Fish Pathology
Heike Schmidt-Posthaus
Dr. med vet, DECVP
Centre for Fish and Wildlife Health
Institute of Animal Pathology
Laenggasstrasse 122
P.O. Box 1866
3001 Bern
Switzerland
Anatomy, Histology
Special features in fish
Anaesthesia / euthanasia
Tricaine methanesulfonate (TMS, MS222®):
– 50 mg/l in tank water for aneasthesia
– 100 mg/l in tank water for euthanasia
– Adjust pH to 7.0 with bicarbonate
Clove oil
– 50-100 ppm (dissolve oil into ethanol 1:10 first) for
anaesthesia
– Double amount for euthanasia
Ethylenglycolmonophenylether (KOI MED Sleep®) for ornamental
fish
– 5 ml/10 l water for aneasthesia
– For euthanasia double dosis for 30 min after anaesthesia
Electricity, blow on head, gill cut
Salmonidae
Fibrous septum transversum
Cyprinidae
Head kidney
Trunk kidney
Swim bladder
Gonads
Fat tissue
Operculum
Heart
Liver
Note: Cyprinids have no intestine
stomach
Integumentary system
Epidermis
Non-keratinizing stratified squamous epithelium
Outermost Malpighian layer retain capacity to divide
Cuticle composed of mucus, mucopolysacharides, immunoglobulins, free
fatty acids
Goblet cells (secrete cuticle), club cells (alarm cells, Schreckzellen),
eosinophilic granular cells, lymphocytes, macrophages
Integumentary system
Dermis
Upper stratum spongiosum, deeper stratum compactum
Stratum spongiosum: chromatophores, mast cells, scale bed, cycloid
scales in salmonids and cyprinids
Cycloid scales: surface layer containing hydroxyapatite and calcium
carbonide, deeper layer composed of mostly collagen, enamel only
in superficial ridges
Integumentary system
Dermis
Scales serve as source of calcium, many fish use this calcium in
preference to the skeleton calcium in times of starvation or
prespawning activity
Chromatophores: Melanophores (M: black; H: black), xanthophores (M:
yellow; H: no colour), erythrophores (M: red; H: no colour),
leucophores, iridophores (M: silver-blue; H: olive green)
Respiratory system
Gills
Four holobranches at each side of pharynx
Each holobranch consists of two
hemibranches
Rows of filaments with cartilagenous core
(primary lamellae)
Two semilunar folds of lamellae
(secondary lamellae)
Respiratory system
Epithelial cells, chloride cells, pillar cells, endothelial lined vascular
channels
Gas exchange, removal of nitrogenous waste, electrolyt exchange
(Cl-/Ca2+ via chloride cells, Na+ via pavement cell)
Pillar cells: regulate blood flow, reticulocyte-like function (antigen
uptake), comparable to intravascular pulmonary macrophages
Respiratory system
Gills
Pseudobranch under dorsal operculum
Single row of filaments
Function probably as thermoregulator, baroreceptor, oxygen supply
to optic choroid and retina
Cardiovascular system
Heart
Only deoxygenated blood
4 chambers
sinus venosus (no inlet valve) atrium (2 sino-atrial valves) ventricle
(valves) bulbus ateriosus (2 valves) ventral aorta gills for
oxygenation
Almost inelastic pericardial sac
Separated from abdominal cavity by fibrous septum transversum
Picture: http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/A/AnimalHearts.html
Bulbus arteriosus
Ventricle
Atrium
Bulbus arteriosus: fibroelastic tissue, some species: smooth
muscle, some species: hyaline cartilage
Cardiovascular system
Heart
Ventricle: outer compact layer, inner spongy layer
Spongy layer: interlacing meshwork of trabeculae, filling functional lumen
of ventricle
Compact and spongy layer separated by thin connective tissue layer
Spongy layer
Compact layer
Cellular components of blood
Blood volume: teleosts: 3-4%, elasmobranchs, other vertebrates: 5-8%
Nucleated erythrocytes
Neutrophils (polymorphnuclear leucocytes)
Granules not necessarily neutral staining
Nucleus may not be multi-lobed
Monocytes
Thrombocytes
Cellular components of blood
Eosinophils
Rare in fish blood
Large eosinophilic cytoplasmic granules
Eosinophilic granular cells
