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C. Brayton September 2007
07AquaPathSumTables
Xenopus
Anura
► Tailless amphibians (only larval stages have tails).
► Head and trunk are fused and toes are webbed.
► 10 vertebrae; ribs are reduced or absent.  positive pressure breathing by suprahyoid muscles (in floor of mouth).
► Do not drink, absorb water through highly permeable, specialized area of skin in pelvic region.
► Skin needs to be kept moist for carbon dioxide and water exchange; largely responsible for respiration via its outer surface.
► Adult anuran lungs have vesicles for some gas exchange.
Xenopus spp
► 6 to 14 species of tongueless, aquatic African frogs (family Pipidae) having small black claws on inner 3 toes of the hind limbs
o Xenopus tongue is completely attached to floor of mouth instead of attached at front of mouth and folded back, as in other anurans
► Hind limbs are large and well adapted to a totally aquatic environment. The much smaller forelegs are used to push food into the mouth
► Normal behavior is for these frogs to spend most of their time lying motionless below water surface.
Interesting X laevis features (phenotypes)
General /Misc
18 chromosome pairs (allotetraploid) ( X tropicalis has 10 pairs (diploid)
Aquatic
Regenerate lost limbs
Large fat bodies attached to each kidney
Mucous layer important for gas/nutrient/waste exchange important
INTEGUMENT
osmoregulatory function
Glands
Mucous + serous secrete immunoprotective substances etc
Agastric (esophagus  intestine); No oral teeth
DIGESTIVE
Teeth
‘teeth’ in maxilla for gripping
Liver
Melanomacrophages
RESPIRATORY
Skin
Critical in osmoregulation & Gas exchange
Gills
In tadpoles
GENITOURINARY
CARDIOVASC
Heart
3 chambers
Dorsal lymph sacs
= paired lymph hearts located dorsally on either side of last vertebrae.
ENDOCRINE
HEMATOPOIETIC
Nucleated RBC’s & heterophils
MISC
Sensual tubercles
Dehydrate rapidly out of water
Initial barrier to infection
Also eliminate most nitrogenous waste
Useful for IV injections
Blood collection by clipping toe web or cardiac
puncture
~ fish Lateral Line organ sensitive to changes in hydrostatic stimuli and sound
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Xenopus
Spontaneous XENOPUS Pathology (phenotypes) NON Neoplastic
XENOPUS Pathology (phenotypes) Non Neoplastic
SYSTEMIC (multisystem
METABOLIC NUTRITIONAL
Water quality
Chlorine/chloramine ammonia, pesticides,
disinfectants, heavy metals
r/o water quality, infections
Lethargy skin slough death
INTEGUMENT
DIGESTIVE
Rectal / cloacal prolapses
RESPIRATORY
Gas bubble disease
ENDOCRINE
GENITOURINARY
Coelomitis
CARDIOVASCULAR
MUSCULOSKELETAL
Often secondary to ascites
Bubbles in skin & foot webs, death usually dt sepsis
(gas) supersaturated water
Coelomitis
r/o iatrogenic (surgical egg harvest)
Spontaneous XENOPUS Pathology (Phenotypes) Neoplastic
XENOPUS Neoplasms
Hematopoietic
Lymphoma
Visceral organomegaly with intense lymphoid cell infiltration/effacement
Sporadic-rare
Various
Viral? sporadic (Balls 1965)
VIRUSES in XENOPUS
Prevalence
Dx/Detection
Virus order /family
Virus
Iridoviridae
Ranavirus
Various spp specific ? ranaviruses
tadpole viruses
NON Viral infectious/infesting agents in Xenopus
Site
Stains
AGENT in XENOPUS
Primary
?
