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The Research Center for
Animal Science (RCAS)
Hiroshima University’s Promising Research
Initiatives
(recognized in 2014)
Education & Research for advanced technological developments
contributing to safe and high quality animal productions
Presented by Dr Yukinori Yoshimura
（Graduate School of Biosphere Science）
Where is Hiroshima University?
Hiroshima is best known as the first city in history to be
destroyed by a nuclear weapon on August 6, 1945, near
the end of World War II.
Peace Memorial Park is a famous place to promise and
learn the peace of the world.
However, Hiroshima is now one of the biggest city in Japan.
The population is more than 1 million. MAZDA that is
automotive manufacture is based on Hiroshima.
Hiroshima University
has two campuses.
Faculties: Hiroshima University
Graduate Schools (12)
Faculties (11)
Faculty of Integrated Arts and Sciences
Faculty of Letters
Faculty of Economics
Faculty of Law
Faculty of Education
Faculty of Science
Faculty of Engineering
Faculty of Medicine
Faculty of Dentistry
Faculty of Pharmaceutical Sciences
Faculty of Applied Biological Science
Students
Undergraduate:11,040
Master course:2,510
PhD course:1,750
Others:500
Foreign students:760 from 57 countries
Graduate Schl of Integrated Arts and Sci
Graduate Schl of Letters
Graduate Schl of Education
Graduate Schl of Social Sciences
Graduate Schl of Science
Graduate Schl of Engineering
Graduate Schl of Biomedical Sciences
Graduate Schl of Health Sciences
Graduate School of Biosphere
Science
Graduate Schl for International
Development and Cooperation
Graduate Schl of Advanced Sciences of
Matter
Staff
Academic : 1,830
Non-academic: 1.440
Departments
DEPARTMENT OF
BIORESOURCE SCIENCE
Animal Science, Aquatic Biology,
Food and Resource Economics,
Field Science
DEPARTMENT OF
BIOFUNCTIONAL SCIENCE AND
TECHNOLOGY
Molecular and Applied Biosciences,
Food Science and Biofunctions
Int’l summer school
AIMS program
DEPARTMENT OF
ENVIRONMENTAL DYNAMICS
AND MANAGEMENT
Modeling, Management and
Assessment of Environmental
Dynamics
Int’l symposium
Short visit Vet Tech students
DEPARTMENT OF BIORESOURCE SCIENCE
Animal Science (Academic staff: 11)
Animal breeding, Reproduction, Nutrition and Feed, Management and
Behavior, Physiology and Histology
Aquatic Biology (Academic staff: 14)
Aquatic Biochemistry, Ecology, Pathology, Neurobiology,
Biology of Aquatic Resources, Aquaculture
Food and Resource Economics (Academic staff: 4)
Food Production, Management, Agricultural Marketing
Field Science (Academic staff: 4)
Animal Farm, Aquatic field studies
gene
Animal Science Division
Graduate School of Biosphere Science
Hiroshima University
The goal of this division is to discover novel functions in animals and applying
such discoveries for the technological advancement of domestic animal
production. The research will contribute to sustainable and efficient production
of safe food for humans derived from animal products. Current research fields
are: breeding and genetics based on modern genomic analysis, reproductive
biology and biotechnology for enhanced reproductive performance, applied
nutritional physiology and feed evaluation for improved nutritional management,
ecology and neurophysiology for better animal management and viability, and
fundamental histology and molecular physiology to enhance veterinary
production and health..
