Aeromonas Salmonicida  Furunculosis of Salmonids  Goldfish Ulcer Disease  Carp Erythrodermatitis  Trout Ulcer Disease

5/6/2010

MAJ Eric D. Lombardini, VMD, MSc., DACVPM, DACVP Chief, Division of Comparative Pathology Armed Forces Radiobiology Research Institute Uniformed Services University of the Health Sciences

Overview:

 Fish research models  Fish necropsy  Research complications  Infectious diseases  Neoplastic diseases  Non‐infectious diseases Photo:10 & 104

Teleosts

 Bony fish  Greater than 18,000 species described

Photo: 11 Photo 12

1 5/6/2010

Extraordinary diversity

Photos: 1-4

Photo: 5

2 5/6/2010

Photo: 6

More Toadfish in Space: NASA will Study Balance in Two Wood’s Hole Toadfish, a Senator and Six Astronauts in Upcoming Shuttle Mission (1996 mission)

Photo: 7

1908 Nobel Prize in Physiology  Phagocytosis after experimenting on the larvae of starfish

Photo: 40

Photo: 41

3 5/6/2010

Historical use of fish in research

 Genetics  Embryology  Renal physiology  Endocrinology  Nerve physiology  Toxicology  Morphology  Nutrition  Pharmacology Photo: 8  Parasitology  Specific neoplastic models

More than just Zebrafish…

 Swordtail‐ Genetics of melanomas in Xiphoporus fishes  Trout‐coding sequences of the MHCII beta chain of homozygous rainbow trout  White perch‐Using white perch to study abnormal hepatic copper metabolism  Japanese medaka‐Freshwater fish in research on radiation biology  Goldfish‐The goldfish visual pathway: Intermediate filament proteins in nerve growth and development  Elasmobranchs‐In vitro metabolism of the pre‐carcinogen aflatoxin B1 by liver preparations of the calf, nurse shark and clearnose skate  Anglerfish‐Simultaneous assessment of prohormone transport and processing in four separate islet cell types  Salmon‐Advantages of using aquatic animals for biomedical research on reproductive toxicology  Eel‐Effects of anaesthesia and surgery on levels of adrenaline and noradrenaline in blood plasma of the eel  Toadfish‐Ichthyotoxins from the oyster toadfish  Etc…

Fish Models

Examples include:  Melanoma  Diabetes Mellitus  Neurofibromatosis  Hepatic copper storage disease (Wilson’s disease)  Oro‐facial‐digital syndrome  Hepatocellular carcinoma  Dehydration studies  Na/K‐ATPase activity in muscle  Renal excretion of xenobiotics  Genotoxic and epigenetic carcinogenesis  Gene regulation and developmental genetics  Mutagenesis

4 5/6/2010

Benefits of transgenic fish versus mammals in research

 External fertilization  Transparent embryos  Ploidy manipulation  Sex manipulation  Rapid growth  Total environment control  Increased availability of standardized genetic stock  Genetic mechanisms correlate well with rodents and humans Photo: 105

Transparent Zebrafish

Cell Stem Cell, Volume 2, Issue 2, 183-189, 7 February 2008

Embryology: Convergent extension movements and ciliary function are mediated by ofd1, a zebrafish orthologue of the human oral‐facial‐ digital type 1 syndrome gene

Glomerular function and structure in ofd1 MO‐injected embryos.

Hum Mol Genet. 2009 January 15; 18(2): 289–303.

5 5/6/2010

Neuroanatomy:

Figure 1 ‐ 4 day old zebrafish embryo labelled with SV2 and acetylated tubulin antibodies showing axon tracts(green) and neuropil(red)viewed from lateral(top) and dorsal(bottom) orientations.

www.ucl.ac.uk/zebrafish‐ group/research/neuroanatomy.php

Carcinogenesis: tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors

Fig. 5. Tumorigenesis features of tp53 M214K mutant zebrafish. (A‐D) When compared with wild‐type zebrafish (A and C), zMPNST development in tp53 mutant zebrafish is identifiable upon external observation because of ocular (B) or abdominal tumor localizations (D). (E‐H) When compared with wild‐type zebrafish (E and G), histopathology staining with hematoxylin/eosin reveals zMPNST in the eye (F) and abdominal cavity (H), as indicated by the stars (×4). (I‐K) Histopathological features of tumors (I) composed predominantly of spindle cells (J) and to a varying degree of epitheloid cells (K) are consistent with the diagnosis of zMPNST. [Bar, 200 μm (E, F, I‐K).]

