in Spontaneous Colitis. 2018, American Association of Immunologists.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Ayyadurai, S., et al., Frontline Science: Corticotropin-releasing factor receptor subtype 1 is a critical modulator of mast cell degranulation and stress-induced pathophysiology. J Leukoc Biol, 2017. 102(6): p. 1299-1312.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Pohl, C.S., et al., Early Life Adversity in Rodent and Porcine Models Induces Enteric Cholinergic System Upregulation and Mast Cell Hyperplasia. The FASEB Journal, 2017. 31: p. 1047.2-1047.2
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Mackey, E., et al., Corticotropin releasing factor receptor 2 exerts global suppression of mast cell degranulation and associated pathophysiology. 2017, American Association of Immunologists.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Li, Y., et al., Chronic social stress in pigs impairs intestinal barrier and nutrient transporter function, and alters neuro-immune mediator and receptor expression. PloS one, 2017. 12(2): p. e0171617.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Pohl, C.S., et al., Early weaning stress induces chronic functional diarrhea, intestinal barrier defects, and increased mast cell activity in a porcine model of early life adversity. Neurogastroenterology & Motility, 2017. 29(11): p. e13118.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Moeser, A.J., C.S. Pohl, and M. Rajput, Weaning stress and gastrointestinal barrier development: Implications for lifelong gut health in pigs. Animal Nutrition, 2017. 3(4): p. 313-321.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Medland, J.E., et al., Early life adversity in piglets induces long-term upregulation of the enteric cholinergic nervous system and heightened, sex-specific secretomotor neuron responses. Neurogastroenterol Motil, 2016. 28(9): p. 1317-29.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser, A., What is" gut health" and how do you quantify/measure it? Journal of Animal Science, 2016. 94: p. 126.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Mackey, E., et al., Heightened Anaphylaxis Reaction in Female Mice is Associated with Increased Synthesis of Mast Cell Secretory Granule?Associated Immune Mediators. The FASEB Journal, 2016. 30: p. 1023.1-1023.1.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Mackey, E., et al., Sexual dimorphism in the mast cell transcriptome and the pathophysiological responses to immunological and psychological stress. Biology of sex differences, 2016. 7(1): p. 1-19.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser, A.J. Environmental influences on gastrointestinal development, function, and disease resistance. in Midwest Swine Nutrition Conference, Indianapolis, Indiana. 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Li, Y., et al., Dietary iron deficiency and oversupplementation increase intestinal permeability, ion transport, and inflammation in pigs. The Journal of nutrition, 2016. 146(8): p. 1499-1505.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Li Y, Hansen S, Borst L, Spears JW, Moeser AJ. Dietary iron deficiency and excess affect intestinal permeability and inflammation in pigs. Proceedings from the American Association Swine Veterinarians (AASV) Annual Meeting. New Orleans, LA February 28, 2016; pp 300301.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Moeser AJ. Optimizing gut function in pigs: the long-term impacts of early weaning. Proceedings from the American Association Swine Veterinarians (AASV) Annual Meeting. Denver, CO 2017; pp 21-23
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Moeser AJ, Pohl CS, and Rajput M. Weaning stress and gastrointestinal barrier development: implications for lifelong gut health in pigs. Proceedings from the Vetagro International Forum: Translating Scientific Discoveries into On-Farm Solutions. Rome, Italy; Pp 17-43. 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Matos K, Li Y, Rajput M, Moeser AJ. Development of intestinal glucose and amino acid transporter function in a porcine model of early life stress (poster). Annual Biomedical Research Conference for Minority Students (ABRCMS), November 2017, Phoenix, Arizona, USA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Moeser AJ. How stress impacts gut health in neonates (Neonatal challenges and disease session). Presentation proceedings at the 35th ADSA Discover Conference, Oct 29Nov1,2018 Itasca, IL
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Rajput M, Li Y, Thelen K, McAulliff A, Maradiaga N, Gerras A, Kessler J, Mackey E, Moeser AJ. Early weaning alters the normal trajectory and long-term function of GI immune function in barrows. Proceedings from the 99th Conference of Research Workers in Animal Diseases (poster). P227. December 2-4, 2018 Chicago IL
  • Type: Journal Articles Status: Under Review Year Published: 2022 Citation: Li Y, Thelen KM, Matos-Fern�ndez K, Nelli RK, Rajput M, Fardisi M, Trottier NL, Contreras GA, Moeser AJ. Developmental Alterations of Intestinal SGLT1 and GLUT2 by Early Weaning Coincides with Persistent Low-Grade Metabolic Inflammation in Female Pigs.
