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Dr. T. Citarasu Dr. T. Citarasu Assistant Professor Assistant Professor Tele-fax: + 91-4652-253078 Tele-fax: + 91-4652-253078 Mobile: +91-9994273822 Mobile: +91-9994273822 Email: Email: [email protected] [email protected] [email protected] [email protected] Website: Website: www.msuniv.ac.in www.msuniv.ac.in www.drcitarasu.com www.drcitarasu.com

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Page 1: Cmst forum

Dr. T. CitarasuDr. T. CitarasuAssistant ProfessorAssistant ProfessorTele-fax: + 91-4652-253078Tele-fax: + 91-4652-253078Mobile: +91-9994273822Mobile: +91-9994273822Email:Email: [email protected]@gmail.com [email protected]@msuniv.ac.inWebsite:Website: www.msuniv.ac.inwww.msuniv.ac.in www.drcitarasu.comwww.drcitarasu.com

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IMPORTANCE OF AQUACULTUREIMPORTANCE OF AQUACULTURE

Lucrative food production industry

Fresh, brackish & Marine based culture

10 % annual growth rate

Global sea food demand-70% in the next 35 years (7 fold)

Capture and culture fisheries

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Shrimp FarmingShrimp Farming

Asia is the largest producer- 80 %Asia is the largest producer- 80 %

India is the second place of Asia & India is the second place of Asia & fifth place in globallyfifth place in globally

>350 shrimp hatcheries 13 billion seed >350 shrimp hatcheries 13 billion seed production/ annum production/ annum

Top 5 Countries M. M tonesChina 34.43India 3.13Vietnam 1.65Thailand 1.39Indonesia 1.30

Aquaculture production (FAO, 2009)

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CURRENT PROBLEMS IN AQUACULTURE CURRENT PROBLEMS IN AQUACULTURE Culture methods

• Extensive: 0-5 m3

• Moderate extensive : 5-15 m3

• Semi intensive : 15-30 m3

• Intensive : 30-60 m3

• Super Intensive : > 60 m3

Significant economic losses

Bacterial diseases - Vibriosis

Viral diseases – WSSV; MBV

Stress Induced diseases

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Finfish DiseasesFinfish Diseases

Freshwater FishesFreshwater FishesMotile Aeromonad septicemia Motile Aeromonad septicemia (MAS)(MAS)

Hemorrhagic septicemiaHemorrhagic septicemiaA. hydrophila, A. sobriaA.caviae, A. veronii, A. salmonicida

Marine FishesMarine FishesVibriosisVibriosis

Pathogenic VibriosPhotobacterium spMicrococcus spFlavobacterium- Gill diseaseEnterobacter- EdwardsiellaPseduomonas Salmonella

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Important Shrimp Diseases in IndiaImportant Shrimp Diseases in India

VibriosisVibriosis RMS or White Patch DiseaseRMS or White Patch Disease

White Spot Syndrome Virus (WSSV)White Spot Syndrome Virus (WSSV) Enterocytozoon hepatopenaei (EHP)

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CURRENT PROBLEMS IN DISEASE CONTROL CURRENT PROBLEMS IN DISEASE CONTROL

Current disease treatment Current disease treatment protocols – difficultprotocols – difficult

Chemicals & Synthetic drugsChemicals & Synthetic drugs

Antibiotics & other chemicalsAntibiotics & other chemicals

MPEDA abandoned more than MPEDA abandoned more than 10 antibiotics10 antibiotics

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Chloramphenicol Metronidazole

Nitrofurans Ronidazole

Neomycin Ipronidazole

Nalidixic acid nitroimidazoles

Sulphamethoxazole Clenbuterol

Chlorpromazine Diethylstilbestrol

Colchicine Sulonamide

Dapsone Floroquinolones

Dimetridazole Glycopeptides

Banned Antibiotics (MPEDA)Banned Antibiotics (MPEDA)

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Herbal MedicinesHerbal Medicines

Storehouses of safer & cheaper compounds

Low cost, effective, no side effects & biodegradable

Various activities such as growth promotion, immunostimulants, anti stress, appetizing, aphrodisiac, antibacterial, antiviral and antifungal etc.