PNAS January 11, 2005 vol. 102 no. 2 407‐412

Melanoma:

Review Article: Genetic and environmental melanoma models in fish Pigment cell and melanoma research; 2010 ‐Gordon‐Kosswig model (Sd‐ Spotted Disease) ‐Xmrk‐2 Oncogene overexpression ‐loss of the Diff tumor suppressor

6 5/6/2010

Toxicology: Identification of a Primary Target of Thalidomide Teratogenicity Takumi Ito, Hideki Ando, Takayuki Suzuki, Toshihiko Ogura, Kentaro Hotta, Yoshimasa Imamura, Yuki Yamaguchi, and Hiroshi Handa Science 12 March 2010: 1345‐1350.

MUTATION In embryos, thalidomide‐induced malformations, bottom, of a fin of a zebrafish, left, and a wing of a chick, right, using mutated cereblon, a protein. NYTimes 17 March 2010

Necropsy:

Fish Necropsy (Generic)

Swim Bladder

Gills

Heart Liver (Atrium;Ventricle; Bulbus arteriosus)

7 5/6/2010

Fish Necropsy (Generic)

Stomach Intestine/rectum

Pyloric ceca ovary

Fish Necropsy (Generic)

Kidney

8 5/6/2010

Pathology General Concepts  Necrotizing versus Granulomatous disease  Variation in leukocytes between species  74 bacterial pathogens of fish, dozens of fungi, hhddundreds of known viruses, thhdousands of parasites, innumerable toxins and miscellaneous or physiological pathologies.  Emerging diseases

Viral Infections  DNA:  Herpesvirus  Iridovirus  Adenovirus  RNA:  Picornavirus  Paramyxovirus  Reovirus  Togavirus  Birnavirus  Rhabdovirus  Retrovirus

Piscine iridovirus  Gross findings:  Range in appearance from miliary pale nodules to a cutaneous mass effect.  Histopathological findings:  Markedly hypertrophic dermal fibroblasts (lymphocystis cells). Cells may measure > 300 µm with vacuolated to granular pale basophilic cytoplasm surrounded by a 10‐ 30µm amphophilic hyaline wall. Occasionally there is the presence of basophilic fibrillar inclusion material.

9 5/6/2010

Lymphocystis

Photo: 20

Lymphocystis

Photo: 46

Lymphocysitis

Photo: 45

10 5/6/2010

Lymphocystis

Photo: 21

11 5/6/2010

 Journal of Fish Diseases, Volume 33 Issue 2 , Pages 93 ‐ 186 (February 2010) Iridovirus infections in finfish –critical review with emphasis on ranaviruses (p 95‐122)

Photo: 114 Liver of redfin perch, found dead 10 days post‐bath inoculation with epizootic haematopoietic necrosis virus. Hepatocytes that surround a focus of hepatic necrosis frequently contain basophilic intracytoplasmic inclusion bodies (arrows) (H & E, bar = 50 µm).

Carp Pox (CyHV‐1)

Photo: 34 & 35

Carp Pox

Photo: 44

12 5/6/2010

KHV  Koi herpesvirus (KHV) is a highly contagious viral disease that may cause significant morbidity and mortality in common carp (Cyprinus carpio) (Hedrick et al. 2000; OATA 2001)  The white patches are due to necrosis (death) of the gill tissue. Gill lesions caused by KHV disease are the most common clinical signs in affected koi. Other external signs of KHV may include bleeding gills, sunken eyes, pale patches or blisters on the skin.

