Progress 12/17/13 to 09/30/18
Outputs
Target Audience:Target audiences include animal scientists, students, pork producers, stakeholder's and related clientele. Processes that deliver science-based knowledge included laboratory instruction and experiential learning opportunities provided to students and outreach activities to related clientele via presentation of research data at professional and scientific meetings including ASAS Midwest Animal Science Meetings (2019). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The activities associated with this experiment have provided training opportunities (applied animal experiments as well as basic bench laboratory experiments) for Graduate students (Joice San Andres-primary, Melanie Trenhaile, and Dana van Sambeek); Technicians (Jeff Perkins, Gail Henderson); and Collaborators (Phillip Miller, Samodha Fernando). How have the results been disseminated to communities of interest?Data from the latest experiment will be reported at the 2019 Midwest ASAS meetings in Omaha, NE. In addition, two full-length manuscripts are in preparation to be submitted to Translational Animal Science and the Journal of Animal Science. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
A healthy gut is comprised of effective digestion/absorption, absence of gut illness, normal/stable intestinal microbiota, effective immune status, and status of well-being. Prebiotics are nondigestible dietary ingredients that beneficially affect the host by selectively stimulating growth/activity of beneficial bacteria. In response to prebiotic consumption, beneficial effects have been observed in pigs including positive changes in gastrointestinal microbota, fermentative end-products, intestinal morphology, nutrient digestibility, and immune function leading to enhancements in overall pig gut health and growth performance. Collectively, effects of prebiotics may be a culmination of host-microbial interactions. The overall goal of this project is to explore how host-microbial interactions may culminate to impact gut health and nutrient metabolism in weaned pigs fed prebiotics and to gather host phenotypic and microbial community data to identify potential probiotic species. During the previous period an additional experiment was designed and conducted to evaluate additional prebiotic feed additives (i.e., fructooligosaccharides, sugar beet pulp, and chicory pulp) with respect to microbial population structure, dynamics and function, intestinal barrier function and integrity. Additional results and outcomes are detailed below; however, the overall conclusion from this latest experiment was that while there was no effect of prebiotic feed additives on growth performance, prebiotics have variable impacts on nutrient digestibility, and postive effects of prebiotics were observed with respect to biomarkers of intestinal injury. Objective 1) Delineate changes in gastrointestinal microbial ecology in weanling pigs fed prebiotics via high-throughput pyrosequencing of microbiome DNA; Objective 3) Compare changes in gastrointestinal microbiology with DNA microarrays and pig performance and digestibility phenotype to identify genes that undergo induction or repression during colonization; and Objective 4) Develop and test optimized prebiotic intervention strategies in production environments (emphasizing non-antibiotic approaches) for enhancing porcine health and food production. Nothing new to report at this time. Objective 2) Evaluate the molecular and metabolic effects of prebiotics and intestinal microbiota on intestinal barrier function and integrity. Based on our previous work indicating that addition of prebiotics to weanling pig diets has a positive impact on growth performance and immune biomarkers, a experiment was designed to evaluate additional prebiotic feed additives. For this experiment, sixty-four pigs (21 d; BW 6±0.5 kg) of mixed sex were used to test 4 dietary treatments with 4 pens/treatment and 4 pigs/pen. Pens were randomly allocated to 1 of the 4 diets following completely randomized design in a 2-phase 28-d feeding trial. Dietary treatments were corn-soybean based diet (Control), Control + fructooligosaccharide (FOS), Control + chicory pulp (PC) and Control sugar beet pulp + (SBP). Prebiotics were incorporated in the diet by partially replacing corn at rate of 0.05% and 0.1% in phase 1 and 2, respectively. Individual BW and feed disappearances were measured weekly from d 0 to 28 for determination of ADG, ADFI, and G:F as indicators of growth performance. Fecal grab samples were obtained in the last week of each phase to evaluate apparent total tract energy and dry matter digestibility. Blood samples were collected from 2 pigs per treatment on d 0, d 14 and d 28 and analyzed for serum biomarkers of intestinal barrier function including tight junction protein 1 (TJP1), intestinal fatty acid binding protein (IFABP) and diamine oxidase (DAO) using porcine specific ELISA kits. Dietary treatments had no effect on growth performance. With respect to nutrient digestibility, pigs fed PC had decreased (P < 0.05) dry matter digestibility compared to FOS and SBP-fed pigs during Phase II. With respect to energy digestibility, pigs fed FOS had decreased (P < 0.05) energy digestibility compared to CTL and PC pigs in Phase I. With respect to serum biomarkers of barrier function, TJP1 concentration increased with days postweaning (P<0.001). DAO and IFABP levels decreased overtime with P<0.001 for both markers. Pigs fed FOS and PC had lower amounts of DAO (P=0.034) and IFABP (P=0.043) than Control and SBP at d 14. Key outcomes for Objective 2: Feeding prebiotics such as fructooliogsaccharides, chicory pulp, and sugar beat bulp to weaned pigs have no effects on growth performance. However, weaned pigs have a variable response with respect to nutrient digestibility when prebiotics are included in the diet. Finally, our results show that serum biomarkers of intestinal barrier function and integrity (i.e., TJP1, DAO and IFABP) vary over the weaning period possibly indicating intestinal recuperation from weaning stress. Moreover, inclusion of prebiotics in the diet impacts biomarkers of intestinal barrier function which implies that addition of prebiotics may have positive impacts on overall gut health.