  • Type: Journal Articles Status: Under Review Year Published: 2022 Citation: Duque-Wilckens N, Sarno E, Stolting F, HKhalid S, Douma A, Maradiaga, Thelen KM, Robison AJ, Moeser AJ. Effects of early life adversity on meningeal mast cells and proinflammatory gene expression in male and female Mus musculus.


Progress 10/01/19 to 09/30/20

Outputs
Target Audience:The target audiences reached by our research efforts during this reporting period are animal health specialists including veterinarians, nutritionists, animal scientists, pharmaceutical/animal health industries, and producers (swine and dairy). Our work understanding the impact of stress on physiology in the pig has relevance to human health professionals as a model. Changes/Problems:COVID19 caused a research shutdown which negatively impacted our progress. What opportunities for training and professional development has the project provided?There were a number of training and professional development opportunities which results from this research including graduate students attending the American Assc of Immunology Meeting to present thier work, and presentation at local university seminars. one graduate student received a MS degree, whle another is working to complete a PhD degree in December 2020. How have the results been disseminated to communities of interest?Results have been presented to the agricultual industry communities including swine veterians, pork producers and industryprofessionals, animal nutritionists, basic science researchers, and University faculty and students What do you plan to do during the next reporting period to accomplish the goals?Current research directions will build upon our new data and published findings and will investigate the role of factors such as biological sex, early life stress, and GI infections on porcine gut barrier and neuro-immune development. Several manuscripts are in preparation that will be completed and submitted for publication. Additional funding will be sought from the federal agencies and industry to further advance our research

Impacts
What was accomplished under these goals? In the past funding year, we have made substantial progress under each of the goals of this research. In 2019-2020, the research focused on how factors such as age and weaning impact mast cells in the gut of pigs and how mast cell products such as histamine, can influence inflammatory gene transcription. A few major findings of our work are: (1) mast cell histamine is elevated in the blood of weaned pigs within 30 minutes of weaning stress and that histamine levels were markedly higher in early weaned (EW) pigs (weaned at 15-16 d of age) compared with later weaned (LW) pigs, (2) intestinal expression of histamine receptor subtypes HR1, HR2 and HR4 were upregulated in the weaned pigs with greater responses in EW pigs, (3) administration of histamine antagonists significantly inhibited several inflammatory markers in the gut including TNF, MPO and IL1 beta (4) histamine receptor subtypes had a differential expression in the gut and were localized to immune cells, enteric neurons and epithelial cells. Together, these studies provide new insight into mast cell activation in the gut of weaned pigs and the potential role and pharmacological interventions, via histamine modulation, that could impact gut development, performance and disease resistance.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ahmad MF, Ferland D, Ayala-Lopez N, Contreras GA, Darios E, Thompson J, Ismail A, Thelen K, Moeser AJ, Burnett R,Anantharam A, Watts SW.Perivascular Adipocytes StoreNorepinephrine by Vesicular Transport. Arterioscler Thromb Vasc Biol. 2019 39(2):188-199. doi: 10.1161/ATVBAHA.118.311720.PMID: 30567483N
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Piliponsky, Adrian Martin, et al. Adrian Martin Piliponsky, Claire Gendrin, Phoenicia Quach, Eric Guga, Gauri Bhise,Siddhartha Saha, Alyssa Brokaw, Adam Moeser and Lakshmi Rajagopa
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Interaction between biological sex and early life adversity on gastrointestinal (GI) immune developmentMahsa Fardisi, Kyan Thelen, Mrigendra Rajput and Adam J MoeserJ Immunol May 1, 2020, 204 (1 Supplement) 92.21
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Kyan Thelen, Neco Wilson, Glorian Berrios-Vazquez and Adam J MoeserMast cell histamine mediates intestinal immune response to early life stress in piglets via histamine 1 receptor (H1R). J Immunol May 1, 2020, 204 (1 Supplement) 92.19
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moeser AJ, Rajput M, Li Y, Thelen KT. Biological Sex and Wean Age Shape Gut Immune Development in the Pig.Proceedings from the 99th Conference of Research Workers inAnimal Diseases (oral). P227. CRWAD,Immunology Section, November 4th, 2019, Chicago, IL
