DIAGNOSIS OF FISH DISEASES

WET MOUNT EVALUATION

Immediately prior to sample collection, the diagnostician should have prepared slides for
biopsy samples by placing one or two small drops of water equidistant on a slide. This will
allow for one or two different biopsy samples to be evaluated per slide. A separate coverslip is
then placed on each biopsy sample with an adequate amount of water to completely flood the
area under the coverslip, and gentle pressure is used to flatten the tissue to allow penetration
of light and easier visualization. For external (i.e., skin, gill, and fin) samples, the use of
dechlorinated freshwater (not distilled water) for freshwater fish and salt water for marine fish,
respectively, is necessary.

During microscopic examination of squash preps/ wet mounts, the clinician should use a
methodical search pattern (e.g., “mowing the lawn”). Scanning each preparation at 40× (i.e.,
low power) will help with orientation and identification of areas of interest. Decreasing the
condenser aperture or use of phase contrast will help increase contrast and facilitate
observation of smaller structures such as protozoan parasites. Most parasites and lesions of
interest are visible at 40×–100× magnification. While movement of some parasites, such as
the flagellates of the genera Spironucleus and Ichthyobodo, may be discerned at 100×,
specific parasite structures are more apparent at 400×. Individual nonmotile spores of
microsporidian parasites are also very small (approximate range 1–5 × 2–8 μm) and are more
easily identified at 200×–400×, although their cysts (sporoblastic vesicles) or xenomas may be
visible at 40×–100×. Similarly, Flavobacterium columnare, the etiologic agent of columnaris
disease, is a long, rod-shaped Gram-negative bacteria that can often be found flexing in
haystack formations that are more easily observed at 200×–400×.
HEMATOLOGY

Venipuncture

A clinically acceptable blood sample can be readily obtained from most fish species. The most
common sampling site to access blood is from the caudal vessels (mixed arterial and venous
sample) using a ventral or lateral approach.

The site for the ventral approach is along the ventral midline of the caudal peduncle halfway
between the tail and anal fin, while the lateral approach is along the lateral midportion of the
caudal peduncle along the lateral line of the fish.

For either of these approaches, the needle is inserted under a scale in a cranial direction and
advanced at a 45° angle to the vertebral column of the fish. The needle is then withdrawn
slightly until blood is obtained. The needle may need to be rotated slightly to allow the beveled
edge of the needle better access to the pooled blood supply.

Cardiac puncture has been utilized for blood sampling in large fish, but there is a higher risk of
death using this technique and it is not recommended for nonlethal sampling.

As a last resort, the caudal peduncle can also be severed to obtain a small amount of blood.
Though generally not acceptable for clinical specimens from pet fish, it provides an alternative
route for sampling.
For most biochemical analytes, plasma is preferred over serum, and
lithium heparin is recommended for use in most assays and with most fish
species. Some clinicians have reported hemolysis of blood samples
collected in EDTA tubes when fish are anesthetized with tricaine
methanesulfonate (MS-222), however, the mechanism for this hemolysis is
unknown. Fish blood often clots very quickly so the needle and syringe
may need to be heparinized prior to blood collection.

A blood volume collection of 0.5%–1.0% of body weight can be safely

taken from most fish. In most cases 1.0 mL can be taken from a 100 g fish
and up to 3.0 mL in larger fish, such as production brood stock and adult
koi. Blood should be used immediately for making blood smears or
transferred to a blood tube containing an anticoagulant to minimize clotting.
The choice of anticoagulant for preservation of cellular morphology is
species-specific with salmonids, cyprinids and sturgeon retaining the best
blood cell morphology with heparin, whereas catfish, bass and tilapia blood
cells are best preserved in EDTA.
Hematology

Complete blood counts (CBCs) and biochemical tests can be valuable

diagnostic tools in determining the cause of disease in many species. However,
normal hematologic parameters, reference interval and interpretation of those
values are not available for most fish species.

Published hematological data exists for economically important food fish species
(e.g., trout, channel catfish, tilapia, striped bass, winter flounder, sturgeon and
yellow perch), but these values are not always applicable to pet and ornamental
fish.

Selected hematology and biochemical reference values for a few common

ornamental and pet species (i.e., koi, goldfish, pacu and zebrafish) can be found in
the literature.

Environmental conditions such as water quality (temperature, pH, ammonia, nitrite,

nitrate, etc.) can significantly affect test results. Capture and handling stressors,
sampling technique, age, gender, reproductive status, photoperiod, diet, activity
level and test methodology are other factors that can affect the results of a CBC
and serum or plasma biochemistry results.
Interpretation of blood results

Interpretation of blood results in fish is difficult when compared to more

common terrestrial species.

