Charles University of Prague

Faculty of Science

Protocol for the

Field Course in Fish Parasitology
for Erasmus Students

Teacher: Libor Mikeš

Student:
Andreea-Monika Lamoly

South Bohemia, Czech Republic

19.10.2023
Contents

I. Fishes...............................................................................................................................................2
II. Parasites...........................................................................................................................................2
1. Khawia japonensis....................................................................................................................2
2. Atractolytocestus huronensis.....................................................................................................3
3. Valipora campylancristrota.......................................................................................................4
4. Glanitaenia osculata (Proteocephalus osculatus)....................................................................4
5. Eudiplozoon niponicum.............................................................................................................5
6. Dactylogyrus vastator...............................................................................................................5
7. Diplostomum spathaceum.........................................................................................................6
8. Tylodelphys clavata...................................................................................................................6
9. Myxidium lieberkuehni..............................................................................................................7
10. Zschokkela nova........................................................................................................................8
11. Argulus japonicus......................................................................................................................8
III. References....................................................................................................................................9

1
 I. Fishes

In 19th of October, we observed a total of 14 species of fish whose taxonomical

classification is:
Class: Actinopterygii
Order: Cypriniformes
Family: Cyprinidae
1) Cyprinus carpio – Common carp
2) Ctenopharyngodon idella – Grass carp
3) Hypophthalmichthys molitrix – Silver carp
4) Abramis brama – Common bream
5) Blicca bjoerkna – Silver bream
6) Rutilus rutilus – Common roach
7) Scardinius erythrophthalmus – Common rudd
8) Tinca tinca - Tench
9) Carassius gibelio – Prussian carp
10) Pseudorasbora parva – Stone moroco
Order: Perciformes
Family: Percidae
11) Gymnocephalus cernuus – Ruffe
12) Sander lucioperca - Zander
Order: Esociformes
Family: Esocidae
13) Esox lucius - Pike
Order: Siluriformes
Family: Siluridae
14) Silurus glanis – Catfish

II. Parasites

After the fish presentation, we started parasites search and identification. We found 11
species of parasites in various developmental stages. I checked more sources for the
taxonomical classification (WORMS data base, Wikipedia , USGS.gov).

Kingdom: Animalia
Phylum: Platyhelminthes
Class: Cestoda
Order: Caryophyllidea
1. Khawia japonensis
o found in the intestines of the common carp (Cyprinus carpio); other
fish reported to harbour K. japonensis are considered to be atypical
host
o its area of distribution is usually Asia and North America (as K.
iowensis – which is morphologically identical), but more cases of

2
 infection are reported in Europe as a result of fish import (Scholz et all,
2011)
o life cycle (Molnár& Székely, 2019):
i. the unembryonated eggs of the parasite are released into water
and then sink to the bottom
ii. these eggs are eated by the tubificid oligochaetes (intermediate
host – eg. Limnodrilus sp.) in which large plerocercoid larvae
can develop
iii. the fish eat the oligochaetes and become infected
o in experimental conditions, in 15 days after the release in the water,
the eggs have fully developed, infective oncospheres (Scholz et al,
2011)

Figure 1 Khawia japonensis

2. Atractolytocestus huronensis
o found in the intestines of the common carp (Cyprinus carpio), for
which demonstrate strict (oioxenous) host-specifity
o life cycle (Oros et al, 2011)
i. as the previous representant of caryophyllid tapeworms, A.
hunonensis need an aquatic annelid as intermediate host (eg.
Tubifex or Limnodrilus)
ii. after the ingestion of the eggs and hatching in the intestines of
the intermediate host, the larvae penetrate from the intestines to
the body cavity using penetration glands
iii. the fully completed development of the plerocercoid (infectious
larvae) takes 58-60 days
iv. the definitive host is infected by the ingestion of intermediate
host harbouring ripe plerocercoids

3
 Figure 2 Atractolytocestus huronensis

Order: Cyclophyllidea
3. Valipora campylancristrota
o found as larva in the gall bladder of a tench (Tinca tinca); can be found
in other cyprinids or fish in the Palaearctic Region which can serve as
the second intermediate host
o life cycle (Jarecka, 1970 and Scholz et all, 2004) is interesting because
it requires 2 intermediate hosts:
i. first intermediate host is the copepods as Eudiaptomus
graciloides or Arctodiaptomus salinus in which the larva is
called metacestodes (“cercoscolex”):
 acystic larva with a cercomer and an invaginated
acetabulate scolex
 it develops in 22 days at room temperature
ii. the second intermediate host are cyprinid fishes in which the
larva is called plerocercus:
 the fish are infected after the ingestion of the copepod
 the metacestode migrate from the intestin to the gall
bladder and become plerocercus
 plerocercus are acystic larva with an evaginated scolex
 an additional envelope develops around the neck region
in 20 days after infection, as a result of reaction of the
host
iii. the definitive host is grey heron (Ardea cinerea); the parasite is
mature after 12-15 days, when the first eggs are relesed; the
longevity is estimated to be about 9 months