Animal Breeding and Genetics
Animal Reproduction
Animal Nutrition and Feeding
Animal Behavior and Physiology
Animal Histophysiology
Animal Farm
Goal of RCAS
Contribution to development of technologies for
safe and high quality animal products
• Progression for animal nutrition, breeding and
reproduction, and health
• Contribution for food safety and security
• Innovation for new agricultural science by crossdisciplinary research collaboration
nutrition
metabolism
health
productivity
breeding
reproduction
well-being
Members of The Research Center for Animal Science
Members
Leader
Yukinori Yoshimura, PhD
Institutes
Grad Schl Biosphere Sci, HU
Majors
Immunodefense in poultry reproductive and digestive
system
Manager
Masayuki Shimada, PhD
Grad Schl Biosphere Sci, HU
Reproductive Molecular Endocrinology
Reproductive technology
Manager
Toshihisa Sugino, PhD
Grad Schl Biosphere Sci, HU
Ruminant metabolism and endocrinology
Masaoki Tsudzuki, PhD
Grad Schl Biosphere Sci, HU
Genome of Japanese native chickens
Masahide Nishibori, PhD
Grad Schl Biosphere Sci, HU
Genome and Polymorphism of Animals and Poultry
Teruo Maeda, PhD
Grad Schl Biosphere Sci, HU
Manipulation of germ cells
Yusuke Sotomaru, PhD
Nat Sci Center Basic Res Dev, HU
Animal developmental engineering
Ikuo Miura, PhD
Grad. Schl Sci, HU
Sex determination and differentiation
Taketo Obitsu, PhD
Grad Schl Biosphere Sci, HU
Ruminant nutrition and feed evaluation
Takashi Bungo, PhD
Grad Schl Biosphere Sci, HU
Animal Behavior and Physiology
Shin-ichi Kawakami, PhD
Grad Schl Biosphere Sci, HU
Animal management and neurology
Naoki Isobe, PhD
Grad Schl Biosphere Sci, HU
Ruminant mastitis and mammary gland immune
system
Yuzo Kurokawa, PhD
Grad Schl Biosphere Sci, HU
Dairy milk production and grassland utilization
Kensuke Kawamura, PhD
IDEC, HU
Grazing management and remote control system
Soichi Tanabe, PhD
Grad Schl Biosphere Sci, HU
Gastro-protection by lactobacilli and bifidobacteria
Noriyuki Yanaka, PhD
Grad Schl Biosphere Sci, HU
Nutritional factors and obesity
Shiro Kushibiki, PhD
Natl Agr Food Res Center
Animal nutrition and metabolism
Tetsuji Okazaki, PhD
Oita Pref Animal Res Center
Porcine reproductive technology
JoAnne S. Richards, PhD
Baylor College of Medicine, USA
Animal reproduction
Masahito Oba, PhD
University of Alberta, Canada
Animal nutrition
8
Research activities in RCAS
Project 1. Hostdefense and genetic resources research
(Example) Creation of livestock disease-resistance
Natl. and prefectural
institutes, Japan
Kasetsart University
（Thailand）
Univ. Gadjah Mada
（Indonesia）
Domestic and international collaborations
Innate immune function for
Genome analysis and
infection-resistance
breeding in Japanese
improvement
native chickens
Excellent genetic
trait of Japanese
native chickens.
Sichuan Agricultural Univ.;
North West Agriculture and
Forestry Univ.（China）
Fundamental studies
on reproduction and
development
Finding of antimicrobial
peptide in reproductive
and digestive organs.
Salmonella E
Analysis of genetic
locus for genome
breeding
New strategy for disease
resistance improvement
in chickens and other
livestock.
Avian β-defensin
Development of new chicken breeds that are
excellent in products quality, disease-resistance and
reproductive performance.
Contribution for sustainable development of
poultry production in Asia and world.
Fundamental
studies on sex
determination,
developmental
engineering, and
reproductive
endocrinology
Research activities in RCAS
Project 2. Reproductive biology, (Example) Breeding technology development
North West Agriculture and
Forestry Univ.（China）
Identification of sperm motility
suppressing factors in semen
Fe3+
Ca2+
ATP
LH
（ovulation
stimli）
Granulosa/
cumulus
cells
NRG1
ovulation
AREG,
Oocyte
maturation
Fertilizing
ability
Ca2+
Fertilizable ovum
Determination of the
fertilization mechanism
Determination of ovulation mechanism and
sperm cryopreservation techniques in pigs
day0
day4
SNP analysis for genetic close-relations in pigs
DNA analysis for determination of parent-child
relationship and identification of individual origins
sire pigs
day6-9
Pig No. 0001
Pig No. 0002
SNP
SNP
SNP No.
Genotype
Genotype
SNP1001
A
SNP1001
A
SNP1002
T
SNP1002
C
↓
↓
↓
↓
SNP1023
G
SNP1023
A
SNP1024
A
SNP1024
G
↓
↓
↓
↓
SNP1192
T
SNP1192
T
Individual Identification using SNP genotypes
SNP No.
