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Theoretical & practical course

INTRODUCTION TO
HELMINTHOLOGY &
FISH PRSITOLOGY
To!"# SCHOL$
&
lai% &e CHM'RIER
(HRTOUM
No)e!*er +,,-
.I%tro&uctio% to Hel!i%tholo/0 & Fish Parasitolo/01
U%i)ersit0 o2 (hartou!3 Su&a% 4No)e!*er +,,-5
To!"# Schol6 To!"# Schol6
INSTITUTE OF PARASITOLOGY
Biology Centre, Academy o Science! o t"e C#ec" Re$%&lic
'e!() B%d*+o,ice, C-EC. REPUBLIC
E/mail0 t!c"ol#1$ar%2ca!2c#
&
lai% &e Cha!*rier lai% &e Cha!*rier
3e$artment o In,erte&rate!
NATURAL .ISTORY 4USEU4
Gene,a, S5IT-ERLAN3
E/mail0 alain2dec"am&rier1,ille/ge2c"
'C(GROUND
Based on good experience from the course organized on 22-23 March 2006
at the Faculty of Science !ni"ersity of #hartoum the topics related to study of
helminth parasites and ichthyoparasitology are de"eloped in more detail to pro"ide a
more comprehensi"e introductory information$
%he course is formed &y a theoretical part 'lectures( and a practical part
'demonstration of helminth parasites and &asic techni)ues applied in helminthology
and fish parasitology($ * manual containing &asic data on the parasites of fish in
*frica together +ith a sur"ey of life cycles of selected representati"es of ma,or
groups of helminths is pro"ided to participants$ %his manual also contains the
information a&out &asic methods used in studies on cestodes parasitic in fish$
%he course should help undergraduate and graduate students researchers
and professionals dealing +ith human parasitology a)uatic animal health and
fisheries in a more effecti"e control of diseases caused &y parasitic animals$
O'7ECTI8ES
%he main o&,ecti"es of the course are-
to pro"ide &asic information on the importance of fish parasitology
to define principal groups of fish parasites +ith focus on their &iology and
control
to o"er"ie+ principal groups of helminths +ith focus on human parasites
TIME SCHEDULE ND PROGRMME
9EDNESDY :+ NO8EM'ER +,,-
.-.-30 /$ I%tro&uctio% '%$ Scholz(
/$/$ Scientific cooperation &et+een Sudan and S+itzerland &
0zech 1epu&lic- sur"ey of results
/$2$ 2rogram and structure of the course
.-30-// 2$ I%tro&uctio% to Hel!i%tholo/0 3 lecture '%$ Scholz(
2$/$ Sur"ey of ma,or groups and their characteristics
2$2$ 4ife cycles
2$3$ 5ctoparasites
2$6$ 5ndoparasites
2$7$ 8uman parasites
2$6$ Fish-&orne helminthoses
2$9$ 0ontrol of helminthoses
/2-/6 3$ De!o%stratio% o2 hel!i%th parasites 3 practical course '%$
Scholz & *lain de 0ham&rier(
- 5ctoparasites- Monogenea
- 5ndoparasites- Flat+orms :ematoda & *canthocephala
THURSDY :; NO8EM'ER +,,-
.-// 6$ Metho&s o2 stu&0 I 3 practical course '*$ de 0ham&rier(
2$/$ Dissectio% o2 2ish 'from #hartoum mar;et(
2$2$ Fi<atio% o2 parasites
/2-/3-30 7$ I%tro&uctio% to Fish Parasitolo/0 3 lecture '%$ Scholz(
/$/$ <efinitions & ma,or groups
/$2$ 8istopathology
/$3$ <inoflagellida & Myxosporea
/$6$ 5ndoparasitic helminths
/$7$ M=:=>5:5*
/6-/6 6$ Metho&s o2 stu&0 II 3 practical course '*$ de 0ham&rier(
3$/$ Staining of +orms and permanent mounts
/6-/6-30 9$ Cesto&es o2 2ish i% 2rica= >here to /o '*$ de 0ham&rier(
6$/$ 2roteocephalidean tape+orms- model for phylogenetic and
zoogeographical studies
6$2$ 5xpeditions to Sudan & 5thiopia
/6-30-/9 ?$ FINL DISCUSSION & FUTURE PLNS '@$ Mahmoud *$ de
0ham&rier & %$ Scholz(

Hu!a%?i%2ecti%/ 2lu@es
LIFE CYCLES OF
HELMINTH PRSITES
DEMONSTRTION OF
HELMINTH PRSITES
4PRCTICL COURSE5
DEMONSTRTION OF HELMINTH PRSITES
DIGENE
Caballerotrema arapaimae 3 intestine of Ara$aima giga!( 'infection +ith
metacercariae in fish(
Dicrocoelium dendriticum 3 gall &ladder of domestic animalsA rarely in man
'infection &y metacercariae in ants(
Fasciola hepatica 3 li"er of domestic animalsA numerous human cases
'infection +ith metacercariae 3 adolescariae 3 on plants and in +ater(
Opisthorchis felineus 3 metacercariae in musculature of fishA adults in the
li"er of man and felids
Schistosoma rodhaini 3 mesenteric arteries of rodents 'male and female(A
'infection +ith cercariae that penetrate through the s;in(
MONOGENE
Anacanthorus spA 3 gills of characiform fish 'piranhas( Colo!!oma
macro$om%m B Piaract%! &rac"y$om%! '0haraciformes(
Dictyocotyle coeliaca 3 &ody ca"ity of rays
Pseudodactylogyrus anguillae + P. bini 3 gills of eelsA pathogen introduced
from 5ast *sia to 5urope and :orth *merica
CESTOD
Bothriocephalus acheilognathi 3 intestine of fishA pathogen introduced from
5ast *sia throughout the +orldA infection +ith copepods 'plan;ton(
Diphyllobothrium dendriticum 3 adults in intestine of fish-eating &irds
'gulls( rarely in manA infection +ith plerocercoids in fish
Diphyllobothrium latum 3 plerocercoids 'lar"ae from the second
intermediate host( in musculature of fresh+ater fish
Echinococcus multilocularis 3 adult from intestine of foxA lar"ae
'al"eococcus or hydatid( in the li"er and other internal organs of rodents
accidentally in man after ingestion of eggs 'fatal cases(
aenia taeniiformis 3 adults in intestine of catsA infection +ith lar"ae
'stro&ilocercus( in rodents
CNTHOCEPHL
Acanthocephalus lucii 3 intestine of fresh+ater fishA infection +ith lar"ae 3
cystacanths 3 in isopods
NEMTOD
Ancylostoma ceylanicum 3 small intestine 'duodenum( of cats rarely in
manA infection +ith third-stage lar"ae after their ingestion or acti"e penetration
richuris suis 3 large intestine 'colon( of pigsA infection after ingestion of
eggs +ith first-stage lar"ae
(e0 to !aBor ta<a o2 !eta6oa% parasites o2 2ish
1a. Body dorsoventrally flattened, not round in cross-section ................................................. 2
1b. Body not dorsoventrally flattened, round in cross-section ................................................. 5
1c. Body possesses appendages for attachment to host ..................................... CRUSTACEA
2a. Body with segmentation or distinct external annulations or rings ...................................... 3
2b. Body without segmentation or distinct external annulations or rings ................................ 4
3a. Anterior attachment organ present which may include hooks and muscular suckers,
bothria or bothridia, posterior attachment organ not present, gut absent ................ CESTODA
3b. Anterior and posterior attachment organs present with well defined posterior sucker, gut
present .................................................................................................................. HIRUDINEA
4a. Posterior attachment organ present which usually includes hooks (hamuli and marginal
hooks! external parasites .................................................................................. MONOGENEA
4b. Posterior attachment organ not present, possesses circumoral and ventral suckers except
for blood dwelling species! internal parasites .......................................................... DIGENEA
5a. Anterior spined proboscis present ................................................. ACANTHOCEPHALA
5b. Anterior spined proboscis not present ........................................................... NEMATODA
2arasites infections and diseases
of fishes in *frica- an update
Ila% PPERN
ECTOPRSITIC PROTO$O 4FLGELLTES ND CILITES5
Species a22ecte&
8ost specific species are associated +ith a +ide range of fish species from most
families$ !&i)uitous or opportunistic species 'Ic"t"yo&odo necator, C"ilodonella spp$ and
some species of Tric"odina, Am&y$"rya and Sco$%lata 6Scy$"idia7 are particularly common
in ,u"enile cichlids and carp$
Geo/raphic ra%/e
%he u&i)uitous ectoprotozoans are cosmopolitan or trans-continentally dispersed "ia
translocation of their cultured fish hosts 'carp and tilapia in particular( 'Ic"t"yo&odo necator,
Cry$to&ia &ranc"iali!, C"ilodonella "e8a!tic"a, C2 $i!cicola, Tric"odina ac%ta, T2
"eterodentata and T2 $edic%l%!, T2 retic%lata, T2 m%ta&ili!, T2 centro!trigata, Tric"odinella
e$i#ootica, - 8offman /.9?A Basson et al2 /.?3A Can *s & Basson /.?9 /.?.A :ati"idad et
al2 /.?6A Shaharom-8arrison & *&dullah /.??A *l&alade,o & *rthur /.?.A Bondad-
1eantaso & *rthur /.?.A Basson & Can *s /..3($
<istri&ution of the more specialised host-specific species follo+s that of their hosts
&ut may also &e more restricted sometimes to only one or a fe+ +atersheds$ %here is
e"idence for the presence of Ic"t"yo&odo, C"ilodonella and in particular trichodinids and
sessilians in a num&er of +ater systems in tropical *frica '4a;e Colta and 5ast *frican la;e
systems D Fryer /.6/A 2aperna /.6?A 2aperna & %hurston /.6?A Fryer & Eles /.92( &ut
taxonomic data are limited to only a fe+ locations '#azu&s;i & 5l %anta+y /.?6A 5l %anta+y
& #azu&s;i /.?6($ %he most comprehensi"e taxonomic data on trichodinids in *frican fish
are from Southern *frica and the @am&ezi ri"er system$ <ata are also a"aila&le from Esraeli
fish 'Basson et al2 /.?3A Basson & Can *s /.?9A Can *s & Basson /.?. /..2($
Descriptio% ta<o%o!0 a%& &ia/%osis
Entegumental ectoprotozoan genera are readily differentiated '#a&ata /.?7A <y;o"a
& 4om /..2( +hile diagnosis of species is difficult and often re)uires special staining$
Most ectoparasitic forms are readily detected in direct microscopic examination of s;in and
gill scrapings from li"e 'or freshly ;illed( fish$ Flagellates may &e further detected in air dried
methanol fixed >iemsa stained smears$
Smears containing ciliates should &e air dried fixed in Bouin for 20 min$ destained in
90F ethanols &rought to +ater stained in a haematoxylin stain and mounted after
dehydration$ %richodinids for specific differentiation should &e impregnated +ith sil"er$ *ir
dried smears should &e placed in 2F sil"er nitrate for 93. min$ in the dar; rinsed in +ater
and exposed to the sun or !C for 73/0 min$
Fla/ellates CMasti/ophora3 (i%etoplasti&aD-
Small trichodinids predominantly from the gills are &ell shaped 'Tri$artiella and
Paratric"odina( and often settle on the tips of the gill lamellae 'see Basson & Can *s /.?.
for generic di"ision of %richodinidae($ En Tric"odinella the ray 'the inner extension of the
denticle( 'Can *s & Basson /.?.( is totally reduced and in the other t+o genera is delicate
or rudimentary '#azu&s;i & 5l %anta+y /.?6A Basson & Can *s /.?9($
>enera of sessile peritrichs are differentiated &y their macronuclei and scopula
'attachment leg( 'Cil,oen & Can *s /.?3 /.?7(-
Cry$to&ia free spindle shaped /0330 G 337 Hm in size 'if C2 &ranc"iali!( or
pyriform +hen attached to the integument +ith t+o flagellae one
+holly or partly ad,unct to the &ody ;inetoplast rod-shaped or round$
Ic"t"yo&od
o
free /3326 G 239 Hm in size or attached to the integumentA +ith four
flagellae$
0iliates I0iliophoraJ-
C"ilodonell
a
rounded to o"al cytostome distinct macronucleus round and cilia on
the conca"e "entral surface are arranged in se"eral conca"e parallel
ro+s-
C2
"e8a!tic"a
size 30367 G 20370 Hm +ith 63? ciliary lines on each side$
C2 $i!cicola 'syn- C2 cy$rini( size 333/00 G 26360 Hm +ith more than /0 ciliary
lines on each side$
Tric"odina cup shaped 203/00 Hm in diameter +ith concentric ro+s of cilia and
a cro+n of denticles$ %he denticle shape is a distinct taxonomic
featureA for differential specific diagnosis of *frican spp$ see Basson
et al2 /.?3A Basson & Can *s /.?9A Can *s & Basson /.?. /..2$
Sco$%lata 6Scy$"idia7 round macronucleus and +ide scopulaA
A$io!oma 6Glo!!atella7 pyriform nucleus small scopulaA
Am&y$"rya 1i&&on shaped macronucleus and +ide scopula$
Stal;ed sessile peritrichs D 8eteropolaria +ith elongate &ody and curled macronucleus
'Foissner et al2 /.?7(A E$i!tili! cup-shaped +ith horseshoe-shaped macronucleus 'Cil,oen &
Can *s /.?3($ Some A$io!oma also de"elop on stal;s$
Suctoria- 'Tric"o$"yra and other genera( D cilia lac;ing "aria&le num&ers of tentacles arise
from the rounded &ody$
Li2e c0cle a%& *iolo/0
Most ectoprotozoans flagellates as +ell as ciliates ha"e simple life histories$ Species
of Cry$to&ia are ectoparasites as +ell as intestinal and "ascular parasites$ Et has &een sho+n
that an ectoparasitic phase occurs in t+o "ascular species 'Koo /.?9($ Both ectoparasitic
flagellates I2 necator and Cry$to&ia spp$ occur either free s+imming or attached to the
integument the former through a cytoplasmic protrusion 'Schu&ert /.6?( and the latter &y
attachment +ith the flagellum '4om /.?0($
1eproduction is usually &y &inary fission$ 0on,ugation is sometimes o&ser"ed in
ciliates$ Sessile species also &ud and gi"e &irth to a free s+imming mo&ile generation
reminiscent of mo&ile peritrichs +hich settle on suita&le su&strates 'fish($
%he sessile suctorians reproduce &y internal and external &udding the detached &uds
are ciliated$ *s the &uds &ecome attached to a ne+ location on the piscine integument cilia
are shed and tentacles appear '8offman /.9?($
Spores or other forms of +aiting stages are un;no+nA the suggested existence of
+aiting stages such as encysted forms of C"ilodonella "e8a!tic"a in the gills '1o+land et
al2 /../( or free cysts 'Bauer et al2 /.6.( has to &e confirmed$
Kater temperatures do not seem to &e an important parameter in spite of reports of
lo+ temperatures &eing more optimal for reproduction of C"ilodonella $i!cicola and some
trichodinids 'Bauer et al2 /.6.($ Massi"e infections +ith I2 necator &oth species of
C"ilodonella and the u&i)uitous trichodinids and sessile species occur in lo+ '/23/9L0( and
high '27330L0( am&ient temperatures in southern *frica and Esrael$ Most fresh+ater
ectoparasitic protozoans disappear in ponds +ith increased salinities 'a&o"e 2000 ppm
chlorinity( only I2 necator and some Cry$to&ia are tolerant and &ecome the predominant
parasites in fish of such ponds$ %here are also halophilic species of Am&y$"rya and
Scy$"idia +hich infect fish 'grey mullet( in estuaries$
* num&er of ciliates 'species of Tetra"ymena, O$"ryoglena, Gla%coma, Col$idi%m
and others D see 8offman /.9?( are facultati"e parasites or opportunists +hich +ill
colonise fish in special circumstances most often +hen fish are stressed or traumatised
'8offman /.9?($ *ll others mentioned a&o"e are o&ligatory parasites +hich +ill apparently
sur"i"e for only a limited time outside their hosts$ :on-parasitic sessile peritrichs are different
