Protozoans of fish

Wesonga Samwel
SNAT/FAS/M/002/20
 Introduction
• The Phylum Protozoa brings together several organisms evolutionarily different
that may act as ecto or endoparasites

• Protozoa is grouped into several phyla; flagellates, Opalines, Amoebae, Coccidia,

Myxosporida, Microspiridia and ciliates

• Today about 50 000 spp of Protozoa lives in water or soil habitats, most of them
are serious pathogens

• They are the causative agents of diseases in the global aquaculture causing,
among other things, damage and reduced growth of the host fish

• Favoring secondary bacterial infections and mortality, all leading to constraints in

 Ciliophora
• These unicellular protozoans possess mobile cilia involving the external
body surface in some stage of their life cycle

• Cytostome, macronucleous and micronucleous present

• Reproduction by binary fission and conjugation

• Main representatives are

• Apiosoma, Balantidium, Chilodonella, Epistylis, Ichthyophthirius multifiliis,

Nyctotherus, Rhynchodinium paradoxum, Tetrahymena, and Trichodinidae
 Ichthyophthirius multifiliis
(Fouquet, 1876)
• Ichthyophthiriasis or white spot disease (ICH)

• Is a holotrichous ciliate and the most important

fish parasites of worldwide

• Distribution compromising skin, fins, gills and eyes

of farmed fish

• Not host specific and any freshwater fish can

potentially transmit the parasite

• Appears as a varying number of classical round and

Source Michael, 2015
slightly raised spots on the fish
 Life cycle
• Monoxenic - involves only a fish to be completed

• its life cycle has three stages as follows:

i. Theront: infective and mobile form; it needs to

find a host otherwise it will die.

ii. Trophont: adult mobile stage found in fish

iii. Tomont: free form of the parasite provided by a

cyst for protection. Attaches to plants and
substrate to divide asexually to produce tomites
that will differentiate in infective theronts
 Transmission

• Its transmission occurs by co-habitation with infested fish or directly from the
theronts

• Fishery utensils used in fish farms and water transport are potential vectors
of ichthyophthiriasis

• The release of theronts into the water is strongly associated with

temperature.

• At water temperatures above 24°C, the life cycle is favored and completed
rapidly. Differently, temperatures below 10°C or above 28°C can inhibit the
parasite life cycle
 Diagnosis
• Ichthyophthiriasis diagnosis is made based on;
• macroscopical observation of trophonts within the host’s skin and,
• microscopical analysis of fresh-mounted material (skin, fins and gills scraps) between a
slide and a coverslip

• Under an optical microscope, the observation of the mobile pear-shaped

theronts and mature trophonts uniformly covered by a layer of external cilia
and with a horse-shoe shaped nucleus confirms the diagnosis

• Histopathological analysis can also reveals the parasitosis

• Because of its similarity with the dinoflagellate Piscinoodinium pillulare,

definitive diagnosis ought to be careful
 Pathogenesis and clinical signs

• The main clinical sign is the presence of white spots on the fish surface
including skin, fins, eyes, buccal cavity and gills

• Others are anorexia, hyperventilation, abnormal swimming behavior, flashing,

discoloration

• The infection challenges the hosts’ osmoregulation and respiration

• The inflammatory reaction of the host with intense epithelial proliferation is

provoked by theront invasion on the epithelium layers with posterior
histophagia stage of trophont
 Treatment and control
• Malachite green, Formalin

• Copper sulphate, methylene blue and potassium permanganate

• Hydrogen peroxide and Hydrogen peroxide releasing products like sodium

percarbonate and peracetic acid

• Lipopeptide secreted as a surfactant from the bacterium Pseudomonas H6 kills

theronts, tomonts and tomocysts

• Total fish removal and repeated transfer to clean tanks may be applied
 Flagellates (phylum
Mastigophora)
• Flagellate protozoans are mainly characterized by the presence of one or
more flagella for movement

• They have one nucleus, or exceptionally more monomorphic nuclei

• The majority of them are ectoparasites while others can be found parasitizing
internal organs

• They reproduce by longitudinal binary fission

• Amyloodinium ocellatum, Piscinoodinium pillulare, Trypanosoma, Cryptobia and
Ichthyobodo are the main representatives.
 Ichthyobodo (Pinto, 1928)

Ichthyobodo necator- Causative agent of ichthyobodiasis

• These obligatory parasites are small biflagellated kinetoplastids found in skin, fins and
gills of wild and farmed marine and freshwater fishes from temperate and tropical waters

• Similar to Cryptobia, it is pear-shaped and when fixed on the host it shows circling or
zigzag movements
 Life cycle
• Ichthyobodo presents monoxenic life cycle
• Are single celled and reproduce asexually via cell
division
• They are able to parasitize fish almost immediately
after division – no larval/developmental stages