Identification of
heat (0h)
AI optimum AI optimum
time
time
Novel artificial insemination
program in pigs
Term possible
for fertilization
Sperm
survival time
Establishment of the novel pig
breeding technology
Contribution for sustainable
development of pig production in
Asia and world.
Research activities in RCAS
Project 3. Endocrinology, Physiology of nutrition, Animal management technology
(Example) Animal management technology development
Kasetsart University
（Thailand）
Nutrient
metabolism in
ruminants
University of Alberta（Canada）
Nutritional factors
causing lifestyle
disease and obesity
Gastro-intestinal
health and
protection by
probiotics
Natl. and prefectural
institutes, Japan
Hiroshima University
(Japan)
Improvement of
Analysis of body
productivity in dairy score by 3D image
milk using advanced and remote sensing
robot system
Work-saving management, High performance, Health
enhancement and Safe products in Animal production
Production and
Tissue regulatory
3D image
health analysis
factors analysis
analysis
Milk amount and
Body shape and
Metabolism regulation
quality, estrus cycle
obesity evaluation by blood factors and
identification etc.
etc.
hormones etc.
Expression of avian β-defensins, a member of
antimicrobial peptides, in hen oviduct
Yukinori Yoshimura
Graduate School of Biosphere Science, Research Center
for Animal Science, Hiroshima University, HigashiHiroshima 739-8528, Japan
[email protected]
Infection of
reproductive organs
ovary
Reproductive disorder
TE
TI
G
Y
Drop in egg production
Disorder of egg-forming
functions such as eggshell
formation.
Contamination of eggs
oviduct
Problems in food safety
L
E
S
Salmonella enteritidis infect
the ovary and oviduct
Increase in mortality of
embryo and chicks
The lower oviductal
segments are the primary
tissues that are infected by
microorganisms colonizing
in the cloaca.
Innate immune system
* First line of defense against infection by various pathogenic microbes
including antibiotics-resistant bacteria and unforeseen microbes.
Virus
flagellin
Lipoprotein
Peptidoglycan
(Gram +ve)
Unmethylated
CpG DNA
ssRNA
dsRNA
LPS
(Gram -ve)
Lipoprotein, peptidoglycan
(Gram positive bacteria )
TLR2
TLR1
lipopolysaccharide; LPS
(Gram negative bacteria )
dsRNA
type 1 & 2
TLR4
CD14
TLR3
Flagellin
TLR5
ssRNA
NF-kB/IkB
MyD88
-dependent,
NF-kB
-independent
pathway
AP-1
type 1 & 2
Bacterial protease
Receptors recognizing
microbe-associated molecular
patterns
Proinflammatory cytokines
Transcription
TLR7
TLR15
Avian Toll-like receptors
TLR21
Unmethylated CpG-DNA
Chemokines
Antimicrobial peptides
(avian β-defensins: AvBDs)
Avian β-defensins (AvBDs)
+ charge
- charge
* Cysteine-rich cationic antimicrobial
peptides
Bacterial cell membrane
* Antimicrobial activity against a wide
spectrum of microorganisms [Grampositive and Gram-negative bacteria,
protozoa, fungi and enveloped
viruses]
Breaking down cell
membrane and killing
bacteria
* Fourteen genes have been identified
M 1
2 3
4
5
AvBDs
6 7 8 9 10 11 12 13 14
Escherichia coli
AvBD9
Fourteen AvBDs genes have been
identified in chicken.
control
AvBD9-AvBD8
Antimicrobial activity of
recombinant AvBD (Gal)-9 and
AvBD-9-AvBd-8 against
Escherichia coli.
(from Ma DY et al. Protein Expr
Purif 58: 284-291. 2008. )
Goal of the study
To determine the mechanism by which AvBDs expression is
induced in response to microbial agents in hen oviduct
Questions
1. Are TLRs and avian β-defensins
(AvBDs) expressed in the oviduct?
2. Do microbial components induce the
expression of AvBDs and cytokines?
3. How is the AvBDs expression
refulated?
