Fish immunology for vaccination - WIDE Project · 2012-12-05 · Problems in fish vaccine • Some vaccines are not highly effective. • Some vaccines are only effective for a short

Fish immunology for vaccination

Hidehiro Kondo

World fisheries production by capture and aquaculture

http://www.fao.org/fishery/statistics/enYear

Am

ount

(kilo

ton)

0

10,000

20,000

30,000

40,000

50,000

60,000

70,000

80,000

90,000

1950 1960 1970 1980 1990 2000 2010

Fish aquaculture Fish capture

World aquaculture production

http://www.fao.org/fishery/statistics/enYear

Am

ount

(kilo

ton)

0

5,000

10,000

15,000

20,000

25,000

30,000

35,000

1950 1960 1970 1980 1990 2000 2010

Freshwater Aquaculture

Diadromous Aquaculture

Marine Aquaculture

Japanese fisheries production by capture and aquaculture

http://www.maff.go.jp/j/tokei/kouhyou/kaimen_gyosei/index.htmlYear

Am

ount

(kilo

ton)

0

2,000

4,000

6,000

8,000

10,000

12,000

14,000

1956 1966 1976 1986 1996 2006

Marine Capture Marine Aquaculture Freshwater Capture Freshwater Aquaculture

Aquaculture in Japan in 2010 (kiloton)

Marine fishYellowtail 138.9Sea bream 67.6Coho salmon 14.8

Freshwater fishJapanese eel 20.5

http://www.maff.go.jp/www/info/bunrui/bun06.html

Yellowtail Sea bream

Japanese fisheries production of marine aquaculture

http://www.maff.go.jp/j/tokei/kouhyou/kaimen_gyosei/index.htmlYear

Am

ount

(kilo

ton)

0

50

100

150

200

250

300

1956 1966 1976 1986 1996 2006

Yellowtail Sea bream Other

Pathogenic problems in aquaculture

•Bacterial diseases

•Viral diseases

•Fungal diseases

•Parasitic diseases

Bacterial diseases in Japanese aquacultureHost

Gram-negative bacteriaAeromonas salmonicida Salmon, Trout, etcEdwardsiella tarda Japanese flounder, Red seabreamFlavobacterium psychrophilum Ayu, Salmon, Trout, etcPhotobacterium damselae subsp. piscicida YellowtailTenacibaculum maritimum Red seabream, etcVibrio anguillarum Salmon, Trout, etc

Gram-positive bacteriaLactococcus garvieae YellowtailStreptococcus iniae Japanese flounder

Acid fast bacteriaMycobacterium spp. YellowtailNocardia seriolae Yellowtail

Antimicrobial agents in Japanese aquaculture

Amoxicillin Pasteurellosis

Ampicillin Pasteurellosis

Erythromycin Streptococcosis

Oxytetracycline Vibriosis

Oxolinic acid Pasteurellosis

Florfenicol Pasteurellosis, Streptococcosis

Fosfomycin Pasteurellosis

Lincomycin Streptococcosis

Sulfamonomethoxine Vibriosis, Nocardiosis

Problems in use of antimicrobial agents

• Impact on environments

• Outbreak of drug resistant bacteria

• Risks to human health

We need to reduce the use of antimicrobial agents!

Viral diseases in Japanese aquaculture

Host

DNA virus

Koi herpesvirus Carp

Red seabream iridovirus Red seabream, Yellowtail, etc

Lymphocystis disease virus Japanese flounder, etc

RNA virus

Infectious pancreatic necrosis virus Yellowtail

Viral hemorrhagic septicemia virus Japanese flounder

Yellowtail ascites virus Yellowtail

Striped jack nervous necrosis virus Yellowtail

Vaccination is one of the solution for controlling infectious diseases

Vaccines work by stimulating the animal’s immune system, which is the ability of an organism to resist a particular infection or toxin by the action of specific antibodies or sensitized white blood cells.

Vaccination is one of the solution for controlling infectious diseases

• Vaccination

Vaccine

Advantages of vaccination against antimicrobial agents

• Vaccine has an effects for long time.

• Vaccine can protect fish not only from bacteria but also from virus.

• Vaccine reduces the use of antimicrobial agents.

Vaccines used in Japanese yellowtail aquaculture

• Streptococcosis

• Vibriosis

• Pasteurellosis

• Iridovirus disease

Changes of the number of the vaccinated yellowtail

The

num

ber o

f irid

oviru

s di

seas

e di

agno

ses

50

100

150

200

250

300

The

num

ber o

f va

ccin

ated

fish

(X m

illio

n)

1

2

3

4

5

1995 2000 2005

Modified from “Aquatic vaccine handbook (2009: Japanese)”.

The effect of multivalent Aeromonas salmonicida/Vibrio vaccines on antimicrobial use in the Norwegian salmon-

farming industry

http://www.hc-sc.gc.ca/dhp-mps/pubs/vet/amr-ram_final_report-rapport_06-27_cp-pc-eng.php

Types of injection methods

• Injection methods

• Intra-peritoneal injection

• Bath immunization

• Oral administration

Intra-peritoneal injection

•High efficacy•Take a time and labor

Bath immunizationOral administration

•Lower efficacy than intra-peritoneal injection•Need less labor than injection

Immersion into vaccine solution

Feed vaccine compound

Types of vaccine

• Types of component

• Inactivated pathogen

• Antigen (component vaccine)

• DNA (DNA vaccine)

Inactivated pathogen• Pathogenic microorganisms are

inactivated by formalin, heat, UV etc.