Publications
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2019
Citation:
San Andres, J.V., D.M. van Sambeek, S.R. Kinstler, M.D. Trenhaile-Granneman, S.M. Winkel, A. Schlageter-Tello, P.S. Miller, and T.E. Burkey. 2019. Potential biomarkers of intestinal injury and inflammation in nursery pigs fed prebiotics. J. Anim. Sci. (Abstr.)
- Type:
Journal Articles
Status:
Other
Year Published:
2019
Citation:
San Andres, J.V., G.A. Mastromano, Y.S. Li, H. Tran, J.W. Bundy, P.S. Miller, and T.E. Burkey. 2019. Effects of prebiotics on growth performance and immune parameters of weaned pigs. Transl. Anim. Sci. (Manuscript in preparation)
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:Target audiences include animal scientists, students, pork producers, stakeholder's and related clientele. Processes that deliver science-based knowledge included laboratory instruction and experiential learning opportunities provided to students and outreach activities to related clientele via presentation of research data at professional and scientific meetings including ASAS Midwest Animal Science Meetings (2017). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The activities associated with this experiment has provided training opportunities (applied animal experiments as well as basic bench laboratory experiments) for Graduate students (Yanshuo Li and Shana Barnett-primary, Melanie Trenhaile, and Dana van Sambeek); Undergraduate students; Technicians (Jeff Perkins, Gail Henderson); and Collaborators (Phillip Miller, Samodha Fernando). How have the results been disseminated to communities of interest?Data from this experiment were reported at the 2017 Midwest ASAS meetings in Omaha, NE. In addition, the bulk of this work comprised the dissertation for one PhD student (Sophie Li) and a manuscript is in preparation to be submitted to the Journal of Animal Science. What do you plan to do during the next reporting period to accomplish the goals?We will continue to evaluate and refine prebiotic and probiotic (as well as there combination-synbiotic) combinations via in vitro and in vivo methods. We plan to conduct an additional experiment in early 2018 to further evaluate mannanoligosaccharides as well as additional prebiotic feed additives by evaluating microbial population structure, dynamics and function, intestinal barrier function and integrity.
Impacts
What was accomplished under these goals?