II. BACTERIA
Aeromonas hydrophila,
Proteus hydrophilus
Pseudomonas hydrophilus etc
Gram Negative septicemia
Skin etc
sepsis
G-
Disease ~ Pathology ~ Phenotypes
Comments
Immunodeficient may be infected
Xenopus model for Frog virus 3 (FV3)
Tropism for Kid prox tubule
Prev
Detection
Disease ~ Pathology ~ Phenotypes
Comments
Common
Cult
Path
hemorrhage congestion swollen frog
+/- serosanguinous exudate
G- colonies in necrotic areas
G- bacteremia
Ubiquitous opportunists + stress 
RED LEG
R/o chlamydiosis mycobacteriosis …
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Xenopus
AGENT in XENOPUS
Site
Primary
Chlamydia psittaci
C pneumoniae
Stains
?
Prev
Detection
Sepsis +/- necrosis of liver, spleen, kidneys,
heart
Basophilic intracytoplasmic inclusions of
coccoid organisms in hepatic and splenic
sinusoidal macrophages and glomeruli
Giemsa
Mycobacterium spp.
M gordonae, liflandii,
marinum, ranae, ulcerans,
xenopi etc
ANYWHERE
Salmonella
sepsis
AF
Disease ~ Pathology ~ Phenotypes
Common
Path PCR
Cult
+/- emaciation, ulcers,
+/- necrosis in viscera
+/- gross nodules (granulomas)
with AFB.
sporadic
Comments
R/o Red leg, mycobacteriosis …
Zoonotic concern
Mycobacterioses transmitted by water
& cannibalism.
Zoonotic via cuts or abrasions  skin
nodules.
Most cold adapted slow growing
Zoonotic concern
III. FUNGI
Batrachochytrium
dendrobatidis
Chytrid Dz
Skin etc
Exophiala sp
Skin etc
Mucor etc
Skin etc
Sporadic
Path
Proliferative dermatitis +mature sporangia up
to 20 u diam with spores & discharge
tubes in stratum corneum
Immature zoosporangia <10u
Granulomas, viscera or skin
Pigmented hyphae; septate branching,
irregular width
Or pigmented yeast forms
Should not need silver or PAS stains to see
them
Ulcers granulomas with Non pigmented
zygomycetes
Significant morbidity/mortality in
susceptible specie/
X – subclinical reservoirs?
Phaeohyphomycosis (hyphae)or
chromomycosis dt pigmented yeasts
Chromomycosis dt pigmented yeasts
fish may become darker, lethargic, or
erratic swimming
-- PROTISTS
Cottony material on skin etc
Broad nonseptate branching hyphae that
produce motile flagellated zoospores in
terminal sporangia.
Saprolegnia
Normal water inhabitants that invade
traumatized skin. Esp with
SUBoptimal (low) temps
IV. PROTOZOA
Costia, Oodinium, Trichodina,
Vorticella etc
Flagellates
Nyctotherus sp
Ciliates
Balantidium xenopodis
Ciliates
Protoopalina xenopodus
Ciliates
Skin
Intestine
Intestine
Intestine
Looks like paramecium
usually in lumen – invasive?
Ciliate with large macronucleus
usually in lumen – invasive?
Looks like opalinid
Pathogenic?
Pathogenic?
Pathogenic?
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Xenopus
AGENT in XENOPUS
V. HELMINTHS
Site
Primary
NEMATO
DES
Stains
?
Prev
Detection
Disease ~ Pathology ~ Phenotypes
Comments
Larval migrations
anywhere
CESTODE
S
?
Proliferative dermatitis with adults & larvae in
skin tunnels – patchy sloughing
Larvated eggs in utero
Pneumonia failure to thrive.
Adult worms in lungs.
Eggs larvae in gut coelomic lymph spaces
Larval nematodes in kidney muscle etc
Cephalochlamys namaquensis
Intestine
?
Adults (+ larvae?) in intestine
African origin Unlikely in clean
quarantined stock ?
African origin?
Unlikely in clean quarantined stock ?
haptor = attachment organ
posterior with 0-2 pair of anchors +
peripheral small hooklets
hemorrhage necrosis
African origin ?