Koi herpesvirus (KHV) disease (CyHV‐3)

Photo: 14

Koi Herpesvirus

Photos: 70 & 71

13 5/6/2010

KHV

Photo: 80

IcHV‐1

Photo: 99 & 100

VHS (Viral hemorrhagic septicemia)

 Affects >50 species of fish (Fresh water and marine)  Multiple strains  Family Rhabdoviridae  Gross: Visceral, cutaneous and muscular petechial hemorrhage  Exopthalmus, ascites, eccymoses around eyes, skin, gills and fins  Histo: Variable necrosis of kidneys (hematopoetic 1st), spleen, liver, and skeletal muscle

en.wikipedia.org/wiki/File:VHS.png

14 5/6/2010

VHS

Photo: 29

VHS

Photo: 30

Spring Viremia of Carp

Photo: 83

15 5/6/2010

Ulcerative Dermatitis in Winter Flounder

Photo: 31

Bacterial Infectious Diseases  In zebrafish, the pneumatic duct acts as the primary point of invasion for systemic fungal and bacterial infections, spreading to the gas bladder  Primarily opportunistic infections of which the 1° agents: Aeromonas sp. and Pseudomonas sp.  Clinical signs of Gram negative septicemia are non‐ specific and include: ascites, exopthalmia, cutaneous hemorrhages (body and fins), cutaneous ulceration

Non‐specific signs of septicemia  Ascites or “Dropsy”

Photo: 43 &45

16 5/6/2010

Non‐specific signs of septicemia  Exopthalmia

Photo: 28

Non‐specific signs of septicemia  Cutaneous hemorrhages

Photo: 19

Non‐specific signs of septicemia  Ulceration

Photo: 32

17 5/6/2010

Aeromonas salmonicida  Furunculosis of salmonids  Goldfish Ulcer Disease  Carp Erythrodermatitis  Trout Ulcer Disease.

 Several other species of AeromonasPhoto: 36 , including: A. hydrophila, A. formicans, A. liquefaciens, and A. hydrophila complex are capable of causing a disease known as “Motile Aeromonas Septicemia” or “Bacterial Hemorrhagic Septicemia”.

Furunculosis

 Aeromonas salmonicida  (4 subspecies: salmonicida, achromogenes, masoucida and smithia  Opportunist resulting in septicemia  Predominantly salmonid fishes, but also affects goldfish and other cyprinids.  “Furunculosis” is derived from the presence of “blisters” or furuncules on the surface of chronically infected fish  Significant resistance to both Terramycin and sulfamerazine

Furunculosis

Photo: 78

18 5/6/2010

Photos: 37 & 91

Koi Ulcer Disease

Photo: 33

Yersinia ruckeri

Photos: 48 & 49

19 5/6/2010

Enteric septicemia of catfish (ESC)

 Edwardsiella ictaluri

Photo: 53

Edwardsiella tarda (Emphysematous Putrefactive Disease EPD)

Photos: 38 & 39

Renibacterium salmonarium  Gross findings: Muscular cavitation; Vesicular dermatitis (Spawning rash); necrotizing nephritis & splenitis.

 Histopathological findings: Granulomatous and necrotizing nephritis, splenitis, hepatitis, endocarditis and pancreatitis; occasionally with giant cell formation. Histiocytes are expanded by intracytoplasmic gram‐positive bacteria

20 5/6/2010

Renibacterium salmonarium

Renibacterium salmonrium

Streptococcus

Photo: 17

21 5/6/2010

Streptococcus  Clinical findings: Ocular hemorrhage, cutaneous hemorrhage, exopthalmia, corneal opacity, dropsy and ulceration.  Gross Findings: Abdominal serosanguinous fluid, splenomegaly, hepatic pallor, endocarditis and nephritis. Many Streps infect the brain and nervous system of fish, explaining the erratic swimming frequently observed in infected fish.  Isolated Streptococcal pathogens in fish:  S. iniae  S. difficilis  S. parauberis  S. milleri  S. shiloi

Mycobacteriosis

 Mycobacterium marinum  Mycobacterium chelonae  Mycobacterium fortuitum

Photo: 89

Mycobacteriosis

Photo: 13 Abdominal cavity: Peritonitis, hepatitis, splenitis, granulomatous, multifocal, severe

22 5/6/2010

Mycobacterium marinum

Mycobacteriosis  Mycobacteria as Environmental Portent in Chesapeake Bay Fish Species, Emerging Infectious Diseases: Volume 13, Number 2–February 2007

 Occurrence of Mycobacterium spp. in ornamental fish in Italy, Journal of Fish Diseases, 2008 Jun;31(6):433‐41.