Nucleated red blood cells and platelets prohibit the use of most automated
analyzers used for mammalian blood counts, though two recent studies
evaluated the use of in-house blood analyzers for fish.

Manual evaluation of stained blood smears and the use of a

hemocytometer are the most common techniques performed in a clinical
setting.

Blood smears can be stained with any variation of a Romanowsky stain

(Wright/Giemsa/Leishman stains).
Blood cell morphology

Most fish blood cell types appear to have a morphology and function similar to
mammalian blood cells.

Erythrocytes are the most numerous blood cell type and are nucleated.
Reticulocytes, frequently present in significant numbers in blood smears, are easily
recognized by their smaller size and slightly basophilic cytoplasm. Thrombocytes
vary in shape depending on their activation state, changing from spiked or oval to
round cells as they activate, and may have a segmented or oval nucleus.
Lymphocytes can be divided into small or large lymphocytes, though functional
differences have not been documented. Neutrophils are one of the largest fish
blood cells. The cytoplasm of the neutrophils may have a grainy appearance, and
the nucleus is open and usually oval to kidney-bean shaped. Heterophils are similar
to neutrophils but with pale eosinophilic to lavender granules in the cytoplasm.
Species of fish usually have either neutrophils or heterophils, although some
species may have both. Functional differences between fish neutrophils and
heterophils have also not been documented. Monocytes are round cells with a deep
blue cytoplasm and a round to horseshoe-shaped nucleus. Eosinophils are
infrequently observed in fish blood, but are the same size or slightly smaller than
neutrophils with distinct eosinophilic granules in the cytoplasm. Basophils are rarely
observed in fish and are similar to mammalian basophils in morphology.
NECROPSY

If a necropsy is to be performed, fish can be euthanized by a variety of humane

methods including immersion, injectable and physical methods. Choice of method
will depend on number of fish, size of fish, drug availability, logistics, cost and
potential interference with sample analysis, as well as carcass disposition.

In addition to the lack of movement, the fish should no longer be responsive to

stimuli and have lost the vestibule-ocular reflex (eyes in normal position, parallel to
body, regardless of position of the body). The clinician may need to gently press the
eye parallel to check this response in fish.

Similar to the initial examination of the fish patient, the fish necropsy should be
conducted in an organized and thorough manner and follow the same routine each
time so that nothing is missed, crosscontamination is avoided, and proper
downstream processing of samples and tissues is ensured, which is paramount for
microbiological, histological, molecular and microscopic examination.

In general, samples collected for bacteriological and fungal recovery are

prioritized, with samples collected for virology and molecular diagnosis following,
whereas collection of internal tissues for histopathology is generally saved until the
end of the necropsy procedure.
MICROBIOLOGICAL SAMPLING

Most of the common bacterial pathogens of fish will grow on nutrient-rich media like
blood agar or tryptic soy agar (TSA) supplemented with 5% sheep’s blood. If
working with brackish or marine water species, it is recommended to also have
media supplemented with NaCl or sea salt, since many of the marine organisms,
like those in the family Vibrionaceae, are halophilic (i.e., salt-loving). There are also
a wide range of fish pathogens that are fastidious and need specialized media and
growing conditions. For example, the Flavobacterium species grow best on
Cytophaga agar, whereas Coomassie blue media can be used to rapidly distinguish
between Aeromonas hydrophila and A. salmonicida.

For nearly three decades, amplification of viral nucleic acid by PCR followed by
Sanger sequencing (first-generation sequencing) has proven invaluable for the
detection and characterization of novel viruses including fish viruses. The strength
of PCR is its rapid ability to detect viruses even in low abundance from a range of
sample types (cell culture pellets or supernatants, fresh tissue, frozen tissue, and
formalin-fixed tissues). However, PCR is a targeted approach requiring previous
sequence knowledge to design primers for amplification. This has proven
challenging given sequence data for fish viruses within public databases is sparse,
with sequences from whole families of RNA viruses lacking altogether (e.g.,
Astroviridae, Filoviridae, Bornaviridae).
INTERNAL ORGAN EVALUATION

After microbiological sampling, the internal organs should be

examined grossly and microscopically as wet mount
preparations.

The stomach should be examined for the presence of food,

hemorrhages, ulcers, granulomas and parasites (i.e., both
metazoan and protozoan). For larger fish species or those with
more fibrous stomachs, the inner mucosal lining of the
stomach should be scraped and examined microscopically. It
should be remembered that some species, for example,
cyprinids, pipefishes, and parrotfishes, lack a true stomach.

The intestinal tract and pyloric cecae should also be checked

for the presence of food or fluid, hemorrhages, ulcers, and
parasites (especially nematodes and flagellates).
The liver should be checked for color, uniformity, fatty consistency, parasites,
pigmented macrophage aggregates and granulomas.

