Journal of Fish Diseases 2006, 29, 573–588
Review
Epitheliocystis in fish
B F Nowak1 and S E LaPatra2
1 School of Aquaculture, University of Tasmania, Launceston, Tasmania, Australia
2 Research Division, Clear Springs Foods, Inc., Buhl, ID, USA
Abstract
Epitheliocystis is a condition affecting the gills and
skin of fish, which has been reported from more
than 50 freshwater and marine species. It is caused
by intracellular Gram-negative bacteria. Mortalities
have been associated with epitheliocystis infections
in cultured fish. This review provides an update of
our current understanding of this condition,
including characterization of the pathogen using
immunohistochemical and molecular studies. In
most fish species the epitheliocystis agent was neg-
ative to an antibody specific for chlamydial genus-
specific lipopolysaccharide antigen. Recently, four
epitheliocystis agents from four different fish species
have been characterized using molecular analysis.
While they all belong to the order Chlamydiales, in
a lineage separate from the Chlamydiaceae, they are
distinct organisms and similarity analysis showed
that they had highest similarity values with other
chlamydia-like bacteria isolated from various sour-
ces, including humans or pig. This confirms the
high diversity and host specificity of the pathogen.
Further molecular analysis should result in an
increased understanding of this condition. To date
the pathogen has not been cultured, making
experimental studies difficult. High stocking den-
sities, presence of nutrients, season, temperature
and fish age have been identified as potential risk
factors for the manifestation of this condition.
Keywords: Chlamydiales, epitheliocystis, fish, gill
pathology, molecular analysis, risk factors.
Correspondence Dr B F Nowak, School of Aquaculture,
University of Tasmania, Locked Bag 1370, Launceston,
7250 Tasmania, Australia
(e-mail: b.nowak@utas.edu.au)
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Introduction
Epitheliocystis disease is a common condition of a
putative infectious aetiology that has been described
in various teleosts (Paperna & Sabnai 1980; Wolf
1988; Turnbull 1993; Fryer & Lannan 1994). The
descriptive term epitheliocystis is derived from the
appearance of epithelial lesions that are manifested
secondary to infection. The disease was first
described in common carp, Cyprinus carpio L., as
mucophilosis (Plehn 1920). Only after it was
reported in other fish species was it correctly
identified as a bacterial disease and named epithe-
liocystis (Hoffman, Dunbar, Wolf & Zwillenberg
1969). Epitheliocystis is a condition common in
many fish species that affects both the gill and skin
epithelium. It is an intracellular infection with
Gram-negative bacteria, resulting in hypertrophy of
host cells. There are no culture methods for the
causative agent.
Microscopically, progressive enlargement of
infected epithelial cells subsequently results in the
formation of spherical cysts that are circumscribed
by an eosinophilic hyaline capsule apparently
composed of retained remnants of the host cell
membrane and cytoplasm (Molnar & Boros 1981).
These enlarged cells can range up to 400 lm in
diameter and are often transparent white to yellow
when observed grossly (Herman & Wolf 1987;
Wolf 1988). Although the gills are the most
common site of infection, the integument (mostly
epidermis but also the epithelial lining of the
orobranchial cavity and nostrils and the mucosa of
oesophagus and pseudobranch) may also be involved
(Hoffman et al. 1969; Crespo, Grau & Padros
 Journal of Fish Diseases 2006, 29, 573–588 B F Nowak and S E LaPatra Epitheliocystis in fish