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Jaron R Lewton, Medhat A Michael, Mrigendra Rajput, Kyan M Thelen, Adam J Moeser, Dale W Rozeboom. Feedingtreated wheat straw to weaned pigs improves feed efficiency and impacts mucosal and system immune parameters.Journal of Animal Science, Volume 97, Issue Supplement_3, December 2019, Pages 123124
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Biological Sex and Wean Age Shape Gut Immune Development in the Pig. CRWAD,Immunology Section, November 4th,2019, Chicago, IL
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Emily Mackey, Kyan M. Thelen,Vedrana Bali, Mahsa Fardisi, Madalyn Trowbridge, Cynthia L. Jordan, and Adam J. Moeser. Perinatal androgens organize sex differences in mast cells and attenuate anaphylaxis severity into adulthood. PNAS first published September 11, 2020 https://doi.org/10.1073/pnas.1915075117


Progress 10/01/18 to 09/30/19

Outputs
Target Audience:The target audiences research by our efforts during this reporting period are animal health specialists including veterinarians, nutritionists, animal scientists, pharmaceutical/animal health industries, and producers. OUr work produced is also of relevance to human health professionals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A number of opportunities for training and professional development were assocaited with this project. Below is a list of example publications and presentations of student trainees during the reporting period: Mackey EM and Moeser AJ. Perinatal Di-(2-ethylhexyl) Phthalate Exacerbates Anaphylaxis in Male Offspring. Society of Toxilogic Pathology Annual Meeting. June 22-27th, 2019, Raleigh,NC Early Life Adversity Programs Mast Cells Toward a Hyperactive Phenotype into Adulthood Nidia CMaradiaga,CalvinPohl,EmilyMackey,Adam J.Moeser. The Journal of ImmunologyMay 1, 2019,202(1 Supplement)54.2 Neco Wilson,Yihang Li,Mrigendra Rajput,Kyan Thelen,Katie KerrandAdam J. Moeser J ImmunolMay 1, 2019,202(1 Supplement)73.11; Weaning and postnatal age influence the early time course and nature of intestinal mast cell activation and mucosal inflammation in a porcine model of early life adversity Yihang Li, Karina Matos, Mrigendra Rajput, Adam James Moeser, 191 Early weaning in pigs induces long-term alterations in intestinal nutrient transporter function and expression partially via beta adrenergic enteric neural receptors,Journal of Animal Science, Volume 97, Issue Supplement_2, July 2019, Pages 112-113. Moeser AJ. How Early Life Stress Impacts Gut Development and Long-Term Health. Proceedings from the 4-State Dairy Nutrition Conference, Duluth IA. AJ Moeser, Yihang Li, Mrigendra Rajput, Kyan Thelen, 115 Negative impacts of early weaning on lifelong gut health in the pig,Journal of Animal Science, Volume 97, Issue Supplement_2, July 2019, Pages 63-64, Emily MackeyandAJ Moeser. Perinatal androgens drive sex differences in mast cell phenotype and severity of mast cell disease. J ImmunolMay 1, 2019,202(1 Supplement)54.15 How have the results been disseminated to communities of interest?Results have been presented to the agricultual industry communities including swine veterians, pork producers and industry professionals, animal nutritionists, basic science researchers, and University faculty and students What do you plan to do during the next reporting period to accomplish the goals?Furture research directions will build upon our recently data and published findings and will investigate the role of factors such as biological sex, early life stress, and GI infections on porcine gut barrier and neuro-immune development. Several manuscripts are in preparation that will be completed and submitted for publication. Additional funding will be sought from industry and federal agencies to further advance our research.

Impacts
What was accomplished under these goals? Aim 1 Cholinergic protein expression is upregulated in peripheral blood monocytes after weaning stress coinciding with previous data on enteric expression including a marked upregulation in the ileal epithelium overlying the Peyer's pathches. This work is currently being prepared as a manuscript for publication. Aims 2 and 3 results: Investigations into the influence of histamine receptor antagonist drugs on intestinal inflammation in weaned pigs were conducted. Preliminary findings indicate that blockade of specificic histamine receptors can influence severity of gut infalmmation induced by weaning stress.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Williams ES, Manning CE, Eagle AL, Swift-Gallant A, Duque-Wilckens N, Chinnusamy S, Moeser AJ, Jordan C, Leinninger G, Robison AJ. Androgen-dependant excitability of mouse hippocampal afferents to nuclueus accumbens underlies sex-specific susceptibility to stress. Biol Psychol 2019 (Accepted).