species from those colonising li"ing organisms$ %richodinids and sessile species found on
a)uatic in"erte&rates comprise different species from those infecting fish 'Can *s & Basson
/.?9A Cil,oen & Can *s /.?3 /.?7($ %here are ho+e"er a fe+ documented exceptions- T2
$edic%l%! &eing reported from &oth hydra and fish and T2 dia$tomi a parasite of a calanoid
copepod +hich temporarily in"aded hatchery gro+n fry of Claria! garie$in%! 'Basson et al2
/.?3A Basson & Can *s /../($
%here are se"eral degrees of adaptation of trichodinids to their piscine hosts-
u&i)uitous species of an opportunistic nature +hich are al+ays found on the fish s;in &ut
ne"er on the gills 'T2 $edic%l%! and T2 ac%ta(A other u&i)uitous species occur &oth on gills and
s;in 'T2 "eterodentata(A additional seemingly u&i)uitous +idespread species appear to ha"e
a "aria&le degree of predilection for one fish family or another 'cichlids or cyprinids($
*mong the latter trichodinids +ith seemingly related morphological characteristics 'e$g$
$edic%l%!-li;e ac%ta-li;e and nigra-li;e( in different geographical regions demonstrate
definite affinities to a particular group of hosts and may in fact comprise di"erse species 'Can
*s & Basson /.?.($ 8ost specific trichodinids are all +ith only a fe+ exceptions gill
parasites- T2 centro!trigata and great num&ers of small trichodinids mainly species of
Tri$artiella are associated +ith 0ichlidaeA T2 retic%lata occurs mainly in goldfish T2 (a#%&!(i
has &een found in South *frican Bar&%! spp$ and T2 no&ili! and T2 (%$ermani mainly in asian
carp 'Basson et al2 /.?3A Can *s & Basson /.?9 /.?.A *&alade,o & *rthur /.?.($
Patholo/0
5ctoparasitic protozoa are "aria&le in their effect on their hosts$ 2athological effects
are density dependent +hen &oth the size of the parasite population and the nature of the
tissue responses are modulated &y the physiological 'clinical( condition of the fish$ 8ostile
en"ironments 'stressful conditions( compromise the fishesM capacity to counteract infection$
Ic"t"yo&odo necator attaches itself to epithelial cells and through an inserted protrusion
consumes their contents 'Schu&ert /.6?( +hereas C"ilodonella spp$ &ro+se the epithelial
surface '2aperna & Can *s /.?3($ 8istopathological changes in the integument follo+ing
infection &y C"ilodonella spp$ and I2 necator are an outcome of t+o counteracting cellular
processes D hyperplasia of the epithelial cells including mucus cells and chloride cells
"ersus a progressi"e cellular destruction$ 0ellular destruction primarily occurs due to direct
action of the parasites and later &y enhanced a&rasion of the peripheral cells after the
depletion of mucus forming cells$ %he production of mucus cells is limited$ *ccelerated
mucus cell production stimulated &y the infection apparently exhausts resources for mucus
production and the infected fish &ecome NdryO$ Some parasites seem to yield cytotoxins or
proteolytic enzymes +hich could &e the cause of spongiosis +hich affects &oth the
proliferated and unchanged epithelial layer '1o&ertson et al2 /.?/A 2aperna & Can *s
/.?3($ Secondary cellular damage due to degeneration necrosis and des)uamation results
in the degradation and disintegration of the epithelial layer$
Cry$to&ia attachment through the flagellum does not induce any pathological or e"en
ultrastructural cellular damage '4om /.?0( contrary to reports of mor&idities associated +ith
this parasite 'Koo /.?9($
*lthough there are a num&er of reports on poor condition and mortalities particularly
of fry coinciding +ith massi"e infestation of trichodinids Tric"odinella e$i#ootica in particular
'4om /.93( and the sessilians A$io!oma, Am&y$"rya and Sco$%lata 'Fi,an /.6/A Meyer
/.90A 2aperna et al2 /.?6A 4ightner et al2 /.??A 2aperna /../( histopathological changes
in e"ents of massi"e infections &y these ectoprotozoans are hardly e"ident if occurring at all
'Fitzgerald et al2 /.?2A 2aperna unpu&lished /.?7($ Tric"odinella e$i#ootica in carp '4om
/.93( and Tri$artiella cic"lidar%m in cichlids '2aperna /../( cause some erosion of the gill
epithelium$ 8o+e"er food "acuoles of trichodinids re"ealed &acteria rather than sloughed
cells '2aperna unpu&lished($ !ltrastructural o&ser"ation on attached A$io!oma did not
re"eal any interference +ith the host cell ser"ing as su&strate '4om & 0orliss /.6?A 4om
/.93A Fitzgerald et al2 /.?2( or peripheral tissue response$ %hus mortalities follo+ing
massi"e colonisation of gills &y sessilians 'Fi,an /.6/( could result from the dense co"er of
sessilians disrupting gas exchange through the respiratory epithelium$ %he only exception
among these infections are the colonies of the stal;ed sessilia .etero$olaria 6E$i!tili!7 +hich
cause lesions 'Nred soreO( at the stal; attachment to the fish s;in these inflamed
haemorrhagic lesions are also contaminated +ith the &acterium Aeromona! "ydro$"ila '5sch
et al2 /.96A Miller & 0hapman /.96($ 1eported localised infection a&o"e the opercular &one
'in cultured tilapia in Esrael( resulted in aggra"ation of the lesion into a +ide '6 mm in diam$(
perforation of the &one '2aperna /../($
Suctorians 'Tric"o$"yra spp$( in certain instances cause cytological damage to the gill
lamellae cells in direct contact +ith the parasites and su&se)uent hyperplasia and
haemorrhages of the gill tissue '8ec;mann & 0aroll /.?7($
Epi6ootiolo/0
%he course of infection &y ectoparasitic protozoans is determined either indi"idually or
&y the interaction of the follo+ing factors-
a$ mo&ility of the fish$
&$ the fishMs capacity to acti"ate its defence systems$
1educed mo&ility facilitates parasite colonisation as +ell as moderating loss through
detachment and drift from the integumental surface$
<efence mechanisms other than epithelial hyperplasia and specific immune
responses to integumental ectoparasites ha"e not yet &een studied 'except in I2 m%ltiilii!
see pp$ 6/362( although spontaneous reco"ery from infection has &een fre)uently o&ser"ed$
Pu"enile fish and fish under stress 'and at &elo+ optimum am&ient temperature( ha"e &oth
limited mo&ility and apparently immunological incompati&ility &eing either nai"e or
immunosuppressed 'Sniez;o /.66A *"talion /.?/($
8ea"y infections &y ectoparasitic protozoans are mainly found in young fish 'less than
one year old( +hen o"ercro+ded and confined to restricted ha&itats and under stress
conditions$ En these circumstances opportunistic and u&i)uitous species are in"ol"ed$
Enfections other+ise in gro+n-up fish are "ery lo+ and host-specific species predominate$
:e+ &orn cichlids as soon as they +ere +eaned from parental care and sometimes &efore
&ecame hea"ily infected &y trichodinids and sessilians of the genera Am&y$"rya and
Sco$%lata$ Enfestation reached its climax le"el in fish /03/2 mm long$ Such infections occur
in natural ha&itats 'la;es( man-made impoundments as +ell as in hatchery installations
'Fryer /.6/A 2aperna et al2 /.?6A 2aperna unpu&lished report /.?7($ 8ea"y infections
ho+e"er +ere not found in all the &reeding ha&itats of the in"estigated la;e system$
0onditions for infestation "aried +ith ha&itat and am&ient conditions and +ere positi"ely
related to the a&undance of fry schools$
4e"el of infection in the fry sharply declined as fish gained in size '2aperna
unpu&lished report($ %he decline in infection also coincided +ith changes in parasite species
composition the u&i)uitous generalists and opportunists 'T2 $edic%l%! IQR T2 migalaJ T2
ac%ta IQRT2 com$acta see Basson & Can *s /.?.J T2 "eterodentata and species of
Am&y$"rya and Sco$%laria( &eing gradually replaced &y species specific to cichlids
'Tri$artiella spp$ Tric"odina centro!trigata and species of A$io!oma( 'Basson et al2 /.?3A
#azu&s;i & 5l %anta+y /.?6A 5l %anta+y & #azu&s;i /.?6A Basson & Can *s /.?9A Can *s
& Basson /.?.($
8ea"y infections +ith u&i)uitous trichodinids 'T2 $edic%l%!( and sessile peritrichs
'mainly Sco$%lata spp$( also occur in carp fry in hatcheries and nursery ponds and li;e+ise
as fish gro+ are replaced &y more specialised species 'such as T2 nigra, T2 m%ta&ili! and
A$io!oma spp$( 'Basson et al2 /.?3A Shaharom-8arrison & *&dullah /.??A *l&alade,o &
*rthur /.?.($
8ea"y infections '&y trichodinids and sessile species( accumulate in fish - small spp$
of Bar&%!, Ale!te! cyprinodontids and ,u"enile cichlids and Claria! spp$ cro+ded into
residual pools in ri"ers drying-out during the dry season$ En larger +ater &odies in *frica
infections +ith &oth trichodines and sessilians in fish other than fry may &e common &ut lo+
'2aperna /.6?A Cil,oen & Can *s /.?7A #azu&s;i /.?6($
4o+ temperature stress plays an important role in epizootic out&rea;s of
ectoprotozoan infections in cichlid fish outside the limits of the tropical en"ironment and of
populations introduced to non-tropical countries such as the southern !S*$
8ea"y infections &y s;in and gill protozoa predominantly of C"ilodonella spp$ are a fre)uent
occurrence in o"er+intering stoc;s of cultured tilapia hy&rids 'Oreoc"romi! a%re%! 8
nilotic%!( in Esrael and O2 mo!!am&ic%! in ponds and dam reser"oirs in southern *frica '<u
2lessis /.72A =lde+age & Can *s /.?9A 2aperna et al2 /.?3A 2aperna /.?6($
En small ponds '/ hectare( fish are not spared e"en in relati"ely mild +inters +ith
minimum temperatures a&o"e /3L0$ Fish in la;es and large reser"oirs on the other hand
&ecome se"erely affected only in extremely cold +inters +ith temperatures declining to /0L0
and &elo+$ Mortalities often occur from the cumulati"e effects of ectoprotozoans dermal
saprolegniases and systemic &acterial diseases all mediated &y stress of lo+ temperature$
En addition to temperature stress o"er+intering tilapia in ponds are often stressed &y
o"ercro+ded stoc;ing and inade)uate feeding$ Entermittence of higher and lo+er am&ient
temperatures characteristic of the Mediterranean type +inters increases the unpredicta&ility
of food demand &y fish and thus complicates feeding schedules$
Enfestation le"els rise &y late fall +ith increased a&undance of trichodinids and C2
"e8a!tic"a$ Fish succum&ing in early +inter +ere predominantly hyperinfected &y C2
"e8a!tic"a$ 4ate +inter and early spring mass mortalities 'e"en +hen temperatures +ere
already rising a&o"e /7L0( +ere associated +ith C2 $i!cicola hyperinfections$ C2 $i!cicola is
a&undant in carp in some ponds already &y early +inter ho+e"er it +ill only infect tilapia at
the end of the cold season +hen they &ecome compromised &y prolonged stress$
Ic"t"yo&odo necator hyperinfections are mor&id to cichlids as +ell as to fish of other families$
Mortalities occur in fish o"ercro+ded in holding tan;s ponds and in &oth +arm and cold
+ater conditions$ :atural infection +as also re"ealed in A$loc"eilic"t"y! gam&iae from a pool
in >hana$ En Esrael Cry$to&ia spp$ occasionally s+arm the gills of tilapia goldfish and sil"er
carp and also in the latter in lo+ saline +aters '?3/0 ppt salt( &ut data from *frica are
lac;ing$
O2 mo!!am&ic%! appears to &e more tolerant to lo+ temperatures in +ater of higher
salinities and also +here most ectoparasites are excluded 'except l2 necator($ Mem&ers of
the genus Tila$ia in Esrael 'T2 #illii( and in southern *frica 'T2 rendalli & T2 !$armanii( are also
less affected in fresh+aters &y lo+ temperatures and are rarely hea"ily parasitised$
Fe+ instances of mortalities coincided +ith hea"y infections concomitantly or exclusi"ely &y
trichodinids sessilians 'A$io!oma( C"ilodonella spp$ and I2 necator in o"er+intering carp
&ut occurred more often in relation to other stress factors such as high le"els of o"ercro+ding
or high nitrite concentrations 'Fi,an /.6/A Sarig /.9/A 2aperna unpu&lished /.?7($
8ea"y infections &y C"ilodonella spp$ seems to ha"e an excluding effect on other
integumental protozoans$ =ther+ise s;in and gill ectoparasites coexist and are e"en
synergistic +ith metazoan ectoparasites 'Gyrodactyl%! and Arg%l%!( and s;in lesions
'epithelioma( '2aperna & #ohn /.66A Sarig /.9/A 2aperna unpu&lished /.?7($ Mass
mortalities of farmed Claria! garie$in%! 'in the 0entral *frican 1epu&lic( +ere associated
+ith mass infestation &y C"ilodonella "e8a!tic"a$
E$i!tili! infections including red sore and opercular perforations only occur
sporadically +ith no particular lin; to o"er+intering$
Co%trol
%reatment +ith formalin is still the only effecti"e means to control massi"e
ectoparasitic infections in all +arm +ater cultured fish species$ En Esraeli fish farms ponds are
sprayed +ith formalin up to concentrations of 27 or 60 ppm 'of the 39F commercial product(
'Sarig /.90A 4aha" & Sarig /.92($ 5fficacy of formalin treatments is affected &y am&ient
temperatures +ater )uality including salinities and parasites treated$ 2roduct )uality is
"aria&le and is particularly affected &y storage resulting in accumulation of polymerised
'paraformaldehyde( sediment$ %richodinids +ere readily eradicated +ith treatment &y 27
ppm +hile elimination of C"ilodonella +as achie"ed after treatment +ith 60370 ppm$ Can *s
et al2 '/.?6( also demonstrated differential efficacy +ith the type of fish treated e$g$ 27 ppm
per 26h +as effecti"e in cleaning infected carp +hile +ith tilapia fry it has &een achie"ed +ith
67 ppm per 26h$
PRSITIC DINOFLGELLIDS
Species a22ecte& a%& /eo/raphical ra%/e
2arasitic dinoflagellids the marine Amyloodini%m ocellat%m and the fresh+ater
Pi!cinoodini%m $ill%lare and P2 limnetic%m are not discriminatory in their choice of piscine
hosts and ha"e &een implicated in mass mortalities of tropical marine and fresh+ater
a)uarium fish 'Paco&s /.66A Schaeperclaus /.76A 2aperna /.?0($ A2 ocellat%m has
infected sea +ater acclimatised Oreoc"romi! mo!!am&ic%! and A$"ani%! di!$ar in inland
salt pansA some strains of this parasite sur"i"e in salinities as lo+ as /0 ppt$ Schaperclaus
'/.76( reports P2 $ill%lare infection in /6 species of tropical ornamental fresh+ater fish of
di"erse families as +ell as in carp and crucian carp$ 5pizootic infections and mortalities +ere
recently reported in farmed cyprinid fish in Malaysia including grass carp
'Cteno$"aryngodon idella( &ighead 'Ari!tic"t"y! no&ili!( Le$to&ar&%! "oe,enii and P%nti%!