4
Figure 3 Larva of Valipora campylancristrota
Order: Proteocephalidea
4. Glanitaenia osculata (Proteocephalus osculatus)
o found in the intestine of a catfish (Silurus glanis)
o life cycle (Scholz, 1999):
i. ripe eggs (containing oncospheres with embryonic hooks) are
released through the uterine pores; if the tapeworms were
placed in water, the eggs are spontaneously released
ii. the intermediate host is a planktonic crustacean (Copepoda) -
Cyclops strenuus who had consumed the floating eggs; in the
body cavity of this crustacean, the metacestode (or procercoid)
develops; especially at G. osculata, the procercoid possesees a
well-developed anterior part (scolex) with four muscular
suckers and perfectly functional
iii. the definitive host can be a fish (specific host of G. osculata is
Silurus glanis), an amphibian or a reptile which has ingested
the infected crustaceans
o the life span has a pronounced seasonality: summer or autumn - the
recruitment of new cestode generations; spring - with the increase of
water temperature, the tapeworms start to grow rapidly and mature,
after overwintering in fish; spring and summer - eggs laiding

Figure 4 Glanitaenia osculata

Class: Monogenea
Order: Diplozoidae
5. Eudiplozoon niponicum
o found in the gills of a common carp (Cyprinus carpio)

5
 o oviparous species with the life cycle (Pečínková et al, 2007 &
Valigurová et al, 2010):
i. oncomiracidium are free swimming larva, hatched in 3-5 days
from oval-shaped eggs with long filaments, which invades a
fish host and then metamorphoses into a post-oncomiracidial
larval stage - diporpae
ii. in the diporpae phase, two new superficial structures appear –
they play a crucial role in the fusion of two diporpae in
permanent copula; the fuse is produced with subsequent
rearrangement and interconnection of their nervous systems
iii. juvenile stage starts after the fusion; this period is characterized
by the development of attachment apparatus and reproductive
organs
iv. the adult phase cand be reached only as a pair; those who did
not find a pair can develop a full attachment apparatus, but fail
to mature; the grown (from egg to sexually mature adult) takes
aproximatly 33 days at 20°C;
Order: Dactylogyridea
6. Dactylogyrus vastator
o found in the gills of a common carp (Cyprinus carpio)
o life cycle (Wikipedia_Dactylogyrus_vastator):
 new hosts were infected by oncomiradium and live attached to
the gills of carp species; in about 10 days, the larva reaches
sexual maturity; the adult lays eggs that are washed out of the gill
cavity into the water; after 3-5 days the embryo is fully developed
 summer is the peak of infestation; late eggs enter diapause
(period of inactivity) and when the water temperaturerise again,
the eggs develop
Class: Trematoda
Order: Diplostomida
7. Diplostomum spathaceum
o found as metacercariae in the eye lens of a common bream (Abramis
brama)
o they can infect numerous species of fish (more than 125)
o life cycle (Capasso & Gutierrez, 2023):
i. adult worms develop in intestine of birds (final host) –
especially gulls and terns, and the eggs are released through
feces
ii. eggs hatch and release free-swimming miracidia; they
penetrated a lymnaeid snail (first intermediate host) tissue
iii. in the snail body, successive generations of sporocysts are
formed
iv. free-swimming cercariae are released into water; the fish
(second intermediate host) are infested via direct penetration
through the skin or gills

6
 v. the parasite forms metacercariae in the crystalline lens, that can
cause cataracts
vi. the birds are infected again by consuming infected fish

Figure 5 Diplostomum spathaceum as

metacercariae
8. Tylodelphys clavata
o found in the vitrous liquid of a roach (Rutilus rutilus)
o the life cycle is similar to Diplostomum sp. (Wikipedia_Tylodelphys)
o the parasite can affect the behavior of the host:
 in the day, Tylodelphys sp. flukes between the lens and retina
but in the night, it settle down and the fish can partialy see
 this strange phenomenon can be explain by the fact that the fish
eating birds are active in the day time, while longfin eels are
active in the night
 so the fish is vulnerable to the final host of the parasite, but
“protected” from other predators

Figure 6 Tylodelphys clavata

Phylum: Cnidaria
Class: Myxozoa
Order: Bivalvulida
9. Myxidium lieberkuehni
o was found in myxospore phase (as plasmodium) in the urinary blader
of a pike (Esox lucius)
o life cycle (Lom & Dyková, 2006) can be describe by the term
myxosporean which covers both life-cycle phases:
i. myxospore phase, resulting in myxospores