EcliptineEcliptine Tinosporone

Ocemine Aegilin

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Percolation ExtractionPercolation ExtractionHerbal powder+ non polar solvent -Herbal powder+ non polar solvent -

Hexane (1:1) at 3 daysHexane (1:1) at 3 days

Filter & centrifugeFilter & centrifuge

Filtrate Filtrate SlurrySlurry

Slurry + Mid polar solvent Slurry + Mid polar solvent (1:1) at 3 days(1:1) at 3 days

Filter & centrifugeFilter & centrifuge

Filtrate Filtrate SlurrySlurry

Slurry + polar solvent (1:1) Slurry + polar solvent (1:1) at 3 daysat 3 days

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Filtrate Filtrate

CondenseCondense

Rotary EvaporatorRotary Evaporator

LyophilizeLyophilize

ScreeningScreening

StorageStorage

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Soxhlet ExtractionSoxhlet Extraction

Herbal powderHerbal powder

Particular solventParticular solvent

16 h extraction16 h extraction

LyophilizeLyophilize

ScreeningScreening

StorageStorage

condensecondense

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Column PurificationColumn Purification

Herbal ExtractsHerbal Extracts

Loaded with silica column with Loaded with silica column with the mobile phase is hexanethe mobile phase is hexane

Elute with mixture of non polar + Elute with mixture of non polar + mid polar solvent 100 hexane; mid polar solvent 100 hexane; 80H + 20EA; 60H+40EA.…… 100 80H + 20EA; 60H+40EA.…… 100 % EA% EA

Elute with mixture of MP + P Elute with mixture of MP + P solvent 80EA+20 solvent 80EA+20 methanol………………100% methanol………………100% methanolmethanol

Collect elution & screeningCollect elution & screening

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ELUTION STEPSELUTION STEPS

HEXANEHEXANE

100100

9090

8080

7070

6060

5050

4040

3030

2020

1010

00

ETHYL ACETATE ETHYL ACETATE

0 0

10 10

20 20

30 30

40 40

50 50

60 60

70 70

80 80

90 90

100 100

ETHYL ACETATE ETHYL ACETATE

100100

90 90

8080

7070

6060

50 50

40 40

30 30

20 20

10 10

00

METHANOL METHANOL

00

10 10

2020

3030

4040

50 50

60 60

70 70

80 80

90 90

100100

Non

Pol

arM

id P

olar

Mid

Pol

arM

id P

olar

Pola

rPo

lar

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Thin Layer Chromatography (TLC)Thin Layer Chromatography (TLC)

Small amount of ExtractsSmall amount of Extracts

Spotted on TLCSpotted on TLC

Placed in a beaker having the Placed in a beaker having the mobile phase of 1:1 Hexane & mobile phase of 1:1 Hexane &

Ethyl acetate Ethyl acetate

Run by capillary actionRun by capillary action

Rf valueRf value

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distance from origin to component spotdistance from origin to component spot

Rf value = ------------------------------------------- Rf value = -------------------------------------------

distance from origin to solvent frontdistance from origin to solvent front

Further purificationFurther purification Lengthy column Lengthy column

100-200 100-200 μm of silica particle sizeμm of silica particle size

Fine separationFine separation

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Partial Characterization of Active compoundsPartial Characterization of Active compounds

UV SpectrometryUV Spectrometry

FT-IR spectroscopy FT-IR spectroscopy

HPLC-MSHPLC-MS

GC-MSGC-MS

LCMSLCMS

NMRNMR

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Antibacterial ScreeningAntibacterial Screening

Piper longumPiper longumMurraya koeniji Murraya koeniji Quercus Quercus infectoriainfectoria Psoralea Psoralea

corylifoliacorylifolia

More than 50 herbals screened against More than 50 herbals screened against aquatic important pathogensaquatic important pathogens

In vitro In vitro antibacterial activityantibacterial activity

Some of them characterizedSome of them characterized

Beta Ocemine

Quercucin PsoralinPiperine

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Mode of Herbal Medicines DeliveryMode of Herbal Medicines Delivery

Artificial Diet PreparationArtificial Diet PreparationIngredients

Diet composition (g.kg -1)C HAD1 HAD2 HAD3 HAD4

Brown fish meal 280 280 280 280 280Shrimp head meal 100 100 100 100 100Squid meal 20 20 20 20 20Squid liver powder 30 30 30 30 30Wheat gluten 60 60 60 60 60Wheat flour 200 200 200 200 200Soy bean meal 100 100 100 100 100Broken rice 102 102 102 102 102Fish oil 20 20 20 20 20Vitamin premix 7.8 7.8 7.8 7.8 7.8Mineral mix 40 40 40 40 40Mixtures of Herbal antibacterial extracts 0 0.2 0.4 0.6 0.8

The basal diet contained 45.1 % protein; 7.2 lipid; 14.6% ash; 7.1 % moisture and 3% fibre.