A healthy gut is comprised of effective digestion/absorption, absence of gut illness, normal/stable intestinal microbiota, effective immune status, and status of well-being. Prebiotics are nondigestible dietary ingredients that beneficially affect the host by selectively stimulating growth/activity of beneficial bacteria. In response to prebiotic consumption, beneficial effects have been observed in pigs including positive changes in gastrointestinal microbota, fermentative end-products, intestinal morphology, nutrient digestibility, and immune function leading to enhancements in overall pig gut health and growth performance. Collectively, effects of prebiotics may be a culmination of host-microbial interactions. The overall goal of this project is to explore how host-microbial interactions may culminate to impact gut health and nutrient metabolism in weaned pigs fed prebiotics and to gather host phenotypic and microbial community data to identify potential probiotic species. During the previous period an experiment was designed to determine the effects of feeding mannanoligosaccharides (MOS) and Lactobacillus mucosae (LM) (alone and in combination) as prebiotic and probiotic sources in weanling pigs under immune challenge. In the previous period we reported key outcomes relative to growth performance, immune biomarkers and small intestinal histolocial anlayses. Additional results and outcomes from this experiment are detailed below; however, the overall conclusion from this experiment is that feeding LM alone improved feed efficiency and ileal morphological structure during the first wk of LPS-challenge; additionally, feeding LM and MOS may have beneficial effects relative to immune biomarkers. Objectives 1) Delineate changes in microbial ecology in pigs fed prebiotics; and 2) Evaluate the molecular and metabolic effects of prebiotics and intestinal microbiota on intestinal barrier function and integrity; and 3) Compare changes in gastrointestinal microbiology with DNA microarrays and pig performance and digestibility phenotype to identify genes that undergo induction or repression during colonization. Nothing new to report at this time. Objective 4) Develop and test optimized prebiotic intervention strategies in production environments (emphasizing non-antibiotic approaches) for enhancing porcine health and food production. Based on our previous results indicating that feeding MOS (Saccharomyces cerevisiae) increased fecal Lactobacillus mucosae (LM) and was positively correlated with circulating IgA concentration, and experiment was designed to investigate the prebiotic and probiotic effects of MOS and LM on growth performance, immune response, and gut health of weanling pigs. To model an enteric infection, we challenged the pigs with E. coli LPS. Weaned pigs (n=100) were randomly allotted to 16 pens with a 2x2 factorial arrangement of treatments. On d 14 post-weaning, all pigs were inoculated with LPS. Cornsoybean-meal based diets (without plasma or antibiotics) with or without 0.1% yeast-derived MOS were randomly assigned to pens and 109 cfu/pig LM broth or an equal volume of control broth was top-dressed daily. Pigs were fed dietary treatments (control, MOS, LM, and MOS+LM) in phase-1 and phase-2 (d 0 to 7 and d 7 to 21 post-weaning, respectively). A common diet was fed to all pigs during phase-3 (d 21 to 35 post-weaning) to investigate the carryover effects of dietary treatments. Lactobacillus mucosae (LM) broth or control broth was top-dressed into feeder troughs according to the treatment arrangement. Individual BW and feed disappearances were measured weekly from d 0 to 35 for determination of ADG, ADFI, and G:F. Weekly blood samples were obtained and additional blood samples were collected on d 1 and 3 post-LPSchallenge. Blood samples were evaluated for immune biomarkers. Tissue (duodenum and ileum) samples and mucosal scrapings were obtained for histological and immune biomarker analyses. Fecal samples (1 pig/pen) on d 0, 14, and 21 were determined for concentrations of VFA. All data were analyzed as a completely randomized design using GLIMMIX procedure. Fecal VFA: On d 0 and d 14, there were no differences (P > 0.10) in concentrations of VFA. On d 21, supplementation of MOS and LM tended (P < 0.10) to have interaction effect in propionate; feeding MOS tended (P < 0.10) to decrease propionate compared to control and MOS+LM group, but was not different (P > 0.10) from LM-fed pigs. Main effects or interaction effects (P > 0.10) were not observed for concentrations of other VFA on d 21. However, a contrast analysis showed that pigs fed MOS tended to have lower (P < 0.10) butyrate compared to control and MOS+LM fed pigs. The amounts of individual VFA in proportion of total VFA were calculated. There were no differences (P > 0.10) on d 0. However, on d 14, pigs fed MOS and LM had interaction effects in percentage of acetate (P < 0.10), butyrate (P < 0.05), and valerate (P < 0.10). Pigs fed MOS had greater (P < 0.05) acetate, but lower (P < 0.05) butyrate and valerate in proportion of total VFA compared to control group. In addition, pigs fed MOS tended (P < 0.10) to have greater acetate compared to MOS+LM group. The proportions of isobutyrate (P < 0.10), butyrate (P < 0.05), and isovalerate (P < 0.10) were lower in MOS vs. MOS+LM group. On d 21, no significant (P > 0.10) main effects or interaction effect were observed, but feeding MOS tended to decrease (P < 0.10) the proportions of propionate and butyrate compared to pigs fed the control diet. Key outcomes for obective 4: Feeding MOS showed lower proportions of propionate, butyrate, and valerate compared to pigs fed the control diet. Because the MOS and LM-fed pigs showed improved immune status during the week of LPS-challenge, we propose that the decreased or moderate change of VFA production in these pigs maybe due to a lower requirement of VFA for intestinal immunity. In addition, the decreased butyrate production in MOS-fed pigs may also attribute to the different gut environmental conditions.