gonionot%! 'Shaharom-8arrison et al2 /..0($ %he presence of P2 $ill%lare has ne"er &een
esta&lished in *frica &ut this u&i)uitous parasite may e"entually &e found$ Ef introduced +ith
culture seed it is li;ely to &ecome esta&lished$
Dia/%osis
%rophonts +hen reaching the final stage of gro+th are "isi&le to the na;ed eye '?03
/00 Hm diameter( as +hite spots 'similar to that seen in ichthyophthiriasis( and turn dar; &lue
+hen exposed to 4ugolMs-iodine$ %hey are o"al +ith a smooth +all and +ith inner aggregates
of glo&ules$ En Malaysian fish clinical signs of P2 $ill%lare infection comprise &oth a rust-
coloured appearance of the s;in indicating the presence of the parasite trophonts '20397 G
/6370 Hm( and a dense co"ering of mucus 'Shaharom- 8arrison et al2 /..0($
Li2e c0cle a%& *iolo/0
%he life cycle of the dinoflagellid fish parasite is comprised of a parasitic feeding stage
'trophont( +hich attaches to integumentary epithelial cells and an encysted di"iding stage
'tomont( +hich is detached from the host$ %he trophonts of P2 $ill%lare deri"e an essential
part of their nutrition from photosynthesis$ %rophonts dislodged at any time during their
trophic stage +ill transform into a di"iding tomont$ <i"isions yield a motile infecti"e stage
'dinospore( +hich attaches to a ne+ host$ %here are se"eral detailed studies of A2 ocellat%m
'2aperna /.?6a( &ut compara&le detailed data on the fresh+ater fish dinoflagellids are
lac;ing$ <ata on P2 limnetic%m gro+th and di"ision 'Paco&s /.66( suggests that parasites
reach a NmaturationO prior to detachment$ P2 $ill%lare trophonts on the gills at 23327L0
de"elop from dinospore to detached tomont in three to four days$ %he tomont then completes
di"ision to the dinospore stage +ithin 70390 hours$ *t /73/9L0 the process of di"ision is
lengthened to // days 'Schaperclaus /.76($
Patholo/0 a%& epi6ootiolo/0
En lar"al fish infections +ere limited to the s;in +hereas in large fish the highest
parasite densities occurred on the gill filaments and in the &uccal-pharyngeal integument$
Fish reco"ering from the epizootic infestation through a gradual decrease in infection could
not &e reinfected '2aperna /.?0($
A2 ocellat%m is attached to and feeds from the host epithelial cell &y means of
rhizoids +hich penetrate the host cell$ %he consumed cell gradually degenerates and
collapses$ <amage to infected cells leads to focal erosion of the epithelium$ 2rolonged
infection exhausts a generation of mucus cells and leads to accelerated des)uamation$
2roliferation of the epithelium causes o&literation of the gill lamellae +hile the inner strata of
the epithelium &ecome spongious and in some cases undergo complete lysis '2aperna
/.?0($ *ttachment and penetration organelles of P2 $ill%lare differ from those seen in A2
ocellat%m in that the host cell is penetrated &y nail-li;e extensions$ 8o+e"er damage to the
host cell is similar '4om & Schu&ert /.?3($ Significant histopathological changes are only
seen in the gills +here most of the infection occurs namely a massi"e proliferation of the
&ranchial epithelium +hich causes fusion of the lamellae &y a confluent cellular mass
'Shaharom-8arrison et al2 /..0($
Pi!cinoodini%m infection in Malaysia initially occurred among ornamental fish &ut it
spread e"entually to pond farmed local and exotic cyprinids causing mortality in fry of
P%nti%! gonionot%! in particular although clinical signs +ere also apparent in a +ider range
of cyprinid fish species 'Shaharom-8arrison et al2 /..0($
Co%trol
A2 ocellat%m is controlled &y continuous application of copper sulphate 0$97 ppm into
infected tan;s$ * further option is a mixture of 7-hydrate copper sulphate +ith citric acid
monohydrate to yield 0$/7 ppm copper ion concentration in the +ater '8ignette /.?/A
#a&ata /.?7($ %he same methodology +ill apparently effecti"ely control Pi!cinoodini%m
infection although concentrations should &e ad,usted to the fresh+ater medium and the fish
targeted for treatment$ En fresh+ater +ith a p8 &elo+ 9$0 'in tropical a)uaculture(
concentrations a&o"e 0$3 ppm may &e lethal to fish 'e$g$ P%nti%! gonionot%! fry($
ICHTHYOPHTHIRISIS
Species a22ecte& a%& /eo/raphical ra%/e
Most species of fresh+ater fish are suscepti&le although some may &e more so than
others$ %he +orld +ide distri&ution of I2 m%ltiilii! '8offman /.90( has apparently &een
facilitated &y the +idespread translocation of cultured and ornamental fish$ %he presence of
this parasite in autochthonous fish in remote areas of the +orld including southern
Cenezuela 'Centura & 2aperna /.?7( and :orthern %rans"aal in South *frica '2aperna
unpu&lished( may suggest ho+e"er that many populations particularly those in the tropics
are comprised of a mix of autochthonous and introduced parasites$ <ata a"aila&le from
*frica are limited to Southern *frica 'in cichlids carp Bar&%! spp$ trout and eels - <u
2lessis /.72A 4om&ard /.6?A Pac;son /.6?A Can *s & Basson /.?6( and !ganda 'on
nati"e Bar&%! am$"igramma and exotic Le&i!te! retic%lat%! from small streams at #a,ansi -
2aperna /.92($ Et is "ery common in Esrael in &oth farmed 'Sarig /.9/A 8ines & Spira
/.93a( and +ild fish including cichlids 'Centura & 2aperna /.?7($
Descriptio% a%& &ia/%osis
>ross signs - +hite spots on the s;in and the gills +hich under microscopic
examination re"eal 'in s;in and gill scrapings( uniformly ciliated organisms +ith a small
cytostome +hich may reach up to / mm in diameter$ Staining +ith either haematoxylins or
>iemsa 'after ade)uate fixation in such as Bouin( re"eals a large crescent-shaped
macronucleus and small micronucleus$ Ic"t"yo$"t"iri%! m%ltiilii! is a monotypic genus of
hymenostomatid ciliates$
Li2e histor0 a%& *iolo/0
%rophonts 'feeding stages( de"elop +ithin the integumentary epithelium al+ays
a&o"e the &asal mem&rane 'Centura & 2aperna /.?7A 5+ing & #ocan /..2($ By maturity
+hich is reached in 2 days at am&ient temperatures of 2732?L0 '336 days at 2/326L0( the
parasite e"acuates the host tissue and settles +ithin 236 hours on a su&strate in the +ater to
form a cyst-encapsulated tomont 'di"iding stage($ 2arasites e"icted from the tissue &efore
the scheduled time for their spontaneous departure fail to de"elop into tomonts and
e"entually die '5+ing & #ocan /..2A 2aperna unpu&lished o&ser"ations($ Kithin the cyst
tomonts undergo successi"e &inary fissions +ith a resulting yield of 27032000 tomites
'infecti"e free s+imming stages( +hich after release +ill see; a suita&le host$ %he di"ision
of tomonts into tomites in am&ient temperatures of 2732?L0 is completed +ithin /7320
hours 'Bauer /.7.A Meyer /.6.A 2aperna unpu&lished o&ser"ationsA 93? hours according
to 8offman /.9?($
En"asion of tomites 'theronts( '30367 Hm long( into the host integument is facilitated
&y the excretion of a stic;y su&stance from su&pellicular crystalline organelles named
mucocysts$ *cti"e penetration causes focal necrosis of the epithelial cells$ Et has &een
suggested that hyaluronidase and other enzymes may &e produced &y the penetrating
parasite '5+ing et al2 /.?7($ En the a&sence of a suita&le host tomites +ill lose their infecti"e
potential +ithin 26 hours at 2632?L0 '5+ing & #ocan /..2($ 8igher temperatures hasten
trophont maturation and tomont di"ision &ut at lo+er temperatures slo+er de"elopment
allo+s the gro+th of larger trophonts '0$?3/$0 mm in 73/0L0 "s 0$730$9 mm in 20326L0(
yielding tomonts +ith higher num&ers of tomite progeny '5+ing et al2 /.?6($ En lo+er
am&ient temperatures the sur"i"al of the tomites is prolonged thus allo+ing more time to
locate a host$ 4o+ temperatures do not interrupt propagation a full cycle is completed at
20L0 in 337 days at /7L0 in 93/6 days and at /0L0 in 2/337 days 'Bauer /.7.A Meyer
/.6.($ <ata on the effect of other en"ironmental parameters is less conclusi"e although it
has &een suggested that dissol"ed oxygen le"els &elo+ / mgSl affect parasite reproduction
'Bauer /.7.($
Patholo/0
Echthyophthiriasis is fatal to fish of all sizes$ 0hronic infection +ill cause serious
damage to the s;in fin and gillsA corneal infection impairs "ision '8ines & Spira /.93a
/.96a($ %he infecti"e stage in"ades the integumentary epithelium and &ecomes esta&lished
in the &asal layer of the epithelium ,ust a&o"e the &asal mem&rane$ 0ellular damage in lo+ to
moderate infections remains restricted to the infected site$ En addition to the damage caused
to epithelial cells &y the feeding and expanding parasites in hea"y infections mass exodus of
parasites from the epithelial layer ha"ing completed their scheduled gro+th causes its
erosion and detachment from the &asal mem&rane$ En some infections parasites cause
+idespread lysis of the inner layer of the epithelium$ 2rolonged infection also induces
epithelial proliferation and haemorrhagic inflammation causing the integument to &ecome
se"erely disintegrated '8ines & Spira /.96aA Centura & 2aperna /.?7A 5+ing et al$ /.?6($
8ines and SpiraMs '/.93& /.96a&( haematological and clinical data from hea"ily infected
fish re"eal e"ident physiological dysfunction resulting apparently from &oth direct
pathological damage induced &y the parasite and as a &y-product of the stress response$
Epi6ootiolo/0
Ic"t"yo$"t"iri%! m%ltiilii! is one of the most common trou&lesome and difficult to
control of fish pathogens$ 5pizootic infections ha"e &een reported in cold +ater salmonid
farms 'also in *frica <u 2lessis /.72A 4om&ard /.6?( and +arm +ater farmed carp eels
Claria! garie$in%! and Ictal%r%! $%nctat%! 'channel catfish( 'Meyer /.90A Sarig /.9/A 8ines
& Spira /.93aA 8ine /.97A Pac;son /.9?A #halifa et al$ /.?3A 2aperna unpu&lished($ Fish
may maintain lo+ su&clinical 'enzootic( infection 'in preimmunity( +hile encysted tomonts
may persist in the ha&itat$ 5nzootic infections in nati"e fish ha"e &een found in Le&i!te!
retic%lat%! in !ganda '2aperna /.92( in cichlids and cyprinids nati"e to the 4a;e #inneret
system in Esrael '2aperna unpu&lished( in glass eels cyprinids and cichlids in nati"e
ha&itats of South *frica 'Pac;son /.9?A Can *s & Basson /.?6( and in a "ariety of nati"e
fish in the southern !nited States '*llison & #elly /.63($ %ransition from nonclinical enzootic
to epizootic clinical infection is usually stress-mediated prompted &y ad"erse gro+th
conditions such as o"ercro+ding poor feeding and excess nitrogenous +aste$ 5pizootic
infection ho+e"er ne"er occurs in o"er+intering tilapia or Claria! garie$in%! in Esrael or
southern *frica &ut rather coincides 'also in southern !S* - Meyer /.90( +ith the +arming
of the +ater in early spring +hen fish are still ;ept in o"ercro+ded conditions after +inter
storage$ En South *frica 6$6F of +ild glass eels in the Southern 0ape are infected$ Cia
these fish infection has &een introduced into eel nurseries +here el"ers especially those not
completely acclimated succum&ed to se"ere infestations '8ine /.97A Pac;son /.9?($
Spontaneous reco"ery has &een o&ser"ed in &oth natural infections in natural
ha&itats and in holding facilities and e"en in experimental infections in a)uaria '2aperna
/.92A 4aha" & Sarig /.93($ %he potential for spontaneous reco"ery "aried +ith fish species$
Enfection in scaled fish nota&ly cichlids regressed faster than in smooth s;inned fish 'eels
Claria! spp$ and other siluriforms mirror and leather carp( '2aperna /.92 and unpu&lished
o&ser"ations($ *fter reco"ery fish +ere refractory to reinfection or retained a merely
su&clinical chronic infection '8ines & Spira /.96cA Kahli & Meier /.?7A 2aperna
unpu&lished o&ser"ations($ %he o&ser"ed interspecific "ariation in suscepti&ility to infection
could ho+e"er also result from differential compati&ility of "arious fish species to man-made
ha&itats and "aria&le "ulnera&ility to stress$
Spontaneous reco"ery from infection and resistance to reinfection of reco"ered fish
indicate that fish are capa&le of de"eloping defence mechanisms against I2 m%ltiilii! '8ines &
Spira /.96c($ Spontaneous reco"ery o&ser"ed in carp at temperatures as lo+ as /0L0
'4aha" & Sarig /.93( implies some protecti"e responses other than "ia humoral anti&ody
production +hich &ecomes suppressed in carp &elo+ /2L0 '*"talion /.?/($
8ines & Spira '/.96c( demonstrated immo&ilisation of free s+imming tomites +ith
sera ta;en from carp after their reco"ery from infection$ %he infecti"e stages +ere also sho+n
to &e una&le to penetrate the s;in of resistant carp$ Emmo&ilisation tests +ith trophonts
sho+ed that in infected trout anti-parasitic acti"ity of the mucus increases )uic;ly after
infection and decreases soon after the infection has disappeared$ %he anti-parasitic acti"ity
of the serum in the same fish increases slo+ly &ut remains at a higher le"el for at least 9
months 'Kahli & Meier /.?7($ Fish immunised +ith Tetra"ymena spp$ de"eloped a
resistance to a challenge of lc"t"yo$"t"iri%! infection '>o"en et al2 /.?/($
0arp +ere immunised follo+ing controlled exposure to the infecti"e tomite 'teront(
stage and sur"i"ed challenges +ith high infecti"e doses &ut lost protection after &eing
immunosupressed &y the administration of corticosteroids$ %hese results could ha"e
simulated a stress mediated situation$ 4e"els of humoral anti&odies in immunosuppressed
fish ho+e"er remained the same as in the immunised group +hich further confirms the
in"ol"ement of other than humoral type immune systems in the protection processes against
l2 m%ltiilii! infections '8oughton & Matthe+s /..0($
Emmunisation +ith ;illed "accines ga"e less satisfactory results although &etter
protection +as o&tained through intraperitoneal inoculation of li"e theronts 'Bur;art et al2
/..0($ Oreoc"romi! a%re%! mothers "accinated through the latter method passi"ely
transferred a protecti"e immunity to their fry 'Su&asinghe & Sommer"ille /.?.($ *dditionally
to immunity passed from mothers "ia eggs demonstra&le &y anti&odies in the solu&le
extracts of fry tissues a protecti"e immunity +as ac)uired directly from the parent mouth
during the &rooding period 'Sin et al2 /..6($
Co%trol
Both trophonts localised &eneath the epithelial layer of the integument and the
encysted tomonts attached to su&strates in the a)uatic ha&itat are resistant to practically all
externally applied usa&le antiparasitic agents$
Enfection can &e effecti"ely controlled only &y destruction or elimination of the free
di"iding tomonts or the tomites they release$ En +arm +ater systems '2632?L0( three to four
daily transfers of fish to clean tan;s +ill effecti"ely reduce infection +hile ena&ling the fish to
de"elop tolerance to reinfections$ %omonts can &e effecti"ely remo"ed from large circulating
tan;s &y repeated &rushing +ith "acuum suction$ Spontaneous reco"ery and transition into a
refractory state +ill &e further promoted &y management techni)ues +hich alle"iate stressing
conditions 'impro"ing +ater flo+ accelerating aeration and reducing stoc;ing densities($
0hemical parasiticides +ill &e effecti"e only through continuous or repeated daily application$
=f the many listed 'Meyer /.6.( the only cost-effecti"e remedy for large scale farming
systems is Malachite green at a dose of 0$07 ppm for continuous application '336 days( or up
to 0$/7 ppm 'depending on the specific fish tolerance +hich "aries +ith species - siluriforms
are particularly suscepti&le($ Formalin +ill dislodge some of the trophonts and is often applied
mixed +ith Malachite green '70 ppm +ith 0$07 ppm( 'Sarig /.9/($
Systemic therapy seems to &e the only means of effecti"e control$ 5limination of
tissue trophozoites +as reported in se"eral species of ornamental a)uarium fish fed
medicated food '%etra M* /00S70( containing Malachite green in a non-+ater solu&le
formulation for 6 days 'neither drug concentration in the food nor daily rations are gi"enA
Schmahl et al2, /..2($ For use in commercial food-fish culture for human consumption the
cost efficiency of Malachite medicated feed formulations and their toxicity to humans must &e
considered$
PROTO$ONS OF THE GUT LUMEN
Species a22ecte& a%& /eo/raphic ra%/e
Enfections &y .e8amita 'S$iron%cle%!( are common in cultured tilapia '4ands&erg
/.?.( as +ell as in commercially reared South *merican cichlids in Esrael &ut thus far ha"e
not &een reported from cichlids in *frica$ =ccasional hexamitoses occur also in farmed
goldfish in Esrael$ =palinids 'Protoo$alina and -elleriella( and ciliates 'Balantidi%m and
Nyctot"er%!( +ere reported from *frican fish of the families Schil&eidae Mochocidae and
0itharinidae 'Fantham /./?A Sandon /.6.($
*moe&ae ha"e ne"er &een reported from *frican fish &ut infections ha"e &een
reported in grass carp 'Cteno$"aryngodon idella( and in a neotropic catfish 'Pimelod%!