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Bioenrichment or BioencapsulationBioenrichment or Bioencapsulation

Artemia naupli/ Pre adutArtemia naupli/ Pre adut

Gut EvacuationGut Evacuation

Enrichment 800mg/lit of herbal Enrichment 800mg/lit of herbal extracts emulsified with Egg extracts emulsified with Egg

YolkYolk

1 hr enrichment1 hr enrichment

Feeding to fishFeeding to fish

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Herbal Antiviral

Phyllanthus niruri Leucus aspara

Ocimum basilicum

Effectively controlled WSSV at earlier stage of infection

Inhibit viral replication

Effective against other shrimp virus such as MBV and YHV

Psidium guajava

Phenol, 2,5- bis Phenol, 2,5- bis (1,1-dimethylethyl)(1,1-dimethylethyl)

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MethodsMethods

5050µg extract+5 µl WSSV suspension (30µg µg extract+5 µl WSSV suspension (30µg protein)protein))- incubate 29)- incubate 290 0 C at 3 hr C at 3 hr

Centrifuged, filtered & LyophilizedCentrifuged, filtered & Lyophilized

Antiviral Herbal ExtractsAntiviral Herbal Extracts

Inject to ShrimpsInject to Shrimps

Monitor survival & external symptoms Monitor survival & external symptoms

PCR amplification PCR amplification

If positive ??If positive ?? If negative ??If negative ??

++

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ImmunostimulantsAegle marmelosAegle marmelos

Picrorhiza kurooaPicrorhiza kurooa

Cynodon dactylonCynodon dactylon

Tinospora Tinospora cordifoliacordifolia

Eclipta albaEclipta alba

A substances that A substances that stimulate the immune stimulate the immune system by inducing system by inducing activation or increasing activation or increasing activity of any of its activity of any of its components. components. Improvement of…… • Hematology• Biochemical• Phagocytosis• Prophenol oxidase activity• Lysozyme activity• Bacterial & viral clearance

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Types of Immunostimulants Types of Immunostimulants

Chitin, Chitoson & Oilgomers Chitin, Chitoson & Oilgomers Shell wasteShell waste

Fucoidon Fucoidon Sulfated polysaccharides –SeaweedsSulfated polysaccharides –Seaweeds

GlucansGlucansYeastYeast

LactoferinLactoferinMilkMilk

LipopolysaccharidesBacterial Cell wallBacterial Cell wall

HerbalsHerbalsW. somniferaW. somnifera, , A. calamos, T. cordifoliaA. calamos, T. cordifolia

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Immunostimulant DrugsImmunostimulant DrugsImmunostimulant molecules recognize PRPs (PPAE) & culminate proteolytic cleavage of proPO to PO

Immunostimulant molecules interaction leading to PO activation

Melanin synthesis ROI, RNI Expression of immune genes& TLR TLR AMP & Phagocytosis

PPAE expression in haemocytes

TLR (DSCAM), MYD88, TRAF6

Immune genes (Caspase, crustin)

Enzyme assay- proPO, Intracellular Superoxide anion & Lysozyme

IMMIMM

SPCSPCPRPsPRPs

PPAEPPAE

ActivationActivation

PO ActivationPO ActivationTLR ActivationTLR Activation

AMP synthesis AMP synthesis & Phagocytosis & Phagocytosis

Immune gene ExpImmune gene Exp& ROI, RNI & ROI, RNI

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““Quasi-Immune Response” Quasi-Immune Response” proved with herbal Immunostimulants proved with herbal Immunostimulants

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Algal Drugs SeaweedsSeaweeds

Broad spectrum of activities

antiviral, anthelmintic, antifungal

and antibacterial

Brominated, aromatics, sterols, dibutanoids, proteins, peptides, sulphated polysaccharides, terpenoids, phlorotannins, acrylic acid, phenolic compounds and steroids.