In skin, hematopoietic and digestive tissue, different from eosinophils
Basophils
No implication in defence mechanisms in fish
Mast cells
Connective tissue
Basophilic and metachromatic granules
Lymphocytes
B and T cells
CD79, CD3, CD4, CD8 identified in
few fish species
IgM-like, IgT and IgA-like antibodies
Delayed and immediate hypersensitivity
Secondary circulation and lymphatics
No lymph nodes, no separate lymph system
Secondary circulation in gills, skin, fins, peritoneum
5% of body weight
Arise from primary circulation
Drain into veins
Restrict access of red blood cells
Physiological function unclear
Probably progenitor of lymphatics of other vertebrates
Reticuloendothelial and hematopoietic systems
Phagozytic cells in endothelial lining of spleen & kidney capillaries,
peritoneum, atrium, gill lamellae
No Kupffer cells in liver sinusoids in teleosts
Melanomacrophage centres in liver, kidney, spleen containing
melanin, haemosiderin, ceroid, lipofuscin
Reticuloendothelial and hematopoietic systems
Thymi paired organs near dorsal commissure of operculum
Hematopoietic tissue in interstitium of kidney, primarily in head kidney
Reticuloendothelial and hematopoietic systems
Thymi paired organs near dorsal commissure of operculum
Hematopoietic tissue in interstitium of kidney, primarily in head kidney
Special features in sturgeon
Subepicardial
haematopoietic tissue
Spleen
One of the two major filtering organs (other kidney)
Remove foreign agents and effete blood cells
Major site of erythropoiesis
Red pulp, white pulp (around arterial vessels, melanomacrophage
centres and ellipsoids), blood vessels, ellipsoids
Urinary system
Retroperitoneally ventral of spine
Posterior portions maybe fused
In higher developed fish two connected lobes
Maybe divided transversely (head and trunk kidney)
Head kidney: hematopoietic tissue
Trunk kidney: hematopoietic tissue and excretory elements
Urinary system
Primary function: osmoregulation (nitrogenous waste mainly
excreted through gills)
Freshwater fish: save ions, excrete water
Saltwater fish: excrete ions, conserve water
Salmonids
Trunk kidney
Tubuloneogenesis
Cyprinids
Thyroid follicles
Glomeruli
Tubuli
Haematopoietic interstitiel tissue
Melanomacrophage centres
Digestive tract
Gastrointestinal tract
Large species differences
Stomach maybe present or absent
Species without stomach: more lymphoid tissue in intestine
In lamina propria and submucosa eosinopilic granular cells
(antimicrobial and vasoactive substances)
Salmonids: pyloric caeca, secrete digestive enzymes
Others: digestive enzymes secreted in liver, pancreas
No small and large intestinal segments, fluent transition between small
and large intestine, no clear border line
Some species: longitudinal foldings of mucosa, few species with
resembling intestinal villi, no distinct crypt compartments no
alterations of villous / crypt ratio (hallmark in mammals)
Digestive tract
Liver
Digestive tract
Liver
No lobular architecture
Many fish hepatopancreas
Hepatocytes often loaded with glycogen or neutral fat
Hematopoietic tissue with melanomacrophage centres around larger
vessels
Fish
Digestive tract
Exocrine pancreas
Exocrine pancreas scattered throughout abdominal mesentery fat between
pyloric caeca, subcapsular in spleen, around hepatic portal vein
Pancreatic duct joins bile duct
Swim bladder
Derived from oesophageal region embryologically
One chamber in salmonids, several chambers in cyprinids
Connection between swim bladder and oesophagus in some fish species
(physostomes) via pneumatic duct
Absent in other fish (physoclists), inflation via gas gland
Absorption of gas always via gas gland
Swim bladder
Epithelium, lam. propria, lam. muscularis mucosae (smooth and/or sceletal
muscle), submucosa with blood vessels
For comparison intestine: mucosa, lam. propria, lam. muscularis mucosae, submucosa, lam.