Monogenetic = single host
Cystacanth
1 report from feral X laevis
Capillaria xenopodis
Pseudocapillaria xenopodis
Skin
Rhabdias spp
Lung
worm
TREMATODES
Monogenetic trematodes e.g.
Protopolystoma xenopodis
Gyrodactylus gallieni
Acanthocephalan
Acanthocepahlus sp
VI. ARTHROPODS
MITES
Crustacea
ns
Common
Direct life cycle no int host
strongyloid lungworms
Migrating Rhabdias etc spp?
??
References / Resources
Kuperman & al. 2004 Parasites of the African Clawed Frog, Xenopus laevis, in Southern California, U.S.A Comparative Parasitology 71 (2) pp. 229–232
http://www.bioone.org/perlserv/?request=get-document&doi=10.1654%2F4112
Tinsley & Kobel ed.’s 1996. The Biology of Xenopus. Oxford
Trans NIH Xenopus initiative http://www.nih.gov/science/models/xenopus/
Xenopus 1 http://www.xenopusone.com/
Xenbase: Xenopus laevis and tropicalis biology and genomics resource http://vize222.zo.utexas.edu/common/
Xenopus Express http://www.xenopus.com/
Xenopus info at http://www.dbsm.uninsubria.it/biocel/xenen.htm
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Danio (Zebrafish)
Zebrafish Danio rerio
Interesting ZFISH teleost features (phenotypes)
General /Misc
25 chromosome pairs
Abundant transparent eggs  transparent embryos – develop in 2-4d
Males = slender = torpedo-shaped, +/- gold on belly, fins
Females have 0-,in gold & are fat when filled with eggs
Not keratinized  gas/nutrient/waste exchange important osmoregulatory
INTEGUMENT
function
Cuticle
Covered with mucus, mucopolysaccharides, immunoglobulins, free fatty acids.
Epidermis
stratified squamous cells, 4-20 cells thick
Scales
calcified plates that originate in dermis
includes melanophores, xanthophores, iridophores
Dermis
leukocytes macrophages immunoglobulins  immune function
Agastric (esophagus  intestine); No oral teeth
DIGESTIVE
Teeth
Pharyngeal teeth in Zebrafish; keratinized appose
Liver
Distinct lobules, portal triads, hepatic cords are not readily apparent.
Pancreas exocrine
Usually in liver near branches of hepatic portal vein.
Physostomous fishes retain open PNEUMATIC DUCT connection between
Pneumatic duct
digestive tract & Swim bladders
RESPIRATORY
Each gill arch  hemibranch  row of Primary lamellae Secondary lamellae 
Gills
large surface area that supports respiratory and excretory functions.
GENITOURINARY
Anterior (head) kidney = primary hematopoietic organ in Zfish – also has
Kidneys
endocrine functions
Posterior kidneys often have osmoregulatory & excretory functions
CARDIOVASC
2 or 4-chamber; 1 atrium 1 ventricle. Blood  sinus venosus  atrium 
Heart
muscular ventricle  fibrous bulbus arteriosus
Skeleton
No marrow
ENDOCRINE
Corpuscles of Stannius
Eosinophilic cells that typically on latero-ventral surface of kidneys
Ultimobranchial Gland
Interrenal cells
Chromaffin cells
Thyroid Gland
Small group of cells centrally on septum separating heart from coelom
Sale eosinophilic cells scattered in hematopoietic tissue near major blood vessels
of anterior kidney
Within interrenal tissue associated with major blood vessels of anterior kidney.
Chromaffin differentiates them from interrenal cells
Thyroid follicles typically in pharyngeal region, but may be anywhere.
Initial barrier to infection
Entry of various fungal, bacterial infections
Also eliminate most nitrogenous waste
~ Parathyroid secrete hypocalcin (teleocalcin)
~ C cells secrete calcitonin (lowers serum
calcium concentrations)
~ adrenal cortical tissue secrete glucocorticoid,
mineralocorticoid.
~ adrenal medullary cells secrete adrenalin,
noradrenalin.