Publications
|
Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Target audiences include animal scientists, students, pork producers, stakeholder's and related clientele. Processes that deliver science-based knowledge included laboratory instruction and experiential learning opportunities provided to students and extension and outreach activities to related clientele via presentation of research data at professional and scientific meetings including ASAS Midwest Animal Science Meetings (2016). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The activities associated with this experiment has provided training opportunities (applied animal experiments as well as basic bench laboratory experiments) for Graduate students (Yanshuo Li and Shana Barnett-primary, Melanie Trenhaile, and Dana van Sambeek); Undergraduate students (Talia Everding and Brady Dierks); Technicians (Jeff Perkins, Gail Henderson); and Collaborators (Phillip Miller, Samodha Fernando). How have the results been disseminated to communities of interest?Data from this experiment will be reported at the 2017 Midwest ASAS meetings in Omaha, NE. What do you plan to do during the next reporting period to accomplish the goals?We will continue to evaluate and refine prebiotic and probiotic (as well as there combination-synbiotic) combinations via in vitro and in vivo methods. In addition, from this most recent experiment, laboratory analyses to complete include microbial community profiling and gene expression (gut barrier function) assays.
Impacts
What was accomplished under these goals?
A healthy gut is comprised of effective digestion/absorption, absence of gut illness, normal/stable intestinal microbiota, effective immune status, and status of well-being. Prebiotics are nondigestible dietary ingredients that beneficially affect the host by selectively stimulating growth/activity of beneficial bacteria. In response to prebiotic consumption, beneficial effects have been observed in pigs including positive changes in gastrointestinal microbota, fermentative end-products, intestinal morphology, nutrient digestibility, and immune function leading to enhancements in overall pig gut health and growth performance. Collectively, effects of prebiotics may be a culmination of host-microbial interactions. The overall goal of this project is to explore how host-microbial interactions may culminate to impact gut health and nutrient metabolism in weaned pigs fed prebiotics and to gather host phenotypic and microbial community data to identify potential probiotic species. During the previous period we determined that, when fed to weanling pigs, dietary mannan oligosaccharides (MOS) increased fecal abundances of Lactobacillus mucosae (LM), which was positively correlated with circulating IgA concentration. Thus, LM was identifyed as a potential probiotic. As a result of those findings, an experiment was designed to determine the effects of feeding MOS and LM (alone and in combination) as prebiotic and probiotic sources in weanling pigs under immune challenge. Results from this experiment are detailed below; however the overall conclusion is that feeding LM alone improved feed efficiency and ileal morphological structure during the first week of immune challenge; additionally, feeding LM and MOS may have beneficial effects relative to immune biomarkers. Objectives 1) Delineate changes in microbial ecology in pigs fed prebiotics; and 2) Evaluate the molecular and metabolic effects of prebiotics and intestinal microbiota on intestinal barrier function and integrity; and 3) Compare changes in gastrointestinal microbiology with DNA microarrays and pig performance and digestibility phenotype to identify genes that undergo induction or repression during colonization. Nothing new to report at this time. Objective 4) Develop and test optimized prebiotic intervention strategies in production environments (emphasizing non-antibiotic approaches) for enhancing porcine health and food production. Based on our previous results indicating that feeding MOS (Saccharomyces cerevisiae) increased fecal Lactobacillus mucosae (LM) and was positively correlated with circulating IgA concentration, and experiment was designed to investigate the prebiotic and probiotic effects of MOS and LM on growth performance, immune response, and gut health of weanling pigs. To model an enteric infection, we challenged the pigs with E. coli LPS. Weaned pigs (n=100) were randomly allotted to 16 pens with a 2x2 factorial arrangement of treatments. On d 14 post-weaning, all pigs were inoculated with LPS. Corn-soybean-meal based diets (without plasma or antibiotics) with or without 0.1% yeast-derived MOS were randomly assigned to pens and 109 cfu/pig LM broth or an equal volume of control broth was top-dressed daily. Pigs were fed dietary treatments (control, MOS, LM, and MOS+LM) in phase-1 and phase-2 (d 0 to 7 and d 7 to 21 post-weaning, respectively). A common diet was fed to all pigs during phase-3 (d 21 to 35 post-weaning) to investigate the carryover effects of dietary treatments. Lactobacillus mucosae (LM) broth or control broth