, Diagnosis an-&& of Shrimp Diseases - Central Institute of ...

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1. Sn~all pieces of ribbon can be put in a water bath containing warm water (or a tissuefloating bath). When the ribbon gets spread due to the high temperature of water, put aclean albumin coated slide underneath the ribbon and just lift the slide in such a way thatthe ribbon sticks to the surface of the slide. Drain the water and keep the slide in a slantingposition on a slide rack free from dust.2. Cut the ribbon into small pieces; keep one or more ribbons over the slide coated withadhesive, put a few drops of water on the slide so as to float the ribbon on the watersurface. Place the slide on a slide-warming table or pass it over a flame of spirit lamp. Twoneedles can be used to spread the ribbon to the maximum, drain the water and keep theslide in a slanting position.Whichever the method we follow, proper care should be talcen to avoid wrinltles in thesection. Improper spreading will interfere with staining and also microscopic observation.After proper drying, slides can be kept in a dust-proof box for some time for adequateadhesion. These slides can also be stored indefinitely.StainingBefore the tissue sections are subjected to staining, the sections should bedeparaffinized thoroughly and dehydrated. Hematoxylin and eosin staining can be employedfor routine histological preparation and this is the best method for histological diagnosis ofviral diseases. Steps involved in staining with H & E are as follows.A. Harris' haematoxylinHaematoxylin crystal5.0 g100% alcohol 50.0 mlAmmonium / potassium alum I00 gDist. Water1000 mlMercuric oxide2.5 gGlacial acetic acid (after cooling) 8 mlDissolve the haematoxylin in absolute alcohol; add the alum, previouslydissolved in hot distilled water. Heat the mixture to boiling points and add themercuric oxide, cool rapidly and filter. This stain is ready for use when cool. Stainingtime 2-3 min.B. Eosin1% aqueous eosin or alcoholic eosin
ProcedureDe-paraffinise the slid'c in sylene 1 h X 2Absolute alcohol15 min X290% alcoholI5 min70% alcohol15 min50% alcohol15 rninWater10 rnin X2Stain in hematoxylin2-5 minWash in wateradequatelyDe-stain in acid alcohol (if needed) adequatelyWash in tap water1 min50 % alcohol 30 mill70 % alcohol 15 minx2Stain in Eosin1 min90 % alcohol15 minx2Absolute alcohol20 min X 2Alcohol + Xylene (I : 1)15 lninXvlene 30 min X 2Mount in DPX and labelObserve under microscopeHistopathological characteristics of viral diseases of shrimpAlthough numerous viral diseases reported form penaeid shrimps, only three areprevalent in India. Following are the account of histopathological characteristics of theseiinportant viral diseases.Monodon baculovirus (MBV) infectionShrimp infected with MBV often appear clinically normal. Hepatopancreas is thetarget organ of this virus. The pathological changes include focaI to extensive necrosis oftubular epithelium of hepatopancreas. Single to multiple intra-nuclear, eosinophilic occlusionEbodies can be detected in the epithelial cells. In case of severe infection, necrotic tubularlumen may be seen filled with the sloughed epithelial cell debris and viral occlusion. Due tothe enlarged multiple occlusion bodies, the nuclei will appear hypertrophied withfragmentation and margination of chromatin material. Very often, hypertrophied nuclei mayappear as 'signet ring' with peripherally displaced and compressed nuclei. Thus, hypertrophy
Page 5 and 6: OVERVIEW OF STRATEGIES FOR PREVENTI
Page 7 and 8: in harmonizing aquatic animal healt
Page 9 and 10: SI-IRXMP DISEASES: GENEIL4L ASPECTS
Page 11 and 12: quantitative level of pathogen is i
Page 13 and 14: presence of prominent eosinophiiic
Page 15 and 16: Disease affected the shrimps of all
Page 19 and 20: BACTERIAL AND FUNGAL DISEASES OF SH
Page 21 and 22: Cause: The epibiotic bacteria stich
Page 23 and 24: PARASiTIC AND NON-INFECTIOUS IIISEA
Page 25 and 26: XOS-INFECTIOUS DISEASESNon-infectio
Page 27 and 28: INVESTIGATING SHRIMP DISEASES: OBSE
Page 29 and 30: lnay be a predisposing factor for b
Page 31 and 32: BACTERIOLOGICAL METHODSS.V. Alavand
Page 34 and 35: In place of ZMA, nutrient agar prep
Page 36 and 37: Interpretation: Positive reaction i
Page 38 and 39: Incubate at 37°C for 1-2 h.Examine
Page 40 and 41: Alkaligens: etc. are resistant. The
Page 42 and 43: Culture Media, Reagents and StainsC
Page 45 and 46: Differential characteristics of Vib
Page 47 and 48: Davidson's Alcohol Formatin Acetic
Page 49: Paraffin cold impregnationXylene an
Page 53 and 54: Biochemistry of Nucleic Acids with
Page 55 and 56: DNA e.rl1i6it.v base equivuleizcc!E
Page 57 and 58: hydrolyzed by dilute alkali. The su
Page 59 and 60: Polymerase Chain Reaction (PCR) is
Page 61 and 62: a number of viral, bacterial and pa
Page 63 and 64: from 15-30 bases. Negative controls
Page 65 and 66: PCR Primers for Indian WSVTwo sets
Page 67 and 68: ANTIOXIDANTS IN MANAGEMENT OF OXIDA
Page 69 and 70: Many potent chemical carcinogens ar
Page 71 and 72: cells. It has been shown that possi
Page 73 and 74: AFB, induced ~nutagenesis in Salmon
Page 75 and 76: FIGURE 2. An overview of oxygen rad
Page 77 and 78: were responsible for production los
Page 79 and 80: for Asiatic region. The participati
Page 81 and 82: charge of the DNA fragment. The neg
Page 83 and 84: European countries have in place th
Page 85: irth to a new concept to treatment

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