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Danio (Zebrafish)
Endocrine Pancreas
HEMATOPOIETIC
Hematology
MISC
Lateral Line organ
islets typically associated with exocrine pancreas in mesentery near small
intestine or pyloric cecae
No marrow  hematopoietic kidney
Hemolymph – Not much in small fish
sensitive to changes in hydrostatic stimuli and sound
Spontaneous ZFISH Pathology (phenotypes) NON Neoplastic
FISH Pathology (phenotypes) Non Neoplastic
SYSTEMIC (multisystem
METABOLIC NUTRITIONAL
Water quality
Chlorine, nitrosamines, plastics, bacteria, etc
die peracutely or gasping with flared opercula near surface;
Ammonia toxicity
May have mucus on gill & or gill hyperplasia (NH3 is irritating)
die peracutely or gasping with flared opercula near surface;
+/- Chocolate brown colored blood and/or gills
Nitrite toxicity (NO2)
 blood methemoglobin levels (Normal concentrations vary with species
Chronic exposure  secondary bacterial infections.
gas emboli in capillaries, vessels in many organs  death dt asphyxiation
emphysematous fish with gas bullae in skin, fins – may see small gas bubbles in
Gas Bubble Disease
skin, fins, and gills.
+/- Exophthalmos dt gas in choroid gland in posterior uvea.
INTEGUMENT
DIGESTIVE
Liver
RESPIRATORY
ENDOCRINE
GENITOURINARY
Eggbound
CARDIOVASCULAR
Cardiomyopathy
Verrucous endocardiosis
MUSCULOSKELETAL
Scoliosis
Overcrowding poor filtration too much food etc
New fish tank disease or Brown blood disease
Increase nitrite levels are often associated with
poor water quality (excessive ammonia levels
Due to water supersaturated with gasses (Not
just oxygen) e.g. pressurized air in water;
rapid temperature rise; algal blooms
Megalocytosis common in some aquaculture situations
Gill diseases
r/o environmental infectious
Abdominal (coelom) distention; abundant ovarian/egg tissue +/- granulomatous
inflammation or fibroplasia
Esp in milked fish or mutants
Dilated atrium or ventricle
Valves or endocardial proliferation
Kyphosis etc r/o spinal malformations, myopathy, vitamin C
r/o xenomas etc CNS muscle infections
Spontaneous ZFISH Pathology (Phenotypes) Neoplastic
Z FISH Neoplasms
INTEGUMENT
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Danio (Zebrafish)
Skin
DIGESTIVE
Liver
Intestine
Pancreas
GENITOURINARY
Testes
ENDOCRINE
Ultimobranchial gland
Thyroid
Sarcoma
Unusual without carcinogen (r/o infections)
Adenocarcinoma, carcinosarcoma
Acinar or ductal adenoma
Increase with carcinogen
Liver = primary target of most carcinogens
Increase with carcinogen, capillariasis
Increase with carcinogen
Seminoma
Prob most common T in young & old fish
Adenomas
Adenoma/carcinoma
Usually in older broodstock
r/o ectopic not tumors
Cholangiocarcinoma, mixed tumors fairly common
VIRUSES in Zebra fish
Virus order /family
Iridoviridae
Lymphocystivirus
Rhabdoviridae
Novirhabdovirus
Rhabdoviridae
Novirhabdovirus
Rhabdoviridae
Novirhabdovirus
Virus
Lymphocystis Disease virus 1
Infectious Pancreatic Necrosis
virus IPNV
Infectious Hematopoietic Necrosis
virus IHNV
Snakehead Rhabdovirus
Viral hemorrhagic septicemia virus
(VHSV)
NON Viral infectious/infesting agents in FISH
Site
Stains
AGENT in FISH
Primary
?