was top-dressed into feeder troughs according to the treatment arrangement. Individual BW and feed disappearances were measured weekly from d 0 to 35 for determination of ADG, ADFI, and G:F. Weekly blood samples were obtained and additional blood samples were collected on d 1 and 3 post-LPS-challenge. Blood samples were evaluated for immune biomarkers. Tissue (duodenum and ileum) samples and mucosal scrapings were obtained for histological and immune biomarker analyses. All data were analyzed as a completely randomized design using GLIMMIX procedure. Growth performance: From d 0 to 14, the LM-treated groups had decreased (P < 0.05) G:F compared to non-LM-treated groups. Immediately after the LPS-challenge, there was an interaction (P < 0.05) in G:F and a tendency of interaction (P < 0.10) in ADG between MOS- and LM-treated pigs. From d 14 to 21, G:F of pigs fed LM was greater compared to MOS+LM (P < 0.05) and control (P < 0.10), but was not different (P > 0.10) from pigs fed MOS. After removal of treatments, G:F of pigs in LM vs. non-LM groups were lower (P < 0.05) from d 21 to 28 and tended to be lower (P < 0.10) from d 28 to 35. In contrast, the LM treatment groups tended to have increased (P < 0.10) ADFI compared to non-LM treatment groups between d 28 and 35. The ADG of pigs were not different (P > 0.10) from d 21 to 35, but G:F of pigs were lower (P < 0.05) in LM vs. non-LM treatment groups. The daily feed intake of pigs recovered within 2 days post-LPS-challenge. Serum and Mucosal immunoglobulins and IL-1: Feeding MOS vs. non-MOS treated diets increased (P < 0.05) serum IgG on d 15 and 17 post-weaning. Serum IL-1 concentration decreased (P < 0.05) from d 0 to 14, increased on d 15 (P < 0.05). There was a tendency of MOS × LM interaction (P < 0.10) in circulating IL-1 concentration. On d 7, 15, and 17, IL-1 of pigs fed MOS+LM was increased (P < 0.05) compared to LM and tended to be greater (P < 0.10) compared to control and MOS groups. On d 14, IL-1 was greater (P < 0.05) in pigs fed MOS+LM vs. MOS and LM, and tended to be greater (P < 0.10) in MOS+LM vs. control. On d 21, pigs fed MOS+LM had the highest (P < 0.05) IL-1 concentration compared to other groups. There was an MOS × LM interaction (P < 0.05) for secretory IgG concentration on d 15; mucosal IgG was increased (P < 0.05) in control vs. MOS and LM groups, but was not different (P > 0.10) from MOS+LM. Small Intestine Histological Analyses: On d 15, feeding LM tended (P < 0.10) to decrease crypt depth in the ileum; pigs fed LM had decreased ileal crypt depth (P < 0.05) compared to control, but were not different (P > 0.10) from the pigs fed MOS or MOS+LM diets. Key outcomes for obective 4: With respect to growth performance, LM showed increased G:F compared control and MOS+LM pigs from d 14 to 21, indicating that supplementation of LM may protect the pigs against acute inflammation due to LPS-challenge. With respect to immune biomarkers, circulating but not secretory IgG antibodies were increased by feeding MOS while pigs fed LM alone had a increase in both secretory and circulating IgA concentrations of pigs. In addition, the LPS-challenge increased serum IL-1 in control and MOS+LM groups, indicating that feeding MOS or LM alone may prevent the acute inflammation caused by LPS challenge. With respect to histological analyses, it was observed that feeding LM decreased ileal crypt depth which may contribute to improved feed efficiency during immune challenge. Taken together, the data indicate that MOS may be used as a prebiotic to promote maturation of IgG production in weanling pigs, while LM can be used as a probiotic to improve immunity especially during acute inflammation.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Li, Y.S., M.D. Trenhaile, M.M. Lima, K.C. Moore, D. M. van Sambeek, T. E. Burkey, and P. S. Miller. 2016. Growth performance and serum IgA concentrations in weanling pigs fed dietary prebiotics. J. Anim. Sci. 94(Suppl. 2):72. (Abstr.) doi:10.2527/msasas2016-155
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Li, Y.S., S. C. Fernando, P. S. Miller, T. E. Burkey. 2016. Alteration of fecal bacterial communities in weanling pigs fed diets supplemented with chicory, mannan oligosaccharides, or chitosan. J. Anim. Sci. 94(Suppl. 2):138. (Abstr.) doi:10.2527/msasas2016-294
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2017
Citation:
Li, Y.S., M.D. Trenhaile, D.M. van Sambeek, K.C. Moore, S.M. Barnett, S.C. Fernando, T.E. Burkey, and P.S. Miller. 2017. Effects of mannan oligosaccharides and lactobacillus mucosae on the growth performance and immune response of weanling pigs challenged with Escherichia coli lipopolysaccharides. J. Anim. Sci. 95(E-Suppl. 2):xx (Abstr.)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2017
Citation:
Li, Y.S. Li, Y.S., M.D. Trenhaile, D.M. van Sambeek, K.C. Moore, S.M. Barnett, S.C. Fernando, T.E. Burkey, and P.S. Miller. 2017. Effects of mannan oligosaccharides and lactobacillus mucosae on the intestinal morphology of weanling pigs challenged with Escherichia coli lipopolysaccharides. J. Anim. Sci. 95(E-Suppl. 2):xx (Abstr.)