claria!( 'additionally to Salmonids in cold+ater ha&itats - Sa+yer et al2 /.97($ %here is also
an unconfirmed report on systemic amoe&iasis epizooty among tilapia cultured in *u&urn
*la&ama !S 'K$ 1ogers per$ comm$($
Descriptio%3 ta<o%o!0 a%& &ia/%osis=
.e8amita 'S$iron%cle%!( I@oomastigophorea <iplomonadidaJ is 93/2 G 63. Hm pear
shaped or round flagellate +ith 3 pairs of anterior flagellae and one pair of posteriorly
pointed flagellae$ %he taxonomic relationships 'at the generic and species le"els( &et+een
parasites found in different hosts are still unresol"ed$ :either examination of li"e flagellates in
fresh smears nor fixed and >iemsa stained can pro"ide sufficient details for taxonomic
differentiation 'Molnar /.96($
Balantidi%m I0iliophora Cesti&uliferia %richostomatidaJ is a large holociliate +ith a
large round macronucleus readily seen in fresh and stained &ouin fixed smears '+ith either
>iemsa or 8ematoxylin( or in histological sections 'Molnar & 1einhardt /.9?($
Nyctot"er%! I2olymenophorea 8eterotrichidaJ are common ciliates in amphi&ia and ha"e a
"ery large cytostome$
=palines I=palinataJ are large 'a&out 270 Hm long( rounded organisms uniformly
co"ered +ith parallel ro+s of short flagellae$ %hey are superficially reminiscent of ciliates &ut
are included among the Sarcomastigophora 'flagellates( 'Sandon /.96A Foissner et al2
/.9.($ %hey ha"e from t+o to many similar nuclei and in this differ from the ciliophorans
+hich ha"e micro and macro nuclei$
%here is a difference of opinion concerning aetiological agents of "isceral granuloma
in tilapia and goldfish regarded as amoe&a ISarcodinaJ &y some '4om & <y;o"a /..2( and
fungi '3ermocy!tidi%m-li;e( &y others 'see 6$6($
Li2e c0cles a%& *iolo/0
Both .e8amita and the gut ciliates di"ide &y &inary fission it is not ;no+n if these
piscine parasites may persist as resting stages in cysts$
%he life history of opalinids has &een studied in species infecting anurans$ Et in"ol"es
se)uences of &inary di"isions and di"isions yielding daughter cells 'tomonts +hich form
cysts and in addition a gamogonous process resulting in a zygocyst$
Patholo/0
8examitosis 'octomitosis( in a)uarium held South *merican cichlids nota&ly
Sym$"y!odon di!c%! and Ptero$"yll%m !calare often coincides +ith poor conditions and
mortalities$ En recent years infection has fre)uently &een diagnosed in tilapia hy&rids cultured
in Esrael$ Massi"e num&ers often congest the posterior digesti"e tract and coincide +ith food
retention$ Et is ho+e"er not yet certain if .e8amita is a primary pathogen or a synergist in
other clinical conditions and &acterial contaminations$ 8ea"y infections in grass carp or
South *merican cichlids cause haemorrhagic enteritis +ith in,uries to the mucosal
epithelium some necrotic changes in the li"er and sometimes haemorrhagic dropsy 'ascitis(
'Molnar /.96($
8ea"y intestinal infection &y Cry$to&ia i%&ilan! in the South *merican cichlids
.eric"t"y! cyanog%tat%! and Cic"la!oma mee(ei caused se"ere inflammation in the entire
digesti"e tract though the epithelial layer remained &y and large intact$ =edema atrophy
and necrosis occurred in the lamina m%!c%lari!A lesions extended to the li"er and the spleen
'<y;o"a & 4om /.?7($ Fish stopped feeding de"eloped dropsy and gradually died
'Be,erano pers$ comm$($ * granulomatous condition in "iscera associated +ith hea"y
proliferation of Cry$to&ia-li;e organisms has &een recently found in tilapia hy&rids in Esrael$
Balantidiosis in grass carp causes haemorrhagic enteritis$ En the terminal stage of the
disease hyperaemia and inflammation extends to the entire gut mucosa$ 4arge num&ers of
parasites may accumulate in the exudate filling the posterior end of the intestine$
8istopathology re"eals loss of the superficial epithelium in many places in"asion of the
lamina $ro$ria &y the parasites and multiple ulcerations 'Molnar & 1eihardt /.9?($
Mortality of a)uarium reared South *merican cichlids 'Sym$"y!odon ae9%ia!ciata(
coincided +ith hea"y infection &y opalinids 'Protoo$alina !ym$"y!odonti!( resulting in
congestion of the digesti"e tract 'Foissner et al2 /.9.($
Epi6ootiolo/0
%here are no data on hexamitosis in *frica$ Species from South *merican and *frican
cichlids seem to &e different and cross infection may not &e possi&le$ Suscepti&ility of
*frican cichlids to Cry$to&ia i%&ilan! has yet to &e examinedA the recent occurrence of
granuloma associated +ith a Cry$to&ia-li;e infection in cultured tilapia could ha"e &een the
conse)uence of cross infection &et+een *merican and *frican cichlids$
Balantidi%m cteno$"aryngodoni! has not &een found thus far in grass carp introduced
to South *frica$
Co%trol
%here are se"eral pharmaceutical products recommended against hexamitoses-
Flagil 5nheptin >a&&rocol 'Farmitalia( *ctinitrazol 'Flu;a($ %he first +as used to control
hexamitosis in South *merican cichlids$ *ll of these are applied "ia medicated feeds '/32 g
per /00 ;g feeds for 339 days - see 1eichen&ach-#lin;e /.?0($ %herapeutic control of
intestinal ciliate infections has not yet &een practised$
COCCIDIOSES
EIMERINE COCCIDI
Species a22ecte&
*mong *frican fish infection &y coccidia has so far &een demonstrated in cichlid fish
in Claria! garie$in%! and in eels 'Ang%illa mo!!am&ica($ =ther fish ha"e not &een
in"estigated$ 0occidia also infect common carp goldfish grass carp and sil"er carp$ =ther
tropical fish found to host coccidia are farmed >ouramies 'Tric"oga!ter tric"o$ter%!( '#im &
2aperna /..3&( and in South *merica cichlids 'Be;esi & Molnar /../A *ze"edo et al2,
/..3( and characids 'Serra!alm%! niger( &oth +ith "isceral coccidia 'Caly$to!$ora spp$($
Geo/raphic ra%/e
Eimeria ,ana!i and Go%!!ia cic"lidar%m occur in cichlid fish in Esrael !ganda and
South *frica '4ands&erg & 2aperna /.?7 /.?9($ E2 ,ana!i has also &een reco"ered from
Oreoc"romi! nilotic%! introduced to %hailand 'from 5gypt "ia Papan($ 0arp and goldfish in
Esrael as else+here '#ent & 8edric; /.?7( are infected &y Go%!!ia car$elli$ * second
species found on 5urasian carp Go%!!ia !%&e$it"eliali! 'Marince; /.93( and Eimeria
!inen!i! in sil"ercarp and &ighead 'Molnar /.96( are a&sent from Esraeli farmed fish$
Entroduced cyprinids in South *frica ha"e not thus far &een examined$ Cisceral tissue
coccidioses are as yet un;no+n in *frican fish ho+e"er li"er and gonadal infections &y
Caly$to!$ora spp$ +ere reported from South *merican hosts 'as +ell as from euryhaline
;illifishes in the southern !S* - ="erstreet et al2 /.?6($ %he only ;no+n coccidia from
Southeast *sian tropical fish is Go%!!ia tric"oga!teri from >ourami 'Sze;ely & Molnar
/..2($
Descriptio% ta<o%o!0 a%& &ia/%osis
2iscine coccidia are intracellular organisms of the epithelium 'of the gut the gall
&ladder the s+im&ladder and the ;idney tu&ules( and tissues 'li"er( of epithelial origin$
<e"eloping intracellular 'endogenous( stages may &e detected +ithin their host tissues &y
microscopic examination of fresh tissue and stained impressions and smears '+ith &uffered
Ito p8 9$2J >iemsa after &eing air dried and fixed in a&solute methanol($ =ocysts of digesti"e
tract-coccidia may &e detected in faeces$ %he oocyst +all of piscine coccidia +ith a fe+
exceptions 'of eels( is soft and fragile and is often lost &y the end of sporulation$ %herefore if
sporulation occurs prior to defecation only the smaller na;ed sporocysts may &e found in
faeces$
0occidia are identified &y the morphometry of their oocysts and sporocysts the site of
endogenous de"elopment and their position in the host cell '<y;o"a & 4om /.?/($
%he sporocystMs hard +all is either &i"al"ed and clea"es &y a longitudinal suture 'in the genus
Go%!!ia 2aperna & 0ross /.?7( or opens at the sporocystMs apex at one pole through a
round pore 'genus E$ieimeria of eels and some Eimeria s$l$( or a short apical suture$ %he
sporocyst +all of the latter type may also form tu&ercules or pro,ections and is further
enclosed in a "eil 'genus Caly$to!$ora - ="erstreet et al2 /.?6($
Li2e histor0 a%& *iolo/0
2iscine coccidia de"elop either in the cytoplasm of the host cell or inside itMs nucleus$
5picytoplasmic coccidia de"elop at the apex of the epithelial cell &elo+ its &rush &order
&ulging as they gro+ together +ith their host cell +all into the space a&out the epithelium
'intestinal s+im&ladder or excretory lumen( '2aperna & 0ross /.?7A Molnar & Bas;a
/.?6($ Sometimes the same coccidium species has cytoplasmic intranuclear and
epicytoplasmic generations 'E2 ,ana!i infecting cichlid fish 4ands&erg & 2aperna /.?9A #im
& 2aperna /..2($
5xtraintestinal coccidia apparently reach their target organ "ia the &lood$ G2
cic"lidar%m also under+ent endodyogenous di"ision &efore &ecoming esta&lished in the
s+im-&ladder epithelium of its cichlid host '#im & 2aperna /..3a($
En the epithelial cell parasites undergo successi"e asexual 'merogonous( di"isions
and a sexual process &y +hich microgametes differentiated from a microgamont fuse +ith a
macrogamont 'macrogamete($ %he zygote thus formed &ecomes li&erated from the host cell
+hile &eing encased in a +all$ %hrough su&se)uent di"isions the zygote di"ides into four
hard +alled sporocysts each of +hich further di"ides into t+o motile sporozoites$
%he pace of de"elopment is fast in intestinal coccidiaA in E2 ,ana!i ? days from infection to
sporulation 'at 26329L0( '#im /..2($ En the s+im&ladder coccidium 'G2 cic"lidar%m(
endogenous de"elopment to sporozoite-containing sporocysts lasted at least 7? days at 233
26L0 '#im & 2aperna /..3a($ %hese differences &et+een intestinal and extraintestinal
species are confirmed &y studies of non-*frican species 'Solangi & ="erstreet /.?0A
Steinhagen /../a($
=ocysts of digesti"e tract and gall &ladder coccidia are e"acuated +ith the faeces
'4ands&erg & 2aperna /.?7 /.?9A 2aperna /..0 /../A #im & 2aperna /..2 /..3&($
Sporulation of most gut coccidia is completed &efore e"acuation in the faeces 'endogenous
sporulation($ Some intestinal species nota&ly G2 car$elli infecting carp and goldfish &ecome
trapped in the gut epithelium +ithin degenerate host cells 'yello+ &odies( '#ent & 8edric;
/.?7($ =ocysts of another carp coccidium G2 !%&e$it"eliali! formed in the epithelium are
displaced &y the regenerating epithelium into the su&-mucosal layer 'Marince; /.93A Molnar
/.?6($ =ocysts of the epicytoplasmic coccidium of eels E$ieimeria ang%illae also infiltrate
into the mucosa rather than &eing e"acuated into the lumen '8ine /.97($ =ocysts of "isceral
and internal ca"ity coccidia accumulate sporulate in the host and +ill &e li&erated only after
the death of the host 'Solangi & ="erstreet /.?0($
<irect transmission &y feeding on e"acuated sporulated oocysts has &een
demonstrated in se"eral intestinal species including those in carp Go%!!ia car$elli
'Steinhagen & #orting /.??( and in cichlids E2 ,ana!i '#im & 2aperna /..2($ Et has also
&een experimentally demonstrated that tu&ificid oligochaetes of the genera T%&ie8 and
Limnodril%! ser"e as paratenic hosts- sporozoites of G2 car$elli +hen ingested &y the
+orms excysted and in"aded their gut epithelial cells$ Such sporozoites remained infecti"e
+hen fed +ith the +orm to carp . +ee;s later 'Steinhagen /../&($ Some eimerine coccidia
'Caly$to!$ora %nd%li( re)uire an o&ligate intermediate host 'grass shrimp( for transmission
'Fournie & ="erstreet /.?3($
%ransmission "ia predation and necrophagy seems to &e the route of transmission not
only for extraintestinal coccidia &ut for endogenously sporulating intestinal coccidia +hile still
in their host gut tissue or in the intestinal lumen 'Molnar /.?6($
Patholo/0
%he economic damage done &y coccidiosis to +arm +ater pisciculture has apparently
&een grossly underestimated$ Since coccidiosis in fish usually manifests itself as a chronic
infection mortality is gradual and is o"erloo;ed in most farms$ 4osses only &ecome e"ident
+hen yields are chec;ed at the end of the gro+th period$
0yprinids and cichlids contract intestinal coccidioses as soon as they hatch$ Enfection
'of G2 car$elli( has &een identified in ?-day-old goldfish +ith mortality occurring 30367 days
later$ G2 car$elli seems to &e more pathogenic to goldfish than to carp '#ent & 8edric;
/.?7($ En cichlids 'cultured Oreoc"romi! hy&rids( intestinal infection 'of E2 ,ana!i( +as
detected in fry &y the end of nursing in their parentsM mouth losses &ecame e"ident +hen
infection reached maximum le"els &y t+o or three +ee;s after hatching '#im /..2($ 8ea"y
infections in carp fingerlings '27370 mm long( ha"e &een found to coincide +ith se"ere
emaciation$ 5maciation also occurred in infected cichlid fry$ Sur"i"ing fish demonstrated
spontaneous reco"ery infection +as lo+ or a&sent in carp and goldfish or cichlids older than
2 months$ :onspecific defence response parameters 'leucocytosis eosinophylia acti"ation
of phagocytes and ele"ation of natural anti&ody titer and of coeruloplasmin( +ere detected in
carp infected +ith G2 !%&e$it"eliali! 'Studnic;a & S+ic;i /..0($
<amage caused &y intestinal coccidioses occurs principally &y the rupture of the
epithelium &y the escaping merozoites and oocysts 'in G2 car$elli and E2 !inen!i! - Molnar
/.96 /.?6A #ent & 8edric; /.?7($ En the intestine of cichlid fry infected +ith E2 ,ana!i most
damage is caused to the mucosal cells &y the de"eloping intracytoplasmic parasites
'4ands&erg & 2aperna /.?9($ 5picytoplasmic infections seem to ha"e less effect on the gut
epithelial layer and the damage induced through consumption of the nuclei &y the
intranuclear generations has not yet &een e"aluated$ Enflammatory changes in intestinal
coccidioses only occur follo+ing disintegration of the mucosal layer and in response to
accumulated cellular de&ris$ En nodular coccidiosis caused in carp &y G2 !%&e$it"eliali! the
accumulation of oocysts in the lamina $ro$ria induces inflammation +ith intense leucocyte
infiltration 'enteritis( 'Marince; /.93aA Molnar /.?6($ %he G2 cic"lidar%m epicytoplasmic
infection leads to an intense des)uamation of the s+im&ladder epithelial lining in cichlids
'4ands&erg & 2aperna /.?7($ 2roliferation of the mucosal epithelial cells o&ser"ed in
intestinal 'Molnar /.?6( and in s+im&ladder infections &loc;s the release of oocysts from
the epithelium resulting in a condition similar to that o&ser"ed in nodular coccidiosis$
S+im &ladders of cichlids +ith late infections turn opa)ue +hite$ %he pathogenicity of the
s+im &ladder coccidiosis to ,u"enile cichlids still needs to &e critically e"aluated$
0occidia of Claria! garie$in%! ha"e not &een studied$ %hey form yello+ &odies similar