• The vaccine contains multiple antigens.

Antigen (component vaccine)DNA (DNA vaccine)

• Inject specific antigenic components(component vaccine)

• Inject DNA fragment encoding antigen (DNA vaccine)

Vaccine elicit specific immunity

PhagocytosisAntigen presentationEnhancement of cell

growth/antibody production

Antibody production

Problems in fish vaccine• Some vaccines are not highly effective.

• Some vaccines are only effective for a short period.

• Injection methods may be improved to be less labor, and less stressful on fish.

We need to know about fish immunity!

Immunity in fish• Innate immune system

• Cellular and humoral mechanisms protect the host from infection by other organisms, in a non-specific manner.

• Adaptive immune system

• Cellular and humoral mechanisms protect the host from infection by other organisms, in a specific manner.

Phylogeny of vertebrates

Agnathans

Fish

Amphinian

Birds

Mammals

500 MYA

450 MYA

370 MYA

310 MYA

Fish is a first animal possessing vertebrate’s immune systems!

Innate immune system in fish

• Prevent the invasion of pathogenic organisms

• Skin mucosa, Scale, Skin, etc

• Kill or inactivate the pathogenic organisms

• Complement components, Phagocytes, etc

• Activate immune cells to develop specific immunity

• Antigen presentation cells, Cytokines, etc

Prevent the invasion of pathogenic organisms

Mucus

ScaleSkin

Kill or inactivate the pathogenic organisms

LysozymeAntimicrobial peptides

Complement components

Phagocyte Cells directly kill the virus infected cells

Activate immune cells to develop specific (adaptive) immunity

Production of signal molecules

(such as cytokines)Presentation of antigenic molecules

Induce specific immunity

Adaptive immune system in fish

• Humoral immunity

• Antibody (Immunoglobulin M)

• Cellular immunity

• Cytotoxic T cell, etc

Humoral specific immunity

AntibodyImmunoglobulin M

Region for antigen recognition

Activate immune cells to develop specific (adaptive) immunity

Activate antibody producing cells

Producing specific antibody

Function of Antibody

Antigen specific antibody

Inactivation of pathogen

Opsonization

Activation of complement

system

Cellular specific immunity

Presentation of antigenic molecules

Virus infected cells

•Cells directly kill the virus infected cells•Activate cytotoxic cells

Phylogeny of vertebrates

Agnathans

Fish

Amphinian

Birds

Mammals

500 MYA

450 MYA

370 MYA

310 MYA

Fish genes involved in immunity are almost similar to those in the

other vertebrates!!

Fish immune system is similar to that of mammals

Are the immune system of these species similar?

Genome;approximately 700Mb in length and comprises 24 pairs of chromosomes

Coding genes: 20,476

Fish and human genome data in public database

Genome;approximately 3,300Mb in length and comprises 23 pairs of chromosomes

Coding genes: 19,686

Investigation in Japanese flounder immune system

Japanese flounder

The numbers of nucleotide sequence information of Japanese flounder in

public database

Japanese flounder genes involved in immunity so far identified

Cytokines Transcriptional factors CD antigensCD antigens Receptor proteins

B cell activation protein BL34CC chemokinesCXC chemokinesFAS ligandG-CSFInterleukin-1 βInterleukin-11Tumor necrosis factor (TNF)TNF superfamily

Interferon regulatory factor-1Interferon regulatory factor-4CCAAT/enhancer-binding protein βCCAAT/enhancer-binding protein εNuclear factor-interleukin-6-βSignal transducer and activator of

transcription 3TNF receptor associated factor

CD3CD8CD11bCD18CD20CD22CD40CD49eCD50CD54CD63

CD71CD82CD83CD102CD116CD178CD180CD183CD183CD197

Immunoglobulin DImmunoglobulin M

membrane formImmunoglobulin light chainFc-γ-1/γ-2 receptorPolymetic Ig receptorLeukotriene β4 receptorT cell receptor αT cell receptor βT cell receptor δT cell receptor γToll-like receptor 2Toll-like receptor 3Transferrin receptor

Cytokine receptors Anti-bacterial and -viral proteins ProteasesProteases Plasma proteins

CC chemokine receptorCXC chemokine receptorG-CSF receptorInterleukin-1 receptorInterleukin-6 receptorInterleukin-8 receptorTNF receptor 1TNF receptor 2

Lysozyme C typeLysozyme G typeMx proteinNK-lysinThymosin β4Thymosin β10

Cathepsin BCathepsin LCollagenase 3Gelatinase BProtease nexinProlycarboxyl peptidase

Cathepsin BCathepsin LCollagenase 3Gelatinase BProtease nexinProlycarboxyl peptidase

C-type lectinComplement C3Complement C7Complement C8βComplement C9Complement regulatory

factorsHaptoglobinImmunoglobulin MTransferrin

Conclusion

• Importance of knowledges on fish immunity is increasing.

• By application of immune system of fish, we will be able to protect fish against pathogenic microorganisms.