Optimization of canola co product utilization in swine

South Dakota State University Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange Electronic Theses and Dissertations 2019 Optimization of Canola Co-Product Utilization in Swine Jung Wook Lee South Dakota State University Follow this and additional works at: https://openprairie.edu/etd Part of the Animal Sciences Commons Recommended Citation Lee, Jung Wook, "Optimization of Canola Co-Product Utilization in Swine" (2019). Electronic Theses and Dissertations.edu/etd/3654 This Dissertation - Open Access is brought to you for free and open access by Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. For more information, please contact michael.

OPTIMIZATION OF CANOLA CO-PRODUCT UTILIZATION IN SWINE BY JUNG WOOK LEE A dissertation submitted in partial fulfillment of the requirements for the Doctor of Philosophy Major in Animal Science South Dakota State University 2019 DocuSign Envelope ID: 2CC1E6E6-A95C-4C32-A47B-EC241052E1CC ii DISSERTATION ACCEPTANCE PAGE Jung Wook Lee This dissertation is approved as a creditable and independent investigation by a candidate for the Doctor of Philosophy degree and is acceptable for meeting the dissertation requirements for this degree. Acceptance of this does not imply that the conclusions reached by the candidate are necessarily the conclusions of the major department. Tofuko Woyengo Advisor Date Joseph P Cassady Department Head Date Dean, Graduate School Date iii This dissertation is dedicated to my parents Kyung Sook Choi and Song Jae Lee. iv ACKNOWLEDGEMENTS First of all, I would like to express my sincerest appreciation to Dr.

Woyengo for granting me this opportunity to pursue a doctorate degree in monogastric nutrition. Without his guidance and support, it would not have been possible for me to complete my doctorate program at SDSU. I am truly honored to have had an opportunity to work with him, learn from him and get involved in various research projects. Second, I would like to sincerely thank my committee members Drs.

Teresa Seefeldt, Benoit St. Pierre, and Joy Scaria for serving on my committee. I am very thankful to the previous and current members of Dr. Woyengo’s laboratory: Emily Scholtz, Casey Zangaro, Kevin Jerez-Bogota, Cristian Sánchez, Jimena Alejandra Ibagón, Dr.

Jinsu Hong, Heeseong Kim, and Samuel Ariyibi for all contributing to my research projects. Last, I greatly appreciate the sacrifices and endless love of my parents, Song Jae Lee and Kyung Sook Choi. v FOREWORD This dissertation was written and prepared in a manuscript format and it is composed of six manuscripts. The six manuscripts were written based on data collected from the five studies that were conducted to test the hypotheses and achieve the objectives of thesis research.

The first manuscript was written based on data obtained from the first in vitro study. The second and third manuscripts were written based on data obtained from the second study. The fourth manuscript was written based on data collected from the third pilot study. The fifth manuscript was written based on results from the fourth study.

The sixth manuscript was written based on data collected from the fifth in vitro study. The first manuscript has been published in Animal Feed Science and Technology, whereas the second manuscript has been published in the Journal of Animal Science. The fifth and sixth manuscripts are in preparation for publication. Authors to the published manuscripts are as follows: first manuscript – J.

Woyengo; second manuscript – J. In this dissertation, the manuscripts were written and prepared in accordance to the guidelines for the Journal of Animal Science manuscript preparation. vi TABLE OF CONTENTS ABBREVIATIONS. ix LIST OF FIGURES.