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Boyd RD, Zier-Rush CE, Moeser AJ, Culbertson M, Stewart KR, Rosero DS, Patience JF. Patience. Invited Review: innvovation through research in the North American pork industry. Animal, 2019
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Early Life Adversity Programs Mast Cells Toward a Hyperactive Phenotype into Adulthood Nidia C Maradiaga, Calvin Pohl, Emily Mackey, Adam J. Moeser. The Journal of Immunology May 1, 2019, 202 (1 Supplement) 54.2
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Neco Wilson, Yihang Li, Mrigendra Rajput, Kyan Thelen, Katie Kerr and Adam J. Moeser J Immunol May 1, 2019, 202 (1 Supplement) 73.11; Weaning and postnatal age influence the early time course and nature of intestinal mast cell activation and mucosal inflammation in a porcine model of early life adversity
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Yihang Li, Karina Matos, Mrigendra Rajput, Adam James Moeser, 191 Early weaning in pigs induces long-term alterations in intestinal nutrient transporter function and expression partially via beta adrenergic enteric neural receptors, Journal of Animal Science, Volume 97, Issue Supplement_2, July 2019, Pages 112113.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moeser AJ. Gastrointestinal immunology: Basic principles and factors that impact immune development and function? Proceedings of the Annual American Association of Swine Veterinarians Meeting, March, 2019, Orlando, FL.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moeser AJ. Gut Health: Whats it Mean and What Dictates It? Proceedings from the Southwest Nutrition Conference February 7-9th, 2019, Chandler, AZ


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The target audiences research by our efforts during this reporting period are animal health specialists including veterinarians, nutritionists, animal scientists, pharmaceutical/animal health industries, and producers. OUr work produced is also of relevance to human health professionals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Research presentations by trainees: Li Y, Rajput M, Matos Fernandez, Moeser AJ, 2018 Early weaning in pigs induces long-term alterations in intestinal nutrient transporter function and expression. FASEB J, 32, Supplement 1. # Pohl CS, Medland JE, Mackey E and Moeser AJ. Biological sex influences trajectories of gut barrier and neuroimmune development and clinical disease in a porcine model of early life adversity. Proceedings of the 12th Annual Meeting of the Organization for the Study of Sex Differences, Atlanta, GA, 2018. # Mackey EM and Moeser AJ. Biological sex influences allergen-induced mast cell activation patterns and severity of anaphylaxis. Proceedings of the 12th Annual Meeting of the Organization for the Study of Sex Differences, Atlanta, GA, 2018 Elizabeth M. Lennon,Jane S. Woodrow,Courtney A. Thomason,Luke B. Borst,Laura E. EdwardsandAdam James Moeser. Mast Cells Play a Protective Role in Spontaneous Colitis, Proceedings from Annual Meeting of the American Association of Immunologists. J Immunol, 2018,200, Supplement 1,pg. 118.20. 873.17. D'Costa S, Ayyadurai S, Gibson AJ, Poopal A, Moeser AJ. Mast cell corticotropin releasing factor receptor CRF2regulates mast cell function in response to psychological and immunological stress (oral presentation) Proceedings from Annual Meeting of the American Association of Immunologists. Journal of Immunology 200; Supplement 1 pg. 105.5, Austin, TX 2018. How have the results been disseminated to communities of interest?Results have been presented to the agricultual industry communities including swine veterians, pork producers and industry professionals, animal nutritionists, basic science researchers, and University faculty and students What do you plan to do during the next reporting period to accomplish the goals?Furture research directions will build upon our recently data and published findings and will investigate the role of factors such as biological sex, early life stress, and GI infections on porcine gut barrier and neuro-immune development. Several manuscripts are in preparation that will be completed and submitted for publication. Additional funding will be sought from industry and federal agencies to further advance our research.

Impacts