to those of G2 car$elli$ <ata on the pathological effect of E$ieimeria ang%illae infections are
a"aila&le from cultured :e+ @ealand eels 'Ang%illa a%!trali! and A2 dien&ac"ii( '8ine
/.97($ =ocysts +hich aggregate +ithin or &elo+ the gut mucosa 'in the lamina $ro$ria(
induce inflammatory infiltration$ En more se"ere infections the condition is reminiscent of
nodular coccidiosis the &asal mem&rane &rea;s and follo+ing the aggregation of oocysts
the su&-mucosal tissue degrades and the loosened epithelial mucosa is sloughed off$ Mature
oocysts and sporocysts are passed out +ith necrotic tissue eels &ecome se"erely emaciated
and die '8ine /.97($ Enfected A2 mo!!am&ica in South *frica passed free sporulated
oocysts and their intestines seemed to &e normal$
8ost response to oocyst aggregation in the li"ers of fish +ith "isceral coccidioses
'Caly$to!$ora %nd%li in *merican ;illifish F%nd%l%! spp$( e"en +hen replacing up to ?7F of
the organMs "olume +as limited to formation of a thin fi&rotic capsule sometimes +ith
collagen or melanin 'Solangi & ="erstreet /.?0A Be;esi & Molnar /../($
Epi6ootiolo/0
En Esrael it +ould &e difficult to find cichlid fry less than 27 mm in length of any
species +ild or cultured from earth ponds or from the hatchery free from infection +ith
Eimeria ,ana!i$ En all these young fish irrespecti"e of species intracytoplasmic
epicytoplasmic and intranuclear forms occur simultaneously$ Enfection declines only after the
sur"i"ing fish reach 60370 mm in length '233 months old($ Enfection in all its three forms also
seems to &e +idespread in South *frican cichlids similarly affecting all local species$ 8ea"y
mor&id infections occurred in larger fish up to 70 mm in length 'O2 mo!!am&ic%! and Tila$ia
!$armanii during +inter($ 2ersistence or recurrences of infection occurred +here defence
responses +ere apparently compromised due to lo+ temperature 'T/3L0( or other stress
conditions$
S+im&ladder coccidiosis infections occur only in Oreoc"romi! spp$ and Sarot"erodon
galilae%!$ <ue to the longer endogenous de"elopment pre"alence of infection extends to a
+ider range of ageSsize classes +hile &eing a&sent in "ery young fish 'less than 2 months
old 'T60370 mm long($ En ponds often all fish e"entually &ecome infected$ Enfection is
retained in O2 mo!!am&ic%! entering marine ha&itats in South *frica$
0occidioses in Esraeli carp and goldfish are not as fre)uent as in cichlids &ut
)uantitati"e data on infections among fish in +arm +ater ha&itats are lac;ing$
Co%trol
*n esta&lished protocol for chemotherapy and pre"enti"e management has not yet
&een formulated$
Cryptosporidium i%2ectio%s
Cry$to!$oridi%m is a common parasite of the stomach in +ild and cultured cichlid fry
'Oreoc"romi! spp$( in Esrael '4ands&erg & 2aperna /.?6($ Meronts and gamonts of this
minute 'a&out 7G3 Hm( coccidium appear as dense spherical or mushroom-li;e structures
located at the &rush &order apices of the stomach epithelium$ %he attached parasite is
encased +ithin the host cell +all +hose rudimentary micro"illi are "isi&le +hen "ie+ed +ith a
scanning electron microscope '2aperna /.?9($ 8ost-cell micro"illi are completely lac;ing in
Cry$to!$oridi%m of other "erte&rates$ * further uni)ue feature to piscine Cry$to!$oridi%m is
the retreat of the mature zygote into the stomach mucosa or su&mucosa +here sporulation
is completed 'into eight na;ed sporozoites( instead of &eing released into the gut lumen as
occurs in the non piscine forms$ *part from cichlids Cry$to!$oridi%m has &een reported to
infect carp 'in 0zechoslo"a;ia - 2"alase; /.?3( and a fe+ marine fish$ Et is difficult to
esta&lish +hether Cry$to!$oridi%m is pathogenic to cichlid fry as this infection occurs
concurrently +ith enteric coccidiosis$ <ata from other infections are insufficient to &e
conclusi"e$ 0ontrol methods are un;no+n$
HEMOPROTO$O
HEMOFLGELLTES
Species a22ecte&
En "arious fresh+ater and marine fishA in *frica- in 0ichlidae Claria! spp$ Bagr%!
spp$ Synodontidae Mormyridae Mugilidae and Proto$ter%! aet"io$ic%!$
Geo/raphic ra%/e
%rypanosomes ha"e &een reported in all ma,or +ater systems of *frica 'Kenyon
/.0?A 8oare /.32A <ias /.72A Ba;er /.60 /.6/( +ith some species apparently distri&uted
as +idely as their hosts 'cf$ Claria! garie$in%!($ %rypanosomes occur in C2 la#era in the
:ear 5ast &ut not in cichlids or cyprinids$ %rypanosomes ha"e also &een reported from
introduced Oreoc"romi! mo!!am&ic%! in Endia 'Mandal /.99($ %rypanosomes 'T2 c2
m%gicola Bec;er & ="erstreet /.9.( are +idespread in grey mullet 'Mugilidae( of the
lagoons and ri"ers of southern *frica$ %here are no reports of "ascular Cry$to&ia from *frica$
Descriptio%3 ta<o%o!0 a%& &ia/%osis
2iscine haemoflagellates s+im freely in the &lood$ Mem&ers of the genus
Try$ano!oma are spindle shaped 273.7 Hm long a single flagellum originating from a
usually apical ;inetoplast is connected longitudinally to the trypanosome &ody &y an
undulating mem&rane$ %he nucleus is usually single except in the course of di"ision and
centrally positioned$
Cry$to&ia also pre"iously called Try$ano$la!ma are reminiscent of trypanosomes in
shape &ut ha"e t+o flagellae connected to a single ;inetoplast one free and one tied
longitudinally &y an undulating mem&rane$ *lthough the t+o are seemingly related in
morphology and in means of transmission "ia leeches there are sufficient &iological
differences to separate the t+o$ Cry$to&ia unli;e trypanosomes are not exclusi"ely "ascular
parasites and 'e"en sometimes the same species( also occur as ectoparasites on the fish
&ody surface and in the digesti"e tract$ %ransmission may also &e direct or &y predation
'Koo /.?9($
Enfections in the &lood are readily detected either &y direct microscopic o&ser"ation of
fresh &lood or more easily from plasma from the surface of the pac;ed erythrocyte layer of
&lood centrifuged in a heparinized haematocrit capillary$ Smears prepared from +hole &lood
or centrifuged material after &eing air dried and immersed in a&solute methanol are stained
in diluted >iemsa '/S/0 in p8 9$039$6 phosphate &uffer($
Morphology and morphometrics are considered insufficient for specific determination
of trypanosomes and comparison of the isoenzymes of in/,itro cultured isolates is currently
the esta&lished taxonomic methodology$ %his methodology is already applied to differentiate
amphi&ian trypanosomes &ut thus far not to those of fish species$
Li2e histor0 a%& *iolo/0
%rypanosome &inary di"ision is rarely o&ser"ed in the &lood$ *t the present state of
;no+ledge it +ill &e difficult to esta&lish if pleomorphism e$g$ small and large forms reported
in non-*frican species '#han /.96( occurs in infections of *frican fish$ Et +as not e"ident in
grey mullet infections$
En the leech ingested trypanosomes in the crop undergo se"eral successi"e
di"isions yielding morphologically di"erse generationsA amastigotes sphaeromastigotes and
epimastigotes +hich migrate to the pro&oscis and transform into metatrypanosomes - the
infecti"e stage +hich +ill enter fish &lood during feeding '#han /.96($
Sphaeromastigotes and epimastigotes +ere re"ealed attached to the crop epithelial
lining of an estuarine piscicolid leech 'a ne+ yet to &e descri&ed species &y =osthuizen
P$8$ !ni"ersity of 2retoria 1$S$*( remo"ed from the mouth of a trypanosome-infected grey
mullet '4%gil ce$"al%!A see also 0hapter /9$( from S+art;op estuary 0ape region South
*frica$
5pimastigotes '0ritidia( +ere also found in Batrac"o&delloide! tricarinata remo"ed
from Proto$ter%! aet"io$ic%! and Bar&%! altianali! from 4a;e Cictoria +here trypanosomes
'T2 m%(a!ai( are common$ 0ichlids and Bagr%! &ayad are additional hosts$ 'Ba;er /.60
/.6/($ %his leech as +ell as trypanosomes fre)uently infect Claria! garie$in%! in Esrael$
%he de"elopment of Cry$to&ia in the leech does not resem&le the se)uential de"elopment of
trypanosomes$ Cry$to&ia ingested +ith &lood &ecomes rounded and follo+ing &inary fission
apparently yields infecti"e forms +hich migrate '+ithin 92 hours compared +ith +ee;s in
trypanosomes( to the pro&oscis$ 1eciprocal transformation of "ascular stages into
ectoparasitic forms also allo+s direct transmission &et+een fish hosts 'Koo /.?9($
Patholo/0 a%& epi6ootiolo/0
:atural infection +ith trypanosomes may &e "ery common particularly +here the
leech "ector is also common 'see 0hapter /9$($ En 4a;e Cictoria 76F of Oreoc"romi!
,aria&ili! and 70F of O2 e!c%lenta +ere infected and 20F of Oreoc"romi! nilotic%! in 4a;e
>eorge 'Ba;er /.60 /.6/($ Enfection in catfish 'Claria! garie$in%! and Bagr%! spp$( also
seems to &e "ery high 'in all four Bagr%! docmac examined in 4$ Cictoria( &ut )uantitati"e
data are lac;ing$ 2re"alence of infection in 0ape ri"erine and estuarine mullets is a&out
70F &ut hea"y infections are rarer 'a&out /7F($ En none of the instances of natural infection
+as there any e"idence of ad"erse effects on the host$ <ecrease in haematocrit and
haemoglo&in le"els and e"idence of accelerated haemopoiesis ha"e &een reported in some
infections '&y T2 m%rmanen!i! #han /.99($ En experimental infections of carp fingerlings
+ith T2 danile:!(yi +here le"els of parasitaemia reached /0
7
per mm
3
some fish de"elop
ascites others &ecome oedematousA infiltrati"e and proliferati"e changes occur in the renal
tissue the pancreas and in "arious connecti"e tissues '4om <y;o"a & Machac;o"a /.?6($
*naemia and anorexia +ere reported in goldfish +ith high le"els of parasitaemia +ith the
same trypanosome ':azrul Eslam & Koo /../ /../a($ %he "ector leech of T2 danile:!(yi
'Pi!cicola geometra( is a cold +ater species '4om /.9.( unli;ely to &e introduced into +arm
+ater carp rearing systems$
Enfections +ith "ascular Cry$to&ia are far more pathogenic causing anaemia
splenomegalia exophthalmia and ascitesA anorexia has also &een reported$ Pu"enile fish
salmonids and carp are particularly suscepti&le &ut all cases are in cold+ater culture
systems '4om et al2 /.?6A Koo /.?9($
Co%trol
*t present the only practical means +hich may &e recommended is en"ironmental
control &y elimination of leeches 'see 0hapter /9$($
HEMOSPORIDI ? Dactylosoma a%& he!o/re/ari%ea
Species a22ecte&
Fresh+ater and marine fish$ En *frica 3actylo!oma has &een found in cichlids
'species of Oreoc"romi! A!tator"eoc"romi! and .a$loc"romi!( and grey mullets 'Mugilidae
-4%gil ce$"al%!, Li#a d%mmerelli and L2 ric"ard!oni($ %he latter t+o species of grey mullets
are thus far the only ;no+n *frican hosts for .emogregarina$
Geo/raphic ra%/e
<ata on piscine hemogregarines and dactylosomes in *frica are scanty$
<actylosomes in cichlids '32 mariae - 8oare /.30A Ba;er /.60( +ere thus far only found in
4a;es Cictoria and >eorge +ith a single record from O2 mo!!am&ic%! in %rans"aal South
*frica +hile 3actylo!oma "anne!i and hemogregarines +ere only found in grey mullets from
Southern 0ape ri"ers 'South *frica($
Descriptio%3 ta<o%o!0
2arasites of the circulating erythrocytes -- 3actylo!oma Isyn$- Ba&e!io!oma,
.aemo"ormidi%mJ or of &oth circulating erythrocytes and of the reticulo-endothelial tissues -
.aemogregarina Iand Cyrilia in South *mericaJ$ =nly asexually di"iding and resting stages
'merogony stages and gametocytes( occur in the piscine host +hile the gamogonous
process ta;es place in the in"erte&rate '"ector( host +hich is a leech$ Enfected cells are
detected from &lood smears and tissue imprints 'touch preparations( air dried a&solute
methanol fixed and >iemsa stained 'same as for 8aemoflagellates($ Kaiting comma-shaped
gametocystes of 3actylo!oma are readily confused +ith young stages of hemogregarines$
Fully gro+n hemogregarine meronts and gametocytes occupy the entire long axis of the
erythrocyte and di"ision is longitudinal$ 2iscine dactylosomes di"ide either into 6 in cross-
li;e or into ? in octagonal formation$
<actylosomes are related to hemogregarines and &oth sho+ affinities +ith
mammalian piroplasms 'Bartha & <esser /.?.($ <actylosomids +ith only )uadruple di"ision
+ere regarded as a separate genus Ba&e!io!oma 'Pa;o+s;a & :igrelli /.76( +hile those
di"iding to ? +ere named .aemo"ormidi%m '#han /.?0( &ut it appears that the same
species had alternating generations forming 6 and ? progeny '2aperna /.?/($
Li2e histor0 a%& *iolo/0
%he definiti"e host and "ector of &oth dactylosomes and hemogregarines are leeches
'4ainson /.?6A Bartha & <esser /.?.A Siddal & <esser /..2($ Enfection is transmitted to
and from the leech during &lood suc;ing$ Engested macrogametocytes +hile &ecoming
associated +ith a microgametocyte 'syzygy process( penetrate the surface of the gut
epithelial cell$ <e"elopment of piscine dactylosomes in the leech is un;no+n$ En the frog
species 3actylo!oma !tadleri gametocytes are isogamous and only one microgamete is
formed$ %he zygote penetrates the epithelial cell +here it di"ides into ? sporozoites$
Sporozoites migrate into the sali"ary cells +here follo+ing a di"ision into four 'a merogony(
infecti"e stages are formed 'Bartha & <esser /.?.($ En piscine hemogregarines '.2
my8oce$"ali and Cyrilia gome!i( four a-flagellate microgametes are formed of +hich one
fertilises the macrogametocyte$ En piscine hemogregarines the zygote undergoes multiple
di"isions producing /6332 sporozoites '4ainson /.?6A Siddal & <esser /..2($ 1eleased
sporozoites reinfect the epithelial cells to undergo a su&se)uent di"ision 'merogony($
=ffspring of this di"ision migrate into the sali"ary cells connected to the pro&oscis 'Siddal &
<esser /..3($
En the fish host merogonous di"ision of dactylosomes occurs only in erythrocytes$
2iscine hemogregarines di"ide in the erythrocytes +hile some 'such as .2 !imondi in Solea
!olea #irmse /.9.( +ill also ha"e a pre-erythrocytic merogony in circulating and tissue
leucocytes 'macrophages($ Furthermore large cyst-li;e &odies containing numerous
hemogregarine merozoites +ere reported in the "isceral organs and muscles of se"eral
thus far only marine fish 'Furguson & 1o&erts /.97A 2aperna /.9.A 2aperna & Sa&nai
/.?2($ Et is not ;no+n if these resting stages are transmissi&le through predation$
Patholo/0 a%& epi6ootiolo/0
%he only pathologically significant infections are the leucocytic hemogregarines +hich
induce proliferati"e lesions thus far reported only from cultured marine fish$ 4esions may &e
comprised either entirely of encapsulated aggregates of merozoites '2aperna /.9.( or of
infected macrophages em&edded in granulomatous tissue 'Furguson and 1o&erts /.97($
Encidence of infection &y 32 mariae in 4$ Cictoria in O2 e!c%lenta is 66F and in O2 ,aria&ili!