CarrageenanCarrageenanPowerful antioxidantsPowerful antioxidants

AlginateAlginate polysaccharaidepolysaccharaide

Laminaria and FucusLaminaria and Fucus

Fucoidon Fucoidon

Powerful antiviral against WSSVPowerful antiviral against WSSV2-(2-hydroxyphenoxy)-1-2-(2-hydroxyphenoxy)-1-

phenylethanolphenylethanol

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2-(2-hydroxyphenoxy)-1-phenylethanol

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Micro AlgaeMicro Algae

Polyketides, amides, alkaloids, peptides and beta carotenoids

antiviral and antibacterial

Antioxidants

Euglena viridisEuglena viridisAgainst fish & Shellfish pathogensAgainst fish & Shellfish pathogens

Chlorella Chlorella AntibacterialAntibacterial

Dunaliella spDunaliella sp

From Halophilic origin From Halophilic origin

Control shellfish virusControl shellfish virus

Powerful antioxidants & Immunostimulants Powerful antioxidants & Immunostimulants

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Marine & Halophilic secondary metabolites

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Metabolite Production & CharacterizationMetabolite Production & Characterization

Bacterial culture @2000 RPM/25Bacterial culture @2000 RPM/2500C/ 7 C/ 7 DaysDays

Spin the Broth with 5000 RPMSpin the Broth with 5000 RPM

Remove the brothRemove the broth

3 times extraction with ethyl 3 times extraction with ethyl acetateacetate

Column purification Column purification

Filter and lyophilizeFilter and lyophilize

Structural elucidationStructural elucidation

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1,10-Biphenyl-3-amine

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Pharmacological Important BiosurfactantsPharmacological Important Biosurfactants

Extracellular Amphiphilic compoundsExtracellular Amphiphilic compounds

Glycolipids, lipopeptides, fatty acids, Glycolipids, lipopeptides, fatty acids,

polymeric surfactants & particulate polymeric surfactants & particulate

naturesnatures

Broad range of applications Broad range of applications

Antibacterial, antiviral, antifungal, anti-Antibacterial, antiviral, antifungal, anti-

adhesive, anti-cancer, antioxidants, adhesive, anti-cancer, antioxidants,

adjuvant etcadjuvant etc

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9-OctadecenamideFatty acid

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Mannosamine- Polymeric natureMannosamine- Polymeric nature

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ProbioticsProbiotics

Probiotics have been defined as “live microbial food supplements which beneficially affect the host by improving the intestinal microflora balance”

Gut & Water ProbioticsGut & Water Probiotics

Competitive exclusion principleCompetitive exclusion principle

Used in Shrimp farms and polluted areasUsed in Shrimp farms and polluted areas

Improved strains of ProbioticsImproved strains of Probiotics

Lactobacillus and Bifidobacterium spLactobacillus and Bifidobacterium sp

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Probiotic Diet making ProtocolProbiotic Diet making Protocol

Mixed substrates Mixed substrates [Rice bran-300 g;[Rice bran-300 g; Wheat bran-300 g; Rice flour-200g &Wheat bran-300 g; Rice flour-200g &

Wheat flour-200 g]Wheat flour-200 g]

600 ml bacterial culture broth600 ml bacterial culture broth

Make small balls & dried @ 40 ºCMake small balls & dried @ 40 ºC

Powdered Powdered

Bacterial countBacterial count

Vacuum packed Vacuum packed

Apply to pondApply to pond

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SPECIFIC PATHOGEN FREE (SPF) SHRIMPSSPECIFIC PATHOGEN FREE (SPF) SHRIMPS

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Specific Pathogen Free (SPF)Specific Pathogen Free (SPF)Development of SPF doubling the production in Development of SPF doubling the production in Litopenaeus Litopenaeus vannamei vannamei in US aqua industry during 1990sin US aqua industry during 1990s

They cross bread between two traitsThey cross bread between two traits

CPF and Oceana Institute develop the SPF by cross bread the CPF and Oceana Institute develop the SPF by cross bread the better characters such as better characters such as salinity, low DO, pH tolerance & disease salinity, low DO, pH tolerance & disease tolerence traits tolerence traits and analyze the characters at 30 generation and and analyze the characters at 30 generation and commercializationcommercialization

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EDIBLE ANTIBODY OR EDIBLE VACCINESEDIBLE ANTIBODY OR EDIBLE VACCINES

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Production of Yolk antibodyProduction of Yolk antibody

Principle

When chickens are faced with a foreign virus or bacterium, they produce antibodies to fight the invader. They pass that immunity on to their offspring, and antibodies wind up in the eggs.