What was accomplished under these goals? Aim 1 Results revealed the cholinergic protein expression is upregulated early after weaning stress with a rapid upregulation of choline acetyltransferase at 8 hours in ileal and colonic mucosa and mesenteric lymph nodes. ChAT staining revealed a marked upregulation in the ileal epithelium overlying the Peyer's pathches. Pigs challenged with Salmonella Typhimurium exhibited upregulated ChAT staining in the Peyer's patch epithelium. Aims 2 and 3 results: Expression of mast cell genes for tryptase, chymase, histamine receptors 1,2,4 are upregulated within 3 hours of weaning stress in early weaned pigs while late weaned pigs exhibit a lower expression. These changes coincided with intestinal SGLT function and breakdown in intestinal barrier function. These new findings are significant as they establish distinct developmental phases for ELA-induced mast cell activity which may define distinct roles for mast cells in both the initiation and perpetuation phases of ELA-induced GI barrier and neuro-immune dysfunction.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: DCosta SD, Ayadurai S, Gibson AJ, Mackey E, Rajput M, Sommerville LJ, Wilson N, Li Y, Kubat E, Kumar A, Subramanian H, Bhargava A and Moeser AJ. Mast Cell CRF2 Suppresses Mast Cell Degranulation and Limits the Severity of Anaphylaxis and Stress-Induced Intestinal Permeability. J Allergy Clin Immunol 2018, in press
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Pohl CS, Lennon EM, DeWilde M, Moeser AJ. S. Typhimurium challenge in juvenile pigs modulates the expression and localization of enteric cholinergic proteins and correlates with mucosal injury and inflammation. Auton Neurosci, 2018, 213:51-59. PMID: 30005740
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Lennon EM, Borst LB, Edwards LL, Moeser AJ. Mast cells exert anti-inflammatory effects in an IL10-/- model of spontaneous colitis. Mediators Inflamm 2018, doi: 10.1155/2018/7817360. eCollection 2018. PMID: 29849494
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Ayyadurai S, Gibson AJ, D'Costa S, Overman EL, Sommerville LJ, Poopal AC, Mackey E, Li Y, Moeser AJ. Frontline Science: Corticotropin-releasing factor receptor subtype 1 is a critical modulator of mast cell degranulation and stress-induced pathophysiology. J Leukoc Biol. 2017, 102:1299-1312 PMID: 28684600; PMC Journal  In Process
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Gulledge TV, Collette NM, Mackey E, Johnstone SE, Moazami Y, Todd DA, Moeser AJ, Pierce JG, Cech NB, Laster SM. J Ethnopharmacol. 2018, 212:166-174. doi: 10.1016/j.jep.2017.10.012. PMID: 29042288


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The target audiences research by our efforts during this reporting period are animal health specialists including veterinarians, nutritionists, animal scientists, pharmaceutical/animal health industries, and producers. OUr work produced is also of relevance to human health professionals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Invited presentations: Moeser AJ. Biological Basis for Optimum Wean Age in Swine. Leman Conference, Sept 19, 2017 Moeser AJ. Early Life Stress, HPA Axis, and Gastrointestinal Disorders Vetagro International Forum 2017, Rome, Italy, May 18-19, 2017. Moeser AJ. Neuroimmune Mechanisms in Early Life Stress-Induced GI Disease,6th Swine in Biomedical Research Conference, Baltimore, MDSept 23,2017 Moeser AJ. Optimizing gut function in pigs: the long-term impacts of early weaning. American Association Swine Veterinarians (AASV) Annual Meeting. Denver, CO February 26, 2017. Moeser AJ. Cargill Biomarker Workshop, "Biomarkers of Intestinal Barrier Function Stress in Early Life" December 7th, 2016. Elk River, Moeser AJ. MSU Neuroscience program seminar "Animal Models of Early Life Adversity". November 17, 2016. East Lansing, MI.MSU GI Group Meeting. "Sex and GI disorders" November 7, 2016. East Lansing, MIAPC.Inc. Research Advisory Meeting. "Gut Health and Integrity". November 9th Ankeny, IA Kansas State University Animal Science/Veterinary Medicine Seminar. "Impact of Stress on Gut Health Disorders. October 4th, 2016. Manhattan, KS Moeser AJ. University of Maryland, College Park Department of Animal Science Seminar. "Pathophysiology of Stress-Induced GI Disorders. September 22, 2016 Moeser AJ. Elanco Gut Health Webinar. "Gastrointestinal Health and Function: Understanding the Impact of Postweaning E. coli on the gut". September 16th, 2016. How have the results been disseminated to communities of interest?Results have been presented to the agricultualindustry communities including swine veterians, pork producers and industry professionals,animal nutritionists, basic science researchers, and University faculty and students What do you plan to do during the next reporting period to accomplish the goals?Furture research directions will build upon ourrecentlydata and published findings and will investigate the role of factors such as biological sex, early life stress, and GI infections on porcine gut barrier and neuro-immune development.Several manuscripts are in preparation that will be completed and submitted for publication.Additionalfunding will be sought from industry and federal agencies to further advance our research.

