sathish t

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DIAGNOSTIC METHODS OF FISH DISEASES : 

DIAGNOSTIC METHODS OF FISH DISEASES Sathish t Fpm 39

INTRODUCTION : 

INTRODUCTION Accurate therapics and avoid indiscriminate use of chemotherapeutics Live fish / shrimp

Importance of case study & clinical signs : 

Importance of case study & clinical signs Water quality Feeding percentage Feed intake Fertilization schedule Time of last Algal bloom Liming details, Treatment details Source of seed,stocking details

Clinical signs : 

Clinical signs

CONVENTIONAL METHODS : 

CONVENTIONAL METHODS Microscopical disease diagnosis Histoligical disease diagnosis Microbioligical disease diagnosis

MICROSCOPICAL METHODS : 

MICROSCOPICAL METHODS By using magnifying glass LIGHT MICROSCOPY Wet mount Lactophenol cotton blue- fungi Squash preparation Smear preparation

Slide 9: 

Direct squash

Electron microscopy : 

Electron microscopy TEM SEM Prefixation Postfixation Infiltration & Embedding Tissue sectioning Staining

Slide 12: 

Fixation Dehydration Embedding Blocking Tissue sectioning Staining Histology

Slide 13: 

Necrotic foci Tumour like alterations in liver

Bioassay : 

Bioassay In Bp virus Pz of P.vanamei is fed with infected parts Pz 3 will exhibit the patent infection

MICROBIOLOGICAL DISEASE DIAGNOSIS : 

MICROBIOLOGICAL DISEASE DIAGNOSIS Culture Tryptone soya agar Martin’s rose bengal medium Staining Simple stain Acid fast Used to distinguish Mycobacterium & Nocardia

Slide 17: 

17 Gram Stain Gram negative Gram positive Heat/Dry Crystal violet stain Iodine Fix Safranin stain Alcohol de-stain

Slide 18: 

Structural stains India ink preparation Detect cryptococcus neoformans KOH preparation MOTILITY TEST

Biochemical test : 

Biochemical test Sugar fermentation test Pheno red + - yellow, - - red MRVP MR – change in PH VP – acetoin production MR + red, - yellow VP + red,- colourless

Slide 20: 

Citrate utilization test Citrate – carbon, monoammonium phosphate -nitrogen + blue,- green Oxidase test Cytochrome oxidase + blue, - colourless Catalse test 3% hydrogen peroxide + effervescence

Immunological test : 

Immunological test Agglutination test Particulate antigen - button formation,+ mat Immunodiffusion test Soluble antigen Opaque line SRID Counter immuno electrophoresis

Slide 23: 

Elisa plates Ag Ab conjugate Ag – Ab complex Substrate Result ELISA

ELISA : 

ELISA

Slide 26: 

Nitrocellulose membrane Ag Blocking Ab conjugate Substrate Result DOT ELISA

Slide 27: 

Latex agglutination test

FAT : 

FAT

Molecular methods : 

Molecular methods PCR Initial denaturation – 4 min at 95*c Denaturation – 1min at 94 *c Annealing – 1 min 54* c Extension – 2 min 72 * c Final extension – 5 min Nested PCR RT PCR

RAPD : 

RAPD Random primers Like PCR RFLP Amplify particular region For example, HindII enzyme cuts at GTGCAC or GTTAAC

Slide 31: 

Shrimp extract Nitrocellulose membrane Specific probe incubation Dot blot hybridisation WSSV and IHHNV Nucleic acid probes

Slide 33: 

Tissue section in buffer Heated at 94 – 95 *c Probe added Incubation result Insitu hybridisation technique

Slide 34: 

In situ hybridisation

Slide 35: 

16 S r RNA SEQUENCING Gold standard for bacterial identification - 1500 bp Ribosomal RNAs in Prokaryotes 5S -120 -Large subunit of ribosome 16S -1500- Small subunit of ribosome 23S- 2900- Large subunit of ribosome

Slide 37: 

Pasteurellosis or pseudotuberculosis- Pasteurella piscicida Photobacterium damselae subsp. piscicida

Slide 38: 

Fatty acid profiling Chain length of structural fatty acids monomers Proteomics and protein profiling: characterization of the entire set of proteins expressed (from the genome) under specific conditions

Slide 39: 

THANK YOU