1990; Crespo, Zarza, Padros & Marin de Mateo

1999). Infected cells contain coccoid or coccobacil-
lary organisms that complete a pleomorphic devel-
opmental cycle with specific morphological
characteristics dependent on the stage of intracel-
lular development (Paperna, Sabnai & Castel 1978;
Paperna & Alves de Matos 1984). Development of
the organism is similar to that of the Chlamydiae
(Ward 1988) and proceeds from small, rigid
infectious forms to larger, pleomorphic non-infec-
tious forms that divide by fission to produce a new
generation of infectious daughter cells (Paperna,
Sabnai & Zachary 1981; Turnbull 1993).
Epitheliocystis has been associated with morbid-
ity and mortality in cultured fish species. However,
the direct link between epitheliocystis and mortal- Figure 1 Developmental cycle I is a typical chlamydial develop-
ities has not been established due to the lack of an mental cycle. Infection is initiated by attachment of an
elementary body to the host cell. After the elementary body
experimental challenge model. As bacteria associ-
enters the host cell, it differentiates into a reticulate body, which
ated with epitheliocystis have not been cultured, later transforms into an intermediate body before becoming
Koch’s postulates have never been fulfilled for this infective elementary bodies. EB, elementary body; RB, reticulate
disease. Recently some progress has been made in body; IB, intermediate body.
identifying the epitheliocystis agent, at least in some
fish species. The aim of this review is to summarize
our current understanding of this condition and
identify potential research needs.

Pathogen
The causative agents of epitheliocystis are intracel-
lular Gram-negative bacteria, which, on the basis of
their ultrastructure belong to Chlamydiaceae (Pap-
erna et al. 1981; Groff, LaPatra, Munn, Anderson
& Osburn 1996; Szakolczai, Vetesi & Pitz 1999) or
Rickettsiaceae (Zachary & Paperna 1977). It has
been suggested that epitheliocystis could be caused
by either chlamydial or rickettsial agents (Nylund,
Kvensenth & Isdal 1998).
The typical chlamydial developmental cycle
includes an infective elementary body, reticulate
body and intermediate body/round cell (Avakyan &
Popov 1984). In some fish species, primary long
cells, intermediate long cells, avacuolated small cells
and vacuolated small cells have also been described.
This led to the suggestion that the infections are
caused by either chlamydial or rickettsial agents or
that the epitheliocystis agent can use two develop-
mental cycles (Crespo et al. 1999). Developmental
cycle I is the typical chlamydial cycle (Fig. 1) and
developmental cycle II (Fig. 2) includes primary
long cells, intermediate long cells and small cells.
Fish age, stress or environmental conditions (Cre-
spo et al. 1999) and type of cell infected (Paperna
Figure 2 Developmental cycle II. Primary long cells and
intermediate long cells are reproductive forms and small cells
are an infective form. It has been suggested that switching between
the developmental cycles I and II occurs in epitheliocystis. PLC,
primary long cell; ILC, intermediate long cell; SC, small cell.
& Alves de Matos 1984) were suggested to be the
factors causing switching between these two devel-
opmental cycles. Cells characteristic for both
developmental cycles can be found in the same fish
species (Paperna et al. 1981; Crespo et al. 1999),
suggesting that the developmental cycle is not
specific for host species or the bacteria.
In addition to cells characteristic for develop-
mental cycle I, head and tail cells were described in
steelhead trout, Oncorhynchus mykiss (Walbaum)
(Rourke, Davis & Bradley 1984); lake trout,
Salvelinus namaycush (Walbaum) (Bradley, New-

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 Journal of Fish Diseases 2006, 29, 573–588 B F Nowak and S E LaPatra Epitheliocystis in fish