The gallbladder, which is located near or surrounded by the liver, should contain a
yellowish to greenish fluid that is normally clear. The size of the gallbladder may
vary and can be relatively large in fish that are anorexic or have not eaten for a
while.

The spleen should be examined for color, size, uniformity, pigmented macrophage
aggregates and granulomas.

The swim bladder should be examined for thickening, hemorrhages, necrosis, fungi
and parasites. The swim bladder in some species (e.g., gar, tarpon, arapaima and
some catfishes) are highly modified and may contain specialized respiratory
tissues.

The kidney in some species of fish has anterior and posterior portions that are
separate, while some are connected and others are merged. Both the anterior and
posterior portions of the kidney should be checked for uniformity, granulomas and
parasites.

The gonads of fish should be checked for gender, maturity, size, granulomas or
hardened structures, which in females is sometimes indicative of degenerating eggs
or past “egg-binding” episodes.
DIAGNOSTIC IMAGING

Survey radiography

Radiography remains perhaps the most useful noninvasive

diagnostic technique in veterinary medicine due to its ease of
use, lower cost compared to other diagnostics, amount of
valuable information provided and quick implementation. It
should be part of all examinations of fish whenever possible.

Fish less than 6 cm can be radiographed effectively with digital

radiography, and multiple exposures allow large fish to be fully
evaluated. Radiographs should also be part of every necropsy,
as the swim bladder and skeletal system are most effectively
evaluated with radiography.
Endoscopy

Endoscopy is often divided into rigid or flexible endoscopy, with the former
having a significant role in fish medicine. Many rigid endoscopic systems
are fully portable for site visits and fieldwork. When endoscopy is
combined with radiology and/or ultrasound, the diagnostic evaluation of the
internal anatomy can equal exploratory surgery with far less risk.

The most obvious use of rigid endoscopy is evaluation and/or biopsy of the
internal organs through smaller incisions compared to exploratory surgery.
Rigid endoscopes are also extremely useful in evaluating macroscopic gill
health and detecting metazoan parasites on the gills. In fish <10 kg, rigid
endoscopes can evaluate the oral and upper gastrointestinal systems in
the same way that flexible endoscopy works in terrestrial animals. Foreign
bodies in the stomach can also be removed safely with endoscopic tools.
In larger fish, reproductive and caudal gastrointestinal structures may be
evaluated through the vent.
Coelioscopy

Visual evaluation of the internal anatomy is often helpful in complicated cases

where hepatic lipidosis, egg binding, ascites or disseminated granulomous disease
may be quickly identified. Coelioscopy may be used to identify the gender of young
fish when secondary sexual characteristics are absent and small gonad size
prevents ultrasonographic identification. Juvenile koi or sturgeon brood stock are
often endoscopically sexed without complication. Rigid endoscopes have also been
used to sterilize male sturgeon while keeping their testicular endocrine function
intact by ligating the vas deferens.

Preparation for the portal of entry is the same as with surgery. The fish is
anesthetized in dorsal recumbency with a fish anesthesia delivery system. Scales
are removed only where necessary, and a topical disinfectant, commonly Betadine,
is applied to the area for several minutes. A local block is recommended to reduce
pain and bleeding (e.g., buffered lidocaine with epinephrine, 1:1 sodium bicarbonate
to epinephrine) and a stab incision is made at the appropriate location with a
scalpel blade.
Most gender identification surgical approaches start at ventral midline or slightly off
midline, cranial to the vent. Incisions are kept small to reduce insufflation gas/fluid
loss, reduce coelomic contamination, reduce skin damage/inflammation and reduce
the number of sutures necessary to close the wound.

Biopsies can be taken for wet mount preparations, cytology, culture, histology or
molecular diagnostics. Carbon dioxide insufflation is recommended over
atmospheric air, which can create prolonged positive buoyancy if not removed. The
nitrogen in atmospheric air can take days to weeks to resolve, whereas residual
carbon dioxide can be usually absorbed within 24 hours.

A radiograph is recommended post-surgery to evaluate the presence of coelomic

gas that should be aspirated prior to recovery. If fluids are chosen for insufflation,
those fluids should be isotonic to the species.
Gill evaluation

Unique to fish medicine is the use of a rigid endoscope to completely

evaluate the gills without the need for biopsy. In addition, the spiracles or
gill slits of most elasmobranchs make standard gill biopsy difficult to
impossible. Endoscopy allows full evaluation of each gill arch compared to
gill biopsy’s small sample and potential risk of hemorrhage. Gill biopsies
may miss organisms or gill pathology that can be readily diagnosed with
endoscopy.