xi LIST OF TABLES. xv CHAPTER ONE.1 Effect of Glucosinolates on Performance, Organ Weights, and Thyroid Hormones .2 Effect of Glucosinolates on Swine Gut Microbiome .3 Glucosinolate Degradation Products .4 Factors Affecting Composition of Degradation Products of Glucosinolates .5 Glucosinolate Metabolism in Swine.6 Strategies for Reducing Toxicity of Glucosinolates .1 Reducing Glucosinolate Content Through Heat Treatment .2 Reducing pH in Hindgut of Pigs .7 Summary and Perspectives .3 MATERIALS AND METHODS .1 Experimental Design and Treatments .2 Porcine In Vitro Digestion.3 Porcine In Vitro Microbial Fermentation .4 Sample Preparation and Feedstuff Analyses .5 Calculations and Statistical Analysis .3 MATERIALS AND METHODS .1 Animals and Housing .3 Experimental Design and Procedure .4 Sample Preparation and Analyses .3 MATERIALS AND METHODS .4 RESULTS AND DISCUSSION .3 MATERIALS AND METHODS .4 RESULTS AND DISCUSSION .3 MATERIALS AND METHODS .1 Animals and Housing .3 Experimental Design and Procedure .4 Sample Preparation and Analyses .Determination of Isothiocyanates Using High-Performance Liquid Chromatography .3 MATERIALS AND METHODS .1 Experimental Design and Treatments .2 Porcine In Vitro Microbial Fermentation .3 Sample Preparation and Analysis. 173 ix ABBREVIATIONS % Percent ± Plus or minus AA Amino acids ADF Acid detergent fiber ADFI Average daily feed intake ADG Average daily gain Ala Alanine AOAC Association of Official Analytical Chemists Arg Arginine Asp Aspartic acid BW Body weight °C Degrees Celsius Ca Calcium cal Calorie CP Crude protein Cu Copper Cys Cysteine d Day DM Dry matter EE Ether extract Fe Iron G:F Gain to feed ratio GE Gross energy x Glu Glutamine Gly Glycine H Hour His Histidine Ile Isoleucine Leu Leucine Lys Lysine Met Methionine NDF Neutral Detergent fiber P Phosphorus P P-value Pro Proline SAS Statistical analysis software Se Selenium SEM Standard error of the mean Ser Serine SBM Soybean meal Thr Threonine Trp Tryptophan Tyr Tyrosine Val Valine Zn Zinc xi LIST OF FIGURES Figure 4. Gas production kinetics of the undigested residue of canola co-products during 72-hour of microbial fermentation………………………………………….1 Microbiome diversity of bacterial population in ileum digesta and fecal samples obtained from pigs fed 4 different diets………………………………….

Gas production kinetics of canola co-products during 72-hour of microbial fermentation……………………………………………………………………….162 xii LIST OF TABLES Table 2. Nutrient composition and energy values of canola co-products (% DM basis)………………………………………………………………………………. Non-starch polysaccharide (NSP) content of conventional canola meal and soybean meal (% DM basis)………………………………………………. Total, aliphatic and aromatic glucosinolate contents in canola co-products for pigs………………………………………………………….

Effects of including canola co-products in diets for pigs on body weight gain and feed intake…………………………………………………………………. Effects of including canola co-products in diets for pigs on organ weights and thyroid hormone concentrations………………………………………………. Analyzed nutrient composition (g/kg dry matter) of canola co-products. Analyzed non-starch polysaccharide (NSP) contents of canola co-products and of enzymatically unhydrolyzed residue…………………………………………62 Table 4.

Coefficients of in vitro digestibility of dry matter (DM) and non-starch polysaccharides (NSP) for canola co-products………………………………………63 Table 4. Fitted kinetics parameters and analyzed VFA concentrations and molar ratios after in vitro fermentation of enzymatically unhydrolyzed residue of canola co- products………………………………………………………………………………64 Table 5. Ingredient composition and analyzed nutrient content of experimental diets (as-fed basis) ………………………………………………………. Analyzed nutrient and glucosinolate content of cold-pressed canola cake (% DM basis) ……………………………………………………………………….

Growth performance of nursery pigs fed increasing levels of cold-pressed canola cake…………………………………………………………………………. Organ weights, complete blood counts and serum thyroid hormones of nursery pigs fed increasing levels of cold-pressed canola cake……………………. Gastrointestinal tract weight (% of live BW) of pigs fed diets containing cold-pressed canola cake……………………………………………………………. Relative abundance of phyla (% of total reads) in ileal digesta and feces of pigs fed cold-pressed canola cake………………………………………………….

Relative abundance of family members (% of total reads) in ileal digesta and feces of pigs fed cold-pressed canola cake……………………………………. Ingredient composition of experimental diets (as-fed basis) …………. Growth performance of nursery pigs fed increasing levels of HA- starch…………………………………………………………. Cecal and colonic pH of nursery pigs fed increasing levels of HA- starch……………………………………………………………………………….

Ingredient composition and analyzed nutrient content of experimental diets (as-fed basis) ………………………………………………………………………. Analyzed nutrient and glucosinolate content of cold-pressed canola cake (% DM basis) ………………………………………………………………………135 Table 8. Growth performance of nursery pigs fed cold-pressed canola cake without or with resistant starch……………. Organ weights and serum thyroid hormone concentrations of nursery pigs fed cold-pressed canola cake without or with resistant starch…………………….