II. BACTERIA
Aeromonas hydrophila,
A sobria
Pseudomonas fluorescens
Skin etc
G-
Gram Negative septicemia
Flavobacterium
branchiophilum
G-
Prevalence
Dx/Detection
Disease ~ Pathology ~ Phenotypes
Comments
Not? Zfish
Skin / fin nodules = hypertrophied
fibroblasts
Each cell is surrounded by a hyaline
capsule.
ICIB, INIB
In cooler months in wild fish
necrosis in pancreas
necrosis of hematopoietic tissue in
kidney, spleen
Exp Dz in
Zfish
Prev
Detection
Common
Cult
Path
Salmonid viruses infect Zebrafish but
not pathogenic
Salmonid viruses infect Zebrafish but
not pathogenic
Exp infections  mortality
Disease ~ Pathology ~ Phenotypes
hemorrhage at base of fins & on skin, shallow
to deep skin ulcers, swollen abdomen,
exophthalmia; Focal necrosis in  muscle,
liver, gonad, pancreas
+/- serosanguinous exudate
Proliferative branchitis esp at distal end of
primary lamellae.
Secondary lamellae may be fused.
Gram NEG long bacilli adhered to gill
Comments
Ubiquitous opportunists + stress 
Motile Aeromonad Septicemia, AKA
Bacterial Hemorrhagic Septicemia
Often affect only 1-2 fish /tank
Environmental Gill Disease
Gliding Bacteria
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Danio (Zebrafish)
AGENT in FISH
Flavobacterium columnare,
Flexibacter columnaris,
Cytophaga columnaris,
Chondrocccus columnaris
Edwardsiella ictaluri.
Site
Primary
Stains
?
Prev
Detection
Disease ~ Pathology ~ Phenotypes
Comments
shallow skin erosions, frayed fins, eroded gills
+/- yellow
Necrosis with long flexing bacilli in
characteristic “haystack”
‘’Columnaris disease’
Ubiquitous, opportunist yellow
pigmented bacteria
(Flavobacterium).
“Hole in the Head Disease” or enteric
septicemia in catfish
E tarda  Emphysematous
Putrefactive Disease of Catfish
Mycobacterioses transmitted by water
& cannibalism.
Zoonotic via cuts or abrasions  skin
nodules.
Most cold adapted slow growing
‘Fin Rot’ Opportunist – common in
water/ environment
Skin gill etc
G-
Skin bone
sepsis
G-
Not ? Zfish
Sepsis, necrotizing ulcerative chain
+/- emaciation, ulcers, spinal curvature,
exophthalmia, loss of equilibrium
+/- gross nodules (granulomas)
with AFB.
Mycobacterium spp.
M marinum, M fortuitum,
M chelonei etc
ANYWHERE
AF
Common
Path PCR
Cult
Pseudomonas fluorescens
etc
Int Var
Sepsis
G-
Cult
See Aeromonas
Sporadic
Path
Granulomas, viscera or skin
Pigmented hyphae; septate branching,
irregular width -Should not need silver or
PAS stains to see them
Phaeohyphomycosis (chromomycosis)
fish may become darker, lethargic, or
erratic swimming
Cottony material on skin gills etc
Broad nonseptate branching hyphae that
produce motile flagellated zoospores in
terminal sporangia.
Normal water inhabitants that invade
traumatized skin. Esp with
SUBoptimal (low) temps
III. FUNGI
Exophiala sp
var
-- PROTISTS
Saprolegnia
IV. PROTOZOA
Cryptosporidium spp
Henneguya
Myxosporidian
Ichthyophthirius multifiliis
Ciliate
Glugea, Loma
Microsporidian
Intracellular extracytoplasmic in apical
epithelium
cysts on skin, fins, gills of fish.
Proliferative-granulocytic branchitis with
elongate myxosporidia in cysts
intestine
Gill Skin
Skin Gill
Various
Common
Large ciliate with C- shape macronucleus
white spots on fish = trophozoite 
 epithelial hyperplasia and necrosis
Zfish?
Infect macrophages etc mesenchymal tissues
 massive hypertrophy + deformity of
viscera (liver, gut, ovaries) +/- muscle
subcutis.
