TrimSize 170mm x 244mm Merrifield ffirs.tex V3 - 07/23/2014 10:35 A.M Page ii TrimSize 170mm x 244mm Aquaculture Nutrition Merrifield ffirs.tex V3 - 07/23/2014 10:35 A.M Page i TrimSize 170mm x 244mm Merrifield ffirs.tex V3 - 07/23/2014 10:35 A.M Page ii TrimSize 170mm x 244mm Merrifield ffirs.tex V3 - 07/23/2014 Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics Edited by Daniel Merrifield School of Biological Sciences, Plymouth University, UK Einar Ringø Norwegian College of Fishery Science, UiT The Arctic University of Norway 10:35 A.M Page iii TrimSize 170mm x 244mm Merrifield ffirs.tex V3 - 07/23/2014 10:35 A.M Page iv This edition first published 2014 © 2014 by John Wiley & Sons, Ltd Registered office: John Wiley & Sons, Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK Editorial offices: 9600 Garsington Road, Oxford, OX4 2DQ, UK The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK 111 River Street, Hoboken, NJ 07030-5774, USA For details of our global editorial offices, for customer services and for information about how to apply for permission to reuse the copyright material in this book please see our website at www.wiley.com/wiley-blackwell The right of the author to be identified as the author of this work has been asserted in accordance with the UK Copyright, Designs and Patents Act 1988 All rights reserved No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by the UK Copyright, Designs and Patents Act 1988, without the prior permission of the publisher Designations used by companies to distinguish their products are often claimed as trademarks All brand names and product names used in this book are trade names, service marks, trademarks or registered trademarks of their respective owners The publisher is not associated with any product or vendor mentioned in this book Limit of Liability/Disclaimer of Warranty: While the publisher and author(s) have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose It is sold on the understanding that the publisher is not engaged in rendering professional services and neither the publisher nor the author shall be liable for damages arising herefrom If professional advice or other expert assistance is required, the services of a competent professional should be sought Library of Congress Cataloging-in-Publication Data Aquaculture nutrition : gut health, probiotics, and prebiotics / edited by Daniel Merrifield and Einar Ringo pages cm Includes bibliographical references and index ISBN 978-0-470-67271-6 (cloth) Fishes – Digestive organs Fishes – Health Fishes – Nutrition Marine animals – Digestive organs Marine animals – Health Marine animals – Nutrition Aquaculture I Merrifield, Daniel, 1983- II Ring?, Einar, 1950QL639.1.A685 2014 571.1′ – dc23 2014015269 A catalogue record for this book is available from the British Library Wiley also publishes its books in a variety of electronic formats Some content that appears in print may not be available in electronic books Cover image: Photos by Daniel Merrifield Set in 10/12pt Times by Laserwords Private Limited, Chennai, India 2014 TrimSize 170mm x 244mm Merrifield ftoc.tex V2 - 07/23/2014 10:36 A.M Page v Contents List of Contributors Preface The Gastrointestinal Tract of Fish Arun Kumar Ray and Einar Ringø 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 Introduction Anatomy of GI tract Stomach and intestinal bulb Pyloric caeca Intestine Endogenous inputs of digestive secreta Luminal pH Passage rate and residence time Acknowledgements References Immune Defences of Teleost Fish Andrew Foey and Simona Picchietti 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 Introduction Innate immunity Antigen-specific adaptive immunity Cytokines drive immune responsiveness Immune tissues Mucosal immunity Common pathogens infecting teleosts: what immune responses are required? Future considerations Conclusion References Gastrointestinal Pathogenesis in Aquatic Animals Jarl Bøgwald and Roy Ambli Dalmo 3.1 3.2 3.3 Introduction Vibrio spp Aeromonas spp xi xv 1 10 10 10 10 14 14 15 18 22 23 32 36 39 40 40 53 53 54 61 