 Husbandry stress exacerbates mycobacterial infections in adult zebrafish, Danio rerio (Hamilton), Journal of Fish Diseases, 2009, 32, 931–941

Gliding bacteria

Photos 18 & 90

23 5/6/2010

Flavobacterium columnare

Photo: 42 & 98

Photo: 50

Histopathology and Ultrastructure of Segmented Filamentous Bacteria– Associated Rainbow Trout Gastroenteritis; Veterinary Pathology 47(2) 220‐230

Photo: 93

24 5/6/2010

Piscirickettsia salmonis

Photo: 81

Protozoae

Ichthyobodo necator Epistylis

Trichodina Glossatella

Microsporidiae  Glugea, Ichthyosporidium, Loma, Pseudoloma, Microgemma, Microsporidium Mrazekia, Nosema, Pleistophora, Spraguea, Tetramicra, and Theragra.

Photo: 54

25 5/6/2010

Microsporidiae

26 5/6/2010

Pleistophora hyphessobryconis

Pseudoloma neurophilia

Photo: 75

Glugea stephani

Photo: 67

27 5/6/2010

Glugea sp.

Photos: 68 & 69

Myxosporidiae

Photo: 51

Henneguya salminicola

Photos: 63 & 94

28 5/6/2010

Henneguya sp. xenomas

Photos: 65 & 66

Myxoma and trematode

Hamburger gill disease

29 5/6/2010

Myxobolus cerebralis  Gross Findings: Severe scoliosis, frequently with black discoloration of the tail, and with chronicity, misshapen head and jaws  Histopathological Findings: Cartilage degeneration and necrosis, granulomatous chondritis with intralesional ovoid to ellipsoidal spores (5‐15µm) with 2 piriform‐shaped polar capsules at the anterior end

Photos: 119

Myxobolus cerebralis

Photos: 76 & 77

Myxobolus cerebralis

30 5/6/2010

Myxobolus sp.

Figure 2—Photomicrograph of an impression smear prepared from a biopsy specimen of the oral cavity mass in the goldfish in Figure 1. Notice a large cluster of epithelial cells (blue) and numerous mature Myxobolus sp spores (red). Modified acid‐fast stain; bar = 25 μm. JAVMA, Vol 236, No. 6, March 15, 2010

Myxobolus sp.

 Figure 3—Photomicrograph of a paraffin‐embedded section of a biopsy specimen of the oral cavity mass in the goldfish in Figure 1. The mass tissue contains a single large, late‐stage, polysporous, histozoic plasmodial pseudocyst (borders of this structure are demarcated by arrows) and evidence of inflammation. H&E stain; bar = 100 μm. JAVMA, Vol 236, No. 6, March 15, 2010

Myxozoa

Renal myxozoanosis in weedy sea dragons, Phyllopteryx taeniolatus (Lacepe` de), caused by Sinuolinea phyllopteryxan. sp. Journal of Fish Diseases 2008, 31, 27–35

31 5/6/2010

 Tetrahymena sp. infection in guppies, Poecilia reticulata Peters: parasite characterization and pathology of infected fish, Journal of Fish Diseases 2009, 32, 845–855  Severe Scuticociliate (Philasterides dicentrarchi) Infection in a Population of Sea Dragons, Vet Pathol 45:546–550 (2008)

Photos: 115 & 116

Tetrahymena corlissi

32 5/6/2010

Tetrahymena corlissi

Ichthyophthirius multifiliis  Gross findings:  Up to 1 mm elevated nodules in a miliary pattern along body, gills and fins  May coalesce or progress to erosion and ulceration due to irritation  Histopathological findings:  Oval holotrich ciliated parasite which forms intraepithelial cysts composed of a 1‐2 µm hyaline wall.  Small circular oral opening, dark granules and a horseshoe/crescent shaped basophilic macronucleus  +/‐ Hyperplasia of neighboring epithelium

Photos: 95 & 96

33 5/6/2010

Ichthyophthirius multifiliis

Piscinoodinium sp.

34 5/6/2010

Piscinoodinium sp.

Ichthyobodo necator

Photos: 118

Coccidiae

Photos: 101 & 103

35 5/6/2010

Hole‐in‐the‐Head Disease Head and Lateral Line Erosion Hexamita sp.?