Impacts
What was accomplished under these goals? Aim 1 Resultsrevealed that ELA in the porcine EWS induces an early and persistent upregulation of the enteric cholinergic system characterized by a persistence of cholinergic enteric neurons, altered expression of markers of acetylcholine synthesis, degradation, and receptor binding, and hypersensitivity of cholinergic secretomotor neurons to neural stimuli. Aim 2 results:These studies revealed that ELA triggers 2 distinct phases of mast activation. A first phase is characterized by a rapid and robust mast cell degranulation upon exposure to ELA and preceding early acute intestinal barrier injury. A second phase is characterized by mast cell hyperplasia, enhanced mast cell-nerve co-localization, and enhanced stimuli-induced mast cell degranulation persisting into adulthood. These new findings are significant as they establish distinct developmental phases for ELA-induced mast cell activity which may define distinct roles for mast cells in both the initiation and perpetuation phases of ELA-induced GI barrier and neuro-immune dysfunction. In Aim 2 we also sought to identify the signaling factors regulating mast cells during stress, focusing specifically on the novel role of mast cell CRF receptors, CRF1 and CRF2. Previous studies, including those by our group showed that the CRF system is activated in stress-related GI disorders and contributes to intestinal barrier disturbances, abdominal pain and altered bowel habits, but the precise signaling pathways were unknown. Graphing mast cells into mast cell-deficient mice, combined with pharmacologic approaches in porcine models, revealed that CRF1 on mast cells potentiates stimuli-induced mast cell degranulation and GI pathophysiology during psychological and immunological stress [57]. This work was the first to demonstrate the role of the mast cell CRF system as a global modulator of mast cell function and our paper was selected as a Frontline Science section article in The Journal of Leukocyte Biology. Aim 3 investigated the impact of ELA on the development of GI barrier function in EWS pigs. Intestinal barrier dysfunction (increased intestinal permeability) is a central factor involved in the initiation of clinical symptoms associated with GI disorders such as IBS and IBD [58]. We showed that EWS triggers an acute disruption of intestinal barrier function within the first 48 h post-weaning which is then followed by an incomplete barrier recovery, with elevated permeability persisting into adulthood. These studies were the first to reveal that ELA irreversibly alters the path of GI barrier development leading to lasting dysfunction. Similar to our 2016 paper on cholinergic dysregulation in the EWS pig, our recent 2017 manuscript reporting ELA-induced mast cell and GI barrier developmental trajectories [12] received recognition for its impact to the field as it too was selected as a journal editorial focus (in press). Additional work which stemmed from Aim 3 included investigating the effects of dietary factors on barrier development and disease susceptibility in ELA pigs and studies in rodents demonstrating the influence of ELA on susceptibility to later life inflammatory and infectious challenge models. Additional projects explored how later life chronic psychosocial stressors regulate barrier and immune function in the pig.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Choi H, Bowen SE, Mio Y, Chan CY, Mioa EA, Abrink M, Moeser AJ, and Abraham SN.Loss of Bladder Epithelium Induced by Cytolytic Mast Cell Granules. Immunity 45:1258-1269, 2016. NIHMS ID: NIHMS830405; PMCID: PMC5177478.
  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Li Y, Song Z, Kerr KA, Moeser AJ. Chronic social stress in pigs impairs intestinal barrier and nutrient transporter function, and alters neuro-immune mediator and receptor expression. PLoS One. e0171617, 2017. PMCID: PMC5295718
  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Moeser AJ, Pohl CS, Rajput M. Weaning stress and gastrointestinal barrier development: Implications for lifelong gut health in pigs. Animal Nutrition xxx (2017) 1-9. (In press). https://doi.org/10.1016/j.aninu.2017.06.003
  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Pohl CS, Medland JE, Mackey E, Edwards LL, Bagley KD, Dewilde MP, Williams KJ, Moeser AJ. Early weaning stress induces chronic functional diarrhea, intestinal barrier defects, and increased mast cell activity in a porcine model of early life adversity. Neurogastroenterology and Motility. Epub ahead of print PMID: 28573751
  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Ayyadurai S, Gibson AJ, D'Costa S, Overman EL, Sommerville LJ, Poopal AC, Mackey E, Li Y, Moeser AJ. Frontline Science: Corticotropin-releasing factor receptor subtype 1 is a critical modulator of mast cell degranulation and stress-induced pathophysiology. J Leukoc Biol. 2017. Epub ahead of print. PMID: 28684600


Progress 03/01/16 to 09/30/16

Outputs
Target Audience:The target audiences research by our efforts during this reporting period are animal health specialists including veterinarians, nutritionists, animal scientists, pharmaceutical/animal health industries, and producers. OUr work produced is also of relevance to human health professionals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? PhD student, Calvin Pohl, worked with Dr. Moeser to prepare and submit and F31 application PhD Student, Calvin Pohl, attended and presented research at CRWAD and at the American Association of Swine Veterinarian Conference Post-Doc researcher, Yihang Li, attended the National Animal Science Meetings and presented his research Dr. Moeser attended several conferences and was an invited speaker at several national and international venues. Conferences included: CRWAD, Midwest Animal Science Meetings, Midwest Swine Nutrition Conference, Experimental Biology, Neurogastroenterology Conference How have the results been disseminated to communities of interest?Dr. Moeser hasgivenseveral invitedtalks at conferences, Industrycompany meetings, and webinars. The topic was on GI health in animals and the target audiences included animal health professionals, animal scientists, nutritionists, producers, academicians, and students. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we will continue our research focus on mechanisms of stress-induced GI health disorders. Specifically we will focus on how stress adversely impacts immune develop and immunocompetence and nutrient utilization and efficiency.