Fish are lightly anesthetized with the animal kept fully submerged to avoid
air pockets that interfere with visualization. Oxygen supplementation is
recommended since opercular movements will be reduced or absent.
Oxygen saturation should be 115%–150% prior to induction and
throughout the examination and recovery. The rigid endoscope is
introduced through the opercular opening, gill slit or spiracle, and the gill is
evaluated from the arch base to the tips of the primary lamellae. Gill
biopsies usually sample only the gill tips to avoid excessive hemorrhage.
Gastric examination

Foreign bodies are common in pet fish. Certain marine

ornamentals (angelfish, Bermuda chubs) often ingest silicone
sealant or cleaning pad fragments, both of which are usually
radiolucent and require contrast radiography or endoscopy for
diagnosis. Rigid endoscopes often have tools (baskets,
forceps) that may facilitate removal of these foreign bodies
through the mouth and avoid surgery. Survey radiographs of
trout often find gravel in their stomach, and a quick gastric
endoscopic evaluation can determine if the stones are causing
inflammation. Similarly, several grouper species suffer gastric
ulceration as they age, which can be quickly diagnosed with
noninvasive gastric endoscopy
FISH PATHOLOGY
 OCULAR DISORDERS
EXOPHTHALMIA

Overview: A nonspecific, unilateral or bilateral condition of the eye,

commonly called “pop-eye,” in which the globe of the eye extends outside
its normal limits.

Etiology: Numerous etiologies have been reported to cause exophthalmia

including infectious (e.g., viral: infectious pancreatic necrosis [IPN],
infectious hematopoietic necrosis [IHN] or viral hemorrhagic septicemia
[VHS]; or bacterial: Aeromonas sp., Flavobacterium sp., Vibrio sp.,
Edwardsiella ictaluri, Renibacterium salmoninarum or Mycobacterium sp.),
parasitic, neoplasia, and noninfectious (e.g., gas supersaturation), or as a
sequela to impaired renal function or increased abdominal pressure from
the accumulation of fluids in the coelomic cavity. Fluid or gas accumulation
in the retrobulbar tissues can also cause protrusion of the eye.
 OCULAR DISORDERS
EXOPHTHALMIA

Clinical presentation: The globe of the eye expands laterally or

circumferentially outside its normal size, or protrusion of the globe outside
the normal recesses of the orbit. The enlarged globe of the eye is
predisposed to a variety of infectious diseases or trauma.
 OCULAR DISORDERS
EXOPHTHALMIA

Pathology: Anything that expands the posterior segment of the globe (e.g.,
infectious etiologies, parasites, tumors, increased coelomic pressure) may
produce a lateral movement of the globe. Both supersaturation of the water
and swim bladder inflammation can cause gas bubbles in the vascular and
choroidal gland, resulting in exophthalmos.

Differential diagnosis: This condition should not be confused with

telescoping eyes found in a number of goldfish varieties or “bubble eye”
goldfish, which have infraorbital lymph-filled sacs of adnexal (i.e., not
ocular) origin.

Diagnosis: Most commonly diagnosed by visual inspection of the fish and

eye(s), though the specific etiology may be more difficult to determine.
Specific diagnostic techniques may include an ophthalmic examination,
tissue or fluid aspirates, impression smears, and biopsies.
 OCULAR DISORDERS
ENOPHTHALMOS

Overview: A condition where the eye decreases in volume and appears retracted or
“sunken” into the skull. Though this condition is not specific for any particular
infectious disease, enophthalmos may occur with several viral diseases (e.g., koi
sleepy disease and koi herpes virus), or bacterial or parasitic (e.g., Myxobolus sp.)
infections. Noninfectious causes may include traumatic injury, social aggression
(e.g., “eye snapping”), emaciation and developmental defects. No matter the cause,
chronic enophthalmos may result in anophthalmos, where the globe is completely
lost and covered by regenerated dermal layers.
 OCULAR DISORDERS
CLOUDY EYE

Overview: A condition where the eye appears to be cloudy or have material in the
anterior chamber that makes it appear cloudy.