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Target audiences include animal scientists, students, pork producers, stakeholder's and related clientele. Processes that deliver science-based knowledge included laboratory instruction and experiential learning opportunities provided to students and extension and outreach activities to related clientele via presentation of research data at professional and scientific meetings including Digestive Physiology of Pigs (2015), ASAS Midwest and National Animal Science Meetings (2015). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The activities associated with first experiment has provided training opportunities (applied animal experiments as well as basic bench laboratory experiments) for Graduate students (Yanshuo Li and Shana Barnett)-primary, Melanie Trenhaile, and Dana van Sambeek;Undergraduate students (Talia Everding); Technicians (Jeff Perkins, Gail Henderson); and Collaborators (Phillip Miller, Samodha Fernando). How have the results been disseminated to communities of interest?Data from the initial experiment was reported at the 2015 Midwest ASAS meetings in Des Moines, IA and the 13th Int. Symp. Dig. Phys. Pigs inKliczkow, Poland. What do you plan to do during the next reporting period to accomplish the goals?In addition to evaluating samples collected from the initial in vivo experiments, we plan to conduct a series of in vitro experiments using a porcine intestinal epithelial cell line (IPEC-J2) to look atmolecular and metabolic effects of prebiotics on intestinal barrier function and integrity. In addition, we hope to conduct an experiment to evaluate a potential prebiotic/probiotic intervention in weanling pigs as an alternative to antibiotics.
Impacts
What was accomplished under these goals?
A healthy gut is comprised of effective digestion/absorption, absence of gut illness, normal/stable intestinal microbiota, effective immune status, and status of well-being. Prebiotics are nondigestible dietary ingredients that beneficially affect the host by selectively stimulating growth/activity of beneficial bacteria. In response to prebiotic consumption, beneficial effects have been observed in pigs including positive changes in gastrointestinal microbota, fermentative end-products, intestinal morphology, nutrient digestibility, and immune function leading to enhancements in overall pig gut health and growth performance. Collectively, effects of prebiotics may be a culmination of host-microbial interactions. The overall goal of this project is to explore how host-microbial interactions may culminate to impact gut health and nutrient metabolism in weaned pigs fed prebiotics and to gather host phenotypic and microbial community data to identify potential probiotic species. During this period we evaluated the effects of prebiotics on growth performance, immune status and fecal bacterial communities. With respect to growth performance, no significant effects were observed. With respect to immune status, circulating immunoglobulin A increased (P < 0.05) over time from 0.148 to 0.438 mg/mL (d 0 to 28 post-weaning), but was not affected by dietary prebiotics. With respect to fecal bacterial communities, dietary prebiotics altered the bacterial community on d 14 and d 28. Specifically, the bacterial community in pigs fed mannanoligosaccharides had increased Lactobacillaceae on d 14 and Streptococcaceae on d 28, while chicory and chitosan increased the bacterial abundances for Ruminoccocaceae on d 14. When evaluating correlations of OTU abundances to phenotypic traits, it was observed that bacteria from the family mucosae were positively correlated with immunoglobulin A and bacteria from the family Christensenellaceae were negatively correlated with pig body weight. Phenotypic and microbial community data will be used to identify potential probiotic bacterial species. Objectives 1) Delineate changes in gastrointestinal microbial ecology in weanling pigs fed prebiotics via high-throughput pyrosequencing of microbiome DNA; and 2) Evaluate the molecular and metabolic effects of prebiotics and intestinal microbiota on intestinal barrier function and integrity; Relative to objectives 1 and 2, to determine the effects of dietary prebiotics on growth performance, immune status, barrier function, and fecal bacterial communities of weanling pigs, 64 crossbred pigs (initial BW = 6.6 kg; d 20 to 22 post-farrowing) were selected and randomly allotted to 16 experimental pens with 4 dietary treatments (2 barrows and 2 gilts per pen, 4 pens per treatment). The treatments were maintained during Phase I (d 0 to 14 post-weaning) and Phase II (d 14 to 28 post-weaning). The control diet was a conventional