Photo: 52

Fungal

Photos: 79 & 92

Saprolegnia

Photo: 22

36 5/6/2010

Saprolegnia

Photo: 23

Parasitic  Nematodes  Cestodes  Trematodes  Pentastomes  Arthropods

Nematodes

Photo: 47

37 5/6/2010

Eustrongylides sp. Eustrongylides ignotus infecting commercial bass, and other fish in the southeastern USA, Journal of Fish Diseases 2009, 32, 795–799

Photos: 15 & 26

Acanthocephalids

Photo: 107

Cestodes

38 5/6/2010

Asian Fish Tapeworm: Bothriocephalus acheilognathi

Photo: 97

Ligula intestinalis

Photo: 24

Digenetic trematodes

Photomicrographs: MAJ Jeremy Bearss

39 5/6/2010

Black spot disease

Photo: 86

Trematode metacercariae

Photo: 27

40 5/6/2010

Monogenean trematodes

Photomicrographs: MAJ Jeremy Bearss

41 5/6/2010

Diplostomum sp.

Photo: 109

Pentastomes

Photo: 16 & 108

Arthropods

Photo: 25 & 117

42 5/6/2010

Neoplastic

Photo: Sarah Johnson & 120

Melanoma

43 5/6/2010

Fibrosarcoma

Photo: 61

Bi‐colored Damselfish Schwannoma

Photo: 73

Neurofibroma

44 5/6/2010

PNST

Peripheral Nerve Sheath tumor

Dermatofibroma of Angelfish

Photo: 113

45 5/6/2010

Walleye Dermal sarcoma

Photo: 87

Walleye Dermal Sarcoma

Photo: 88

Lymphoma

Photo: 85 & 110

46 5/6/2010

Lymphoma

Basal cell carcinoma

47 5/6/2010

Branchial carcinoma

Seminoma

Photo: 58

Seminoma

Photo: 59

48 5/6/2010

Nephroblastoma

Ependymoma

49 5/6/2010

Neuronal Embryonal Tumors in Fish; Vet Pathol published online 31 December 2009

Photo: 92

Hepatocellular carcinoma

Photo: 102

Cholangiocarcinoma

Photo: 60 112

50 5/6/2010

Papilloma (Cyrinid herpesvirus 1)

Photo: 72

Viral papillomas of Brown bullhead catfish

Photo: 111

Reported Zebrafish tumors  Seminoma (Most common spontaneous  Myxoma/myxosarcoma tumor)  Fibrosarcoma  Ovarian papillary adenocarcinoma  Chordoma  Adenocarcinoma  Hemangioma (Retrobulbar hemangioma)  Small cell carcinoma of intestine at Ampulla of  Rhabomyoma/rhabdomyosarcoma Vater  Chondrosarcoma  Hepatocellular adenoma/carcinoma  Osteoma/osteosarcoma  Biliary carcinoma/Cholangiocarcinoma  PNST  Hepatoblastoma  Melanoma  Nephroblastoma  Complex‐compound odotoma of the  Renal cell carcinoma pharyngeal teeth  Thyroid adenocarcinoma  Neuroblastoma/neuroectodermal tumor  Ultimobranchial tumor  Medulloepithelioma  Gas bladder adenoma/adenocarcinoma  Ganglioglioma  Pancreatic acinar cell adenoma/carcinoma  Branchioblastoma  Pancreatic ductal adenoma  Epidermal papilloma  Islet cell adenoma  SCC  Lymphoma  Teratoma  Acute lymphoblastic leukemia

51 5/6/2010

Miscellaneous

Figure 1. Internal view of stomach prolapse Figure 2. Adult female sand tiger shark with in a sand tiger shark. Note also the spleen stomach prolapse through the right third gill (arrow) is pulled up into the ventral throat slit. region. Journal of Fish Diseases 2008, 31, 311–315; Gastric prolapse in sand tiger sharks

Pericardial effusion and dilated cardiomyopathy

Photo: 55

Nephrocalcinosis

52 5/6/2010

Hepatic megalocytosis

Photo: 56

Gas Bubble Disease

Photo: 57

Gas Bubble Disease: Supersaturation

Photo: 84

53 5/6/2010

Goiter

Cardiomyopathy syndrome

Photo: 82

54 5/6/2010

 Knockdown of Bicaudal C in Zebrafish (Danio rerio) Causes Cystic Kidneys: A Nonmammalian Model of Polycystic Kidney Disease  Bouvrette et al.  Comparative Medicine, Volume 60, Number 2, April 2010 , pp. 96‐106(11)

Photo: 106

Dilatation of the pronephric ducts (arrows) and cysts which form in the single glomerulus (arrowheads)