Esp Carp; tang, cichlids
Usually opportunist
2 polar capsules +long tail-like
extension
Assoc with proliferative gill disease
Trophozoite  tomont in
environment divisions  theronts
 break out of tomont  infect
burrow into fish epithelium
life cycle may require 7-14 days
cysts (xenomas) filled with 1 - 2
micron spores +/-hypertrophy of
infected cells usually in muscle or
CNS
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Danio (Zebrafish)
AGENT in FISH
Pseudoloma neurophila
Microsporidian
Pleistophora
Microsporidia
Site
Primary
CNS
Skeletal
muscle
Skin – bone
etc
V. HELMINTHS
NEMATODE
S
Pseudocapillaria tomentosa
Intestine
CESTODES
TREMATODES
Digenetic trematodes e.g.
Diplostomum
Monogenetic trematodes e.g.
Gyrodactylus, Dactylogyrus
VI. ARTHROPODS
? Spp
? var
PCR
Giemsa
Myxosporidian
Flagellate
Prev
Detection
Muscle
Myxobolus cerebralis
Spironucleus (Hexamita) spp
Stains
?
Eye fluke
gill skin
Common
Disease ~ Pathology ~ Phenotypes
Emaciated, ataxic, or spinal malformations
Cysts (xenomas) filled with 1 - 2 u spores +/hypertrophy of infected cells usually in muscle
or CNS
 myofibers filled with organisms (spores).
No inflammation.
Spores in cartilage & bone esp skull & gill
arches -10-u oval spore with 2 pyriform polar
capsule
Binucleate pyriform Flagellate 6 anterior + 2
posterior flagella – skin ulcer  bone +/enteritis, coelomitis granulomas with
flagellates
Comments
problems for neurobehavioral studies,
phenotyping
Neon tetra disease
‘Whirling disease’ in Salmonid
Feeds on cartilage;
Tubifex worms = intermediate host
Hole in the Head Disease in gouramis
etc aquarium fish, salmonids
Capillarid enteritis + increase intestinal
tumors esp in distal intestine
Unlikely in clean quarantined stock ?
Some metacercaria are very specific with
respect to location (organ/tissue) on fish
host.
cercaria encyst in fish  metacercaria
Usually not serious
Digenetic trematodes require several
hosts
Adult usually in gut of a fish-eating
bird  egg  free swimming larva
(miracidium)  snail / clam  freeswimming larva (cercaria)  encyst
in fish  metacercaria – eaten by
bird
haptor = attachment organ
posterior with 0-2 pair of anchors +
peripheral small hooklets
hemorrhage necrosis
Monogenetic = single host
May see these on eggs etc
Environment – management issue
On Eggs etc – looks like a shrimp
attaches by “anchor” structure that develops
below skin, in muscle.
The “worm” trails outside the fish
Environment – management issue
MITES
Crustaceans
? Spp
Lernea (Copepod)
Skin-muscle
Anchor worm = adult female
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Danio (Zebrafish)
AGENT in FISH
Ergasilus (Copepod)
Site
Primary
gill
Stains
?
Argulus (Branchiuran)
Prev
Detection
Disease ~ Pathology ~ Phenotypes
‘’Tongs’ attach to gills
flattened disk-like on external surface of
often under operculum, may  extensive
skin necrosis.
Comments
Fish louse
D. OTHER
References / Resources
Alestrom & al. 2006. Zebrafish in functional genomics and aquatic biomedicine. Trends in Biotechnology. 24(1): p. 15.
Baker, D.G. Ed. 2007. Flynn's Parasites of Laboratory Animals, 2nd Edition, Blackwell
The Zebrafish book http://zfin.org/zf_info/zfbook/zfbk.html
Zfin Zebrafish Model Organism Database http://zfin.org/cgi-bin/webdriver?MIval=aa-ZDB_home.apg
Acknowledgments
Jan Spitsbergen, Shannon Fisher, Andy MacCallion
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