nursery diet formulated with corn, soybean meal, dried whey, fish meal, plasma, and supplements to meet or exceed the 2012 NRC requirements. The additional 3 diets were formulated to contain 0.1% chicory, 0.1% mannan oligosaccharides (MOS), and 0.02% chitosan, respectively. Pigs were given ad libitum access to feed and water in an environmentally-controlled room. From d 0 to 28 post-weaning, feed disappearance and individual BW were measured weekly for determination of average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F). Blood samples were collected weekly for measurements of serum IgA concentrations as an indication of immune status and gut barrier function. Data were analyzed as a completely randomized design using the MIXED procedure of SAS. Overall, dietary prebiotics did not affect (P > 0.10) BW (average 16.7 kg; d 28), ADG (361 g), or ADFI (513 g). However, from d 7 to 14, pigs fed MOS had lower (671 g/kg; P < 0.05) G:F compared to pigs fed control (830 g/kg), chicory (851 g/kg), and chitosan (871 g/kg) diets. For Phase I, G:F of pigs fed MOS tended to be lower (656 g/kg; P < 0.10) than pigs fed control (791 g/kg) and chitosan (783 g/kg), but was not different from chicory (755 g/kg; P > 0.10). For Phase II, G:F of pigs fed control (673 g/kg), chicory (686 g/kg), MOS (695 g/kg), and chitosan (695 g/kg) were not different (P > 0.10). There were no time × treatment interactions (P > 0.10) affecting circulating IgA concentrations. Serum IgA increased (P < 0.05) over time from 0.148 to 0.438 mg/mL (d 0 to 28 post-weaning), but was not affected by dietary prebiotic. For bacterial community analysis, fecal samples from 2 pigs/pen were collected on d 0, 14, and 28 post-weaning to evaluate bacterial community composition using 16S rRNA tag sequencing. Database independent operational taxonomic unit (OTU) approach was used for bacterial community analysis. The α-diversity estimate (Chao 1) was different (P < 0.05) over time, but was not affected (P > 0.10) by treatment. Additionally, β-diversity changed over time (P < 0.05). On d 14, β-diversity of the bacterial community in control pigs were different (P < 0.05) from chicory and chitosan, but were not different (P > 0.10) from MOS. Linear discriminate analysis (LDA) by LEfSe was used for pair-wise comparisons of OTU abundances in the bacterial communities ( LDA score > 2 was considered significant). On d 14, control and MOS had 26 and 12 OTUs, respectively more abundant in the 3 comparisons with other treatments, in which the majority of these OTUs belonged to family Lactobacillaceae. Additionally, 25 and 17 OTUs, respectively that predominantly belonged to family Ruminococcaceae and Christensenellaceae were associated with chicory and chitosan. Using multivariate association with linear models (MaAsLin; P < 0.05, q < 0.05), 3 OTUs belonging to Lactobacillaceae family were positively correlated to MOS (coeff. > 0.012). Mocosae spp. was associated (coeff. = 0.092) with IgA. One OTU belonging to family Christensenellaceae was negatively correlated (coeff. = -0.01) to BW. Family level classification of OTUs was analyzed using ANOVA1 of MATLAB. On d 14, Lactobacillaceae decreased (P < 0.05) in chicory (18.4%) compared to control (46.5%) and MOS (41.7%); whereas, Ruminoccocaceae increased (P < 0.05) in chicory (15.4%) and chitosan (15.0%) compared to control (10.6%) and MOS (10.3%). Chicory had greater (P < 0.05) Christensenellaceae (5.6%) compared to the other 3 treatments (< 1.8%). On d 28, Streptococcaceae increased (P < 0.05) in control (27.2%) compared to chicory (8.7%) and chitosan (9.5%). Key outcomes for Objectives 1 and 2. With respect to growth performance, immune status, and barrier function, with the exception of subtle decreases in feed efficiency, prebiotic supplementation had no effect on growth performance or serum IgA concentrations in weanling pigs. With respect to fecal bacterial community analyses, the most intriguing finding was the positive correlation of 3 different Lactobacillaceae OTUs (a potential probiotic) with the mannanoligosaccharide treatment. This data will be fundamental in developing and implementing probiotic and prebiotic/probiotic (symbiotic) combinations that can be directed to specific phenotypic traits. Objectives 3) Compare changes in gastrointestinal microbiology with DNA microarrays and pig performance and digestibility phenotype to identify genes that undergo induction or repression during colonization; and 4) Develop and test optimized prebiotic intervention strategies in production environments (emphasizing non-antibiotic approaches) for enhancing porcine health and food production. Obectives 3 and 4: Nothing to report at this time.