Cardiac dissecting hemorrhage

Figure 1 Dissecting haemorrhage of the ventricle in rainbow trout. (a) Cut surface of the formalin‐fixed ventricle showing haemorrhage along the lateral wall. (b) Sickle‐shaped haemorrhage between compact and spongious ventricular myocardium (H&E, bar = 1 cm). (c) Detail showing haemorrhage and compact myocardium with focus of inflammation to the left (H&E, bar = 50 lm). Journal of Fish Diseases 2009, 32, 1041–1043; Dissecting haemorrhage in rainbow trout

Photography credits:  65. http://www.maine.gov/ifw/fishing/health/vol4issue5.htm  66. http://ag.ansc.purdue.edu/courses/aq448/images/henneguya1.jpg  1.en.wikipedia.org/wiki/File:Leafydragon.jpg  67. http://upload.wikimedia.org/wikipedia/commons/3/3e/Glugea_stephani.jpg  2.en.wikipedia.org/wiki/File:Pterois_volitans_Manado‐e_edit.jpg  68.http://www.wildaboutbritain.co.uk/archive/data/517/glugea_anomala_stickleback.jpg  3. en.wikipedia.org/wiki/File:Georgia_Aquarium_‐_Giant_Grouper_edit.jpg  69.http://tolweb.org/onlinecontributors/app?page=ContributorImagesPage&service=external&sp=3682  4. en.wikipedia.org/wiki/File:White_shark.jpg  5. en.wikipedia.org/wiki/File:Lamprey_mouth.jpg  70.http://www.koihealth.org/images/Interocean_KHV/images/Allen_Hsu_KHV%20272_jpg  6. en.wikipedia.org/wiki/File:GambianMudskippers.jpg  71.http://www.koihealth.org/images/Interocean_KHV/images/Allen_Hsu_KHV%20080_jpg  7. en.wikipedia.org/wiki/Oyster_toadfish  72.http://www.koihealth.org/images/Interocean_KHV/pages/Allen_Hsu_KHV%20180_jpg.htm  8. blogs.nature.com/nm/spoonful/zebrafish.jpg  73. http://ehp.niehs.nih.gov/docs/1995/103‐3/niehsnewsfigfish2.GIF  9. dx.doi.org/10.1371/journal.pbio.0020148  74. http://zebrafish.org/zirc/health/Disease_images/ultimobranchialTumors4.jpg  10. www.genome.gov/pressDisplay.cfm?photoID=95  11. www.realscience.us/blog/wp‐content/uploads/2007/06/zebrafish‐embryo.jpg  75. http://zebrafish.org/zirc/health/diseaseManual.php  12.images.google.com/imgres?imgurl=http://dreamsforpeace.files.wordpress.com  76. http://en.wikipedia.org/wiki/File:Deformed_Brook_Trout.jpg  13.http://edis.ifas.ufl.edu/LyraEDISServlet?command=getImageDetail&image_soid=FIGURE%201&document_soid=VM055&documen  77. http://en.wikipedia.org/wiki/File:Whirling_disease_pathology.jpg t_version=42850  78. http://www.fws.gov/midwest/lacrossefishhealthcenter/Index.htm  14. http://edis.ifas.ufl.edu/vm113  15.http://edis.ifas.ufl.edu/LyraEDISServlet?command=getImageDetail&image_soid=FIGURE%2012&document_soid=FA091&docume  79. http://biology.unm.edu/ccouncil/Biology_203/Summaries/Protists.htm nt_version=1943678729  80. http://www.fao.org/docrep/009/a0192e/A0192E07.htm  16.http://edis.ifas.ufl.edu/LyraEDISServlet?command=getImageDetail&image_soid=FIGURE%204&document_soid=FA090&docume  81. http://www.cdc.gov/ncidod/eid/vol3no2/fryer.htm nt_version=37638   17.http://edis.ifas.ufl.edu/LyraEDISServlet?command=getImageDetail&image_soid=FIGURE%201&document_soid=FA057&document 82. http://www.marlab.ac.uk/Delivery/standalone.aspx?contentid=2015 _version=48516  83.http://www.lethbridgecollege.ab.ca/grasscarp/images/stories/projects/Spring%20Viremia%20in%20Carp/D  