comer & Maxwell 1988); and Atlantic salmon,
Salmo salar L. (Draghi, Popov, Kahl, Stanton,
Bronw, Tsongalis, West & Frasca 2004). This type
of cell was suggested to be an abnormal form of
reticulate body, which can develop under adverse
conditions (Avakyan & Popov 1984; Crespo et al.
1999). This morphology does not, however, pre-
clude the bacteria from being Chlamydiae, as
demonstrated for Rhabdochlamydia (Corsaro &
Venditti 2004). Despite attempts, the pathogen
has not been cultured in vitro (Bradley et al. 1988;
A. Meijer, unpublished data).
The presence of chlamydial antigen was demon-
strated within cytoplasmic inclusions in white
sturgeon, Acipenser transmontanus Richardson, by
the peroxidase-antiperoxidase (PAP) immunohisto-
chemical technique using 11B5 antibody specific
for the chlamydial genus-specific lipopolysaccharide
(LPS) antigen (Groff et al. 1996). Labelling of
membranes and the peripheral cytoplasm of reticu-
late bodies and intermediate body inclusions using
mouse monoclonal antibody anti-chlamydial LPS
was reported in the cases of Atlantic salmon
epitheliocystis in Norway and Ireland (Draghi et al.
2004). However, in epitheliocystis in most fish
species there was no positive reaction with group
specific chlamydial antibody (Table 1). This sug-
gests that there is a wide taxonomic variation
between the pathogens causing epitheliocystis in
different fish species. Furthermore, even infections
in one fish species can give different results. For
example, epitheliocystis in rainbow trout from
Idaho reacted positively whereas epitheliocystis
from the same fish species from Washington and
Montana was negative. Additionally, monoclonal
antibody-immunofluorescence test for chlamydial
antigen was negative for leafy sea-dragon, Phycodu-
rus eques (Günther), affected by epitheliocystis
(Langdon, Elliott & Mackay 1991). However, cysts
in the sections from the same block were reported
to stain very weakly positive with monoclonal
antibodies against Chlamydiaceae LPS (Meijer,
Roholl, Ossewaarde, Jones & Nowak 2006).
Additionally, staining was positive for epitheliocys-
tis in yellowtail from Ecuador but negative for
epitheliocystis in yellowtail kingfish, Seriola lalandi
Valenciennes, from Australia (Table 1). This could
be due to inherent susceptibility amongst different
strains of a particular fish species, developmental
stage of the chlamydia tested or even infection of
the same host species with two different species of
bacteria. This is further supported by two types
of host response, proliferative and non-proliferative
in the same host species from the same location.
Examples of this include sea bream, Sparus aurata
L. (Paperna 1977; Padros & Crespo 1995; Crespo
et al. 1999), and Atlantic salmon in Ireland in 1995
and 1999 (Draghi et al. 2004).
Epitheliocystis agents in different species of fish
show ultrastructural differences (Paperna 1977;
Paperna et al. 1981). This variation has been
confirmed by molecular techniques (Table 2). To

Table 1 Immunohistochemical testing of

epitheliocystis specimens (methods Chlamydia
immunohisto-
described in Groff et al. 1996)
Species Location chemical result

Rainbow trout, Oncorhynchus mykiss (Walbaum) Idaho +

Yellowtail larvae, Seriola lalandi (Valenciennes) Ecuador +
Yellowtail, S. lalandi (Valenciennes) Australia )
Japanese yellowtail, Seriola quinqueradiata Japan )
(Temminck & Schlegel)
Leopard shark, Triakis semifasciata (Girard) California )
Arctic char, Salvelinus alpinus (L.) West Virginia )
Silver perch, Bidyanus bidyanus (Mitchell) Australia )
Rock cale, Crinodus lophodon (Günther) Australia )
Pacific herring, Clupea pallasii (Valenciennes) Alaska )
Sea bream, Sparus aurata L. Spain )
Amber jack, Seriola dumerili (Risso) Spain )
Brown bullhead, Ameirus nebulosus (Leseur) New York )
Marine catfish, Arius sp. (Valenciennes) Ecuador )
Steelhead, O. mykiss (Walbaum) Washington )
Spring chinook, Oncorhynchus Washington )
tshawytscha (Walbaum)
Rainbow trout, O. mykiss (Walbaum) Washington )
Lake trout, Salvelinus namaycush (Walbaum) Wisconsin )
Rainbow trout, O. mykiss (Walbaum) Montana )
Walleye, Sander vitreus (Mitchill) Montana )

