CHAPTER 5 Exposure of gnotobiotic Artemia franciscana larvae to abiotic stress promotes heat ...

190 Pages·2008·3.06 MB·English

Promoters: Prof. dr. ir. Peter Bossier
Department of Animal Production, Faculty of Bioscience Engineering, Ghent
University.
([email protected])

Prof. dr. Patrick Sorgeloos
Department of Animal Production, Faculty of Bioscience Engineering, Ghent
University.
([email protected])

Dean: Prof. dr. ir. Herman Van Langenhove

Rector: Prof. dr. Paul Van Cauwenberge

Members of the Examination and Reading Committee

Prof. dr. ir. Nobert De Kimpe
(Chairman)
Department of Organic Chemistry, Faculty of Bioscience Engineering, Ghent University.
([email protected])

Prof. dr. Els J. M. Van Damme
(Secretary)
Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent
University.
([email protected])

Prof. dr. Thomas H. MacRae
Department of Biology, Dalhousie University, Canada.
([email protected])

Prof. dr. James S. Clegg
Bodega Marine Laboratory,University of California Davis, USA.
([email protected])

Prof. dr. Eric Cox
Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine,
Ghent University.
([email protected])

Prof. dr. Frank Pasmans
Department of Pathology, Bacteriology and Poultry Disease, Faculty of Veterinary Medicine,
Ghent University.
([email protected])
Yeong Yik Sung

Protection of Artemia from vibriosis by heat shock and
heat shock proteins

Thesis submitted in fulfillment of the requirements for the degree of Doctor (PhD) in Applied
Biological Sciences
Dutch translation of the title: Bescherming van Artemia tegen vibriose door
warmtebehandeling en hitteschokproteїnen

Cover page: Artemia nauplii (figure adapted from www.osc.mun.ca/ardf/artemia.html)

To cite this work:

Yeong, Y. S. (2008) Protection of Artemia from vibriosis by heat shock and heat shock
proteins. PhD thesis, Ghent University, Belgium.

The author and the promoters give the authorisation to consult and to copy parts of this work
for personal use only. Every other use is subject to the copyright laws. Permission to
reproduce any material contained in this work should be obtained from the author.

ISBN number: 978-90-5989-224-8

This work was funded by the University Malaysia Terengganu, UMT (formerly known as
University College of Science and Technology Malaysia, KUSTEM) through a doctoral grant
to YYS and the Belgian Foundation for Scientific Research (FWO) for research funding.
Table of contents

List of Abbreviations and Units ii

List of Tables v

List of Figures vii

Chapter 1 Introduction 1

Chapter 2 Literature overview 13

Chapter 3 Heat shock proteins in aquatic organisms: Induction, stress
tolerance and application 21

Chapter 4 Non-lethal heat shock protects Artemia franciscana against
pathogenic Vibrios 45

Chapter 5 Exposure of gnotobiotic Artemia franciscana larvae to abiotic
stress promotes heat shock protein 70 synthesis and
enhances resistance to pathogenic Vibrio campbellii 61

Chapter 6 DnaK, the 70 kDa bacterial heat shock protein, protects
Artemia larvae from Vibrio infection 77

Chapter 7 Ingestion of DnaK producing bacteria attenuates Vibrio
infection of Artemia franciscana larvae 95

Chapter 8 Discussion and conclusions 109

References 115

Summary / Samenvatting 141

Curriculum Vitae 147

Acknowledgements 151

i
List of abbreviations and units

°C Degree Celsius
% Percentage
β-glucans Beta-glucans
µE/m2s Micro Einstein per second and square meter
µg Microgram
µm Micrometer
µl Microlitre
λ Wavelength
g Relative centrifugal force or G force
± Approximately
/ Per
AMPs Antimicrobial peptides
ANOVA Analysis of variance
CCAP Culture Collection of Algae and Protozoa Department, Scotland
cDNA Complementary deoxyribonucleic acid
cfu Colony forming unit
CHO Chinese hamster ovary
cm Centimeter
CTR Control treatment
d day
Da Dalton
DAB Diaminobenzidinetetrahydrochloride dihydrate
DNA Deoxyribonucleic acid
ELISA Enzyme-linked ImmunoSorbent Assay
exp Exponential growth phase
FAO Food and Agricultural Organization of the United Nations
FASW Filtered and autoclaved seawater
FT Falcon tube
g Gram
GART Gnotobiotic Artemia test system
GenBank Genetic sequence database of the National Institute of Health, USA
ii
GH Growth hormone
GR glucocorticoid receptor
g/l Gram/liter
h Hour
HS heat shock
Hsc Heat shock cognate
HSF Heat shock factor
Hsps Heat shock proteins
IGF Insulin-like growth factor
IL1 Interleukin 1
IL Individual length
ISSCAAP International standard statistical classification for aquatic animals and
plants