TrimSize 170mm x 244mm vi Merrifield ftoc.tex V2 - 07/23/2014 10:36 A.M Page vi Contents 3.4 3.5 3.6 3.7 3.8 3.9 3.10 3.11 3.12 Yersinia ruckeri Edwardsiella spp Piscirickettsia salmonis Pseudomonas anguilliseptica Photobacterium damsela subsp Piscicida (Pasteurella Piscicida) Streptococcosis ‘Candidatus arthromitus’ Mycobacterium spp Conclusion References The Gut Microbiota of Fish Jaime Romero, Einar Ringø and Daniel L Merrifield 4.1 4.2 4.3 4.4 4.5 Introduction The importance of the microbiota Composition of the microbiota in early life stages Factors that influence microbiota composition Conclusion References Methodological Approaches Used to Assess Fish Gastrointestinal Communities Zhigang Zhou, Bin Yao, Jaime Romero, Paul Waines, Einar Ringø, Matthew Emery, Mark R Liles and Daniel L Merrifield 5.1 5.2 5.3 5.4 5.5 5.6 5.7 Culture-dependent approaches Molecular techniques Fluorescence based methods Electron microscopy Microbial activity and functionality Summary Acknowledgements References Indigenous Lactic Acid Bacteria in Fish and Crustaceans Daniel L Merrifield, José Luis Balcázar, Carly Daniels, Zhigang Zhou, Oliana Carnevali, Yun-Zhang Sun, Seyed Hossein Hoseinifar and Einar Ringø 6.1 6.2 6.3 6.4 6.5 6.6 6.7 6.8 Introduction Lactic acid bacteria Salmonidae Gadidae Clupeidae Anarhichadidae Acipenseridae Percidae and sciaenidae 63 63 64 65 65 66 66 66 68 68 75 75 84 86 88 93 94 101 102 106 115 115 117 120 120 120 128 129 130 130 141 143 143 143 144 TrimSize 170mm x 244mm Merrifield ftoc.tex V2 - 07/23/2014 Contents 6.9 6.10 6.11 6.12 6.13 6.14 6.15 6.16 6.17 6.18 6.19 6.20 6.21 Moronidae Sparidae Pleuronectiformes Cyprinidae Channidae Siluriformes Cichlidae Serranidae Rachycentridae Mugilidae Coastal Fish Shellfish Summary References Probiotics and Prebiotics: Concepts, Definitions and History Hélène L Lauzon, Arkadios Dimitroglou, Daniel L Merrifield, Einar Ringø and Simon J Davies 7.1 7.2 7.3 7.4 7.5 10:36 A.M Page vii vii 145 145 146 146 149 150 150 151 151 152 152 153 156 156 169 Introduction The probiotic concept and history The prebiotic concept and definition Synbiotics Summary References 169 170 174 180 180 180 Probiotic Modulation of the Gut Microbiota of Fish Daniel L Merrifield and Oliana Carnevali 185 8.1 8.2 8.3 8.4 8.5 8.6 8.7 Introduction Bacillus spp Lactic acid bacteria (LAB) Other probionts Probiotic colonization? Conclusion and future perspectives Acknowledgements References Probiotic Applications in Cold Water Fish Species Hélène L Lauzon, Tania Pérez-Sánchez, Daniel L Merrifield, Einar Ringø and José Luis Balcázar 9.1 9.2 9.3 9.4 9.5 Introduction Salmonidae Gadidae Pleuronectiformes Percidae 185 187 192 206 210 213 214 214 223 223 225 234 240 245 TrimSize 170mm x 244mm viii Merrifield ftoc.tex V2 - 07/23/2014 10:36 A.M Page viii Contents 9.6 Conclusion References 10 Probiotic Applications in Temperate and Warm Water Fish Species Oliana Carnevali, Yun-Zhang Sun, Daniel L Merrifield, Zhigang Zhou and Simona Picchietti 10.1 10.2 10.3 10.4 10.5 10.6 10.7 10.8 10.9 10.10 Introduction European sea bass (Dicentrarchus labrax L.) Gilthead sea bream (Sparus aurata L.) Probiotic applications in sole spp Groupers Tilapia Carps Zebrafish (danio rerio) Catfishes General conclusions References 11 Probiotic Applications in Crustaceans Mathieu Castex, Carly Daniels and Liet Chim 11.1 11.2 11.3 11.4 11.5 Introduction Main microorganisms evaluated and used as probiotics in crustacean aquaculture Probiotic modes of action Related benefits in crustacean aquaculture Conclusion References 12 Can Probiotics Affect Reproductive Processes of Aquatic Animals? Giorgia Gioacchini, Elisabetta Giorgini, Lisa Vaccari and Oliana Carnevali 12.1 12.2 12.3 12.4 12.5 12.6 12.7 Introduction The fish reproductive system Broodstock reproductive dysfunctions Reproduction and metabolism The effects of probiotic applications on fish reproduction Concluding remarks Acknowledgements References 13 Issues with Industrial Probiotic Scale-up Mathieu Castex, Henri Durand and Bernadette Okeke 13.1 13.2 13.3 Introduction Scaling-up guidelines Mode