1. Chickens are more apt than mammals to make high-avidity antibodies

2. A single chicken can produce an enormous amount of antibody, upto 3 grams of IgY per month, which is 10-20 times the amount of a rabbit

3. chickens produce antibody much quicker-high-titre antibody is available from eggs as early as day 25

4. Storing of eggs (antibodies) is very easy

5. It is cheaper to feed and house chickens than rabbits

6. Effective against drug-resistant bacteria.

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IgY IgY production & production & purificationpurification

YolkYolk

DDHDDH22O WashO Wash

Cut Open Cut Open

Isoproponanl wash 1: 3 – 2 times Isoproponanl wash 1: 3 – 2 times

Acetone wash- 1 timeAcetone wash- 1 time

Fitered & storeFitered & store

Complete removal of Complete removal of LipidsLipids

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Anti WSSV IgY (37 k Da)

M C IgY IgY- Adj

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VACCINE DEVELOPMENTVACCINE DEVELOPMENT

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Vaccine Generations Vaccine Generations

First Generation of vaccineLive, attenuated and killed forms

killerCellular immune responses

Second Generation of vaccineSubunit vaccines T Helper cells immunity

Third Generation of vaccineDNA vaccinesHumoral & cellular immunityStrong & long lasting

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Vaccines in AquacultureVaccines in Aquaculture

Whole Cell VaccineWhole Cell Vaccine

Bacterial CellsBacterial Cells

Viral ParticlesViral Particles

Inactivated formsInactivated forms

Virus- Grown in cell lines/ eggsVirus- Grown in cell lines/ eggs

Demerit- revert Demerit- revert

ECP VaccineECP Vaccine

ECP – haemolysin, virulent factorsECP – haemolysin, virulent factors

Need adjuvantNeed adjuvant

Improved survivalImproved survival

Partial purified forms Partial purified forms 52 kDa52 kDa

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Subunit VaccineSubunit Vaccine

Proteins Proteins

Envelop / OMPs (37kDa) Envelop / OMPs (37kDa)

Strong ImmunityStrong Immunity

Need adjuvantNeed adjuvant

Hematological & immunological improvementHematological & immunological improvement

Bio Film Vaccine Bio Film Vaccine

Matrix of Protein, carbohydrate & DNA Matrix of Protein, carbohydrate & DNA

90 % Bacteria secretes 90 % Bacteria secretes

Difficult to treat by antimicrobialsDifficult to treat by antimicrobials

Modulate cytokine synthesis and interrupt production of antibodies

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DNA VACCINEDNA VACCINE

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DNA VaccineDNA Vaccine

Immunization with a circular piece of DNA that code for an antigen

Plasmids consist of strong viral promoter to drive the in vivo transcription and translation

When injected into an individual, the plasmid is taken up by the host cells and its genetic information is translated into the immunizing protein.

Multicistronic vectors are sometimes constructed to express more than one immunogen, or to express an immunogen and an immunostimulatory protein.

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Plasmids consist of strong viral promoter (SV-40/CMV ) to drive the in vivo transcription and translation of gene interest.

Intron A may sometimes be included to improve mRNA stability and hence increase protein expression.

Plasmids also include a strong polyadenylation/transcriptional termination signal, such as bovine growth hormone or rabbit beta-globulin polyadenylation sequences.

Multicistronic vectors are sometimes constructed to express more than one immunogen, or to express an immunogen and an immunostimulatory protein.

Vector DesignVector Design

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Recombinant plasmids enter to the host cell

Gene of interest is transcribed by the RNA polymerase II & synthesis of messenger RNA (mRNA)

Translated into the corresponding protein in the cytoplasm of the host cells

T- Cell as well as B-Cell immunity will developed & produced antibody against the proteins

When pathogen (antigen) enters to the host cell, the T-Cells as well as B cells recognize and killed.

How DNA vaccine plasmids Stimulate immune responses?

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Immune system Aquatic species

Shell Fishes

Innate immune system

No clear responses

No antibody production

Pathogen persists

Infectious to others

Humoral immunity AMP, PO & Coagulation

Cellular immunity Encap, nodule & pago..

Adaptive immune system

Specific responses

Antibody produced

Pathogen eliminated

May or may not

Humoral immunity B cells

Cellular immunity T cells

Fin Fishes

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DNA VACCINE DEVELOPMENT PROTOCOL DNA Vaccine ConstructDNA Vaccine Construct

PCR amplification of desired gene

Design & amplify the gene by specific primers

5’-GGATCCAATCATGGATCTTTCTTTCAC-3’-BamHI

5’- GAATTCTTACTCGGTCTCAGTGCC-3’- EcoRI Cloning to suitable vectors

Purify the PCR products Clone to “T” Easy vectors Ligation @ 4 or16° C for 16 hrs/ Over Night Transformation to DH5 α Sequencing by using M13+ and M13- primers

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DNA Vaccine Construct by cloning

Plasmid extraction from positive clonesRestriction of REC plasmid by respective enzymesLigate to suitable DNA vaccine vector such as DNAVAX or pCDNA 3.1 @ ratio of 1: 3

Transformation

Recombinant plasmids & comp. cells will be mix and keep on ice 30 mts

The mixture will be put in water bath at 42°C for 90 seconds

Snap chill the and add 400 µl LB broths

10 µl of this culture will be plated on LB agar in presence of antibiotics.