90F$ 32 "anne!i infection in grey mullets of the 0ape coastal ri"ers is sporadic only
occurring in one fish species '42 ce$"al%!( +here the le"el of parasitaemia reached 3$2F
and re"ealed di"iding stages$ En the remaining fish parasitaemia +as &elo+ 0$06F and
infection +as comprised predominantly of non-di"iding stages$ Enfection &y hemogregarines
in the same fish +as rare$
MYEOSPORE
Species a22ecte&
1epresentati"e of many fish families "ery common in 0ichlidae 0yprinidae and in
&rac;ish +aters in Mugilidae$
Geo/raphic ra%/e
<ata &eyond single species descriptions are a"aila&le from 5gypt 'Fahmy et al$
/.97( 5ast *frica 'Ba;er /.63A 2aperna /.93( >hana '2aperna /.6? /.93( the
0ameroons 'Fomena et al$ /.?6S7A Fomena & Bouix /.?9( and Esrael 'in cichlids and
Claria! la#era 4ands&erg /.?7 /.?6($ En some of these reports differentiation to species is
lac;ing or incomplete '2aperna /.6? /.93A Fahmy et al$ /.97($ Enfections &y 4y8o&ol%!,
T"elo"anell%! and .enneg%ya occur in &oth 5ast *frican and Kest *frican +aters$ %hey are
apparently host specific and as +idespread as their respecti"e hosts$ ="arian infections of
cichlids are thus far ;no+n from the >reat 4a;es .a$loc"romi! spp$ '2aperna /.93($
Descriptio% ta<o%o!0 a%& &ia/%osis
Myxosporea cause histozoic 'e$g$ in tissue( and coelozoic 'in internal ca"ities e$g$ in
gall and urinary &ladders( infections$
>ross signs of histozoic infection are +hitish cysts +ith a mil;y su&stance containing
microscopic '7-7 Hm longer axis( spores$ 4arge cysts are readily traced small cysts in
tissues inside "iscera and connecti"e tissue and muscles are detecta&le +hen tissue
samples are pressed &et+een slides or in histological material$ Spores are readily detected
in a)ueous methylene &lue-stained smears$ %he so called cyst is a parasite-origin
plasmodium +hich forms a specialised mem&ranous ,unction of pinocytotic "esicles 'canals(
+ith the surrounding host cells '0urrent & Pano"y /.96($
Spores hard-+alled +ith one to six polar capsules are differentiated &y sporogenesis
+ithin the plasmodium$ %he polar capsule contains a coiled filament +hich may &e extruded
under pressure or other stimulants including fixation +ith a&solute methanol$
0oelozoic myxosporea of the urinary and &ile ca"ities ha"e a small plasmodium and produce
fe+ spores often only t+o$ 2remature plasmodia are attached &y pseudopodia to the
epithelial lining of the &ladder '2aperna et al$ /.?9($ Small diplosporic plasmodia
'NpseudoplasmodiaO( are characteristic of 4y8idi%m and S$"aero!$ora +hich de"elop from
+ithin the ;idney tu&ule into the lumen '<y;o"a & 4om /.?2( and also in the gut lamina
$ro$ria '<iamant /..2($
>enera are identified &y spore configuration$ <iagnosis to species le"el is &ased
mainly on measurements of fresh unfixed spores their polar capsule dimensions as +ell as
the length of the extruded polar filament '4om & *rthur /.?.($ Separation of the genus
4y8o!oma from 4y8o&ol%! &y the presence or a&sence of a iodophilus "acuole does not
appear to &e taxonomically "alid 'Kalli;er /.6?($ !ltrastructural studies 'scanning(
demonstrate interspecific and intergeneric "ariation in the surface configuration '+hich
consists of /36 "al"es ,oined &y sutures( '4om & 8offman /.9/($ 4ocation of the
cystsSplasmodia in the host may &e of diagnostic "alue in site specific species +hile others
are not fastidious in their site preferences$
Li2e c0cle a%& *iolo/0
%he life histories of myxosporeans in *frican fish are not ;no+n$ Et has &een sho+n
that transmission of 4y8o!oma cere&rali! of trout and se"eral other myxosporean parasites
of other fish in"ol"es tu&ificid oligochaetes as intermediate hosts 'Kolf & Mar;i+ /.?6A 5l-
Mat&ouli & 8offmann /.?.($ >oldfish &ecame infected +ith 4y8o&ol%!, -c"o((ella and
T"elo"anell%! after &eing fed on tu&ificids infected &y actinosporeans 'Uo;oyama et al2
/../($ Et has also &een demonstrated that an early migratory proliferati"e phase precedes
later sporulating plasmodia in tissues or of S$"aero!$ora in the ;idneys '4om & <y;o"a
/.?6( or the gills '4om et al2, /.?3($ Such di"iding plasmodia occur in &lood '0sa&a or 0 -
&odies( and in the s+im&ladder tissue '#orting /.?2A 0sa&a et al2, /.?6A Molnar & #o"acs-
>eyer /.?6( in carp and in the ;idney haematopoietic tissue of trout '2#V cells #ent &
8edric; /.?7($ Myxosporean multinucleate formations preceding spore formation are
generated &y endogenous di"isions +here cells are formed +ithin cells$ %his process occurs
in the early proliferati"e stages and during the process of sporogenesis$ 2rimary cells from
the plasmodia generate &y endogenous di"ision secondary inner cells +hich further di"ide
&y mitosis$ %heir progeny 'the sporogenic cells( produce &y endogenesis tertiary cells +ithin
them$ %he se)uences of di"isions +hich lead to spore formation also include meiotic di"ision
'4om & <y;o"a /.?6($ %he spore is formed from en"eloping 'pericytes( and inner cells
transforming into shell "al"es polar capsules and to sporoplasm$
Spores of coelozoic forms are e"acuated +ith the &ile and the urine$ 0ysts in the s;in
gills and the digesti"e tract release spores &y rupture$ En grey mullets entire cysts in the
digesti"e tract +all are discharged into the gut lumen in the form of a +hite &all$ %he only
other alternati"e for the release of myxosporean spores is after their hostMs death$
Patholo/0
Se"eral myxosporean infections of cultured fish +ere reported to &e pathogenic$ Most
notorious is the +hirling disease of trout manifested &y s;eletal deformities +hich is also
claimed to ha"e &een introduced +ith rain&o+ trout into South *frica 'Can Ky; /.6?($ En
farmed carp 4y8o&ol%! spp$ caused locomotory distur&ances coupled +ith emaciation and
sun;en eyes in &rain infections '<y;o"a et al2 /.?6( anaemia and haemorrhagic dropsy and
mortality in a hea"y cardiac infection 'Bauer et al2 /../( and circulatory disfunctions in
infections at the &ases of the gill lamellae '#o"ac->eyer & Molnar /.?3($ 8ea"y infection of
carp gills +ith 42 (oi caused fusion through epithelial hypertrophy and congestionA rupture of
cysts caused inflammation$ <amage to the gills &y dense infestation resulted in respiratory
pro&lems fish +ere s+imming near the surface +ith distended operculi '1u;yani /..0($
Se"ere disaggregation of the respiratory epithelium is caused &y S$"aero!$ora spp$
infections of carp and goldfish gills '4om et al2, /.?3($
Se"eral myxosporeans cause renal pathology in carp and goldfish- S$"aero!$ora
renicola proliferating in the renal tu&ules damages the renal tu&uli epithelium and the
released spores congest excretory passages '<y;o"a & 4om /.?2($ 2resporulating forms
named .oerell%! +ith a postulated identity +ith species of 4itra!$ora 'Molnar et al2, /.?6(
cause #idney 5nlargement <isease &y infecting the epithelial cells of the renal tu&ules
'Uo;oyama et al2, /..0($ 2roliferati"e stages of S$"aero!$ora cause proliferati"e hypertrophy
of the ;idney in salmonids '#ent & 8edric; /.?7( and grey mullets '2aperna /../($
2roliferati"e presporulating stages of S$"aero!$ora renicola +ere demonstrated to &e the
aetiological cause of s+im&ladder inflammation in carp '#orting /.?2A 0sa&a et al2, /.?6A
Molnar & #o"acs->eyer /.?6($
0ysts of histozoic myxosporidians pro"o;e inflammation only after their disintegration
and in most instances it is +hen myxosporan infections &ecome pathogenic to their host and
cause clinical conditions 'examples- gill infections of *merican catfish +ith .enneg%ya e8ili! -
Mac0raren et al2 /.97A C"anna $%nctata +ith .enneg%ya :altairen!i! - #ala"ati &
:arasimhamuri /.?7 and carp +ith 4y8o&ol%! (oi - 1u;iani /..0($ 1upture of cysts also
leads to haemorrhaging sometimes resulting in considera&le loss of &lood 'Schulman /.79A
#ala"ati & :arashimhamurti /.?7($
En some infections host response may lead to tumour-li;e proliferations incorporating
parasite plasmodia and the capillary net-+or; ':igrelli & Smith /.3?($ 1upture of dermal and
&ranchial cysts caused intense haemorrhaging and facilitated in"asion of secondary
opportunistic pathogens '2aperna & ="erstreet /.?/($
En fish in *frica histozoic myxosporidians occur as s;in infections +ith cysts formed
in the dermis under the scales extending to the surface of the head 'face lips( or onto the
fins '4y8o&ol%! spp$ in ,u"enile cichlids .enneg%ya lateroca$!%lata in Claria! la#era( and in
the gill filaments '4y8o&ol%! spp$ in 0yprinidae siluroids 0haracidae 3i!tic"od%! ro!trat%!
.eteroti! nilotic%!A T"elo"anell%! in cyprinids and .enneg%ya in Late! al&ertian%! as +ell as
in citharinids cyprinids and siluroids( '2aperna /.93A *&olarin /.96A 4ands&erg /.?6A
Fomena et al2 /.?6S7($ 4y8o&ol%! cysts occur in the pharyngo-&ranchial ca"ity 'of
Cteno$oma spp$( the interior organs muscles and "iscera$ Such infections are &est ;no+n
from cichlids &ut also occur in fish from other families '2aperna /.93($ S$"aero!$ora occur
in ;idneys of Claria! la#era '4ands&erg /.?6( and >rey mullets '2aperna /../($
Enfections 'e"en hea"y infections( often occur in fish other+ise in good condition$
:onetheless multiple cysts and particularly large cysts in "ulnera&le organs such as the gills
may compromise fish health$ 4arge 4y8o&ol%! cysts caused &ody deformity or cur"ature of
Bar&%! lineat%! and cichlids '2aperna & %hurston /.6?($ ="arian infection in female cichlids
from 4$ Cictoria remained focal in the interstitial tissue +hile that 'a different species(
occurring in cichlids '.a$loc"romi! ang%!tiron! and .2 elegan!( from 4$ >eorge enlarged
the o"ary +hile displacing the entire o"igerous tissue e"idently causing castration$
%he only reported coelozoic infection in fresh+ater *frican fish is that of 4y8idi%m clariae
from C2 la#era gall &ladder '4ands&erg /.?6($
1upture of mature cysts leads to host-tissue responses as +ell as phagocytosis of
spores and their capsule residues$ Spores accumulate in melanomacrophage centres in the
;idney and the spleen '=ga+a et al2 /..2($ Spores encountered in the spleen are therefore
residues of past infections else+here 'Ba;er /.63A 4ands&erg /.?7($
Epi6ootiolo/0
2re"alence of infection is "aria&le$ En the 5ast *frican 4a;es "isceral 4y8o&ol%!