Hindgut pH and cecal glucosinolate degradation metabolite of nursery pigs fed cold-pressed canola cake without or with resistant starch…………. Cecal volatile fatty acid concentrations of nursery pigs fed cold-pressed canola cake without or with resistant starch……………………. Analyzed nutrient and glucosinolate content of canola co-products (% DM basis)…………………………………………………………. Analyzed glucosinolate degradation products and fermentation medium pH after in vitro fermentation of canola co-products……….

Fitted kinetics parameters and analyzed VFA concentrations after in vitro fermentation of canola co-products………………………….161 xv ABSTRACT OPTIMIZATION OF CANOLA CO-PRODUCT UTILIZATION IN SWINE JUNG WOOK LEE 2019 Canola co-products have a high content of fiber and glucosinolates. Fiber reduces nutrient utilization in pigs, whereas glucosinolates are degraded to toxic products, which interfere with liver, kidney and thyroid functions. Negative effects of fiber can potentially be alleviated by fiber-degrading enzymes, whereas negative effects of glucosinolates in pigs can potentially be alleviated through reduction in hindgut pH. However, there is a lack of information on effects of supplemental fiber- degrading enzymes on digestion and fermentation characteristics of canola co- products for pigs.

Also, there is limited information on effects of reducing hindgut pH on toxicity of glucosinolates in pigs. Four experiments were conducted to fill these gaps in knowledge. The first experiment investigated effects of supplementing canola co-products with fiber-degrading enzymes on porcine in vitro digestion and fermentation of canola co-products. Supplemental fiber-degrading enzymes increased in vitro digestibility of canola co-products.

The second experiment investigated effects of increasing levels of cold-pressed canola cake (CPCC) in diets for pigs from 0 to 40% on growth performance, organ weights, blood thyroid hormone levels. Growth performance, metabolic activity in liver and thyroid functions were negatively affected by dietary inclusion of CPCC at 40%. The third experiment investigated effects of reducing hindgut pH through dietary inclusion of high-amylose cornstarch (HA-starch) on the fore-mentioned response criteria and cecal concentration of glucosinolate degradation products in pigs fed diets that contained 40% CPCC. Dietary xvi CPCC increased thyroid gland weight of pigs fed HA-starch-free diet, but not of pigs fed HA-starch-containing diet.

Inclusion of HA-starch in CPCC-based diets increased isothiocyanate production in cecal digesta of pigs. However, nitriles were undetected in cecal digesta of pigs fed CPCC-based diets. Thus, the fourth experiment was conducted to determine effects of reducing pH on composition of glucosinolate degradation products in canola co-products using porcine in vitro fermentation technique. Reduction in fermentation medium pH from 6.2 increased production of indole- 3-acetonitriles.

In conclusion, the results demonstrated that fiber-degrading enzymes can be supplemented to canola co-products-based diets for pigs to improve efficiency of nutrient utilization and that toxicity of canola glucosinolates can be alleviated through reduction in pH of hindgut of pigs. 1 CHAPTER ONE GENERAL INTRODUCTION Canola co-products are the second most commonly used sources of amino acids (AA) after soybean meal (SBM) in swine diets (Woyengo et al. Generally, oil is extracted from canola seeds by solvent extraction, expeller or cold pressing method. Solvent extraction is the most widely used method of oil extraction followed by expeller pressing, and then cold pressing.

Thus, solvent-extracted canola meal (SECM) is the most widely fed canola co-product followed by expeller-pressed canola meal (EPCM). However, cold-pressed canola cake (CPCC) is increasingly becoming available for livestock feeding because cold pressing does not involve the use of thermal or chemical treatment, and the demand for natural food products such as cold-pressed oil has increased over the past few years (Siger et al. Inclusion of canola co-products in diets for pigs can be limited by their high content of fiber and glucosinolates, which reduces dietary nutrient utilization (Bell, 1993). Thus, there is a need to alleviate the negative effects of fiber and glucosinolates in canola co-products in order to optimize their utilization in the formulation of swine diets.

The negative effects of fiber can be alleviated through supplementation with fiber-degrading enzymes that target fiber present in canola co-products. Supplementation of SECM-based diets for broilers with multi-enzyme that targets most of the fiber in canola did not affect nutrient digestibility (Kocher et al., 2000; Meng and Slominski, 2005). However, supplementation of full-fat canola seed-based diets for broilers with multi-enzyme that targets most of the fiber in canola improved nutrient digestibility (Meng et al.