of administration 245 246 253 253 255 258 262 266 269 272 275 277 279 279 290 290 293 300 308 318 319 328 328 329 331 332 333 341 341 341 347 347 349 354 TrimSize 170mm x 244mm A B Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page C E E Lp E Lp Bb D E E Plate Histological characterization of intestinal mucosal immune cells Immune cells of the teleost intestinal mucosa are less well organized and more diffusely arranged than in mammals This figure shows the distribution of some of these immune cells in the mucosa and the lamina propria of bony fish in a series of five images (A/B) Predominance of CD8α-positive cells in both the epithelium and the lamina propria of the European sea bass intestine (CD8α RNA in situ hybridization) Scale bar = 50 μm The CD8α+ cells aggregated in the lamina propria are shown at higher resolution in the insert (B), where the scale bar = 10 μm (C) May-Grümwald-Giemsa staining of the gilthead sea bream (Sparus aurata) intestinal mucosa, showing lymphoid cells (arrows) and acidophilic granulocytes (arrowheads) Scale bar = μm (D) Semi-thin section of the gilthead sea bream intestinal mucosa housing numerous basolateral lymphocytes (arrows) and an intraepithelial macrophage (star) Scale bar = μm (E) Transmission electron microscopy of the gilthead sea bream intestinal mucosa showing a goblet cell (star) amongst enterocytes and basolateral lymphocytes (arrows) Scale bar = μm Key: Ep, epithelium; Lp, lamina propria; Bb, brush border For further information see text near Figure 2.1 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page Mucosal folds Blood capillaries IMMUNE FATE?: CD4+ Th1 CD4+ Treg Tolerance! CD4+ Th2 CD4+ Th17 CMI: Humoral: CMI: Intracellular pathogens Extracellular pathogens fungal protection Key: Epithelial cell M-like cell Goblet cell Mucus CD4+ Theg CD8+ Tc / IEL CD4+ Thp B cell slg DC CD4+ Th2 Macrophage CD4+ Th17 + CD4 Th1 Commensal Pathogen Defensin Plate Mucosal immune system of the gut This figure shows the current understanding that exists for the gut mucosal immune system of the teleost fish This tissue exists in folds, but cannot be described as villi due to the lack of lacteal/lymphatics The immune defences exist at many levels of the layers of the mucosal tissue, namely commensal organisms, mucus, epithelial cell layer and sub-epithelial mucosa Unlike the mammalian GALT, teleosts not exhibit Payer’s patches or lymphoid follicles Instead, their lymphoid tissue is rather diffuse but contains all the immune cells which give the teleost GALT the capability to either tolerize immune mechanisms (Treg ) or initiate immune mechanisms directed at the clearance of intracellular-resident (CD4+ Th1 , CD4+ Th17 and CD8+ Tc ) and extracellular-resident pathogens (CD4+ Th2 and B cells) One area of controversy in this diagram is the existence of dendritic cells (DCs); it is not understood whether DCs, specialized macrophages or specialized M-cell-like epithelial cells capable of antigen transcytosis act as APCs to contextualize MHC-restricted T-cell-driven adaptive immune responses Broad-spectrum pathogen defence is associated with barrier function (commensal organisms, mucus secreted by goblet cells and trapping of sIg-bound pathogens, intact tight junctions and epithelial cell secretion of anti-microbial products such as defensins) and innate immune responses (macrophage and granulocyte activity mediated by cytokines and anti-microbial compounds) Through the induction of expression of teleost cytokine homologues, immune responses can be directed towards the desired effector response to pathogens and non-pathogens Such teleost cytokines include TGFβ and IL-10 (Treg ), IL-12 (Th1 ), IL-4 (Th2 ) and IL-23 (Th17 ), thus directing mechanisms of mucosal tolerance, cell-mediated immunity, humoral immunity and anti-fungal responses, respectively For further information see text near Figure 2.2 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page As Lp E L Plate Immunohistochemical staining, using rabbit anti-A salmonicida LPS antiserum diluted 1:1000, biotinylated goat