Etiology: Most commonly the condition is due to a local or systemic infection but
may also be due to rapid changes in water quality (i.e., ammonia, nitrites, pH,
salinity or osmolality). Several protozoan parasites such as Ichthyophthirius
multifiliis, Cryptocaryon irritans, and Tetrahymena corlissi have been known to
directly infest the epithelium of the cornea, causing it to become cloudy.
Occasionally monogeneans (e.g., Neobenedenia sp.), turbellarians, and copepods
(e.g., Lernaea spp. and Argulus spp.) may directly parasitize the cornea and
surrounding tissues of the eye. Lymphocystis, a generally benign viral disease, has
also been reported to affect the cornea and retrobulbar tissues of the eye. Larval
digenetic trematodes (e.g., Austrodiplostomum spp.) can invade the anterior and
posterior chambers of the eye. Dietary deficiencies, such as vitamin A, thiamin or
riboflavin, may also contribute to the development of cloudy eye in fish.
 OCULAR DISORDERS
CLOUDY EYE

Clinical presentation: The condition may be due to corneal edema, corneal opacity
or material in the aqueous humor of the eye, all of which may cause the cloudy eye
appearance and compromise vision.
 OCULAR DISORDERS
CLOUDY EYE

Pathology: Both ulcerative keratitis and nonulcerative keratitis can result in corneal
edema and a cloudy appearance to the eye. Superficial abrasions of the eye can
rapidly result in corneal ulcerations, which left unchecked can lead to rupture of the
globe and subsequent enophthalmos . Non-ulcerative keratitis can result in edema
of the cornea, accumulation of cellular infiltrates and fibroplasia in the anterior
chamber, giving the eye a cloudy appearance.

Differential diagnosis: The most common differential would be cataracts. Cloudy

eye or corneal edema should not be confused with corneal opacification that
commonly occurs postmortem, especially with fish that are chilled.

Diagnosis: Most commonly diagnosed by visual inspection of the eye, though the
etiology may be more difficult to identify. The diagnosis should include examination
of the water parameters and various systemic diseases.
 OCULAR DISORDERS
UVEITIS

Overview: Uveitis is inflammation of the uvea, which includes the iris, choroidal
gland, ciliary body, and associated blood vessels of the eye. As in mammals,
anterior uveitis (e.g., anterior chamber and iris) is most commonly diagnosed in fish.
This may be caused by a primary infection (i.e., bacterial, viral or fungal) of the eye,
the secondary extension of another systemic disease, or a noninfectious (i.e.,
neoplastic) problem. This condition is often described as a panophthalmitis. In
addition, a granulomatous uveitis associated with vaccination has been reported in
Atlantic salmon.
 OCULAR DISORDERS
HYPHEMA

Overview: A condition where there is blood in the anterior chamber of the

eye. This is usually the result of direct trauma, uveitis, panophthalmitis, or
an infectious systemic disease such as with enteric redmouth disease, a
bacterial infection of the kidney of salmonids caused by Yersinia ruckeri,
which often has secondary consequences of hyphema.
 OCULAR DISORDERS
CATARACTS

Overview: A condition of the eye in which the lens becomes cloudy or

opaque, resulting in impaired vision or complete blindness.

Etiology: Numerous etiologies have been reported to cause unilateral or

bilateral cataracts in both wild and captive fish, including nutritional,
infectious, intralenticular parasites, trauma, excess ultraviolet light,
changes in water temperature and hereditary factors. In addition, poor
water quality can cause osmotic changes in the lens.

Clinical presentation: Gradual increasing cloudiness and opacity of the

lens of the eye. Behavioral alterations may include a lack of response to
predators or shadows/movement. In addition, the fish may lose weight due
to the inability to feed or compete for food.
 OCULAR DISORDERS
CATARACTS

Pathology: The histologic changes seen in cataracts of fish include hydropic

swelling of the lens fibers, lysis of fibers, epithelial hyperplasia and intralenticular
migration of surface epithelium.

Differential diagnosis: Need to distinguish cataracts from corneal edema or corneal

opacity, which cause cloudy eye. Cataracts should also not be confused with lens
opacification that normally occurs postmortem.

Diagnosis: Most commonly diagnosed by visual inspection of the eye along with
behavior suggesting decreased vision in one or both eyes.
 OCULAR DISORDERS
GAS BUBBLE DISEASE

Overview: A noninfectious condition associated with the supersaturation of

dissolved gases, most commonly nitrogen or oxygen, in the water column
causing bubbles to form in the eyes and other tissues of the fish.

Etiology: Numerous etiologies are known to cause supersaturation of

gases within the water column including leaks in pumps, valves or pipe
connections; overaeration of water; sudden extreme temperature
gradients; heavy algal blooms; use of spring or well water that has not
been sufficiently degassed; and being in close proximity to plunge pools of
dams or waterfalls where increased levels of dissolved gases may occur in
the water column.
 OCULAR DISORDERS
GAS BUBBLE DISEASE

Clinical presentation: Clinical signs and mortality vary with species, age,
and degree and duration of gas supersaturation. Fish can be affected by
acutely or chronically increased levels of gas saturation that manifest in a
variety of clinical signs including air bubbles in the anterior chamber of the
eye, air emboli in the capillaries of the gills and tissues, and dermal bulla in
the skin and fins.
 OCULAR DISORDERS
GAS BUBBLE DISEASE

Pathology: Retrobulbar gas bubbles can push the globe forward causing
exophthalmos, which can result in keratitis, uveitis, panophthalmitis, and
cataract formation. Supersaturation of the water can cause emboli to
accumulate in the anterior chamber, vasculature, and adnexal structures of
the eye.

