The study on the effect of a mixture of plant essential oils and organic acids on gut health of broiler chickens infected with avian pathogens

分分分分 分分分分分 10019 分 分分 分 分分 LB20173040007 分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 The study on the effect of a mixture of plant essential oils and organic acids on gut health of broiler chickens infected with avian pathogens 分 分 分分PHAM VAN HIEU 分 分 分 分分分分 分分分 分分分分分分分分分分分分分 分分 分分分分分分分分分分分分 分 分 分 分分分分分分分分分分分分分分分分 分 分 分 分分分分分分分 分分分 2021 分 分 分分分分分分分 分分分分 分 分 分 分 分 分 分 分 分 分 分 分 分 分分 分分分分分分分分分分 China Agricultural University Thesis The study on the effect of a mixture of plant essential oils and organic acids on gut health of broiler chickens infected with avian pathogens Ph.D Candidate: PHAM VAN HIEU Advisor: Associate Professor Zhong Wang Classification of Degree: Doctoral Degree of Agriculture Major: Animal Nutrition and Feed Science Research Field: Poultry immunity, animal nutrition and feed science College: College of Animal Science and Technology June, 2021 *A��ffi¥���X�ft�A{±��ffi��illfi���I�&�������o �ftpfr� �7x$1�1.1ittm��t3F5ifO�iM�±fuJJ9� itx$BEA8f}Đxt u mBA$oo*rtmwfilfflutt MoftI$*ffiffMBlx$7#*7 atfsJ: 2021 iƠ Yl 02 B *A��7m$OO��*��*�m,�ffl�w�x���o*A��$00�� * *�B n fDtfl1���itx��ftmt#&fD � -=r- #& 5ti!fitxft�1mto111m; t)(f*tt&!D] oo *Aã$oo**wx$Đx$3A($OOfflw x xttffl$)Đx($00ftwxxttffl$)illfiili#& *ffl*o * ( f*w1iritxrÊmw § m.51� utt1}tSl) BtfsJ: 2021 i¥ Yl 02 B Declaration of Originality I declare that the thesis submitted is my own research work and achievements under the guidance of my supervisor As far as I know, the thesis does not include the research results that have been published or written by others, nor the materials used to obtain the degree or certificate of China Agricultural University or other educational institutions, except for the special notes and thanks in the thesis Any contribution made by my colleagues to this research has been clearly explained and expressed in the thesis Student’s Name分 Date分2021 Y M 02 D Authorized Use Agreement I fully understand the regulations of China Agricultural University on retaining and using dissertations I agree that China Agricultural University has the right to keep and send the thesis and electronic versions of the thesis to the relevant departments and institutions of the state, allowing the thesis to be consulted and borrowed; I agree that China Agricultural University will authorize the compilation of all or part of the contents of this thesis into the full text database of Chinese doctoral dissertations or the full text database of Chinese excellent Master Dissertations for publication and enjoy relevant rights and interests (The confidential dissertations are subject to this agreement after decryption) Student’s Name分 Supervisor’s signature分 Date分2021 Y M 02 D Date分 Y M D 分分 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分 分分分分分分分分分分分分分NE分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分(EOA)分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分 分 2×2 分分分分分分 288 分 分分分分 分分分分分分分分分分分分分分分分分 分 500 mg/kg EOA 分分分分分分分 分分分分 分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分 EOA 分分分分分 分分分分分 EOA 分分分分分分分 分分分分分分分分分P < 0.01分分分分分分分分分分分 /分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分FITC-D分分分分分 分P < 0.05分分分分分分 EOA 分分分 分分分分分分分 EOA 分分分分分分分分 Claudin-1 分 分分分分分分分分分2分IGF-2分 分 mRNA 分分分分分分分P < 0.05分分A20 mRNA 分分分分分TNF 分分分分分分 (TRAF6)分分分分分分分分分分分分 15分TNFSF15分 分 Toll 分分分分分分分Tollip分 mRNA 分分分分分分分P < 0.05分分 分分分分分分分分 EOA 分分分分分分分分分分分分 分分 EOA 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 分分分 分 EOA 分分分分分分分分分分 EOA 分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分NE 分 分分分分分 EOA 分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 EOA 分分分分分分分分分分分分分分分分分分分分分分分分分分 NE 分分分 分分分分分分分分分分分分分分分分分EOA分分分分分分分分分分分分分分 分分分AGPs分分分分分分分分分分分分 分分分分分分Clostridium perfringens – (C Perfringens)分分分分分NE分分分分分分分分分分分分分分分分分 432 分分分分分分 分分分分分分分分分分 分分分分分 NE 分分分分分分A分分分分 NE 分分分分分 分D分分分分 NE分分分分分分分分分分分 250 mg/kg 分分分分分分分分分分BMD分 I 分分 15%分分200分500 分 800 mg EOA/kg 分分分分分分E分F分G 分 H 分分分分 AGPs 分分分分分 200 mg/kg 分 500 mg/kg EOA 分分分分分分分分分分分FCR分分分分分分 EOA 分分分分分分分分分分 200 mg/kg 分 500 mg/kg EOA 分分分分分分分分分分分 分分分分分分分分分分分分分分分分分 AGPs 分分 分分分分分分分分分 NE 分分分分 分分分分分分分分分分分分分分分分分分分分分分分分 EOA 分分分分分分分分分分 分分-1分分分分分分分分-2分GLP2分分分分分分分分分分-2分IGF-2分mRNA 分分 分分分分分分分 1分分分分分分分分分-1分ZO-1分分Toll 