anti-rabbit Ig and streptavidin biotinylated alkaline phosphatase, and Fast red/Naphthol AS-Mx as the substrate, in analysis of the spotted wolffish intestine from fish infected with waterborne atypical A salmonicida Large numbers of bacteria (As) were observed in the lamina propria (Lp) Sections obtained from non-infected controls were negative for bacteria Key: E, epithelium; L, lumen Scale bar = 10 μm (Source: Dag Vollstad.) For further information see text near Figure 3.5 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page Lm Cm Mu G L 50 μm Plate Light microscopy (H& E staining) transverse section of the intestine of zebrafish with clinical signs of chronic mycobacteriosis A lesion (granuloma, G) in the submucosa is indicated by the formation of fibrous connective tissue around a small (∼50 μm) focus of acellular material Key: arrow, epithelium; Cm, circular muscle; L, lumen; Lm, longitudinal muscle; Mu, muscularis Scale bar = 50 μm (Source: Kuehlwein et al., unpublished.) For further information see text near Figure 3.6 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page Phylum reported in salmonids Deinococcus-Thermus Tenericutes Fusobacteria Bacteroidetes Actinobacteria Firmicutes Proteobacteria Number of reports 10 12 Plate Bacterial phyla observed in the gut microbiota of salmonids (Source: Nayak 2010 Reproduced with permission of John Wiley & Sons.) For further information see text near Figure 4.2 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page A B Plate 10 Epifluorescence micrographs of the indigenous gut microbiota of fish (A) In situ hybridization of rainbow trout gut microbiota with green fluorescein labelled probe EUB338-mix (detecting all bacteria) (Waines and Merrifield unpublished data) Despite the clearly identifiable cocci (arrows) and rod-shaped bacteria (arrowheads) present within the sample, reliable enumeration is difficult due to the dense nature of the gut contents which leads to poor differentiation between individual cells in some cases In addition, non-specific hybridization and fluorescence of background material (digesta and/or mucus) is present (five-pointed stars) (B) DAPI staining (blue) and in situ hybridization of common carp gut microbiota with green fluorescein (FITC) labelled probe VA (detecting Aeromonas and Vibrio spp.) (Source: Mark Rawling.) For further information see text near Figure 5.1 TrimSize 170mm x 244mm A Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page B E Lp C D Gb Ga Plate 11 (A) Immunohistochemistry of Ig+ intestinal lymphocytes in 99 dph sea bream fry The polyclonal antiserum ORa immunostained Ig+ cells seed both the mucosal epithelium and the lamina propria (probiotic group A) (B) May-Grümwald-Giemsa staining of posterior intestine in 99 dph sea bream fry showing lymphoid cells (arrows) and acidophilic granulocytes (arrowheads) in the mucosal epithelium (E) and lamina propria (Lp) (probiotic group A) (C) Electron microscopy of sea bream granulocytes housed in the intestinal mucosa This subpopulation (Ga) is characterized by a rounded shape and cytoplasmic granules The granules have electron dense paracrystalline cores (D) Electron microscopy of intestinal sea bream granulocytes housed in the lamina propria (Gb) Gb granulocytes have an elongated shape, a distinct granule type and autolysosomes (arrows) Scale bars: A = 20 μm; B = μm; C = 500 nm; D = μm (Source: Simona Picchietti.) For further information see text near Figure 10.1 TrimSize 170mm x 244mm Merrifield bplate.tex EXTERNAL STIMULI (temperature, photoperiod, etc.) INTERNAL STIMULI (metabolic state, etc.) +/‒ +/‒ Brain + Pituitary 10:26 A.M Page 10 GnRH GtH Egg FSH LH + Ovary V3 - 07/23/2014 Sex Steroids Plate 12 Diagrammatic presentation of the actions of the HPG axis in the control of reproductive function The hypothalamus, integrating internal and external stimuli, releases the gonadotrophin releasing hormone (GnRH) Under the influence of the GnRH, the pituitary secretes follicle stimulating hormone (FSH) and luteinizing hormone (LH) which act upon the ovary, regulating oogenesis