infections in Oreoc"romi! +ere "ery pre"alent '?.3/00F( +hile in .a$loc"romi! spp$ they
+ere only exceptionally a&o"e 27F$ S;in and gill infections in ,u"enile as +ell as larger
cichlids are also not too pre"alent although "ery hea"ily infected in"ididuals +ere collected$
Enfection le"els seem to change +ith time 'Fryer /.6/A Ba;er /.63A 2aperna /.93($ >ill
infection '4y8o&ol%!, T"elo"anell%! and .enneg%ya( of cyprinid fish and siluroids seems to
&e "ery common in 4a;e Colta in the 1uaha ri"er %anzania and in the 0ameroons &ut
)uantitati"e data are limited '2aperna /.6?A Fomena et al2 /.?6S7( or non existent
'2aperna /.93($ 5pizootic infection has &een reported in grey mullets farmed in fresh+ater
ponds in Esrael 'Sarig /.9/($
="arian infections leading to castration in .a$loc"romi! spp$ occurred in less than
6F of female fish and only in 4$ >eorge$ En 4$ Cictoria only the mild o"o-mesenteric infection
occurred$
:either in Esrael +here tilapia are intensi"ely farmed nor else+here in *frica has the
epizootic form of infection e"er &een o&ser"ed except in one farm in Mala+i +here fish
de"eloped large su&dermal deforming cysts$ <etailed information on this case is ho+e"er
lac;ing$
=f all pathogenic myxosporeans reported in carp or goldfish none ha"e thus far &een
reported in carp farmed in *frica despite the fact that s+im&ladder inflammation and ;idney
infections +ith S$"aero!$ora renicola occur in carp reared in Esrael '4ands&erg /.?.($ %he
carp gill infecting 4y8o&ol%! (oi has &een recently reco"ered on goldfish stoc; imported to
Esrael from 0hina$
Co%trol
%o this day no readily effecti"e and easy-to-use drug is a"aila&le to treat myxosporean
infections$ %rials +ith Fumagillin 'also used to control microsporidial infections( produced
encouraging results &ut at the same time the tolerance of the fish to therapeutic doses +as
"aria&le and some +or;ers reported side effects of intoxication follo+ing treatment schedules
'Sit,a-Bo&adilla & *l"ares-2ellitero /..2A Molnar /..3($
Sphaerosporosis of trout '2#<( and carp and 3icentrarc"%! la&ra8 hoferellosis of
goldfish and trout +hirling disease +ere treated +ith medicated food containing 0$73/ g
Fumagilin ;g
-/
fed at /$7F &ody +eight 'or 0$73/ g ;g
-/
fish &ody mass( for 93? +ee;s
'8edric; et al2 /.??A Uo;oyama et al2 /..0A 5l Mat&ouli & 8offmann /../A Sit,a-Bo&adilla &
*l"ares-2ellitero /..2A Molnar /..3($ Fumagillin +hen applied to 4y8idi%m giardi infected
el"ers arrested de"elopment &ut failed to eliminate the parasite +hich resumed de"elopment
+hen medication +as ended 'Sze;ely et al2 /.??($ %he cost of Fumagilin is ho+e"er
economically prohi&iti"e for use in food-fish farming 'tilapia catfish and carp($
%he efficacy of other tested drugs such as furazolidon and toltrazuril remains inconclusi"e
'Uo;oyama et al2 /..0A Molnar /..3($
MICROSPOR
Species a22ecte&
%hus far only reported in cichlids from *frica and the :ear 5ast$
Geo/raphic ra%/e
Microsporidian infection has &een descri&ed from many 8olarctic marine and
fresh+ater fish$ %here are "ery fe+ reports of infections in fish in the tropics$ Microsporidian
infections +ere reported from 4a;e >eorge 5ast *frica 'S+im&ladder of .a$loc"romi! spp$(
'2aperna /.93( and in the 1epu&lic of Benin 'in gills and "iscera of Tila$ia and
Sarot"erodon spp$( 'Sa;iti & Bouix /.?9($ Microsporidian spores +ere also detected in
;idneys of Oreoc"romi! a%re%! and its hy&rids in Esrael$ Pli!to$"ora 'syn$ Plei!to$"ora(
infection has &een reported from glass eels and el"ers in the !mtata and #eis;ama ri"ers in
%rans;ei 'southern *frica( 'Pac;son /.9?($
Descriptio%3 ta<o%o!0 a%& &ia/%osis
Microsporidia are o&ligate intracellular parasites$ Enfected cells usually enlarge to
accommodate the proliferating parasite$ Such enlarged cells are termed xenomas$ Kithin
these xenomas parasites undergo merogonous and sporogonous de"elopment +hich
culminates in the production of spores '0anning & 4om /.?6A 4om & <y;o"a /..2($ %he
spores are thic; +alled and contain a characteristic coiled polar filament and one
'2leistophoridida( or t+o nuclei ':osematidida( 'Keiser /.?7($ Enfection is readily detecta&le
&y the presence of spores in tissue smears 'fresh or >iemsa stained( or in histological
section$ 8ypertrophic infected cells may reach macroscopic sizes and often yield
characteristic gross pathological signsA multiple +hitish nodules in tissues or in case of the
s+im&ladder a significant thic;ening of the +alls$ <ifferentiation from other aetiological
agents inducing cellular hypertrophy '4ymphocystis 5pitheliocystis and Myxosporea( is
achie"ed &y demonstration of spores in such tissues$
>eneric and specific differentiation is often aided &y ultrastructural data '0anning &
4om /.?6($ #eys for generic differentiation +ere prepared &y Keiser '/.?7($
Li2e histor0 a%& *iolo/0
*ll acti"e stages of the microsporidians de"elop in the host cell$ %+o microsporidia
+ere reported to de"elop +ithin the host cell nucleus 'in 1odlet cells - Modin /.?/ and in
haematopoietic cells - 8edric; et al$ /../($ %he spores are the infecti"e stage transmissi&le
to ne+ hosts$ En the in"asi"e stage the uni- or &inucleate sporoplasm of the spore is
released through the hollo+ed polar filament$ <e"elopment of the parasites often ta;es place
in tissues far remo"ed from the site of hatching +andering macrophages and other non-
differentiated mesenchymal cells pro&a&ly aid their distri&ution or ser"e as hosts$ Enfected
host cells often &ecome grossly hypertrophic 'xenoma( +ith concurrent enlargement of the
nuclear mass 'remaining single or &ecoming fragmented( and multiplication of nucleoli$
Microsporidia are unusual in lac;ing mitochondria and their ri&osomes are unusually large$ En
some microsporidia nuclei appear single 'WisolatedX( +hereas in others nuclei are paired$
<e"elopment +ithin the xenoma proceeds through a merogonous and a sporogonous
process$ %he sporo&lasts formed from sporonts differentiate into spores '2utz & Mc4aughlin
/.90A Keiser /.?7A 0anning & 4om /.?6($
Patholo/0 a%& epi6ootiolo/0
%he effect of microsporidian infection on the piscine host is "aria&le- fish seem to
sur"i"e infections in spite of the presence of huge xenomata often pressing on and
displacing important organs +hile infection &y some microsporidians undou&tedly has a
mor&id effect on the fish '2utz & Mc4aughlin /.90A Morrison & Sprague /.?/($ Entranuclear
infection of haematopoietic cells +as associated +ith acute anaemia '5lston et al$ /.?9($
En Pli!to$"ora infected .a$loc"romi! ang%!tiron! and .2 elegan! the s+im&ladder +alls
+ere thic; and +hite$ Microscopic examination re"ealed most +all tissue to &e loaded +ith
inclusions 'pansporo&lasts( containing up to 6? spores$ 2re"alence of infection in 4$ >eorge
+as "ery lo+ '&elo+ /F out of 302 fish examined from &oth species( '2aperna /.93($
Enfections &y No!emoide! tila$ia in Tila$ia #illii, T2 g%inen!i! and Sarot"erodon
melanot"eron +ere common '/3330F pre"alence in 4a;e :a;oue and 2orto :o"o lagoon(
+ith some fish demonstrating numerous xenomata on the gills 'some reaching 760 G ?00 Hm
in size( as +ell as in the mesenteries the gut +all and in the li"er 'up to 603/00 Hm in size(
&ut +ithout apparent clinical effect on the fish 'Sa;iti et Bouix /.?9($ 2re"alences of infection
+ith Pli!to$"ora 'R( in glass eels and el"ers in %rans;ei ri"ers +ere 6$?3.$7F and 2$6F
respecti"ely$ Microsporidial infection of ;idneys in Oreoc"romi! spp$ are rare and do not
induce any detecta&le pathological changes$
Co%trol
%here is no routine treatment$ Fumagilin <08 used to control No!ema infections in
&ees has &een tested for efficacy in treatment of microsporidial infections in fish$ %he drug
administered in food 0$/ gS;g food at /$7F &ody +eight daily ratio for four +ee;s pre"ented
mortalities and relie"ed Enterocyto#oon-infected chinoo; salmon from clinical signs '8edric;
et al$ /../($ 1ecurrences of infection occur in treated Pli!to$"ora infected eels '#ano &
Fu;ui /.?2($ Kith ele"ation of doses and prolongation of treatment toxic side-effects
'haematological and stress-lin;ed( occur 'Sit,a-Bo&adilla & *l"ares-2elleteiro /..2($
%oltrazuril 'Beyer product - 2$7F *$E$( administered &y a /h &ath of 20 HgSml 'or a 6h &ath of
7 HgSml( repeated six times at t+o day inter"als damaged de"elopmental stages of the
microsporidian Gl%gea anomala in stic;le&ac;s &ut did not affect spores 'Schmahl &
Mehlhorn /.?.($
La*orator0 Tech%iFues 2or the Stu&0 o2 Cesto&es
CESTODES PRSITIC IN FISH
''othriocephali&ea Car0oph0lli&ea Nippotae%ii&ea
Proteocephali&ea and Spathe*othrii&eaA lar"ae of
Diph0llo*othrii&ea and 0yclophyllidea- Gr0porh0%chi&ae5
Ro!a% (UCHT & To!"# SCHOL$
Enstitute of 2arasitology Biology 0entre of the 0zech *cademy of Sciences
BraniYo"s;Z 3/ 390 07 [es;\ Bude,o"ice 0zech 1epu&lic
';rtte;]yahoo$com & tscholz]paru$cas$cz(
lai% &e CHM'RIER
<epartment of En"erte&rates :atural 8istory Museum
2$=$ Box 6636 08-/2// >ene"a 6 S+itzerland
'alain$decham&rier]"ille-ge$ch(
^ Ge%eral co!!e%ts
%ape+orms of the a&o"e-mentioned orders mature in the intestine of
fresh+ater and marine fish in particular teleosts$ %hey are "aria&le in size
measuring from a fe+ millimetres 'Arc"igete! :ippotaeniidea( up to almost / metre
'some &othriocephalideans($ 8ot fixation has &een pro"en to pro"ide the &est
material suita&le for morphological and &iometrical studies histological sections and
scanning electron microscopical o&ser"ations 'S5M($ Some specimens or at &est
parts of each +orm should &e al+ays fixed also for molecular analysis '<:*
se)uencing($ Ef possi&le small pieces of the scolex and stro&ila should &e fixed for
ultrastructural o&ser"ations 'transmission electron microscopy( too$
Metacestodes of some diphyllo&othriideans the adults of +hich mature in
tetrapods '#uchta et al$ 200?( proteocephalideans &othriocephalideans and
gryporhynchids '0yclophyllidea( infect internal organs of fish in particular muscles
mesenteries li"er gall &ladder and intestinal lumen$ %heir fixation may slightly differ
as recommended for adult cestodes &ut in general the procedure descri&ed &elo+ is
"ery similar to that for adults$
E$ Fiel& collectio% !etho&s a%& host trac@i%/
.o!t!
%he condition of the fish host +hen examined is the most important factor
+hich influences considera&ly the )uality of tape+orms found$ %he &est option is to
examine freshly ;illed hosts &ecause this guarantees that li"e cestode specimens
suita&le for su&se)uent studies are o&tained$ Ef fresh hosts cannot &e examined they
should &e placed on ice &ut %e)er 2ro6e%$ %ape+orms from frozen hosts are almost
al+ays unsuita&le for morphological histological and ultrastructural studiesA pro&lems
ha"e &een found e"en +ith se)uencing of cestodes from frozen hosts$ %he pro&lem
of freshness of fish hosts is most important in the case of marine fish that usually do
not sur"i"e for a long time after their catching and in tropical countries +ith high
temperature e$g$ in *mazonia$ Similarly fish from mar;ets are generally unsuita&le
for parasitological examination &ecause cestodes are rapidly decomposed and
autolysed including destruction of the tegument detachment of microtriches and
sclerotized structures 'hoo;s or hoo;lets( on the scolex and "acualization of internal
organs$
Fish should &e al+ays &issecte& as soo% as possi*le a2ter the hostGs
&eath$ Ef the num&er of fish to examine is too high they can &e maintained ali"e for
some time in large tan;s +ith +ater from the place of origin 'or +ith dechlorinated
+ater($ 8o+e"er the inter"al &et+een catch of fish and their dissection should not &e
too long &ecause tape+orms can disappear from their hosts especially from
star"ing fish 'Scholz /../a($
0orrect identification of the fish host is an important part of parasitological
examination and data on the hosts should &e ta;en including photos of hosts
examined and samples of their tissues for su&se)uent <:* analysis$
3i!!ection
For dissection standard e)uipment 'utensiles( such as scissors 'small and
regular( t+eezers 'soft and hard( dissecting needles 'soft and hard( and &rushes is
re)uired$ %he intestine of the host including other parts of the alimentary tract such
as the li"er and gall &ladder should &e remo"ed from the host and placed in a 2etri
dish +ith a small amount of saline 'or sea +ater in the case of marine fish if saline is
not a"aila&le( to pre"ent drying of the organs$ %he intestine is opened &y cutting its
+all longitudinally +ith small scissors prefera&ly from the posterior part to a"oid
cutting of the scolex$
%ape+orms found should &e remo"ed carefully from the intestine &y soft
'entomological( t+eezers dissecting needles or &rush$ Special attention should &e
paid to isolation of the scolex that may &e "ery small and deely em&edded +ithin the
intestinal epitheliumA it may e"en penetrate through the intestinal +all in some cases
'A&ot"ri%m Penetroce$"al%!($ En fish +ith pyloric caeca such as salmoniform or
perciform fish the scolex is often firmly attached to the apical part of the caecum &ut
its stro&ila is freely +ithin the lumen of the intestine$ %he tape+orms remo"ed from
the intestine are placed in a small 2etri dish +ith saline and gently +ashed to clean
them from the intestinal content andSor host tissues$ 4arger +orms such as
E%&ot"ri%m and other &othriocephalideans are often surrounded &y mucous
contents of the intestine +hich is difficult to remo"e mechanically$ En this case it is
possi&le to put the +orms on a filter paper for a +hile$ Ef the infection is hea"y the
+orms can &e cleaned &y gentle decantation and repeated changes of saline in a
dish or a small &ea;er$
%he >or!s shoul& *e 2i<e& as soo% possi*le after their isolation from the
host intestine &ecause their internal organs and surface structures are getting
decomposed "ery fast e"en if they are ;ept in refrigerator$
<issecting microscope may help considera&ly during dissection and
processing the +orms &ut it is not necessary unless tape+orms are too small and
thus can &e o"erloo;ed &y na;ed eyes +hich may &e the case of metacestodes
such as gryporhynchids$
EE$ Fi<atio%
*s already mentioned a&o"e o%l0 tape>or!s 2ro! 2reshl0 @ille& 2ish hosts
are suita*le 2or ta<o%o!ic !olecular a%& ultrastructural stu&ies$ Et is
recommended to use specimens for &oth morphological 'and histological and S5M(
and molecular '<:* se)uencing( e"aluation$ %his is possi&le if the +orms are large
and thus it is easy to cut off the last segments to fix them in ethanol for genetic
analyses$ En the case of monozoic +orms '0aryophyllidea( it is necessary to cut the
+orm to three parts and fix the central part of the &ody in ethanol +hereas the
anterior part +ith the scolex and the posterior part +ith an o"arian complex cirrus-
sac and genital pores should &e fixed +ith a hot fixati"e$ Ef the num&er of apparently
conspecific +orms in the same host is high some specimens can &e fixed entire for
morphology +hereas others put to ethanol$ 8o+e"er e"en in this case it is ad"isa&le
to get samples for &oth procedures 'hot formalin and ethanol( from at least a fe+
+orms of such a sample$
For ultrastructural studies '%5M( small pieces of the scolex and mature and
gra"id segments should &e cut to small pieces 'prefera&ly 233 mm($
%he fixation methods descri&ed &elo+ ha"e &een pro"en to &e "ery simple
fast and resulting in good-)uality material suita&le for taxonomic and phylogenetic
studies 'see e$g$ de 0ham&rier and Caucher /...A de 0ham&rier and Scholz 200?A
de 0ham&rier et al$ 200?A Scholz et al$ 200?($
*$ Fi<atio%
/$ +hole mounts 'morphology( and histology-
a$ place li"e +orms 'after their gentle +ashing( in a small amount of
saline in a &ea;er or 2etri dishA large plastic "ial can also &e usedA
&$ heat 6F formaldehyde solution 'further the term NformalinO is used(
until &u&&les are lea"ing from the &ottom and then to simply add
fixati"e to the +orms in saline 'proportion &et+een the amount of
fixati"e and saline should &e at least /0 - / to ensure fixation +ith
heat li)uid(A
c$ transfer the fixed +orms +hich should &e relaxed if fixati"e +as
sufficiently hot and the +orms ali"e +hen fixed to a "ial +ith 6F
formalin for su&se)uent storage 'at least /32 +ee;s(
d$ add la&el +ith pertinent data 'see EC(A
e$ for longer storage transfer the +orms to 90F ethanolA similarly the
+orms stored in formalin should &e transferred to 90F ethanol at
least a fe+ days &efore staining and mountingA
f$ if fixation +ith formalin is not possi&le &oiled +ater 'or saline( can
&e used insteadA the procedure is the same as descri&ed a&o"e
'/&(A fixed +orms should then &e transferred to 6F formaldehyde
solution and pure ethanol for <:* analyses as descri&ed &elo+
'2&($
g. !etacesto&es 'cestode lar"ae from fish( should &e fixed &y in the
same +ay except for plerocercoids of species of 3i$"yllo&ot"ri%m
and related genera +hich should &e ,ust place in a "ial +ith cold 6F
formalinA metacestodes of gryporhynchids +hich possess rostellar
hoo;s on the scolex should &e fixed as lo+er monogeneans 'see
&elo+ 3 EEE$8( i$e$ they should &e press in a drop of +ater under
co"erslip to ;eep the hoo;s in one plane and then the corners
should &e fixed +ith :oyer sealantA thereafter fixati"e 'mixture of
glycerin and ammonium picrate or picric acid( is addedA these
preparations are semipermanent &ut can &e remounted to 0anada
&alsam '5rgens /.6.A see also Cidal-Mart_nez et al$ 200/A Scholz
et al$ 2006($
2$ molecular study-
a$ +ash +orms in salineA
&$ place ali"e +orm or their pieces to pure 'non-denaturated( .63..F
ethanol or special fixati"es for maintaining <:* undamaged
'<:*later etc$(A
c$ it is necessary to put the field num&er into the "ial
d$ if possi&le ;eep samples in cold 'refrigerator or freezer($
B$ Fi<atio% 2or electro% !icroscop0
/$ Scanning 5lectron Microscopy- use the procedure descri&ed a&o"e for
material to &e studied as +hole mounts and histological sections '*/($
2$ %ransmission 5lectron Microscopy-
a$ +ash the +orms in salineA
&$ cut them to small pieces '&est a&out 233 mm( in diameterA
c$ place the pieces in freshly prepared cold '6L0( 2$7F glutaraldehyde
in 0$/M cacodylate &uffer 'p8 9$2( 'for more details see 4e"ron et
al$ 2006($
0$ Fi<atio% in situ
Fixation in !it% should &e used only +hen it is not possi&le to isolate all
tape+orms from the host intestine due to shortage of time or hea"y infection of the
host$ Et is recommended to ta;e some samples for <:* analyses 'put them to
ethanol 3 see *2( &efore hot formalin is used for fixation of the +orms attached to the
intestinal epithelium$
EEE$ Processi%/ o2 2i<e& !aterial
*$ Staining specimens- Schu&ergXs hydrochloric carmine 'for details see
Scholz and 8anzelo"Z /..? and de 0ham&rier 200/( +as found to gi"e
good results +hen staining fish cestodesA destaining is made using Nacid
ethanolO i$e$ .6F ethanol +ith a fe+ drops of hydrochloric acid '80l($
B$ Mounting specimens- after dehydration in a series of graded ethanol and
clearing in clo"e oil 'eugenol( stained +orms are mounted as permanent
slides in 0anada &alsam 'see Scholz and 8anzelo"Z /..? and de
0ham&rier 200/(A a "ery important step is to put tape+orms on a paper
'cartoon( and co"er +ith a glass &efore transfer to .6F ethanol to ensure
that they remain straight not rolled$
0$ S;inning specimens- %his method ine"ita&le +hen studying many
cyclophyllidean cestodes especially large-sized taeniids is usually not
necessary for fish cestodes$
<$ Sectioning- 8istological sections 'cross longitudinal and sometimes
sagittal( are almost al+ays necessary to study internal morphology$
Formalin-fixed material is +ell suita&le for sectioning using standard
histological procedure 'dehydration using a graded ethanol series and
toluol em&edding in paraffin or paraplast sectioning 'thic;ness of sections
/03/2 `m( and staining +ith haematoxylin-eosinA for more details see
Scholz and 8anzelo"Z /..? and de 0ham&rier 200/($
5$ Scanning electron microscopy-
/$ 2ost-fixation- :ot used in the groups maturing in fish 'see Scholz et
al$ /..?($
2$ <ehydration- Samples are dehydrated in a graded series of ethanol
and /00F acetone 'amylacetate($
3$ <rying- 0ritical-point method 'acetone +ith 0=
2
( is +idely used also
for fish cestodes$
6$ Mounting- <ehydrated samples are mounted on stu&s +ith a
dou&le-tape$
7$ Sputtering- %he samples are sputtered +ith gold 'or paladium(
&efore their examination$
F$ %ransmission electron microscopy-
/$ rinse samples o"ernight in 0$/ M sodium cacodylate &uffer at p8
9$2A
2$ postfix them in cold '6L0( /F osmium tetroxide in the same &uffer
for / hA
3$ dehydrate the samples in a graded ethanol series and propylene
oxideA
6$ em&ed the material in 5pon and then let polymerize at 60L 0 for 6?
