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      Graded Eimeria challenge linearly regulated growth performance, dynamic change of gastrointestinal permeability, apparent ileal digestibility, intestinal morphology, and tight junctions of broiler chickens

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          Abstract

          This study was conducted to evaluate graded Eimeria challenge on growth performance, apparent ileal digestibility, gastrointestinal permeability, intestinal morphology, gene expression of tight junction protein, and intestinal lesion scores in broiler chickens. There were 5 groups in this study, including a control and 4 different Eimeria treatment doses. A mixed Eimeria spp. solution with 50,000  Eimeria maxima, 50,000  Eimeria tenella, and 250,000  Eimeria acervulina per milliliter was prepared for the high-dose challenge treatment. The 2-fold serial dilution was used to make the medium-high (25,000  E. maxima; 25,000  E. tenella; 125,000  E. acervulina), the medium-low (12,500  E. maxima; 12,500  E. tenella; 62,500  E. acervulina), and the low challenge dose (6,250  E. maxima; 6,250  E. tenella; 31,250  E. acervulina). A total of three hundred sixty 13-day-old male broiler chickens were randomly allocated into 5 treatments with 6 replicated cages. Growth performance was calculated from 0 to 6 D postinfection (DPI). Intestine lesion was scored on 6 DPI. Gastrointestinal permeability was measured on 3, 5, 6, 7, and 9 DPI. The results indicated significant linear reduction in growth performance, intestinal villi height, and ileal nutrient digestibility in response to the increase of Eimeria challenge dose. Furthermore, gene expression of tight junction protein was linearly upregulated by the increasing challenge doses. Significant linear increases of gastrointestinal permeability were found on 5, 6, and 7 DPI ( P < 0.01). On 9 DPI, the gastrointestinal permeability was recovered back to normal level in the challenge groups. In conclusion, the higher Eimeria doses birds received, the more severe intestine damage was observed in several gastrointestinal health parameters. The medium-low or medium-high levels of mixed Eimeria oocysts is suggested as an optimum Eimeria-challenge dose to establish a subclinical challenge model for future studies evaluating nutritional strategies. Moreover, it is recommended to measure gastrointestinal permeability on 5 DPI with higher oocysts doses and 6 DPI when using the lower oocysts doses.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            The intestinal crypt, a prototype stem cell compartment.

            Due to its intense self-renewal kinetics and its simple repetitive architecture, the intestinal epithelium has become a prime model for studying adult stem cells in health and disease. Transgenic mouse models allow in vivo visualization and genetic lineage tracing of individual intestinal stem cells and their offspring. Fluorescently marked stem cells can be isolated for molecular analyses or can be cultured to build ever-expanding "mini-guts" in vitro. These studies are filling in the outlines of a robust homeostatic self-renewal process that defies some of the classical definitions of stem cell behavior, such as asymmetric division, quiescence, and exhaustion. Copyright © 2013 Elsevier Inc. All rights reserved.
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              Regulation of tight junction permeability by intestinal bacteria and dietary components.

              The human intestinal epithelium is formed by a single layer of epithelial cells that separates the intestinal lumen from the underlying lamina propria. The space between these cells is sealed by tight junctions (TJ), which regulate the permeability of the intestinal barrier. TJ are complex protein structures comprised of transmembrane proteins, which interact with the actin cytoskeleton via plaque proteins. Signaling pathways involved in the assembly, disassembly, and maintenance of TJ are controlled by a number of signaling molecules, such as protein kinase C, mitogen-activated protein kinases, myosin light chain kinase, and Rho GTPases. The intestinal barrier is a complex environment exposed to many dietary components and many commensal bacteria. Studies have shown that the intestinal bacteria target various intracellular pathways, change the expression and distribution of TJ proteins, and thereby regulate intestinal barrier function. The presence of some commensal and probiotic strains leads to an increase in TJ proteins at the cell boundaries and in some cases prevents or reverses the adverse effects of pathogens. Various dietary components are also known to regulate epithelial permeability by modifying expression and localization of TJ proteins.
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                Author and article information

                Contributors
                Journal
                Poult Sci
                Poult Sci
                Poultry Science
                Elsevier
                0032-5791
                1525-3171
                17 June 2020
                September 2020
                17 June 2020
                : 99
                : 9
                : 4203-4216
                Affiliations
                [1]Department of Poultry Science, University of Georgia, Athens, GA, USA
                Author notes
                [1 ]Corresponding author: wkkim@ 123456uga.edu
                Article
                S0032-5791(20)30309-6
                10.1016/j.psj.2020.04.031
                7598010
                32867964
                bd0ab7a2-37e4-4f82-8f9f-837e6a787570
                © 2020 Published by Elsevier Inc. on behalf of Poultry Science Association Inc.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 4 December 2019
                : 22 April 2020
                Categories
                Immunology, Health and Disease

                coccidiosis,eimeria,gastrointestinal permeability,fluorescein isothiocyanate-dextran,nutrient digestibility

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