by activating steroidogenesis For further information see text near Figure 12.1 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 11 BRAIN MATURATION PITUITARY GROWTH LH FSH ESTRADIOL MIH MPF LIVER GVBD VITELLOGENIN THECA CELLS GRANULOSA CELLS Plate 13 Control of follicle growth and maturation in teleosts The action of follicle stimulating hormone (FSH) and luteinizing hormone (LH) is depicted in teleost oocytes during growth and maturation phases A teleost follicle contains an oocyte, one layer of granulosa cells and one layer of theca cells The basement membrane separating the granulosa and thecal cells and the zona radiata are omitted in the Plate Under the influence of FSH, granulosa cells aromatize testosterone to estradiol, which initiates vitellogenin production in the liver, leading to follicle growth Under the influence of LH, granulosa cells produce MIH, leading to the oocyte maturation (Source: The model is based primarily on the work of Clelland and Peng 2009.) For further information see text near Figure 12.2 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 12 A PV V 100nm B PV V 100nm Plate 14 Representative micrographs of ovaries from (A) zebrafish females fed on an Lb rhamnosus supplemented diet and (B) control fed females, revealing an increase in vitellogenic follicles in ovary sections from fish treated with the probiotic PV = pre-vitellogenic follicles; V = vitellogenic follicles Scale bars = 100 μm (Source: Gioacchini et al 2011.) For further information see text near Figure 12.3 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 13 A Correlation 0.800 B 0.753 0.709 0.662 0.619 0.572 C 0.526 0.481 0.436 0.391 0.345 Plate 15 Compare (A) correlation map in the range 1800–1480 cm−1 on an ovary section from zebrafish with (B) VTG and (C) LV representative spectra The correlation maps obtained suggest that vitellogenin is present mainly in the outer zone of IV class oocytes, with lipovitellin homogeneously distributed in the inner (Source: Carnevali et al 2009.) For further information see text near Figure 12.5 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 14 A 10 μm Absorbance 4000 D Wavenumber/cm−1 800 4000 Absorbance C Absorbance B Wavenumber/cm−1 800 4000 Wavenumber/cm−1 Plate 16 Compare (A) correlation map in the range 1800–1480 cm−1 on an ovary section from zebrafish with representative spectra from (B) class I-II, (C) class III and (D) class IV oocytes (Source: Carnevali et al 2009.) For further information see text near Figure 12.6 800 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 15 QUALITY CONTROL QUALITY CONTROL Centrifugation −80 °C Strains library Amplification QUALITY CONTROL Fermentation H2O H2O Packaging Blending Grinding Freeze drying Plate 17 Schematic representation of a probiotic bacteria production process For further information see text near Figure 13.1 TrimSize 170mm x 244mm Merrifield bplate.tex V3 - 07/23/2014 10:26 A.M Page 16 A B Plate 18 (A) Intestine of turbot larva fed with rotifers with bioencapsulated Phaeobacter 27-4 Counterstaining with haematoxyline (blue) and immunohistochemistry staining with primary antiserum against Phaeobacter 27-4 (red) The arrows point to partially digested rotifers with positive immunostaining, enlarged in (B) (Source: Planas et al 2006 Reproduced with permission of Elsevier.) For further information see text near Figure 16.1 WILEY END USER LICENSE AGREEMENT Go to www.wiley.com/go/eula to access Wiley’s ebook EULA ... Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics and other organ systems to adapt to changes in the contents of the GI tract (amounts and types of nutrients, pH, ionic composition etc.) and. .. Cataloging-in-Publication Data Aquaculture nutrition : gut health, probiotics, and prebiotics / edited by Daniel Merrifield and Einar Ringo pages cm Includes bibliographical references and index ISBN 978-0-470-67271-6... De Silva and Anderson 1995) A stomach is defined as a portion TrimSize 170mm x 244mm Merrifield c01.tex V3 - 07/23/2014 10:30 A.M Page Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics