Effects of dietary methionine on feed utilization, plasma amino acid profiles and gene expression in

175 Pages·2015·5.77 MB·English

Effects of dietary methionine on feed utilization,
plasma amino acid profiles and gene expression in
rainbow trout (Oncorhynchus mykiss)
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Written by Marine Rolland
Defended 26 June 2014
Effects of dietary methionine on feed utilization, plasma amino acid profiles and
gene expression in rainbow trout (Oncorhynchus mykiss)
Ph.D. Thesis by
Marine Rolland
March 2014
Technical University of Denmark
National Institute of Aquatic Resources
Section for Aquaculture
The thesis was founded by BioMar A/S
and
Danish Ministry of Higher Education and Science
Colophon
Effects of dietary methionine on feed utilization, plasma amino acid profiles and gene expression in rainbow
trout (Oncorhynchus mykiss)
By Marine Rolland
PhD thesis
Defended on the 26th of June 2014
DTU Aqua, National Institute of Aquatic Resources
Reference: Rolland, M. (2014) Effects of dietary methionine on feed utilization, plasma amino acid profiles
and gene expression in rainbow trout (Oncorhynchus mykiss). PhD thesis. DTU Aqua, National Institute of
Aquatic Resources, Technical University of Denmark, 173 p.
Cover picture: Marine Rolland
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Preface
The work presented in the thesis was undertaken during my enrolment as an industrial PhD student at
BioMar A/S, Denmark and the National Institute of Aquatic Resources, Section for Aquaculture, Technical
University of Denmark (DTU). The PhD was co-financed by BioMar A/S and granted by the Danish
Ministry of Higher Education and Science.
The thesis, in its entirety, took place over three busy years. During these years I was given the chance to
learn at my own pace thanks to the patience and support of my supervisors and colleagues. This thesis
represents a mosaic of collaborations; every person that I went to at some point surely took part in the final
piece as it stands today. I can surely not express on other’s behalf how participating in the work felt like, but
from where I have been standing, it was a great adventure. I “loveded” it.
The thesis could not have been achieved without the great help from my main supervisor at the DTU, Dr.
Peter V. Skov. Thank you for your constant and precious support; not a single day has gone by, even
Christmas, where your door was not open. I would also like to thank my second supervisor from DTU, Dr.
Anne Johanne Dalsgaard, largely involve in this project, and always providing conscientious advice. I
appreciated greatly the support from Dr. Bodil K. Larsen, who spent countless hours in the lab to teach me
the methodology and patience needed when working on the HPLC. Thanks to Ulla, Brian and Dorthe for
being so helpful during my time in the lab and fast with any assistance I required.
I would like to thank M.Sc. Jørgen Holm form BioMar A/S, who after being my supervisors for six month
during my master thesis, accepted to supervise me for another three years and gave me a chance, together
with BioMar R&D manager at the time, Ole Christensen, to be part of the BioMar R&D team. I surely learnt
a lot and beneficiated greatly from the productive and welcoming atmosphere. I would also like to thank my
colleagues from the BioMar R&D team and from the BioMar research station in Hirtshals for their support
and all the great moments we shared. Special thanks to Dr. Kim S. Ekmann for taking time to help and
advise me during the trial periods.
Heartfelt thanks to all my dear friends for never letting me down even when I became the most absent friend
in the world. A special thought to Dr. Jordan P. Feekings, a precious friend and co-worker. Many thanks to
my sisters, Eline, Lucile and Solene, for giving me strength back when they went missing. Last but not least,
thanks to Juliette for putting up with me, even during the numerous difficult moments, never failing to
support me and providing me with valuable time to reenergize.
Hirtshals, March 2014
Marine Rolland
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Table of contents
Preface ............................................................................................................................................................... 3(cid:3)
Table of contents ............................................................................................................................................... 5(cid:3)
List of papers ..................................................................................................................................................... 7(cid:3)
List of abbreviations and biochemical terms ..................................................................................................... 9(cid:3)
Dansk resumé .................................................................................................................................................. 13(cid:3)
Abstract ........................................................................................................................................................... 15(cid:3)
Objectives of the thesis .................................................................................................................................... 17(cid:3)
Introduction and background ........................................................................................................................... 19(cid:3)
1. Current practices in commercial feed formulation for salmonids ........................................................... 21(cid:3)
1.1. Formulation of aquafeeds to the optimal digestible protein and digestible energy content ............. 21(cid:3)
1.2. Dietary amino acid supply based on the “ideal protein” concept ..................................................... 22(cid:3)
2. Dietary parameters affecting amino acid availability .............................................................................. 23(cid:3)
2.1. The limitation of using digestibilities to estimate amino acid availability ....................................... 23(cid:3)
2.2. Effect of crystalline amino acid supplementation on plasma amino acid profile ............................. 24(cid:3)
3. Effect of dietary changes on amino acid role as substrates molecules and signalling factors ................. 26(cid:3)
3.1. A method to measure gene expression: real time quantitative PCR ................................................. 27(cid:3)
3.2. Amino acid regulation of gene expression through the TOR pathway ............................................. 28(cid:3)
3.3. Regulation of growth by amino acids: measurement of production variables and molecular
mechanism ............................................................................................................................................... 31(cid:3)
3.3.1. Regulation of the somatotropic axis as a major factor of growth .............................................. 32(cid:3)
3.3.2. Regulation of protein turnover by amino acids ......................................................................... 34(cid:3)
3.4. Adaptation of intermediary metabolism to dietary changes: regulation of gene expression by amino
acid and measurement of metabolic indicators ........................................................................................ 37(cid:3)
3.4.1. Dietary factors affecting the utilization of amino acids for energy production ......................... 39(cid:3)
3.4.2. Regulation of de novo synthesis of glucose by amino acids ...................................................... 40(cid:3)
3.4.3 Regulation of lipid metabolism .................................................................................................. 43(cid:3)
Main results and discussion ............................................................................................................................. 47(cid:3)
Conclusion and future perspectives ................................................................................................................. 49(cid:3)
References ....................................................................................................................................................... 51(cid:3)
Paper I .............................................................................................................................................................. 61(cid:3)
Paper II ............................................................................................................................................................ 87(cid:3)
Paper III ......................................................................................................................................................... 121(cid:3)
Paper IV ......................................................................................................................................................... 149
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List of papers
Paper I: Rolland M, Larsen BK, Dalsgaard J, Holm J, Skov PV (2014) Effect of plant proteins and
crystalline amino acid supplementation on postprandial plasma amino acid profiles and
metabolic response in rainbow trout (Oncorhynchus mykiss). Aquaculture International
Paper II: Rolland M, Feekings JP, Dalsgaard J, Holm J, Skov PV (2014) Modelling the effects of dietary
methionine level and form on postprandial plasma amino acid profiles in rainbow trout
(Oncorhynchus mykiss). Submitted, Aquaculture Nutrition
Paper III: Rolland M, Dalsgaard J, Holm J, Gómez-Requeni P, Skov PV (2014) Dietary methionine level
affects growth performance and hepatic gene expression of GH-IGF system and protein
turnover regulators in rainbow trout (Oncorhynchus mykiss) fed plant protein-based diets.
Comparative Biochemistry and Physiology, Part B
Paper IV: Rolland M, Larsen BK, Dalsgaard J, Holm J, Gómez-Requeni P, Skov PV (2014) Dietary
methionine level affect plasma methionine profile, ammonia excretion and the expression of
genes related to the hepatic intermediary metabolism in rainbow trout (Oncorhynchus mykiss)
fed plant based diets. Manuscript

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List of abbreviations and biochemical terms
Acetyl-CoA: Acetyl coenzyme A, key intermediate of metabolic pathways
Akt: Tyrosine protein kinase is involved in the transduction of insulin or IGF signal
ALT1: Alanine transferase, a transaminase which catalyses the transfer of an amino group from
alanine to (cid:302)-ketoglutarate, resulting in the synthesis of pyruvate
ANF: Antinutritional factor
ATP: Adenosine triphosphate carries energy at a cellular level from a catabolic to anabolic
reactions
BCAAs: Branched-chain amino acid, isoleucine, leucine and valine
CAA: Crystalline amino acid
Capn: Calpain, proteolytic enzyme of the calpain cascade, existing in two genes 1 and 2 (Capn 1
and Capn 2, respectively)
CAST: Calpastatin, inhibitor of the calpain protease, existing in long and short sequences (CAST-L
and CAST-S, respectively)
cDNA: complementary Deoxyribonucleic acid, produce of the reverse transcription step
CPT1a: Carnitine palmitoyltransferase 1 isoforms a, enzyme of the (cid:533)-oxidation
CT: Cycle threshold, number of cycle necessary to obtain a threshold level of fluorescence in the
RT-PCR analysis
DE: Digestible energy
D-L isomers: Dextrorotary and levorotary are the chiral isomers resulting from the presence of an
asymmetric carbon in molecule
DM: Dry matter
DNA: Deoxyribonucleic acid
DNase: Deoxyribonuclease, enzyme responsible for DNA degradation
DP: Digestible protein
E: Efficiency of a RT-PCR, E= 10-1/slope
EAA: Essential amino acid
EER: Energy efficiency ratio, unit of bodyweight gain per unit of dietary energy intake
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