分分分 分TLR-4分分分分分 分分 1β分IL-1β分分分分分 γ分IFN-γ分分TNF 分分分分分分 6分TRAF-6分分分分 分分分分分 分分分分 15分TNFSF15分分 Toll 分分分分分分分Tollip分分分分分分分 分P < 0.05分分16S 分分分分分分分 200 mg/kg 分 500 mg/kg EOA 分分分分分分分分 分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分P < 0.05分分分分分分分 EOA 分分分 NE 分分分分 分分分分分分分分分分分分分分分分分分分EOA 分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分EOA分分分分分分 O78 分分 分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分 228 分 分分分分分分分分分分 分分分分分分分分 分分分分分分分分分分分分分 分分分 EOA分500 mg/kg 分分分分分分分 分分分分分分 EOA 分分分分分分分分分分 分分分分分分分分分分 O78 分分分分分 EOA 分分分分分分分分分分ADG分分分分分 分分分分 EOA 分分分分分分分分 EOA 分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分VH/CD分 分分分分分分分分分分分分分分分分分分 分分 EOA 分分分分分分分分分分分分分分分分分分分分分分分分 EOA 分 CD3 分 CD4 分分分分分分分分分分分分分分 EOA 分分分分分EOA 分分分 IgG 分分分分分 分分分分分分分 A分IgA分分分分分分分 M分IgM分分分分分分分分分分分分分 EOA 分分分分分分分分分分分分分分分分分分分 EOA 分分分分分分分分分分分分分分分 分 分分分分分分分分分分分 EOA 分分分分分分分分分分分分分P < 0.05分分分分分 分分分分分分分分分分分分分分分分分分分分 EOA 分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分分 分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分分 II Abstract The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis (NE) due to C perfringens In the first study, we investigated the effects of dietary supplementation with a blend of encapsulated essential oils and organic acids (EOA) on growth performance and gut health using a coinfection model of NE in broiler chickens Two hundred and eighty-eight one-day-old male Arbor Acres broiler chicks were randomly assigned using a 2 × 2 factorial design into two groups fed either or 500 mg/kg dietary EOA and co-challenged (or not challenged for the control) with Eimeria spp./C perfringens Infected birds fed the EOA-supplemented diet exhibited an improved feed conversion ratio throughout the trial (P < 0.01), a higher villus height and villus height/crypt depth ratio, and reduced intestinal C perfringens counts, liver C perfringens carriage, gut lesion scores and serum fluorescein isothiocyanate dextran (FITCD) concentrations at days-post-infection compared with those of birds without EOA supplementation (P < 0.05) NE-infected birds fed EOA exhibited significantly upregulated claudin-1 and insulin-like growth factor-2 (IGF-2) mRNA levels (P < 0.05), increased A20 mRNA expression and significantly downregulated TNF receptor-associated factor (TRAF-6), tumor necrosis factor superfamily member 15 (TNFSF15) and Toll-interacting protein (TOLLIP) mRNA levels in the jejunum at days-post-infection compared with those in birds without EOA supplementation (P < 0.05) Compared with the uninfected and untreated birds, the uninfected birds fed EOA displayed increased relative abundances of Lactobacillus and Coprococcus but reduced Rikenellaceae levels Compared with the unsupplemented NE- challenged birds, infected birds fed EOA showed an increased relative abundance of unclassified_Lachnospiraceae and a significantly decreased relative abundance of Erysipelotrichaceae EOA supplementation improved growth performance and gut health in NE-infected broiler chickens by strengthening the intestinal barrier function, positively modulating the gut microbiota community and differentially regulating intestinal immune responses Our results also suggested that adding EOA effectively controlled NE infections after experimental Eimeria and C perfringens coinfection In the second study, we evaluated the impacts of a blend of encapsulated organic acids with essential oils (EOA), as alternatives to antibiotic growth promoter (AGPs) on growth performance and gut health of Eimeria spp./C perfringens co-infected (NE) broilers A total of 432 day-old male Arbor Acres chicks were randomly distributed into treatment groups: non-infected negative control (A); NE-infected positive control (D) and NEinfected birds fed a basal diet supplemented with 250 mg/kg bacitracin methylene disalicylate (BMD, 15% purity), 200, 500, and 800 mg EOA /kg of diet (E, F, G and H group), respectively Feeding EOA at 200 mg/kg and 500 mg/kg significantly improved feed conversion ratio (FCR) In addition, increased villous height to crypt depth ratio and goblet cells counts were observed in NE-infected birds received EOA at 200 mg/kg and 500mg/kg as compared to the single NE-challenged groups without EOA supplementations (P < 0.05) Beside, feeding EOA at 200 mg/kg and 500 mg/kg significantly reduced gut lesions, serum fluorescein isothiocyanate dextran (FITC-D) level and C perfringens load in the cecum and liver of the NE-infected birds which was similar to AGPs In addition, upregulated claudin-1, glucagon- like peptide-2 (GLP-2), and insulin-like growth factor-2 (IGF-2) mRNA gene expression, and downregulated occludin, zonula occludens-1 (ZO-1), Toll-like receptor (TLR-4), interleukin (IL-1β), interferon γ (IFN-γ), TNF receptor-associated factor (TRAF-6), tumor necrosis factor superfamily member 15 (TNFSF15), and Tollinteracting protein (Tollip) genes expression in the jejunum were observed in NE-infected birds received EOA at 200 mg/kg and 500mg/kg as compared to the single NEchallenged groups without EOA supplementations (P < 0.05) 16S analysis showed that EOA supplemented with 200 mg/kg or 500 mg/kg enriched relative abundance of Lactobacillus, unclassified_Lachnospiraceae and Enterococcus as well as carbohydrate metabolic pathways but suppressed unclassified_Erysipelotrichacease and the organismal systems involving in immune the system (P < 0.05) In conclusion, feeding EOA could alleviate NE-induced gut impairment and growth depression and modulate cecal microbiota composition, which has potentials as antimicrobial alternatives In the third study, we conducted to evaluate essential oil and organic acids (EOA) supplementation on growth performance, intestinal morphology, immunity and gut microbiota of broilers challenged with E coli O78 The 228 1-day-old male broiler chicks were randomly distributed into treatments (with six replicates per group): experiment birds were fed from hatch with a diet supplemented with EOA (500 mg/kg feed), or with a non- supplemented control diet, and either uninfected or orally challenged with E Coli O78 The EOA supplementation failed to improve average daily weight gain (ADG) However, jejunum morphology was apparent in EOA-fed chickens as illustrated by 分分分分分分分分分分分分 References 165 Broom LJ, Kogut MH Deciphering desirable immune responses from disease models with resistant and susceptible chickens Poult Sci 2019;98:1634–42 166 Edelblum KL, Turner JR The Tight Junction in Inflammatory Disease: 156 分分分分分分分分分分分分 References Communication Breakdown Curr Opin Pharmacol 2009;9(6):715–20 167 Jeong E, Lee JY Intrinsic and extrinsic regulation of innate immune receptors Yonsei Med J 2011;52(3):379–92 168 Segain J, Blétière DR De, Bourreille A, Leray V, Gervois N, Rosales C, et al Butyrate inhibits inflammatory responses through NF kB inhibition : implications for Crohn ’ s disease Gut 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DM, Baird AW Cytokine regulation of epithelial permeability and ion transport Gut 1999;44(2):283–9 250 Wei C, Hengmin, C, Xi P Effect of Vanadium on Splenocyte Apoptosis in Broilers Med Chem (Los Angeles) 2012;02(03):57–60 251 Pedersen J, LaCasse EC, Seidelin JB, Coskun M, Nielsen OH Inhibitors of apoptosis (IAPs) regulate intestinal immunity and inflammatory bowel disease (IBD) inflammation Trends Mol Med 2014;20(11):652–65 252 Xavier RJ, Podolsky DK Unravelling the pathogenesis of inflammatory bowel disease Nature 2007;448(7152):427–34 253 Al-Sadi R, Boivin M, Ma T Mechanism of cytokine modulation of epithelial tight junction barrier Front Biosci 2009;14(7):2765–78 254 Yan W, Sun C, Yuan J, Yang N Gut metagenomic analysis reveals prominent roles of Lactobacillus and cecal microbiota in chicken feed efficiency Sci Rep 2017;7(45308):1–11 164 分分分分分分分分分分分分 Acknowledgments Acknowledgments I would like to express my deep thanks to Associate Professor Wang Zhong at State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University who enthusiastically guided me in the process of implementing and writing this thesis His experiences, research-oriented suggestions, research styles and problem conclusions are very helpful to me I also to thank Associate Professor Wang Zhong's graduate students for their assistance in the analysis, testing, and execution of this experiment I would like to thank all of the teachers and students in State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, for their help and guidance during the experiment I also extend my gratitude to the Farm of Zhuozhou (Hebei, China) staff team for their help and guidance during the implementation of these experiments I acknowledge