muscularis (circular, longitudinal muscular layer)
Primary function: buoyancy
Sound and pressure reception, sound production
Absent in bottom-living species and some fast-swimming fish
Reproductive system
Testis
Paired organs alongside or below swim bladder
Tubules or blind sacs (seminiferous tubules), lined by spermatogenic
epithelium
Sertoli cells, interstitial cells (testosterone production)
Vas deferens conducts mature spematozoa to excretory meatus at urinary
papilla
Reproductive system
Ovary
Simple cluster of ovarian follicles (ova and surrounding granulosa cells) in
lower teleosts
Different stages of development
Higher teleosts: oviduct
Complex organ in viviparous species (egg production, spermatozoa store,
vagina, uterus)
Primitive species (eg. salmonids): eggs passed into fold of mesentery,
which rupture, release eggs in abdominal cavity, evacuation via genital
opening
Endocrine system
Thyroid gland
Thyroid follicles distributed bilaterally symmetrical throughout
connective tissue of pharyngeal area
Around eye, ventral aorta, hepatic veins, renal hematopoietic tissue
Thyroid hormones influence carbohydrate metabolism, mobilization
of lipid reserves
Endocrine system
Adrenals (interrenal and suprarenal)
Interrenal cells (mammalian adrenal
cortex): Pale eosinophilic cuboidal cells
associated with major blood vessels in
anterior kidney
Glucocorticoid and mineralocorticoid
hormones
Chromaffin cells (mammalian adrenal
medulla), clustered with interrenal cells
Epinephrine, norepinephrine,
catecholamines
Regulation of heart rate, cardiac
contraction, respiratory function
Endocrine system
Ultimobranchial body
Ventral to oesophagus in septum separating heart from abdominal
cavity
Calcitonin (serum calcium level)
No parafollicular or C cells in thyroid gland
Endocrine system
Corpuscles of Stannius
Paired organs, island of eosinophilic granulated cells arranged in
folded cords
Ventral surface of kidney
Hypocalcin (teleocalcin) (calcium metabolism)
Parathyroid gland absent
Endocrine system
Endocrine pancreas (islets of Langerhans)
Associated with exocrine pancreas
Sometimes grossly visible (Brockman bodies)
3 cell types:
• Glucagon-production (gluconeogenesis, incorporation of amino
acids in liver)
• Insulin-production (inhibition
of gluconeogenesis, turn-over
of liver proteins)
• unknown function
Lateral line system
Basic sensory unit = neuromast: bundle of sensory and supporting cells
whose projecting hairs are encased in a gelatinous cap
Neuromasts occur
singly
in small groups = pit
organs
in rows within
grooves or canals,
referred to as lateral
line system
Lateral line system runs along sides of body onto head, where it
divides into three branches, two to snout and one to lower jaw
Lateral line system
Neuromasts continuously send out trains of nerve impulses
Pressure waves cause gelatinous caps to move, bending enclosed hairs
change in frequency of nerve impulses
Swimming fish sets up pressure wave detectable by lateral line
systems of other fishes
Also bow wave in front of itself, near-field differences are registered by
own lateral line system
Sound waves = waves of pressure, the lateral line system detect very
low-frequency sounds
Modified groups of neuromasts = ampullae of Lorenzini, (sharks and
certain bony fishes), electroreceptors
Most animals emit electric field in seawater
Wound, even scratch, markedly alter these electrical fields
Lateral line system
Pathology
Some special features in fish
Histopathology
10% buffered formalin
Bouin’s fluid
Use 10x volume of fix : fish
After 24 hrs transfer to 70% ethanol
Decalcification
Paraffin embedding
Staining: hematoxylin /eosin (HE)
Additional stainings according to special requests
Inflammation
Acute inflammation: initial response
Characterized by pain (dolor), heat (calor), redness (rubor), swelling
(tumor), and loss of function (functio laesa)
Cells involved: granulocytes (eosinophilic granular cells, eosinophils,
neutrophils, basophils), macrophages
Mechanism of innate immunity
Temperature influence: highest on T helper cells, less on non-specific
defense mechanisms, like complement pathways, non-specific
cytotoxic cells, respiratory burst
Inflammation
Chronic inflammation: Prolonged inflammation, simultaneous
destruction and healing
Cells involved: mononuclear cells (macrophages, lymphocytes, plasma
cells), fibroblasts
In contrast to mammals fish very efficient in regeneration
also myocard, cns, retina
temperature dependant
Hypertrophy / Hyperplasia
Hypertrophy: increase in cell size
Hyperplasia: increase in cell number
Common response in fish
Common as preneoplastic response, morphological normal cells
Neoplasia
Autonomous division of cells
No coordination with surrounding normal tissues
Growth persists in same excessive manner even after cessation of
stimuli
Classification based on mammalian criteria
Benign, pre-malignant (carcinoma in situ) or malignant
Benign Malignant
Slow growing Variable, may be rapid
Few mitoses Variable, may be many mitoses
Usually resemble tissue of origin Variable, may only poorly resemble
tissue of origin
Nuclear morphology normal Nuclear morphology variable, can
be very abnormal
Usually well circumscribed Poorly defined or irregular
or encapsulated
Necrosis rare Necrosis common
Never invade May invade surrounding tissues
Never metastasize May metastasize
Fish: many signs of malignancy, but rarely metastases
Microscopic examination of tissue in order to study the manifestations
of disease
etiology / cause
pathogenesis
Summarize in morphologic diagnosis
Type of process (neoplasia, inflammation, etc)
Location and tissues / cells involved
Duration (acute-chronic)
Severity