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 Journal of Fish Diseases 2006, 29, 573–588
Table 2 Molecular identification of the epitheliocystis agent
Epitheliocystis organism
(GenBank accession no.) ISH Fish species
Candidatus Piscichlamydia +ve Atlantic salmon, Salmo salar L.
salmonis (AY462244)
CRG18 (AY013394) +ve Silver perch, Bidyanus bidyanus
(Mitchell)
CRG 98 (AY013474) +ve Barramundi, Lates calcarifer
(Bloch)
CRG20 (AY013396) )ve Leafy sea dragon,
Phycodurus eques (Günther)
Near full length 16S rDNA was sequenced for Candidatus Piscichlamydia salmonis, 210–223 basepair region of 16S rRNA Chlamydiales signature sequence
was used for CRG18, CRG20 and CRG98. Epitheliocystis was diagnosed histologically in sections from the same blocks (Langdon et al. 1991; Draghi
et al. 2004; Meijer et al. 2006).
ISH, in situ hybridization; CRG, Chlamydia Research Group.
date four cases of epitheliocystis-associated bacteria
have been identified using polymerase chain reac-
tion (PCR) of highly conserved regions of 16S
rDNA. Molecular analysis confirmed that all four
epitheliocystis agents belonged to the order
Chlamydiales, in a lineage separate from the family
Chlamydiaceae (Draghi et al. 2004; Meijer et al.
2006). Two of the three positive agents from
Australian fish could be confirmed with in situ
hybridization using a Chlamydiales 16s rRNA-
specific oligoprobe (Meijer & Ossewaarde 2002;
Meijer et al. 2006). On the basis of their partial
sequences, each of these epitheliocystis agents had
higher similarity values with sequences from chla-
mydia-like bacteria isolated from humans, pigs or
water storage than with each other (Meijer et al.
2006). Current results confirm high diversity and
host species-specificity of the epitheliocystis agents.
This suggests that transmission between species is
limited. Diagnostic real-time PCR has been devel-
oped for ÔCandidatus Piscichlamydia salmonisÕ (T.M.
Steinum, personal communication), which should
improve our understanding of the presence of the
pathogen, its relationship with pathology and
disease dynamics, at least in Atlantic salmon.
While the four epitheliocystis agents are different
in each species, they all belong to Chlamydiales,
despite the bacteria showing different developmen-
tal cycles in these species. Primary long cells,
intermediate long cells and small cells were des-
cribed in silver perch, Bidyanus bidyanus (Mitchell)
(Frances, Tennent & Nowak 1997), and primary
long cells and intermediate long cells were described
in barramundi, Lates calcarifer (Bloch) (Anderson &
Prior 1992), these cells are characteristic for cycle II
(Crespo et al. 1999). Cells characteristic for devel-
opmental cycle I were shown for the other two
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B F Nowak and S E LaPatra Epitheliocystis in fish
Origin Environment Reference
Culture Norway Marine Draghi et al. (2004)
Culture Australia Fresh water Meijer et al. (2006)
Culture Australia Fresh water Meijer et al. (2006)
(but originally
marine hatchery)
Aquarium Australia Marine Meijer et al. (2006)
species; intermediate body/round cell in leafy sea
dragon (Langdon et al. 1991) and reticulate body,
intermediate body/round cell and elementary body
in Atlantic salmon (Nylund et al. 1998). This
suggests that all epitheliocystis agents may be from
the order Chlamydiales.
Diagnosis
The disease cannot be diagnosed through routine
microbiology screening, because the bacteria cannot
be cultured. As a result, if histology is not used in
diagnosis, the disease can go unreported. For
example, epitheliocystis was not included in the
two published lists of diseases of Japanese yellowtail,
Seriola quinqueradiata Temminck & Schlegel
(Egusa 1983; Kusuda & Kawai 1998), however,
after using histology it was found in 100% of gill
specimens sampled from Japanese yellowtail cul-
tured in Japan (B.F. Nowak, unpublished data).
Epitheliocystis has been reported in experimental
fish for example in silver perch (Frances et al.
1997), yellowtail kingfish (Mansell, Powell, Ernst
& Nowak 2005) and common carp (B.F. Nowak,
unpublished data). Lack of a correct diagnosis may
have adverse effects on the experimental results
when the researcher assumes that the experimental