LPS
Poly (I:C)
Flagellin
CpG ODN
TLRs
pathogen
LPS
TLR4
NFκB/AP-1
Transcription
IL receptor
Proinflammatory
cytokines
AvBD cells
NFκB/AP-1
Transcription
AvBDs
AvBDs
pathogen
Estimated pathway from TLR stimulation to
synthesis of AvBDs and cytokines
TLRs
M
1-1
ligands
1-2
2-1
2-2
3
4
5
7
15
21
peptidoglycan
Expression of TLR in vaginal cells
M
Expression of TLR adaptor molecules
and transcription factors in vaginal cells
Expression of Toll-like receptors and
transcription factors in the vaginal cells
cloaca
vagina
M
1
2
AvBD3
protein,
isthmus
L
C)
3
4
5
6
AvBD3 protein, uterus
L
AvBDs
7
8
9
10
AvBD3 protein, vagina
L
12
13
14
kDa
E
E
37
LP
TG
11
E
TG
LP
LP
25
20
15
AvBD3 mRNA,
vagina
Identification of avian β-defensins
(AvBDs) in the vagina
10
Western blot of
AvBD-3 in the
uterus
3
AvBD1
8
1.5
4
1
2
0.5
0
102
10
103
104
0
0
2
AvBD4
1.5
1.5
1
1
0.5
0.5
0
0
0
10
102
103
104
Fold change
5
4
AvBD10
b
ab
3
2
Fold change
6
2
25
AvBD3
2
0
Fold change
2.5
a
a
a
1
0
0
10
102
103
LPS (ng/mL), 3h
104
7
6
5
4
3
2
1
0
102 103
10
20
10
102
103
104
AvBD12
104
b
ab
ab
ab
a
0
10
102
103
104
LPS (ng/mL), 3h
Effects of lipopolysaccharide on the expression of avian
β-defensins (AvBDs) in the cultured vaginal cells
c
bc
bc
10
5
0
AvBD5
0
IL1β
15
Fold change
Fold change
10
6
5
4
3
2
1
0
ab
a
0水
2
3
10
10
104
10
100 10
1000 10000
LPS (ng/mL)
ab
IL6
ab
b
a
00
a
4
3
2
10
10 10
100 10
1000 10
10000
LPS (ng/mL), 3h
Fold change
4
AvBD1 b
b
3
2
2
00
Fold change
M
ab
4
a
0
100
102
1000
103
AvBD4
a
0
00
2
0.8
100
102
1000
103
Expression of IL1
and IL-6 receptors
in the vaginal cells
AvBD5
1.5
0.6
1
0.4
0.5
0.2
0
00
Fold change
b
AvBD3
6
1
1
8
1.2
1
0.8
0.6
0.4
0.2
0
100
102
1000
103
AvBD10
0
102
103
0IL1β100
1000
(ng/mL),
3h
0
1.2
1
0.8
0.6
0.4
0.2
0
00
2
10
100
3
10
1000
AvBD12
00
102
100
103
1000
IL1β (ng/mL), 3h
Effects of IL1β on the expression of
AvBDs in the cultured vaginal cells
Conclusions
2. The innate immune system may
attack and prevent infections
from a wide spectrum of
pathogenic microbes including
those to which vaccinations are
not performed, those resistant to
antibiotics, and other unforeseen
microbes.
AvBD8
TE
Y
AvBD12
1. Innate immune system formed by
TLRs and avian β-defensins is
expected to play the first line of
host defense in the oviduct. That
immunity is essential not only for
the health of individuals but also
for the production of hygienic
eggs.
Largest yellow follicle
TI
G
20
m
TE
TI
G
20 m
Y
Immunohistochemistry
For the future
AvBD3
Outer layer
AvBD3
L
E
Inner layer
TG
LP
20 μm
AvBD12
kDa
50
37
L
E
25
20
15
TG
20 μm
10
AvBD3
kDa
50
37
25
20
15
AvBD12
Non-specific
Non-specific
10
LP
AvBDs in the uterus
Are AvBDs transmitted
from oviduct to the eggs?
AvBDs in the eggshell
For the future
AvBDs
bp
M 1
2
3
4
5
6
7
8
9
10
11
14
12 13
500
100
mucosal fold of the proventriculus
E
LP
proventricular gland
L
F
G
GL
For production of healthy chickens
Do antimicrobial peptide play roles in the defense in
the gastrointestinal mucosa?
Are probiotics and other natural substances useful to
enhance the synthesis of them?