7
  hard-shelled, with two (sometimes more) shell valves
and are produced in small or large sporogonial
plasmodia; the “cell-in-cell condition” is typical - the
endogenously produced (secondary) cells persist inside
the primary (mother) cells
 the actinospore, discharged from an annelid into water,
randomly encounters the fish (intermediate) host and
upon the contact with the skin or gill epithelium the
spore shell valves open and the sporoplasm is released
 after that, a very complicated process follow:
presporogonic stages -> sporogonic stage (a
plasmodium or a pseudoplasmodium) -> plasmodial
stages (histozoic – in tisues or coelozoic – in organs) ->
sporogony, resulting in myxospores -> the myxospores
are released
ii. actinospore phase, resulting in actinospores
 involve a sexual process that takes place in annelids,
rarely sipunculids (the definitive host)
 after the ingenstion of the myxospore, another very
complicated process follow: the sporoplasm initiates the
merogony phase -> the enveloping cells have eight pairs
of gametic cells (α and β cells) resulted by meiosis ->
this cells fuse and produce eight zygotes -> the zygotes
become eight triradiate actinospores after cell division
and differentiation -> the actinospores are released

Figure 7 Myxidium lieberkuehni as plasmodium

(myxospore phase)
10. Zschokkela nova
o was found in myxospore phase (as spores) in the gall blader of a grass
carp (Ctenopharyngodon idella)
o the life cycle is very similar to the one of Myxidium lieberkuehni,
which is a closely phylogenetic related species (Lom & Dyková, 2006)
Phylum: Arthropoda
Subphylum: Crustacea
Class: Maxillopoda
Order: Arguloida

8
 11. Argulus japonicus
o dioecious ectoparasite found on the common carp (Cyprinus carpio)
and pikeperch (Sander lucioperca)
o life cycle (USGS_Argulus japonicus)
i. the matting occurs on the body of the host fish an then females
lay many eggs on hard substrate, which they cover in
gelatinous excretions
ii. the eggs hatch between 15 (at 35 ºC) and 61 (at 10ºC) days; in
the winter eggs may stay dormant until warmer temperatures
occur
iii. the larvae survive 1-2 days from their yolk sacs, but then they
have to consume host’s epithelial cells and mucus;
iv. their live cycle (excluding eggs laying) is 30-100 days, during
which they are attached to the fish

Figure 8 Argulus japonicus

III. References
1. Capasso, S.; Gutiérrez, J. S. Diplostomum spathaceum. Trends in Parasitology 39, no.
11 (2023): 969-970.
2. Jarecka, L. Life cycle of Valipora campylancristrota (Wedl, 1855) Baer and Bona
1958-1960 (Cestoda-Dilepididae) and the description of Cercoscolex--a new type of
Cestode larva. Bulletin de l'Academie polonaise des sciences. Serie des sciences
biologiques 18, no. 2 (1970): 99-102.
3. Lom, J.; Dyková, I. Myxozoan genera: definition and notes on taxonomy, life-cycle
terminology and pathogenic species. Folia Parasitologica 53 (2006): 1–36.
4. Molnár, K.; Székely, C.; Láng, M. Field Guide to Warmwater Fish Diseases in
Central and Eastern Europe, the Caucasus and Central Asia. FAO Fisheries and
Aquaculture Circular SEC/C1182 (2019).
5. Oros, M.; Král'ová-Hromadová, I.; Hanzelová, V.; Bruňanská, M.; Orosová, M.
Atractolytocestus Huronensis (Cestoda): A new invasive parasite of common carp in
Europe. In Carp: Habitat, Management and Diseases, 63-94. 2011.
6. Pečínková, M.; Matějusová, I.; Koubková, B.; Gelnar, M. Investigation of
Paradiplozoon homoion (Monogenea, Diplozoidae) life cycle under experimental
conditions. Parasitology International 56, no. 3 (2007): 179-183.
7. Scholz, T. Life cycles of species of Proteocephalus, parasites of fishes in the
Palearctic Region: A review. Journal of Helminthology 73, no. 1 (1999): 1-19.

9
8. Scholz, T.; Bray, R. A.; Kuchta, R.; Řepová, R. Larvae of gryporhynchid cestodes
(Cyclophyllidea) from fish: a review. Folia Parasitologica 51 (2004): 131–152.
9. Scholz, T.; Brabec, J.; Kráľová-Hromadová, I.; Oros, M.; Bazsalovicsová, E.;
Ermolenko, A.; Hanzelová, V. Revision of Khawia spp. (Cestoda: Caryophyllidea),
parasites of cyprinid fish, including a key to their identification and molecular
phylogeny. Folia Parasitologica 58, no. 3 (2011): 197-223.
10. Valigurová, A.; Hodová, I.; Sonnek, R.; Koubková, B.; Gelnar, M. Eudiplozoon
nipponicum in focus: monogenean exhibiting a highly specialized adaptation for
ectoparasitic lifestyle. Parasitology Research 108 (2011): 383–394.
11. USGS.gov [October 2023]
12. Wikipedia [October 2023]
13. WORMS data base [October 2023]

10