Menon Animal Nutrition Technology Co Ltd., Shanghai, China, for providing the protected essential oil and organic acid products In particular, I would like to express my deep thanks to my beloved wife, my parents and children who have suffered so many hardships to help me complete this thesis Finally, I also want to thank brothers and sisters in my family and close friends who have always supported and encouraged me to strive to complete this thesis PHAM VAN HIEU 2021 分 分 165 分分分分分分分分分分分分 Resume Author’s Resume Personal Information: Full name : Pham Van Hieu Date of birth : Mar 07, 1981 Place of birth : Bac Ninh Province, Vietnam Gender : Male Nationality : Vietnamese Health : Excellent Address : Song Cong city, Thai Nguyen province Contact No : +86 188 1132 0681 and + 84 988 302 446 Email Address : phamhieudhnltn@gmail.com Language Spoken : Vietnamese, Chinese and English Educational Attainment: I M M ution aj i o n T V A et i M ni sc N ed ie gu T ici nc V i et M N ed gu ici D D e u g r G r d20 i 00 p l20 M20 a 09 st 20 Research Experience: - Pathologists strongyloidiasis (Strongyloides) in cattle in some localities in Thai Nguyen province and treatment method - Scientific works published: Prevalence of strongyloides Papillosus roundworm of cattle in Thai Nguyen and treatment method (2011) Journal of Animal Husbandry Sciences and Technics, (9), 19-23 166 分分分分分分分分分分分分 Resume Working Experiences: - Working as the technician and manager at the poultry farm and pig farm - Teaching and working in veterinary subjects in Thai Nguyen University of Agriculture and Forestry, Vietnam - Training employees at the trade center of Thai Nguyen Labor and Social Welfare Service - Working as a consultant of Care for development biosecurity model for small scale poultry production in Thai Nguyen province, Cao Bang province, Vietnam Interests: Research in chicken nutritions, immune response, essential oils and organic acids Publications Van Hieu Pham, Liugang Kan, Jinyu Huang, Yanqiang Geng, Wenrui Zhen, Yuming Guo, Waseem Abbas and Zhong Wang*, Dietary encapsulated essential oils and organic acids mixture improves gut health in broiler chickens challenged with necrotic enteritis; Journal of Animal Science and Biotechnology; (2020) 11:18 doi 10.1186/s40104019-0421-y Impact factor (2019): 4.393 Van Hieu Pham, Waseem Abbas, Jinyu Huang, Qiang He, Wenrui Zhen, Yuming Guo, and Zhong Wang*, Evaluation of the blend of encapsulated essential oils and organic acids as antibiotic growth promoter alternative on growth performance and intestinal health in broilers challenged with necrotic enteritis, Animal Feed Science and Technology, under review, impact factor (2020): 2.582 Wenrui Zhen, Yujing Shao, Yuanyuan Wu, Lianhong Li, Van Hieu Pham, Waseem Abbas, Zhang Wan, Yuming Guo, Zhong Wang, Dietary yeast β-glucan supplementation improves eggshell color and fertile eggs hatchability as well as enhances immune functions in breeder laying hens; International Journal of Biological Macromolecules; 159 (2020) 607–621 Impact factor (2020): 5.162 DOI: 10.1016/j.ijbiomac.2020.05.134 Liugang Kan, Fangshen Guo, Yan Liu, Van Hieu Pham, Yuming Guo, Zhong Wang; Probiotics Bacillus licheniformis Improves Intestinal Health of Subclinical Necrotic Enteritis-Challenged Broilers by Regulating Intestinal Microbiota, Mucosal Immune 167 分分分分分分分分分分分分 Response and Resume Barrier Function; Front Microbiol 10.3389/fmicb.2021.623739; impact factor (2020): 4.235 Computer Skills Microsoft PowerPoint Microsoft Office Publisher Microsoft Word, Microsoft Excel Communication Skills Excellence in written & oral communication Excellence in Interpersonal Communication Others Problem-solving Teamwork 168 12:623739 doi: ... China Agricultural University Thesis The study on the effect of a mixture of plant essential oils and organic acids on gut health of broiler chickens infected with avian pathogens Ph.D Candidate:... Impact of OAs on antioxidant enzyme activity The data from a study by A. M Abudabos [111] demonstrated that the blend of organic acids enhanced the total antioxidant capacity (TAC) concentration... Impact of EOs, OAs and EOAs on antibacterial activity and intestinal lesions 27 3.3.1 Impact of EOs on antibacterial activity and intestinal lesions 27 3.3.2 Impact of OAs on antibacterial