PARASITOLOGY

HONOURS
PROTOZOA
 PROTOZOA

Species affected:
All species, especially fry and fingerlings.

Gross signs:
• Changes in skin (colouration).
• Scattered haemorrhages, red sores, eroded scales and
excessive mucus production.
• Infected fish often respond by “fleshing” (rubbing).

Cause:
• Ectoparasitic flagellates of the genera Costia, Colponema &
Bodomonas.
• Ciliates of the genera Trichodina, Epistylis, Ambiphrya,
Apiosoma, Chilodonella and Ichthyophthirius.
• Suctorians of the genus Trichophyra.
Diagnosis:

• Fish should be examined fresh.

• Make skin and gill scrapings and follow fixation
and staining techniques.

Morphology:
• Differential diagnosis key:

Sarcodina – Amoebida – neither flagella nor cilia

present, “amoeboid” body shape, and movement by
pseudopodia.

Genera: Acanthamoeba,
Entamoeba
  Flagellata (Mastigophora) – one or more flagella:

• Genus Ichthyobodo (Costia)

 Four flagella, two long and two short, free-swimming
forms are rounded, sessile form are oval.
Ichthyobodo (formerly Costia)
Ichthyobodo (Costia) is a single cell
microscopic parasite that attacks skin of
fish. It is a flagellate. It destroys the skin; in
mild cases the skin appears cloudy. In
severe cases skin is open and bleeding. It
usually attacks fish in a poor state of health
or living in poor water conditions or in
overcrowded conditions. Secondary
bacterial and/or fungal infections can occur
at open sore areas. Costia can only survive
on fish. If it is without fish for 1 hour or
more it dies. It cannot live in temperatures
greater than 86°F.
• Genus Colponema
Two flagella, blepharoplast rod-shaped.
• Genus Bodomonas
Two flagella, blepharoplast rounded.
 Ciliata – cilia present, covering part of the entire
body.

• Genus Ichthyophthirius
Entire body covered by cilia, cytostome at anterior
end, not visible in large individuals, macronucleus large
and horseshoe-shaped.
• White spot trophonts seen at low power wet mount of a skin
scrape.
• Note the light horseshoe shaped nucleus and variation in size.
• Histological preparation of gill tissue with embedded
trophonts.
• Note the distortion to gill tissue and severe
hyperplasia.
• Genus Chilodonella

Cytostome distinct, ciliation complete, and cilia

arranged in several parallel rows on concave ventral
surface, macronucleus round.
Class Kinetophragminophorea de Puytorac et al., 1974
Order Cyrtophorida Faure-Fremiet in Corliss, 1956
Family Chilodonellidae Deroux, 1970
Genus Chilodonella Strand, 1926

• Many free-living species and two species infesting

freshwater fishes.
• Both parasitic species have cosmopolitan distributions,
occurring in estuarine and brackish waters as well.
• Both species appear to also infest most, if not all, teleost
fishes.
• They cause the well-known chilodonellosis, a disease
affecting the skin and gills, especially in fish cultures.
Chilodonella piscicola (Zacharias, 1894) Jankovski,
1980
Syn. C. cyprini (Moroff, 1902, see Shulman and
Jankovski, 1984)
1. Asymmetrically oval body, with clear notch in
posterior margin
2. 55x43 µm (range30-80 µm x 20-62 µm)
3. Mostly 8-11 (range 7-15) kineties in right
arched ciliary band), and 12-13 (range 8-14) in
straight left band.
4. Cytopharynx curved at inner end, reinforced
by 14-16 nematodesmata.
5. Two contractile vacuoles present, one
anteriorly at the right, other posteriorly at left.
6. Infests fingerlings more than adult fish.
Chilodonella hexasticha (Kiernik, 1909)
Kahl, 1931
1. Absence of a notch at posterior body
margin.
2. Has less numerous & more loosely
spaced kineties (5-7 in right, and 7-9 in
left ciliary band)
3. Smaller (30-65 µm x 20-50 µm).
4. Distribution less widespread than C.
piscicola.
5. Prevails on older fish.
• Genus Trichodina

Cup-shaped, with ventrally located ring of denticles as

an attachment organelle.
• Genus Epistylis

 Goblet or conus-shaped, cilia limited to wider dorsal

mouth zone (oral zone), several individuals sprout
from a common stalk.
 Macronucleus elongated or ribbon-shaped.
• Genus: Apiosoma

 Similar in shape to Epistylis, but individuals are solitary

without stalk.
 Macronucleus compact, conical or ellipsoidal.
• Genus: Ambiphrya

 Solitary organisms cylindrical rather than conic in shape,

with a widened attachment base at adoral end, cilia
located around mouth area, permanent equatorial ciliary
fringe of cilia.
 Macronucleus long, thin and ribbon-like.
• Suctorians

 Cilia absent, group of tentacles arises from the body,

genus Trichophyra.
Life history:

• Reproduction: binary fission.