kDa kilo Dalton
kg Kilogram
kW Kilowatt
l litre
LMG Laboratory of Microbiology of the Ghent University (Culture Collection)
LPS Lipopolysaccharides
LRRs Leucin-rich repeats
LVS… Strain … isolated by Laurent Verschuere
M Molar
MA Marine agar 2216
mg Milligram
MHC Major histocompatibility
min Minute
ml Milliliter
mm Millimeter
mM Milimolar
mnn… Mutants with deleted genes involved in the extension of
mRNA Messenger ribonucleic acid
MTT -3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide
n Number of replicates
ng Nanogram
iii
NLHS Non-lethal heat shock
NSS Nine salt solution
nm Nanometer
NO Nitric oxide
OD Optical density
oPRL ovine prolactin
p protein
p Statistical p-value obtained
PCR Polymerase chain reaction
pH Measure of the acidity of a solution
ppt Part per thousand
proPO Prophenoloxidase
PUFA Polyunsaturated fatty acids
PVDF Polyvinylidene difluoride
R2 Coefficient of determination
r Coefficient of correlation
RNA Ribonucleic acid
rpm Rotations per minute
RNA Ribonucleic acid
s Second
SDS-PAGE Sodium dodecyl sulfate polyacrylamide gel electrophoresis
sHsps Small heat shock proteins
TBT Tributyltin
TLR Toll like receptor
TNF Tumor necrosis factor
US$ United states currency (Dollar)
USA United States of America
v/v Volume per volume
VC Vibrio campbellii
VP Vibrio proteolyticus
w/v Weight per volume
WT Wild type

iv
List of Tables

Chapter 1 - Introduction Page

Table 1.1. Top ten ISSCAAP species groups for aquaculture production
of aquatic animals (excluding aquatic plants) for 2004 2

Table 1.2. Common disease associated with fish and shrimp culture 6

Table 1.3. Mortalities caused by luminescent vibrios in different shrimp
species 7

Chapter 2 – Literature overview

Table 2.1. Major Hsp families 17

Chapter 3 - Heat shock proteins in aquatic organisms: Induction, stress
tolerance and application

Table 3.1. Factors that influence Hsp production in aquatic organisms 30

Chapter 4 - Non-lethal heat shock protects Artemia franciscana against
pathogenic Vibrios

Table 4.1. Average survival (%) of Artemia nauplii after 24 h challenge
test using 107 cells/ml of V. campbellii in relation to
different heat shock (HS) temperature for 30 min exposure
with subsequent 6 h and 24 h recovery periods 53

Table 4.2. Average survival (%) of Artemia nauplii after 24 h challenge
test using 107 cells/ml of V. campbellii in function of
different recovery periods with non-lethal heat shock
temperature of 37°C for 30 min 54

Table 4.3. Average survival (%) of Artemia nauplii after 24 h challenge
test using 107 cells/ml of V. campbellii in function of different
heat shock duration with non-lethal heat shock temperature
of 37°C and 6 h recovery period 54

Chapter 5 - Exposure of gnotobiotic Artemia franciscana larvae to abiotic
stress promotes heat shock protein 70 synthesis and enhances
resistance to pathogenic Vibrio campbellii

No table

v
Chapter 6 - DnaK, the 70 kDa bacterial heat shock protein, protects Artemia
larvae from Vibrio infection

Table 6.1. Transformed bacterial strains and plasmids 81

Table 6.2. Nutritional value of bacterial strains 85

Table 6.3. Nutritional value of bacterial strains YS1 and YS2 89

Chapter 7 - Ingestion of DnaK producing bacteria attenuates Vibrio
infection of Artemia franciscana larvae

Table 7.1. Bacterial strains 98

Table 7.2. Feeding with heat stressed bacteria increases Artemia
resistance to V. campbellii 102

Table 7.3. Nutritional value of bacteria 105

vi