Publications
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2016
Citation:
Li, Y.S., S. C. Fernando, P. S. Miller, T. E. Burkey. 2016. Alteration of fecal bacterial communities in weanling pigs fed diets supplemented with chicory, mannanoligosaccharides (MOS), or chitosan. J. Anim. Sci. (Abstract)
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2016
Citation:
Li, Y.S., M. Trenhaile, K. Moore, M. Monteiro, D. M. van Sambeek, T. E. Burkey, and P. S. Miller. 2016. Growth performance and serum IgA concentrations in weanling pigs fed dietary prebiotics. J. Anim. Sci (Abstract)
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Progress 12/17/13 to 09/30/14
Outputs
Target Audience: Target audiences include animal scientists, students, pork producers, stakeholder's and related clientele. Processes that deliver science-based knowledge included laboratory instruction and experiential learning opportunities provided to students and extension and outreach activities to related clientele via presentation of data at the Midwest ASAS Meetings (2014) Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The activities associated with first experiment has provided training opportunities (applied animal experiments as well as basic bench laboratory experiments) for Graduate students (Gabriel Mastromano)-primary, Huyen Tran, and Justin Bundy; Undergraduate students (Talia Everding); Technicians (Jeff Perkins, Gail Henderson); Collaborators (Phillip Miller, Samodha Fernando). How have the results been disseminated to communities of interest? Data from the initial experiment was reported at the 2014 Midwest ASAS meetings in Des Moines, IA What do you plan to do during the next reporting period to accomplish the goals? A second experiment has been designed and will be conducted in January, 2015 with the goal of addressing the Obj. 1 (Delineate changes in gastrointestinal microbial ecology in weanling pigs fed prebiotics via high-throughput pyrosequencing of microbiome DNA).
Impacts
What was accomplished under these goals?
A healthy gut is comprised of effective digestion/absorption, absence of gut illness, normal/stable intestinal microbiota, effective immune status, and status of well-being. Prebiotics are nondigestible ingredients that beneficially affect the host by selectively stimulating growth/activity of beneficial bacteria. Beneficial effects have been observed in pigs fed prebiotics including positive changes in gastrointestinal microbota, fermentative end-products, intestinal morphology, nutrient digestibility, and immune function. Collectively, effects of prebiotics may be a culmination of nutrient/microbial interactions. The first (preliminary) experiment conducted for this project was designed with the goal of determining the appropriate concentration of a specific prebiotic (inulin from chicory) that could be included in pig diets. The specific objectives of this experiment were to to investigate the effects of dietary prebiotics (inulin from chicory) in nursery pigs by evaluating growth performance and immune biomarkers. The overall conclusion of this experiment was that inclusion of prebiotics in nursery pig diets has positive effects on growth performance and may have immunomodulatory effects (in vitro) on cells isolated from prebiotic-fed pigs. A second experiment, to be conducted in January, 2015, has been designed to address Objective 1 (Delineate changes in gastrointestinal microbial ecology in weanling pigs fed prebiotics via high-throughput pyrosequencing of microbiome DNA) for the current project.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Mastromano, G.A., Y.S. Li, H. Tran, J.W. Bundy, P.S. Miller, and T.E. Burkey. 2014. Prebiotic effects on growth performance and immune health in nursery pigs. J. Anim. Sci. 92(E-Suppl. 2): 342 (Abstr.)
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