Differential diagnosis: Though the condition is fairly straightforward to

diagnose, the cause can often be difficult to determine.

Diagnosis: Most commonly diagnosed by visual inspection of the eyes,

gills and skin for the presence of gas bubbles in the anterior chamber of
the globe of the eye (or in the lamellae of the gill or under the epithelium of
the skin).
 OCULAR DISORDERS

NEOPLASIA

Overview: Various types of neoplasia have been reported from a number of

different species of fish, most commonly in older captive fish.

Etiology: Most occur spontaneously, but there are a few reported to be

caused by genetic mutations, environmental factors and the Oncorhynchus
mason virus (i.e., Herpesvirus salmonis type 2). Some of the eye tumors
reported in the literature include melanosarcomas, fibrosarcomas,
adenocarcinomas, retinoblastomas, neuronal embryonal tumors,
glioneuromas and epitheliomas.

Clinical presentation: Neoplastic conditions of the eye usually present as

exophthalmia and/or blindness.
 OCULAR DISORDERS

NEOPLASIA

Pathology: Most present as a space-occupying lesion of the various

tissues of the eye. As the tumor enlarges, the retrobulbar tissues or globe
increase in size, causing exophthalmia.

Differential diagnosis: This condition needs to be differentiated from other

causes of exophthalmia including infections, parasites and
supersaturation.

Diagnosis: Determination of neoplastic conditions of the eye are primarily

diagnosed by enucleation of the globe or at necropsy by histopathology.
 SKIN AND FIN DISEASES

FURUNCULOSIS

Overview: Furunculosis is caused by an infection with Aeromonas

salmonicida subspecies salmonicida, which can cause a bacterial
septicemia in salmonids.

The term furunculosis stems from the boil-like lesions on the skin and in
the musculature of infected fish. Even though the disease is named after
the raised liquefactive muscle lesions, development of “furuncles” are the
exception rather than the rule and generally only occur in older fish
suffering from a chronic infection.

The distribution is practically worldwide, but absent from salmonid

aquaculture in Australia, New Zealand and Chile.
 SKIN AND FIN DISEASES

FURUNCULOSIS

Etiology: Typical Aeromonas salmonicida subspecies

salmonicida is a Gram-negative, nonmotile, facultative,
anaerobic rod. An atypical strain of A. salmonicida has been
isolated from a wide range of cultivated and wild fish species
including cyprinids, marine flatfish, and non-salmonids as well
as salmonids, inhabiting freshwater, brackish water and
marine environments.
 SKIN AND FIN DISEASES

FURUNCULOSIS

Clinical presentation: Acute and chronic furunculosis can occur and is contingent on water
temperature, age of the fish and pathogenicity of the organism.
Outbreaks are prompted by stressors including sudden changes in water temperature,
handling, crowding and poor water quality.
Occurrences of acute furunculosis result in rapid fish death with few or no prior signs of
disease, and hence pathological changes are infrequent.
For chronic infections the fish show lethargy, inappetence, pale gills and darkening of the
skin, although such clinical signs are also reported for other bacterial septicemias. Ventral
hemorrhage may be seen near the base of the pectoral, pelvic and anal fins in addition to
exophthalmia.
Liquefactive, hemorrhagic “boil” lesions occur in the superficial muscle, and raised,
fluctuating lesions that may rupture can be seen on the skin surface. Although such
furuncles are characteristic, they are not always present in diseased fish and are not regarded
as a diagnostic characteristic.
Gross pathology includes ascites, splenomegaly, subcapsular hemorrhage involving
the liver and pyloric ceca. The intestine is devoid of food and often contains exudates
of blood, mucus and cellular debris. A carrier state may become established in surviving
fish after an infection.
 SKIN AND FIN DISEASES

FURUNCULOSIS

Differential diagnosis: Tissue

sections stained with
hematoxylin and eosin highlight
colonies of bacteria in the
heart, kidney, muscle, pancreas
and spleen. Motile Aeromonas
species (e.g., A. hydrophila)
have been implicated as the
causative agents of various fish
septicaemias. Other bacterial
infections should also be
considered as part of a
differential diagnosis including
those attributed to Vibrio spp.
infections.
 SKIN AND FIN DISEASES

FURUNCULOSIS

Diagnosis: Sophisticated techniques are available including

molecular methods, but primary isolation of the pathogen and
hence confirmation are readily achieved from the kidney and
other organs using media such as tryptic soy agar (TSA) or
brain heart infusion agar (BHIA) incubated at 22°C. Most
strains of A. salmonicida are nonmotile, aerobic, and oxidase-
positive and produce a distinct water-soluble brown pigment
on media containing tryptone. Diagnosis is based upon gross
and histopathological lesions and isolation of the causative
agent.
 SKIN AND FIN DISEASES

AEROMONAS

Overview: An opportunistic bacterial skin and fin disease of all freshwater

and occasionally marine fishes.