• Flagellates attach themselves to integumentary cells via
flagella, Trichodina via oral denticles, attachment of
Childonella is aided by ventral cilia, in Ambiphrya and
Apiosoma the adoral end develops into an attachment
organelle, while in Epistylis the stalk is the attachment
organelle.
• Some Epistylis species are non-obligate parasites.
Ambiphrya and Apiosoma utilize the host only as a
substrate for attachment. Epistylis, Ambiphrya and
Apiosoma are filter feeders, where as suctorians are
predators. Trichodina, Chilodonella and parasitic
flagellates - more specialized parasites and feed from host
tissues.
• Site preference varies with the different species.
Pathology:
• Costia:
 Results in acute hyperplasia of gill epithelium.
 These cellular changes cause fusion of gill lamellae and
proliferation of mucous cells.

• Skin infections in catfish results in:

 excessive mucus production,
 degeneration and necrosis of epithelial cells and
 hyperaemia of dermis.

• Pathological changes in infections with Trichodina and

Chilodonella include:
 irritation of skin integument and gill epithelium resulting in
 hyperplasia and
 later degeneration and
 necrosis of epithelium lining and
 proliferation of mucous cells (excessive mucus production).
• In some Trichodina infections of gills:
 desquamation and
 erosion of epithelium takes place.
• Mechanical interference with gas exchange is suggested as the
cause of morbidity.
• In Epistylis infections:
 irritation caused by the attached stalked parasites cause large
haemorrhagic lesions,
 with erosion of scales, surface of dermal bones, and hard fin rays.
 Lesion begins as a small hyperplastic protrusion of epithelium,
which later erodes and becomes haemorrhagic.
• Additional pathological changes may be superimposed by
secondary bacterial and fungal infections resulting in severe
cutaneous ulcers and even bacterial septicaemia.

Control:
• Protozoa can successfully be controlled by 15-25 ppm formalin.
 CILIOPHORA

7200 species

CHARACTERISTICS
• Multi-nucleated.
• Cilia in its ciliary organelles.
• Cytostome present.
• Asexual reproduction: binary fission.
• Sexual reproduction: conjugation & autogamy.
CLASSIFICATION
Kingdom Protozoa
Phylum Ciliophora
Class Oligohymenophorea
Subclass Peritrichia
Order Peritrichida

Suborder 1: Sessilina
Family 1: Epistylidae
Genus Apiosoma
Epistylis
Family 2: Operculariidae
Genus Propyxidium
Family 3: Scyphidiidae
Genus Riboscyphidia
Ambiphrya
Family 4: Ellobiophoridae
Genus: Caliperia
Suborder 2: Mobilina
Family 1: Urceolariidae
Genus Urceolaria
Leiotrocha
Polycycla
Family 2: Trichodinopsidae
Genus: Trichodinopsis
Family 3: Trichodinidae
Genus Trichodina
Trichodinella
Tripartiella
Paratrichodina
Dipartiella
Hemitrichodina
Trichodoxa
Semitrichodina
Pallitrichodina
Vauchomia
CHARACTERISTICS OF OLIGOHYMENOPHOREA

• Oral apparatus – hollow infundibulum on ventral surface

of body with cytostome at the base.
• Cytopharynx not visible.
• Conjugation temporary.
• Mostly free living symbionts.

CHARACTERISTICS OF PERITRICHIA / PERITRICHIDA

• Body cup-shaped.
• Mouth (buccal ciliature) anti clock-wise.
• Scopula not present.
• Ciliated infundibulum leads to cytostome.
• Widely distributed – free living, commensialists, parasitic.
SUBORDER: SESSILINA

• Sessile organisms, permanently attach to substratum.

• Oral ciliature well developed.
• Scopula well developed.
• Pedicle striations present.
• Telotroch larvae in life cycle.

 Large number of sessile organisms, but mostly they

occur on a non-living substrata like rocks, soil or other
animals and plants.
 Only some are associated with fish, although real
parasitism is not involved.
Key to Sessiline genera epizoic on fish
(According to Lom & Dykova, 1992)

1a. Ciliophorans attached to substrate directly by scopula,

mostly circular, often of large diameter, sometimes
lobed, occasionally split into long projections…………….2
1b. Ciliophorans attached by semicircular outgrowth of
scopula joined to form a circle around the gill
lamellae……………………………………………..………….Calliperia

2a. Locomotory fringe of cilia exists only in telotrochs……….3

2b. Locomotory fringe of cilia permanent, occurs also in
attached ciliophorans…..…..…………………….……Ambiphrya
3a. MA compact, conical or ellipsoidal, cell shape usually
elongated conical……….……………….................Apiosoma
3b. MA sausage-formed, cell shape mostly cylindrical
……….………………………………………………………Riboscyphidia

4a. Non-contractile stalk is branched, bearing a small

colony of several zooids….…………………..………....Epistylis
4b. Non-contractile stalk short, bears a spoon-like shield
sheltering a single zooid……………………..…….Propyxidium
4c. Stalk contractile, unbranched, bearing colonies of many
zooids, opportunistic epibionts of genera ……………...…….
..……………………Vorticella, Zoothamnium & Carchesium
Families associated with fish:

FAMILY: EPISTYLIDIDAE

• Scopula produces non-contractile stalk, either simple,

bearing solitary zooid, or branched, bearing many
zooids.
• Retractile peristomial lip, more or less divergent,
encircles wide, slightly elevated epistomial disc.
• Great numbers of species exist, free-living and epizoic.
• Species of 2 genera occur on fish.