18. http://zebrafish.org/zirc/health/Disease_images/TailRot4.jpg SC01878.JPG  19. http://zebrafish.org/zirc/health/Disease_images/Erythema.jpg  84. http://www.advancedaquarist.com/2010/3/fish  20. http://vp4.afip.org/wsco/do_slideshow2.php?pic_series=082303&cYear=2008  85. http://www.ramp‐alberta.org/ramp/gallery.aspx?galleryimage=493  21. http://vp4.afip.org/wsco/do_slideshow2.php?pic_series=082303&cYear=2008   22. http://www.frw.ca/albums/Correcting‐Stormwater‐Mistakes 86. http://www.ramp‐alberta.org/ramp/gallery.aspx?galleryimage=503  23.http://media.photobucket.com/image/fish%20fungus/aquaticquotient/disease/fungus.jpg  87. http://www.ohio.edu/news/pix/FISH.JPG  24. http://apscience.org.au/projects/APSF_04_2/apsf_04_2.htm  88. http://i244.photobucket.com/albums/gg22/way2phat/walleyefungus2.jpg  25. http://apscience.org.au/projects/APSF_04_2/apsf_04_2.htm  89. http://ag.ansc.purdue.edu/courses/aq448/images/tangmycobact.jpg  26. http://apscience.org.au/projects/APSF_04_2/apsf_04_2.htm  90. http://www.fishdoc.co.uk/photos/Fin_rot_02.JPG  27. http://apscience.org.au/projects/APSF_04_2/apsf_04_2.htm   28. http://www.focusonfishhealth.org/signs‐of‐disease.php 91. http://www.koi ‐fish.co .uk/acatalog/ulcer .jpg  29. http://www.focusonfishhealth.org/signs‐of‐disease.php  92. http://kentsimmons.uwinnipeg.ca/16cm05/16labman05/lb2pg3_files/saprolegnia4.jpg  30. http://www.focusonfishhealth.org/signs‐of‐disease.php  93. http://vet.sagepub.com/content/47/2/220.full.pdf+html  31. www.mwra.state.ma.us/harbor/omsap/graphic/omsap_050811_flounder.ppt  94. http://www.scielo.br/scielo.php?pid=S0074‐02762005000200011&script=sci_arttext  32. http://www.blueridgekoi.com/images/categories/C76.jpg  95. http://nippyfish.net/Ich4.jpg  33. http://www.ponddoc.com/koihospital/Diagnostic/Bacterial/AdvancedUlcer.jpg   34. http://www.backyardpuddle.ca/images/healthpix/smapril02‐4.jpg 96. http://www.vet.cornell.edu/PUBLIC/FishDisease/AquaticProg/highlights/Ich/ICH1.JPG  35.http://www.spfdwatergardensoc.bizland.com/sitebuildercontent/sitebuilderpictures/pox.jp36.http://www.cnr.vt.edu/fisheries/afs/  97. http://www.cnr.vt.edu/fisheries/afs/AFS_education_fisheries_techniques_visuals_chap_14_add.htm AFS_education_fisheries_techniques_visuals_chap_14_add.htm  98. http://www.cnr.vt.edu/fisheries/afs/fisheries_techniques  37.http://www.aquacultuur.wur.nl/NL/Visteelt/Productie+en+teelt/Ziekten/Bacteri%C3%ABn  99. http://www.cnr.vt.edu/fisheries/afs/fisheries_techniques/Chapter14/Channel%20catfish%20virus%202.jpg  38.http://www.aquacultuur.wur.nl/NL/Visteelt/Productie+en+teelt/Ziekten/Bactari%C3%ABn  100. http://www.cnr.vt.edu/fisheries/afs/fisheries_techniques/Chapter14/Channel%20catfish%20virus%201.jpg  39. http://www.time4fish.net/FDB.html  40. http://phlogthat.wordpress.com/2007/07/16/two‐star‐fish‐making‐love‐in‐the‐sand‐at‐morro‐bay‐ca/  101. http://www.alaineknipes.com/parasite_photos/index.html  41. http://www.odec.ca/projects/2007/sank7b2/figure1.jpg  102. http://www.nature.com/nprot/journal/v5/n3/thumbs/nprot.2010.8‐F5.jpg  42. http://www.blueridgekoi.com/Disease‐Fish‐Health‐sc‐17.html  103. http://www.alaineknipes.com/parasite_photos/index.html  43. http://www.flippersandfins.net/Dropsy.htm  