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Plasmid extraction

Plasmid will be purified from the positive clones using standard methods

Steps 1. Alkaline lysis2. Phenol chloroform Extraction3. Ethanol precipitation

Plasmid will be suspended to PBS or sterile saline and store to -20° C

Diluted to sterile saline for delivery

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Vaccine Delivery

Small volume of DNA vaccines is enough 0.1 or 0.2 ml volume is enough 10 µg DNA is enough (5+ 5= 2 doses or 10= 1 dose) Disadvantage: Cant’ apply for small fishes

Methods of DeliveryGenerally injectionLarvae- injection impossibleAlternative methods are

1. The DNA vaccine transformants will be coated to artificial fed using fish oil and feed them.

2. Bioencapsulation may be a preferred method of oral delivery (rotifers, brine shrimp)

3. DNA vaccine transformant (Bacterial mode)4. Conjugate with Chitoson / sodium alginate

complex

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Tissue Distribution studies

RNA extracted from different tissues

2µg of total RNA will be primed with 1µg oligodT(18) and reverse-transcribed into cDNA with 200U MMLV reverse transcriptase

PCR amplification with specific gene primers & product confirmation

q-RT PCR for gene copy

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RNA INTERFERENCE (RNAi)RNA INTERFERENCE (RNAi)

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RNA Interference (RNAi) against Viral diseaseRNA Interference (RNAi) against Viral disease

““A process in which the introduction of double-stranded A process in which the introduction of double-stranded RNA into a cell RNA into a cell inhibits the expressioninhibits the expression of genes” of genes”

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Post Translational Gene Silencing Post Translational Gene Silencing (PTGS) Pathway(PTGS) Pathway

ds RNA in the cytoplasm triggers the ds RNA in the cytoplasm triggers the multi domain multi domain ribonuclease II enzyme ribonuclease II enzyme DICER which cleaves the ds RNA in to si DICER which cleaves the ds RNA in to si RNA which are RNA which are 21 to 23 21 to 23 nucleotide nucleotide fragments.fragments.

These si RNA is recognized by the These si RNA is recognized by the RNA RNA induced Silencing complex (RISC)induced Silencing complex (RISC), a , a multi enzyme unit that brings about multi enzyme unit that brings about separation of two si RNA stands.separation of two si RNA stands.

The antisense Si RNA stand remains The antisense Si RNA stand remains bound to RISC while the sense strand is bound to RISC while the sense strand is released. Finally the antisense and released. Finally the antisense and RISC complex bind the target mRNA RISC complex bind the target mRNA allowing the nuclease activity and allowing the nuclease activity and degraded the target gene. degraded the target gene.

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Crustaceans such as penaeid shrimp, which can be infected by more than twenty different viruses.

Bunyaviridae, Herpesviridae, Picornaviridae, Parvoviridae, Reoviridae, Rhabdoviridae, Togaviridae, Iridoviridae or a new virus family, the Nimaviridae

Culture practices leading to stress induced diseases, bacterial and viral infections leading to severe economic losses

White spot syndrome virus (WSSV), Yellow head virus (YHV), and Taura syndrome virus (TSV)

Role of RNA Interference in aquatic disease control Role of RNA Interference in aquatic disease control

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CONCLUSIONS & FUTURE DIRECTIONS CONCLUSIONS & FUTURE DIRECTIONS

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Improved health and wealthImproved health and wealth

Environmental spoilage can be avoidedEnvironmental spoilage can be avoided

Preventing emergence of resistant Preventing emergence of resistant

strainsstrains

Biomagnifications can be controlledBiomagnifications can be controlled

I am I am HealthyHealthy

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Consumption of antibiotic treated Consumption of antibiotic treated

shrimp/ fish in humans can be prevented shrimp/ fish in humans can be prevented

Consignment rejection can be controlledConsignment rejection can be controlled

The product will be of great benefit for The product will be of great benefit for

the farmersthe farmers

Improved economy to the farmers Improved economy to the farmers

Affected farms due to indiscriminate Affected farms due to indiscriminate

usage of antibiotics can be reutilized.usage of antibiotics can be reutilized.

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