Etiological agent: The disease, a condition often called “motile aeromonad

septicemia,” is caused by motile aeromonads of the Aeromonas hydrophila
complex: A. hydrophila, A. sobria and A. veronii.

Clinical presentation: The most recognizable clinical signs of disease are

hemorrhagic fin and tail erosions, cutaneous hemorrhage and skin
ulcerations.
 SKIN AND FIN DISEASES

AEROMONAS

Pathology of disease: A number of virulence factors have been shown to be involved with the
pathogenicity of the organism. Acute infections may result in a generalized septicemia with
few clinical signs or the fish may exhibit non-specific signs of erythema of the skin,
exophthalmia and accumulation of coelomic fluid. Skin infections result in dermal ulceration
with focal hemorrhage and inflammation with the underlying musculature eventually becoming
necrotic.
 SKIN AND FIN DISEASES

AEROMONAS

Differential diagnosis: Skin and fin lesions caused by other infectious

agents such as bacteria (e.g., A. salmonicida, Flavobacterium columnaris,
Renibacterium salmoninarum, Edwardsiella spp., Mycobacterium spp.,
Pseudomonas spp., Vibrio spp.) and protozoan ectoparasites must be
differentiated from A. hydrophila infections. As an opportunistic pathogen,
A. hydrophila may also exist as a co-infection with other pathogens.

Diagnosis: The motile, Gram-negative bacilli can be isolated on a number

of bacteriologic media and can be identified by standard biochemical tests,
direct or indirect fluorescent antibody techniques, ELISA, or polymerase
chain reaction (PCR) assay.
 SKIN AND FIN DISEASES

EDWARDSIELLOSIS

Overview: Several species of Edwardsiella, in addition to E. ictaluri, are

pathogenic to finfish. Agents of edwardsiellosis previously classified as E.
tarda affect a wide range of fish taxa including teleosts as well as
elasmobranchs in tropical and temperate freshwater and marine
environments worldwide.

Etiology: Edwardsiella tarda, E. piscicida and E. anguillarum are Gram-

negative, motile, short rod bacteria. Following the establishment of the
genus Edwardsiella in the family Enterobacteriaceae, fish pathogenic
Edwardsiella other than E. ictaluri were considered to be representatives of
the species E. tarda.
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EDWARDSIELLOSIS

Clinical presentation: Edwardsiellosis is commonly described as a systemic disease, but mild

infections in channel catfish (Ictalurus punctatus) have been reported to manifest as small
cutaneous lesions located on the posterior-lateral surfaces of the body. External lesions
including petechial hemorrhages and ulcers may also occur on the mouth, isthmus, operculum
and abdomen. As the disease progresses in channel catfish and certain other fish, deep
ulcers and abscesses develop within the skeletal musculature.

The abscesses are often visible from the fish surface as convex, depigmented swollen areas
and are filled with malodourous gas and necrotic tissue remnants.

External clinical signs observed in various species include exophthalmos and swelling around
the eyes, eye opacity, ecchymosis and congestion of the skin and bases of the fins, increased
mucus production, fin erosion, abdominal swelling associated with ascites, and swelling and
hemorrhage around the vent with occasional rectal prolapse. In some fish species, reddening
of the head associated with subcutaneous hemorrhages has given rise to the name “red-head
disease.” Fish with edwardsiellosis may also show loss of equilibrium and either lie on their
sides at the bottom of a tank, or float on the surface due to swim bladder hyperinflation.
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EDWARDSIELLOSIS
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EDWARDSIELLOSIS

Differential diagnosis: Skin lesions caused by other bacterial pathogens

such as Aeromonas hydrophila, Pseudomonas anguilliseptica, Vibrio
anguillarum and Mycobacterium spp. may appear similar to those caused
by edwardsiellosis.

Diagnosis: Identification of the causative bacterium is usually

accomplished by isolation on standard media such as brain-heart infusion
agar (BHIA) or tryptic soy agar (TSA) at 26°C–30°C for 24–48 h.
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PISCIRICKETTSIOSIS

Overview: This is an acute, subacute or chronic systemic bacterial disease

affecting a variety of teleost species farmed in brackish and seawater in
different locations of the world. The disease is endemic in salmonids
cultured along the shores of southern Chile.