Impacts
What was accomplished under these goals? Gastrointestinal (GI) disorders continue to be a major cause of economic losses to the agricultural animal industries. Economic losses associated GI disorders range from subclinical reductions in feed efficiency to morbidity and mortality associated enteric infections. It has long-been known that stressors in production play a major role in GI disease susceptibility in animals. Stressors include psychological, environmental, and physical stressors such as weaning, crowding and transport stress, nutritional, and environmental (heat or cold stress). Despite the well-established role of stress in disease pathogenesis, our understanding of the biological mechanisms by which stress predisposes animals to disease remains limited. Because of this lack in knowledge, limited therapeutic or alternative management strategies are available to combat the impacts of stressors in animal production. This project is centered upon understanding the biological mechanisms by which stress impacts intestinal health in agricultural animals, with a specific emphasis on swine production, efficiency, and disease and therefore, this project aligns with the mission of AgBioResearch. In the past year, we have made significant progress towards our objectives and our research has revealed new findings that have significant implications to animal and human gut health and disease resistance. Specifically during the past year we have focused on understanding how early life stressors such as early weaning and chronic stress in adult animals impact theGI immune, nervous and epithelial barrier functions and its implications to animal health. Objective 1: Determine the mechanism by which early life stress triggers long-term amplification of enteric cholinergic nervous system activity. Major activities completed / experiments conducted: Completed experiments studying the impact of early weaning stress in pigs on development of the cholinergic nervous system in pigs Prepared manuscript which was accepted to the Journal of Neurogastroenterology and Motility (Medland et al., 2016) Data collected: Electrical physiology data on Ussing chambers to evaluate cholinergic function Measured expression of neuronal markers by qPCR, Western blotting and ELISA Summary statistics and discussion of results: Results from this research demonstrates that early weaning stress in pigs induces long-term, heightened ileal secretomotor responses in sex-specific manner. Furthermore, EWS induced a marked upregulation of the cholinergic ENS characterized by persistence and elevated ChAT+ neurons and mucosal AchE activity. These findings may represent an important mechanistic link between early life stressors and life-long GI disease susceptibility. Key outcomes or other accomplishments realized. Current early life production practices such as early weaning in pigs have both immediate and long-term consequences on animal health through the altered developmental trajectory of the GI system Objective 2: Investigate the mechanisms by which cholinergic signaling triggers persistent mast cell activation. Major activities completed / experiments conducted: Completed experiments characterizing the impact of early weaning stress on intestinal mast cells localization, numbers and activity Manuscript is in preparation Data collected: Histochemical analysis of mast cells in the pig intestine: mast cell numbers, localization, and gene expression as influenced by early weaning and biological sex Summary statistics and discussion of results: Results from this research demonstrates that early weaning stress in pigs induces long-term, heightened increases in intestinal mast cell numbers Mast cells from early weaned pigs were elevated in the intestinal mucosa, submucosa, and longitudinal muscle No differences were observed for biological sex Mast cell activity was associated with increased expression of cholinergic nervous system markers Together, these data indicate that mast cells and the cholinergic nervous system is upregulated and that this may represent a central pathophysiology in stress-induced alterations in gut health in animals Key outcomes or other accomplishments realized. In stressed conditions there is a marked expansion of intestinal mast cells in the porcine intestine which drive intestinal permeability and aberrant immune function. The connection between stress and nerve-immune development in the pig stressors is new and future investigations are expected to yield new insight into how stress adversely impacts gut health and animal disease susceptibility SA3; Understand the impact of early life stress and cholinergic nerve-mast cell signaling on alterations in epithelial tight junctions Major activities completed / experiments conducted: Focused on analyzing intestinal barrier function disturbances in pigs subjected to early weaning stress and chronic mixing and crowding stress Manuscript is in preparation Data collected: Ussing chamber analyses of intestinal permeability Expression of immune and stress markers in the intestine Summary statistics and discussion of results: Results from this research demonstrates that early weaning stress in pigs induces long-term, heightened increases in intestinal permeability (measured as FD4 flux rates) and that females are more susceptible Chronic mixing and crowding stress results in increased intestinal permeability Both mixing and crowding stress and early weaning increase intestinal permeability and mast cell activation but downregulate inflammatory cytokines. Together, these data indicate that impairment in intestinal barrier function is a common mechanism of GI injury in stressed pigs and that this is driven by mast cell activation. Interestingly, despite the elevated mast cell activation and GI permeability, the immune system is suppressed. Key outcomes or other accomplishments realized. Under chronic