Transmission
• The parasitosis is horizontally transmitted by direct contact among diseased
and healthy fish.
• Free-swimming infective parasites are responsible for disease dissemination
as well as contaminated fishery utensils.
 Pathogenesis and clinical signs
Diagnosis
• In the pathological alterations - destruction of
• Fresh-mounted smears of skin and gills
the epithelial cells due to parasite attachment
viewed under microscope is the main
technique for routine diagnosis • Attaches via a disk like structure that cuts into
the skin then extend microtubules into the
fish’s cell
• Histopathological analysis also presents
an efficient tool for diagnosis of
• Clinical signs are non-specific indicating
ichthyobodiasis but non-sensitive in respiratory difficulty and altered skin and gill

low parasitism intensity color

 Treatment and control
1. Salt baths (ParaSalt): safe for gill damage or difficulty breathing (72hrs), to avoid
stress start with 3% salt

2. Formalin: careful dosing is recommended due to its toxicity to the fish, short
baths (2hrs) in solution with 50 ppm or 5ml/10 gallons

3. Paraziquantel: gentle chemical used in less serious cases

4. Quarantine & inspection: is the primary prevention, use microscope to check for
costia in skin or water

5. Improve water quality: activated carbon captures and neutralizes costia and
pollutants enabling it. Aqua-plants naturally help to filter and oxygenate water
 Amoebae (Phylum Rhizopoda)
• Their cell has a simple structure which a naked plasmalemma

• Move using pseudopodia or a simple protoplasmic flow

• Flagella when present are usually restricted to developmental or sexual stages

• There are very few specific endocommensals of fish (spp of genera Entamoebae or
Schizamobea)

• All other amoebae infecting fish are amphizoic species; free-living forms (which under certain
conditions can colonize fish)

• These are found in families Hartmanellidae, Mayorellidae, Acanthamoebidae and

Vahlkamphiidae
• Often the relationship between amoeba and fish diseases is not clear
(commensals and parasites)

• Not associated with the diseases on surfaces they colonize

Diseases Caused by Amoebae

a. AGD (amoebic gill disease) associated with Neoparamoeba perurans -
M
b. NGD (nodular gill disease) caused by different species of amoebae
involved – F
c. Systemic granulomatous disease associated with he archamoebic species
Endolimax piscium
 AGD (amoebic gill disease)
• AGD is nowadays one of the most relevant disease in Atlantic salmon (Salmo salar)
farming in different geographical locations

• Causes high mortality (sometimes up to 85% in a total production cycle) and a

substantial reduction in growth

• The causative agent of AGD is the amphyzoic amoebae Neoparamoeba perurans

(formerly known as Paramoeba perurans)

• AGD is typically a chronic disease, but with a relatively fast development

• Affects not only gills but also causes a progressive systemic involvement of other
organs due to their effects on respiratory, osmoregulatory and circulatory functions
 Pathenogenesis
• It is characterized by the development of a progressive hyperplasia of epithelial
gill cells in the primary and secondary (lamellae) filaments

• This leads to an increase of the thickness of the gill epithelia, progressive

formation of synechia and lacunae between filaments and finally total
obliteration of the interlamellar space

• These alterations in gill structure lead to an impairment in respiration and

osmoregulation
Clinical signs and symptoms
• Affected fish become lethargic, with reduced swimming speed, feeding rates
and growth, and display signs of respiratory distress (increased rate of
ventilation)
 Myxozoans
Myxobolus cerebralis – the “whirling disease”
• Odd, exclusively endoparasites

• Worldwide distribution (salmon and trout spp)

• Multicellular during adult life

• Attacks cartilage (younger fish have more cartilage)

• Infections in the spine can cause the fish’s tail to turn black and the spine to curve

• Infections in the head cartilage create head and jaw deformities, while infections in the
auditory capsule cause young trout to become disoriented and chase their tails in a
A) Infected trout are whirling (photo by Drew Mitchel) Spinal curvature (Photo by Glenn Hoffman)

Heavy infections can kill fish before clinical signs have a chance to develop
 Life cycle
• Spores can be shed from infected live fish as well as from dead and
decomposing fish.

• The spores are very resistant (15yrs dessication)

• The spores also can be spread via bird feces.

• Spores are ingested by an annelid worm intermediate host, Tubifex tubifex,

which lives in the bottom mud

• The spores develop into actinosporeans that penetrate fish or injested

• Plasmodia develop in the fish’s cartilage and eventually produce the
characteristic spores.
 Diagnosis
a) Remove gill arch, grind and allow to settle then check for spores

b) Flourescent Antibody Test (FAT)

c) Pepsin – trypsin digestion/centrifuge

Transmission
• Direct during the first year

• Indirect via annelid or contamination

 Prevention
• Not stocking trout fry in infested waters until they are older than 6 months

• Raising fish only in concrete tanks or raceways

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