Etiology: Piscirickettsiosis is caused by Piscirickettsia salmonis, a Gram-

negative, non-motile, pleomorphic, facultative intracellular bacterium that
multiplies in macrophages and several other cell types of the host.
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PISCIRICKETTSIOSIS

Clinical presentation: Clinical signs are nonspecific, and include lethargy, anorexia, skin
darkening in the dorsal area of the fish, respiratory distress, coelomic swelling and abnormal
swimming. Skin abnormalities are not always present, but in some cases are the only gross
lesions shown by fish infected with piscirickettsiosis. Areas with raised scales are usually
observed, as well as small nodules that progress to shallow ulcerations. Petechial and
ecchymotic zones of the skin can occur either accompanying the lesions already mentioned or
by themselves. In some field outbreaks, diseased fish show skin ulcers that are extensive and
deep. It has been experimentally demonstrated that P. salmonis can enter through the skin in
salmonid fish. Skin sites exposed to P. salmonis show progressive bacterial penetration,
inflammation and necrosis. Integument lesions in nonsalmonid species affected with
piscirickettsiosis are rare, and when they occur are usually mild. The septicemic infection
causes anemia, reflected by gill paleness and dyspnea, and ascites. Vascular and
perivascular necrosis is common, and the internal organs can be eventually affected showing
inflammation, hemorrhage, degeneration and necrosis. Internal lesions are nonspecific, but
some fish may show mottled livers with white to yellowish, circular foci that appear solid or as
ring-shaped formations that are indicative of the presence of this disease.
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PISCIRICKETTSIOSIS
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PISCIRICKETTSIOSIS

Differential diagnosis: A variety of septicemic infections of fish can share some clinical and/or
pathological features with piscirickettsiosis, but due to its closer similarity, francisellosis is
probably the most important disease to be considered in a differential diagnosis. Other
infectious diseases (e.g., bacterial, parasitic or mycotic) can cause granulomas or nodular
abscesses in the liver and require investigation to differentiate these from piscirickettsiosis.

Diagnosis: Diagnosis is based on the identification of the bacteria in fish having clinical and
pathological manifestations of the disease. A presumptive diagnosis is obtained by the
observation of organisms morphologically and tinctorially compatible with P. salmonis in tissue
smears or imprints stained with Gram, Giemsa, Giménez, Pinkerton’s method or toluidine
blue. Histopathology is helpful to support the diagnosis. Bacterial isolation provides a
definitive diagnosis, but this may be difficult as it requires the use of enriched axenic media or
fish cell cultures.
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VIBRIOSIS

Overview: Diseases caused by Vibrio spp. affect a wide range of wild and
farmed fish species around the world, and occur mostly in marine and
brackish water environments, although disease has also been reported in
freshwater environments. Most outbreaks are associated with relatively
warm water temperatures (>15°C), and therefore occur mainly in the
summer in temperate climates.

Etiology: Several Vibrio species and closely related bacteria cause finfish
diseases. The etiologic agents of “classical” vibriosis are Vibrio (Listonella)
anguillarum and V. ordalii.
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VIBRIOSIS

Clinical presentation: Vibrio spp. generally cause disseminated systemic

infections. External macroscopic signs are considered nonspecific and
may include lethargy and weight loss, dark swollen skin lesions that may
develop into bleeding ulcers and abscesses, hemorrhages at the bases or
other areas of the fins and on the body (especially on lateral and ventral
surfaces, at the vent, and around and in the mouth), hemorrhages of the
gills, eye opacity, ulceration and exophthalmos, and abdominal distension
associated with ascites. Winter ulcer initially presents as small, raised skin
lesions that progress to rounded or oval ulcers that expose underlying
muscle, and are characterized by a white demarcation zone separating the
lesion from normal tissue. Fish affected by winter ulcer may exhibit lesions
for long periods of time and may recover with increasing water
temperature.
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VIBRIOSIS
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VIBRIOSIS

Differential diagnosis: Skin and fin lesions associated with bacteremia and
viremia caused by pathogens such as Aeromonas salmonicida and other
Aeromonas spp. and viral hemorrhagic septicemia virus (VHSV) should be
excluded. For suspected cases of winter ulcer, lesions caused by atypical
Aeromonas salmonicida infection should also be excluded.

Diagnosis: Preliminary diagnosis of vibriosis is made by detection of

curved, motile, Gram-negative rods in the kidney, spleen or blood samples
from marine, estuarine or anadromous fish. Culture at 15°C–25°C on
standard media such as brain-heart infusion agar (BHIA), tryptic soy agar
(TSA) or thiosulfate-citrate-bile salts-sucrose (TCBS) agar may require
addition of 0.5%–3.5% NaCl for growth of vibrios.
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