104. http://upload.wikimedia.org/wikipedia/commons/7/75/F_de_Castelnau‐poissonsPl12.jpg  44. http://www.blueridgekoi.com/Disease‐Fish‐Health‐sc‐17.html  105. http://www.sciencedaily.com/images/2009/05/090507121953‐large.jpg  45. http://www.drpez.net/panel/showthread.php?t=167187  46. http://www.drpez.net/panel/showthread.php?t=167187  106. http://renalfellow.blogspot.com/2009/05/zebrafish‐model‐of‐cystic‐kidney.html  47. http://www.fishbase.org/diseases/DisPic/NemtodD5.jpg  107. http://www.mybay.umd.edu/parasites.html  48. http://web.abo.fi/instut/fisk/Fig/yr1.jpg  108. http://edis.ifas.ufl.edu/pdffiles/FA/FA11400.pdf  49. http://www.daff.gov.au/_media/images/animal‐plant/aquatic/aquatic‐cd/enteric‐redmouth1.jpg  109. http://www.thefishsite.com/articles/102/invasion‐of‐the‐eye‐flukes\  50. http://en.wikipedia.org/wiki/Columnaris  110. http://www.seagrant.umn.edu/fisheries/img/parasites/lymphosarcoma_b.jpg  51. http://en.wikipedia.org/wiki/File:Henneguya_zschokkei.jpg  52. http://en.wikipedia.org/wiki/File:Oscar2.jpg  111. http://www.glfc.org/tumor/f2b.gif  53.http://coursewares.mju.ac.th/section2/fb406/0002lecture/htm/003allchapter3/image/image003_5/edwardsiella.gif  112. http://www.glfc.org/tumor/f5a.gif  54. http://zebrafish.org/zirc/health/diseaseManual.php  113. http://www.ultimatebettas.com/index.php?showtopic=8863  55. http://zebrafish.org/zirc/health/Disease_images/BigHeartGross4.jpg  114. http://www3.interscience.wiley.com/cgi‐bin/fulltext/123227302/PDFSTART  56. http://zebrafish.org/zirc/health/diseaseManual.php  115. http://vet.sagepub.com/content/45/4/546.full.pdf+html  57. http://zebrafish.org/zirc/health/Disease_images/GasBubbleDis4.jpg  58. http://zebrafish.org/zirc/health/Disease_images/GrossSeminoma4.jpg  116. http://vet.sagepub.com/content/45/4/546.full.pdf+html  59. http://zebrafish.org/zirc/health/Disease_images/seminoma24.jpg  117. http://www.state.me.us/ifw/fishing/health/vol2issue2.htm  60. http://zebrafish.org/zirc/health/Disease_images/LiverTumorsDMBA4.jpg  118. http://www.drpez.net/panel/showthread.php?t=330799  61. http://zebrafish.org/zirc/health/Disease_images/SpincleCellGrossB4.jpg  119. http://www.forumaquario.com.br/portal/tx_doencas.html  62. http://zebrafish.org/zirc/health/Disease_images/SpindleCellEye4.jpg  120. http://www.epa.gov/glnpo/aoc/presque/catfish‐tumors.jpg  63.http://en.wikipedia.org/wiki/File:Henneguya_salminicola_in_flesh_of_coho_salmon,_BC,_Canada.JPG  64. http://www.virtualslides.psu.edu/zebrafish.jsp?compare1=08B98770‐2FC7‐5057‐42CF‐8695A7F9C030&

55 5/6/2010

References  Histology:  1. http://zfatlas.psu.edu/reference.php  2. http://training.fws.gov/CSP/fish/histo1.html  3. http://aquaticpath.umd.edu/fhm/  4. http://www.vspo.us/vspo  Neoplasia:  5. DrD . JanJ SpitbergenSitb POLA notest 2008  6. Registry for Tumors in Lower Animals  General:  7. Edward Noga, Fish Diseases, Diagnosis and treatment, 2000  8. Ronald Roberts, Fish Pathology, 2003  9. Greg Lewbart, Ornamental fish (Self assessment colour review), 1998  10. Gary Ostrander, The laboratory fish, 2000  11. Erwin Amlacher, Textbook of fish diseases, 1970  12. Fish diseases vol 1 and vol 2, (Ed.) J Eiras et al., 2008