control fish are uninfected and healthy. Sometimes
epitheliocystis can be seen grossly and in wet
preparations (Fig. 3), however, histopathology is
used to provide a definitive diagnosis (Fig. 4).
Host species
Epitheliocystis has been described in more than 50
species of fish from marine and freshwater environ-
ments (for review see Paperna & Sabnai 1980; Fryer
 Journal of Fish Diseases 2006, 29, 573–588
(a)
(b)
(c)
Figure 3 Wet preparations of fish gills showing epitheliocystis:
(a) hyperinfection in gills of cultured largemouth bass showing
numerous cysts (arrow), this infection resulted in mortalities (see
Table 5); (b) higher magnification of gills of largemouth bass
infected with epitheliocystis showing one filament with large
number of cysts (arrow); (c) high magnification showing
epitheliocystis in threadfin shad, Dorosoma petenense (Günther)
showing infiltration with inflammatory cells (arrow). Figure
courtesy of Dr Andy Goodwin.
& Lannan 1994) and in both cultured and wild
fish. In contrast to the claim that no clear data exist
on chlamydia-like infections in non-teleosts (Cor-
saro & Venditti 2004), epitheliocystis has been
reported in white sturgeon (Groff et al. 1996) and
in a leopard shark, Triakis semifasciata Girard
(Table 1).
The prevalence of epitheliocystis is generally
greater in cultured fish than in wild fish (Table 3).
The mean maximum prevalence was 28% for wild
species (ranged from 1% to 91%) and 73% for
cultured species (ranged from 20% to 100%).
Atlantic salmon farmed in Tasmania was the only
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B F Nowak and S E LaPatra Epitheliocystis in fish
cultured fish species to show a low prevalence of
infection (Table 3). Chum salmon, Oncorhynchus
keta (Walbaum), exhibited the greatest prevalence
of epitheliocystis of all wild fish examined. The wild
fish showing greatest prevalence of epitheliocystis
were generally all marine demersal species. How-
ever, this could be a result of a sampling bias
because many fish disease surveys focus on demersal
fish.
Epitheliocystis hyperinfection was reported in the
early developmental stages of cultured yellowtail,
Seriola mazatlana (Valenciennes), but not in floun-
der Paralichthys adspersus (Steindachner) and Par-
alichthys woolmani Jordan & Williams, cultured at
the same facility (Venizelos & Benetti 1996). While
epitheliocystis was found histologically in several of
the flounder larvae, there was no increase in
mortalities and no pathological changes as seen in
the yellowtail. It was suggested that the difference
in epitheliocystis outbreaks within the same facility
was related to the differences in the biology and
physiology of these host species.
When epitheliocystis was reported in the same
species from the same geographical area on two
different occasions about 10 years apart, the results
were similar, despite very low sample sizes. For
example, epitheliocystis prevalence in American
plaice Hippoglossoides platessoides (Fabricius), was
reported to be 20% by Morrison & Shum (1983)
and 17% by Lewis, McLaughlin, Bodammer &
Sawyer (1992). This suggests that this condition can
be stable and may persist in some wild populations
of fish. However, larger variation was reported in
prevalence of epitheliocystis in pilchards, Sardinops
sagax (Jenyns), from the same locations in Australia
and New Zealand (0–30%, fish were repetitively
sampled every few days, Whittington, Jones, Hine
& Hyatt 1997), this was most probably due to
small sample sizes (three to 10 individuals on each
occasion).
While there are some fish species which were
reported not to be infected by epitheliocystis, the
confidence in these results is often low. When the
apparent (or observed) prevalence of epitheliocystis
was reported to be 0, the true prevalence (at 95%
confidence, assuming test sensitivity 90% and test
specificity 95%) ranged from 3% to 29% (Table 4).
This is mostly due to the limited sample size.
Furthermore, in most cases, a negative result was
based on an examination of a limited number of
histological slides, often only a single section from
one gill arch. To date, no reproducible challenge
 Journal of Fish Diseases 2006, 29, 573–588 B F Nowak and S E LaPatra Epitheliocystis in fish