h or em&ed it in SpurrXs epoxy resin 'or in *raldite and 5pon($
>$ 5gg preparations- Since en"elopes of intrauterine eggs in +hole mounts
may &e collapsed deformed or in"isi&le egg morphology should &e
studied in distilled or saline +ater in temporary preparations of eggs
li&erated from the uterus in ethanol-fixed samples placed 7 minutes in
distilled +ater &efore o&ser"ation$
8$ =ther specialized preparations- For a study of em&ryonic 'oncospheral(
hoo;s eggs should &e o&ser"ed under pressure of a co"erslip in
temporary mounts$ En some species +ith the scolex armed +ith hoo;s
'2roteocephalidea 3 >angesiinaeA Bothriocephalidea 3
Polyonc"o&ot"ri%m Senga Tetracam$o!( some scoleces may &e strongly
pressed to ena&le hoo; o&ser"ationA in this case a method used for
Nlo+erO monogeneans i$e$ fixation +ith >*2 '3 parts of glycerin - / part of
ammonium picrate 3 see Malm&erg /.79( or +ith Berlese fluid used for
some cyclophyllideans '15F$( should &e applied$ %hese preparations are
temporary &ut can &e re-mounted follo+ing the procedure of 5rgens
'/.6.($
8$ Couchering specimens for molecular study-
Et is ad"ised to al+ays ;eep a part of the specimens used for <:*
analyses prefera&ly those parts of the +orm that ena&le its specific
identification$ *s already recommended a&o"e 'EE( the &est option is to fix
the same specimen for morphological as +ell as molecular studies$
<eposition of these "oucher specimens in internationally accessi&le
collections is strongly recommended to ma;e it possi&le to chec;
identification of samples used for molecular studies$
EC$ La*els
5ach "ial +ith a +orm must contain la&els +ith pertinent data &est +ritten &y a
pencil or printed on a paper put into "ialA it is not recommended to put data &y a
mar;er only onto the external surface of the "ial &ecause letters may disappear
especially in "ials +ith ethanol$ %he follo+ing data should &e al+ays pro"ided-
a$ name of the parasiteA
&$ name of the host 'prefera&ly +ith 2iel& %u!*er(A
c$ locality and date of collectionA
d$ fixati"eA
e$ name of collectorA
Ef there is not enough space for all this information put at least the field
num&er$ Et is essential to fill a field &oo; +ith all data$
C$ 4ong-term storage of specimens
Specimens fixed +ith formalin should not &e stored for a long time and should
&e transferred to ethanol 'prefera&ly 90F($ Samples for <:* analyses should &e
maintained in lo+ temperature 'refrigerator of freezer( and e"aporation should &e
a"oided &y chec;ing the content of ethanol in "ials regularly$
CE$ Maintenance of li"ing cestode specimens and cestode life cycles
*$ Maintenance of free-existing stages
Et is impossi&le to ;eep free-li"ing stages such as eggs or coradicia for a
longer timeA their sur"i"al depends mainly on the temperature and thus their
maintaining in lo+ temperature can prolong their life$ 5ggs released from gra"id
segments can &e ;ept for some time in lo+ temperature +ith daily changing of +ater
that should &e dechlorinated 'see Scholz /../a /..9A #uchta et al$ 2006(A
anti&iotics may &e added to pre"ent gro+th of &acteria$
B$ Maintenance of host animals 3 in"erte&rate hosts
%ape+orms parasitic as adults in marine and fresh+ater teleost fish use
plan;tonic crustaceans 'copepods 3 Bothriocephalidea 2roteocephalidea and
:ippotaeniidea( &enthic crustaceans 'amphipods 3 Spathe&othriidea( and
oligochaetes 'tu&ificids 3 0aryophyllidea( as intermediate hosts 'Be"eridge 200/($
%hese hosts can &e maintained in the la&oratory for some time especially tu&ificids$
=n the contrary sur"i"al time of copepods in la&oratory cultures is usually rather
short and mortality is high +hich ma;es experimental infections +ith these
in"erte&rates difficult$ >ammarid amphipods can &e ;ept in la&oratory conditions for
a longer time than copepods pro"ided that cold +ater aeration and ade)uate food
are a"aila&le$
$ 0$ Maintenance of host animals 3 "erte&rate hosts
En general most fresh+ater fish can &e maintained in la&oratory conditions
especially in a)uaria or tan;s +ith dechlorinated +ater and oxygen supplyA lo+
temperature may help in maintaining the hosts for longer time$ Ef +ell fed infected
fish may har&our tape+orms e"en for a long period$
Many species of fresh+ater fish are also easy to reproduce or to gro+ from fry$
En contrast it is extremely difficult to maintain marine fish in the la&oratory for longer
time +ithout special conditions a"aila&le in marine a)uaria$ %his limits the possi&ility
of experimental studies including elucidation of life cycles of tape+orms parasitic in
these hosts$
<$ Manipulation of li"e material
4ar"al stages 'metacestodes( of fish tape+orms 'procercoids plerocercoids or
merocercoids 3 see 0her"y 2002( are usually small especially stages in copepods
+hich measure much less than / millimetre '#uperman /.93A Scholz /../& /...($
%herefore their handling is often difficult$ Manipulation +ith infected intermediate
hosts and isolated lar"al stages should &e made +ith a 2asteur pipette to a"oid
damage of metacestodes$ *dult tape+orms can &e handled using pipettes soft
t+eezers or small pieces of hard paper 'cartoon( +ith sharped terminal end$
F$ Enfection of hosts
Methodology of host infection depends on the type of life cycle of the gi"en
tape+orm$ Briefly intermediate hosts are infected either +ith free-s+imming lar"ae
'copepods challenged +ith coracidia of some &othriocephalideans and +ith floating
eggs +ith a hyaline outer en"elope of proteocephalideans and nippotaeniideans( or
+ith eggs containing an oncosphere infecti"e for the intermediate host 'tu&ificids +ith
operculated eggs of caryophyllideans amphipods 3 gammarids +ith eggs of
spathe&othriideans and pro&a&ly &enthic copepods +ith unoperculate eggs of some
&othriocephalideans($ %he infecti"e stages of tape+orms are ;ept in the +ater +ith
potential intermediate hosts for some usually short time until the stages are
s+allo+ed 'consumed($ %he in"erte&rate hosts ate then examined in regular inter"als
to detect parasite lar"ae and to o&ser"e their morphogenesis$
<efiniti"e hosts fish can &e infected &y ;eeping them +ith infected
intermediate hosts that should &e ingested &y these hosts +hich is relati"ely easy
mainly +ith tu&ificid oligochaetes 'Scholz /../a($ Sometimes it is necessary to use
syringe for peroral infection of fish 'Killemse /.69(A this procedure is difficult +ith
small fish &ecause their stomach and intestine can &e damaged &y the syringe$
CEE$ *ppendix of 1eagent recipes 'fixati"es stains mounting media(
Some data on reagents especially stains and mounting media can &e found
e$g$ in Scholz and 8anzelo"Z '/..?( de 0ham&rier '200/( and Cidal-Mart_nez et al$
'200/($
Re2ere%ces
Be"eridge E$ 200/$ %he use of life-cycle characters in studies of the e"olution of cestodes$ En-
4ittle+ood <$%$P$ Bray 1$*$ '5ds$( Enterrelationships of the 2latyhelminthes$ %he
Systematics *ssociation Special Colume Series 60 %aylor & Francis 4ondon and :e+ Uor;
pp$ 2703276$
de 0ham&rier *$ 200/$ * ne+ tape+orm from the *mazon Ama#otaenia y,ettae n$ gen$ n$ sp$
'5ucestoda- 2roteocephalidea( from the siluriform fishes Brac"y$laty!toma ilamento!%m and
B2 ,aillanti '2imelodidae($ 1e"ue Suisse de @oologie /0?- 30333/6$
de 0ham&rier *$ Scholz %$ 200?$ %ape+orms '0estoda- 2roteocephalidea( of fire+ood catfish
Sor%&imic"t"y! $lanice$! 'Siluriformes- 2imelodidae( from the *mazon 1i"er$ Folia
2arasitologica 77- /932?$
de 0ham&rier *$ Sane *$ Mahmoud @$ Mariaux P$ Scholz %$ 200?$ 1edescription of Sandonella
!andoni '4ynsdale /.60( '0estoda- 2roteocephalidea( an enigmatic and morphologically
uni)ue parasitic in the osteoglossiform fish .eteroti! nilotic%! in *frica$ Pournal of 2arasitology
.6- 20232//$
de 0ham&rier *$ Caucher 0$ /...$ 2roteocephalidae et Monticelliidae '5ucestoda-
2roteocephalidea( parasites de poissons dXeau douce du 2araguay a"ec descriptions dXun
genre nou"eau et de dix espaces nou"elles$ 1e"ue Suisse de @oologie /06- /673260$
0her"y 4$ 2002$ %he terminology of lar"al cestodes or metacestodes$ Systematic 2arasitology 72- /3
33$
5rgens 1$ /.6.$ %he suita&ility of ammonium picrate-glycerin in pregparing slides of lo+er
Monogenoidea$ Folia 2arasitologica /6- 320$
Freze C$E$ /.67$ E!!ential! o Ce!todology$ Col$ C$ Proteoce$"alata in Fi!", Am$"i&ian! and Re$tile!$
Mos;"a- EzdatelXst"o N:au;aO 73? pp$ 'En 1ussian- 5nglish translation Esrael 2rogram of
Scientific %ranslation /.6. 0at$ :o$ /?73$ " B 7.9 pp($
#uperman B$E$ /.93$ %ape+orms of the >enus Triaeno$"or%! 2arasites of Fish$ 5xperimental
Systematics 5cology$ :au;a 4eningrad 20? pp$ 'En 1ussian$(
#uchta 1$ Shinn *$2$ 8anzelo"Z C$ Scholz %$ 2006$ * comparati"e study of the egg morphology in
four species of E%&ot"ri%m '0estoda- 2sedophyllidea( +ith comments on their early
de"elopment$ En"erte&rate Biology /27- /-?$
4e"ron 0$ Brubans;Z M$ #uchta 1$ Freeman M$ Scholz %$ 2006$ Spermatozoon ultrastructure of
the pseudophyllidean cestode Paraec"ino$"all%! +a$onic%! a parasite of deep-sea fish
P!eno$!i! anomala '2erciformes 0entrolophidae($ 2arasitology 1esearch /00- //7-/2/$
Malm&erg >$ /.79$ =m fcre;omsten a" >yrodactylus pa s"ens;a Fis;er$ S;r$ scd$ S"er$ Fis;Fcr$
*rss;r$ /.76 pp$ /.396$
Scholz %$ /../$ <e"elopment of ;"a:ia !inen!i! 8sd /.37 '0estoda- 0aryophyllidea( in the
definiti"e host$ Folia 2arasitologica 3?- 2273236$
Scholz %$ /../$ 5arly de"elopment of ;"a:ia !inen!i! 8sd /.37 '0estoda- 0aryophyllidea($ Folia
2arasitologica 3?- /333/62$
Scholz %$ /..9$ 4ife-cycle of Bot"rioce$"al%! cla,ice$! a specific parasite of eels$ Pournal of
8elminthology 9/- 26/3272$
Scholz %$ /...$ 4ife cycles of species of Proteoce$"al%! Keinland /?7? '0estoda-
2roteocephalidae( parasites of fresh+ater fishes in the 2alearctic region- a re"ie+$ Pournal of
8elminthology 92- /3/.$
Scholz %$ de 0ham&rier *$ #uchta 1$ 200?$ 1edescription of the tape+orm 4onticellia ama#onica
de 0ham&rier et Caucher /..9 '0estoda- 2roteocephalidea( a parasite of Calo$"y!%!
macro$ter%! 'Siluriformes- 2imelodidae( from the *mazon 1i"er in Brazil and 2eru$ *cta
2arasitologica 73- 30337$
Scholz %$ <rZ&e; 1$ 8anzelo"Z C$ /..?$ Scolex morphology of Proteoce$"al%! tape+orms
'0estoda- 2roteocephalidae( parasites of fresh+ater fish in the 2alaearctic 1egion$ Folia
2arasitologica 67- 29363$
Scholz %$ 8anzelo"Z C$ /..?$ %ape+orms of the >enus Proteoce$"al%! Keinland /?7? '0estoda-
2roteocephalidae( 2arasites of Fishes in 5urope$ Studie *C [1 :o$ 2S.? *cademia 2raha
//. pp$
Cidal-Mart_nez C$M$ *guirre-Macedo M$4$ Scholz %$ >onzZlez-Sol_s <$ Mendoza-Franco 5$F$
200/$ *tlas of the 8elminth 2arasites of 0ichlid Fish of Mexico$ *cademia 2raha /66 pp$

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