stress, mast cell-intestinal barrier disturbances are central to GI dysfunction and performance reductions. Our new data also suggest that the immune response in these animal is markedly suppressed which could explain increased disease susceptibility observed in chronically stressed animals. Further investigation into the link between GI permeability and mast cell activation and regulation of immune functions could reveal the mechanism of stress-induced disease susceptibility in agricultural animals, which has remained poorly understood. SA4: Determine whether dietary modifications can influence intestinal barrier function in stressed pigs. Major activities completed / experiments conducted: Completed the analysis and write up of a project focused on understanding the impact of dietary Fe levels on GI health in pigs Manuscript was prepared and recently accepted for publication in the Journal of Nutrition (Li et al., 2016) Data collected: Intestinal barrier function Intestinal inflammatory markers via q PCR and ELISA Intestinal oxidative stress measures Compared with control diets containing 100 ppm Fe, Low Fe diets (50 ppm) and high Fe diets (500 ppm) fed to grower pigs resulted in impaired performance and increased intestinal inflammation, oxidative stress and permeability. Together, these data show that current dietary Fe levels in swine diets can be impairing gut health and function. Key outcomes or other accomplishments realized. This is the first study looking into the influence of dietary Fe level commonly found in swine diets on GI health and function. These studies revealed a new understanding of how dietary factors such as Fe can have a dramatic impact on animal gut health.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Dietary iron deficiency and over-supplementation increase intestinal permeability, ion transport, and inflammation in pigs. Li Y, Hansen SL, Borst LB, Spears JW, Moeser AJ. J Nutr. 146:1499-505, 2016
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Medland JE, Pohl CS, Edwards LL, Frandsen SF, Bagley K, Li Y, Moeser AJ. Early life adversity in piglets induces long-term upregulation of the enteric cholinergic nervous system and heightened, sex-specific secretomotor neuron responses. Neurogastroenterol Motil. 28:1317-29, 2016.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Sexual Dimorphism in the Mast Cell Transcriptome and the Pathophysiological Responses to Immunological and Psychological Stress for consideration in Biology of Sex Differences Journal
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. Environmental Influences on Gastrointestinal Development, Function, and Disease Resistance. Proceedings from the Midwest Swine Nutrition Conference. Indianapolis, IN, September 8-9, 2016
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Mackey EM, DCosta S, Laster SM, and Moeser AJ. Heightened anaphylaxis reaction in female mice is associated with increased synthesis of mast cell secretory granule-associated immune mediators. Abstract presented at the Experimental Biology Meeting 2016, San Diego, CA. Abstract selected for oral presentation at the Experimental Biology 2016 Trainee Advisory Committee Cross-Sectional Symposium Sex Differences in Health and Disease
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. What is Gut Health and How Do You Measure It?. Midwest Animal Science Meetings. Des Moines, IA, March 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. The relationship between stress, gut physiology and function: Implications for feed efficiency. American Association Swine Veterinarians (AASV) Annual Meeting. New Orleans, LA February 28, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. The relationship between stress, gut physiology and function: Implications for feed efficiency. International Conference for Feed Efficiency in Swine (Physiology and Feed Efficiency Symposium). Omaha, NE, October 21, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Pohl CS, Medland JM, Edwards LL, Moeser AJ. Early weaning stress in pigs alters postnatal enteric neuro-immune development, inducing long term disease susceptibility. Abstract presented at the Conference for Research Workers in Animal Disease (CRWAD), Chicago, IL, December 6-7, 2015. *1st place student award, poster session
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. Elanco Gut Health Webinar. Gastrointestinal Health and Function: Understanding the Impact of Postweaning E. coli on the gut. September 16th, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. Kemin Swine Summit. Stress Effects on Gut Health and Performance August 29-30, 2016, Duluth, MN.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. MSU Respiratory Research Initiative. The impact of stress on intestinal immune and epithelila barriers: role of the CRF-mast cell axis. April 30,2016. East Lansing, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. Elanco Canada Animal Health Symposium. Gastrointestinal Health and Function: Understanding the Impact of Postweaning E. coli. April 13th, 2016, Toronto, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. MSU Department of Microbiology and Molecular Genetics. Breaking Down Barriers: The Influence of Stress on Gastrointestinal Epithelial and Immune Function. February 24, 2016. East Lansing, MI
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. MSU Animal Science Club Seminar: My Path from Animal Science Student to Clinician Scientist. February 17, 2016. East Lansing, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. MSU Department of Food Science and Human Nutrition Seminar. Early Life Origins of Gastrointestinal Disease. February 10, 2016. East Lansing, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Moeser AJ. South Dakota State University, Department of Veterinary and Biomedical Sciences. Pathophysiology of Stress-Induced Gastrointestinal Disease. February 19, 2016. Brookings, SD.