(a) (b)

(c) (d)

Figure 4 Histopathology of epitheliocystis: (a) hyperinfection in the gills of largemouth bass showing numerous cysts (arrow), this is the
same outbreak as in Fig. 3(a,b); (b) higher magnification of gills of largemouth bass infected with epitheliocystis showing cysts (arrow)
and host response resulting in lamellar fusion; (c) two cysts in the gills of cultured kingfish, note the lack of host response; (d) one cyst in
gills of cultured Atlantic salmon, note the host response, in particular infiltration with inflammatory cells. Figure 4(a,b) courtesy of
Dr Andy Goodwin.

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Table 3 Maximum prevalence of epitheliocystis reported in different fish species

Fish species Prevalence, % (n) Origin Reference

Brown bullhead, Ictalurus nebulosus (Lesueur) 6 (56) Wild freshwater Desser et al. (1988)
American plaice, Hippoglossoides platessoides 20 (11) Wild marine Morrison & Shum (1983)
(Fabricius)
Atlantic cod, Gadus morhua L. 34 (215) Wild marine Lewis et al. (1992)
Fourspot flounder, Paralichthys oblongus (Mitchill) 27 (20) Wild marine Lewis et al. (1992)
Ocean pout, Macrozoarces americanus 47 (15) Wild marine Lewis et al. (1992)
(Bloch & Schneider)
Spotted hake, Urophycis regia (Walbaum) 15 (75) Wild marine Lewis et al. (1992)
Summer flounder, Paralichthys dentatus 25 (209) Wild marine Lewis et al. (1992)
Weakfish, Cynoscion regalis (Bloch & Schneider) 39 (59) Wild marine Lewis et al. (1992)
Windowpane, Scophthalmus aquosus (Mitchill) 40 (144) Wild marine Lewis et al. (1992)
Winter flounder, Pleuronectes americanus (Walbaum) 31 (950) Wild marine Lewis et al. (1992)
Yellowtail flounder, Pleuronectes ferrugineus (Storer) 9 (44) Wild marine Lewis et al. (1992)
Red morwong, Cheilodactylus fuscus (Castelnau) 12 (60) Wild marine Nowak (1996)
Rock cale, Crinodus lophodon (Günther) 48 (60) Wild marine Nowak (1996)
White ear parma, Parma microlepis (Günther) 7.5 (120) Wild marine Tricklebank (1997)
Jack mackerel, Trachurus declivis (Jenyns) 21 (81) Wild marine Nowak et al. (2004)
Chinook salmon, Oncorhynchus tshawytscha 15 (82) Wild marine Kent, Traxler, Kieser,
(Walbaum) Richard, Dawe, Shaw,
Prosperi-Porta, Ketcheson
& Evelyn (1998)
Chum salmon, Oncorhynchus keta (Walbaum) 91 (32) Wild marine Kent et al. (1998)
Coho salmon, Oncorhynchus kisutch (Walbaum) 23 (48) Wild marine Kent et al. (1998)
Pink salmon, Oncorhynchus gorbuscha (Walbaum) 8 (12) Wild marine Kent et al. (1998)
Pacific herring, Clupea pallasi (Valenciennes) 21 (38) Wild marine Kent et al. (1998)
Shiner perch, Cymatogaster aggregata (Gibbons) 41 (32) Wild marine Kent et al. (1998)
Pacific cod, Gadus macrocephalus (Tilesius) 1 (98) Wild marine Kent et al. (1998)
Pacific hake, Merluccius productus (Ayres) 12 (25) Wild marine Kent et al. (1998)
Walleye pollock, Theragra chalcogramma (Pallas) 5 (20) Wild marine Kent et al. (1998)
Pacific tomcod, Microgadus proximus (Girard) 2.5 (40) Wild marine Kent et al. (1998)
Lingcod, Ophiodon elongatus (Girard) 14 (111) Wild marine Kent et al. (1998)
Sablefish, Anoplopoma fimbria (Pallas) 12 (33) Wild marine Kent et al. (1998)
Canary rockfish, Sebastes pinniger (Gill) 67 (12) Wild marine Kent et al. (1998)
Darkblotched rockfish, Sebastes crameri (Jordan) 30 (10) Wild marine Kent et al. (1998)
Redbanded rockfish, Sebastes babcocki (Thompson) 40 (10) Wild marine Kent et al. (1998)
Redstripe rockfish, Sebastes proriger (Jordan & Gilbert) 55 (31) Wild marine Kent et al. (1998)
Rougheye rockfish, Sebastes aleutianus 61 (13) Wild marine Kent et al. (1998)
(Jordan & Evermann)
Silvergrey rockfish, Sebastes brevispinis (Bean) 25 (11) Wild marine Kent et al. (1998)
Yellowtail rockfish, Sebastes fladivus (Ayres) 33 (21) Wild marine Kent et al. (1998)
Arrowtooth flounder, Astheresthes stomia 27 (15) Wild marine Kent et al. (1998)
(Jordan & Gilbert)
Greater pipefish, Sygnathus acus L. 23 (30) Wild marine Longshaw, Green
& Feist (2004)
Pilchard, Sardinops sagax (Jenyns) 30 (25) Wild marine Whittington et al. (1997)
Amberjack, Seriola dumerili (Risso) 46 (15) Wild marine Grau & Crespo (1991)
Sea bream, Sparus aurata L. 70 (13) Cultured marine Paperna (1977)
Red sea bream, Pagrus major (Temminck & Schlegel) 100 (100) Cultured marine Syasina et al. (2004)
Red sea bream, P. major · black sea bream, 10 (50) Cultured marine Syasina et al. (2004)
Acanthopagrus schlegeli hybrid
Atlantic salmon, Salmo salar L. 20 (20) Cultured marine Nowak & Clark (1999)
Dentex, Dentex dentex (L.) 88 (19) Cultured marine Company, Sitja-Bobadilla,
Pujalte, Garay,
Alvarez-Pellitero &
Perez-Sanchez (1999)
Yellowtail kingfish, Seriola lalandi (Valenciennes) 86 (82) Cultured marine B.F. Nowak, unpublished data
Japanese yellowtail, Seriola quinqueradiata 100 (10) Cultured marine B.F. Nowak, unpublished data
(Temminck & Schlegel)
Channel catfish, Ictalurus punctatus (Rafinesque) 46 (46) Cultured freshwater Zimmer, Ewing & Kocan (1984)
Silver perch, Bidyanus bidyanus (Mitchell) 75 (105) Cultured freshwater Frances et al. (1997)