Genera: Apiosoma

Epistylis
Genus: Apiosoma

• Solitary, with circular scopula, of large diameter in some spp.

• Secretes short disc in shape of pad of fibrillar material.
• MA usually conical, apex pointed to scopula, or ellipsoidal.
• 66 spp on surface of freshwater fish.
• Live single.
• Often with stalk, non-contractile, transparent and not
branched.
• Body cylindrical, broad at peristome and tapers towards
scopula.
• Peristomial disc flat.
• Groove in centre of body.
• Micronucleus oval shaped, situated below macronucleus.
Fig. 1 Light microscopic images of
Apiosoma piscicola.
a. Living specimens stained with
Neutral Red, showing the strong
peristomial cilia (pc) of this
peritrich and the food particles (fp)
that were filter-fed. Scale bar = 20
μm.
b. & c. Specimens stained with
Ehrlich’s haematoxylin, showing
the general form, peristomial disc
(pd), adoral cilia (pc), transverse
ciliary band (tcb and nuclei (ma
macronucleus, mi micronucleus).
Genus: Epistylis
• Colonial, with bell-shaped or elongated cylindrical or conical
body.
• Non-contractile stalk bears several to many zooids.
• Vaulted epistomial disc slightly elevated above peristome.
• Macronucleus horse-shoe-shaped or very long, sausage-
shaped.
• Numerous spp. free-living, or epizoic mostly on invertebrates, 5
spp. on surface of freshwater fish.
FAMILY: OPERCULARIIDAE
• Have a narrow epistomial disc (different from former
family), operculum-like, elevated on narrow neck above
rigid, non-retractile peristomial lip.
• Numerous spp., free-living and epizoic.
• Single species in freshwater fish.

Genera: Propyxidium
• Solitary zooids on short stalk.
• Horse-shoe-shaped MA.
• Many free-living or epizoic spp.
FAMILY: SCYPHIDIIDAE

• Solitary, attached to substrate directly by scopula.

• Scopula may be very large.
• Body cylindrical or cylindrical-conical.
• Adoral cilia makes one turn around slightly elevated
epistomial disc.
• Two genera on fish.
Genus: Riboscyphidia

• Scopula extremely large, with thin flat border.

• Body mostly cylindrical, sometimes conical.
• MA sausage-shaped, often sinuous in oral half of cell.
• Surface of epistomial disc slightly vaulted above
peristomial lip.
• Exclusively on skin and gills of freshwater and marine fish.
• Approximately 18 spp.
Genus: Ambiphrya

• Similar to Riboscyphidia, but with permanent equatorial

ciliary fringe.
• MA long, thin and sinuous ribbon.
• Gills and skin of brackish and freshwater fish.
• Four spp.
FAMILY: ELLOBIOPHRYIDAE

• Solitary, fastened to substrate by means of 2 outgrowths

from either side of scopula, that encircles gill filament of
host and grow together to form a firm attachment ring.
• Two genera, one on marine fish.

Genus: Caliperia
• Body cylindrical, slightly contractile.
• Attachment outgrowths have a distinct central axis.
• Three spp. on fish.
Mobilina
SUBORDER: MOBILINA
Characteristics:
• Mobile forms, conical, cylindrical.
• Aboral disk = attachment structure (unique).
• Aboral disk made up of 3 rings:
• Denticular ring with denticles.
• Denticles consist of ray + central part + blade.
• Adoral side of blade is striated membrane and consist of
number of radial pins.
• Outside of striated membrane is thin border membrane.
• Ventral of velum – aborally of velum – is groove with long
cilia (ciliary girdle).
• Trichodines display two types of movement:
 (a) glide action with aboral-side in contact with host, and
 (b) rotating action in one place.
• Asexual reproduction: binary fission.
• Sexual reproduction: conjugation.
GENERA: Trichodina (skin & gills)
Tripartiella (only gills)
Trichodinella (only gills)
SPECIES DIAGNOSIS:

1. Form of body.
2. Structure of attachment disk.
3. Form of denticles.
4. Dimensions of denticles.
5. Number of denticles.
6. Diameter of denticles ring.
7. Form, shape & dimensions of border membrane on
outside.
8. Nucleus apparatus.
9. Adoral zone (spiral degree).
10. Velum.
The End