Only species for which a minimum of 10 individuals were examined are included.

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Table 4 Fish species where epitheliocystis was reported undetectable, i.e. reported negative (0% prevalence) for epitheliocystis (when 10
or more than 10 individuals were examined)
Number of fish
Fish species examined
Spiny dogfish, Squalus acanthias L. 10
Spotted ratfish, Hydrolagus colliei (Lay & Bennett) 16
Skate, Bathyraja abyssicola (Gilbert) 12
Pink salmon, Oncorhynchus gorbuscha (Walbaum) 15
Threespine stickleback, Gasterosteus aculeatus L. 12
Rex sole, Glyptocephalus zachinus (Lockington) 11
Petrale sole, Eopsetta jordani (Lockington) 10
Pacific halibut, Hippoglossus stenolepis (Schmidt) 11
Bigfin eelpout, Lycodes cortezianus (Gilbert) 11
Sand flathead, Platycephalus bassensis (Cuvier) 122
Red cod, Pseudophycis bachus (Forster) 84
Smallmouth flounder, Etropus microstomus (Gill) 41
Prevalence estimates from a survey (observed prevalence or apparent prevalence) can be misleading due to small sample size and imperfect diagnostic tests
used. True prevalence is calculated taking sample size as well as false-positive and false-negative results into account. True prevalence was calculated at 95%
confidence, assuming test sensitivity 90% and test specificity 100% using Pooled Prevalence Calculator (http://www.ausvet.com.au).
method has been developed for this disease. Thus, it
is difficult to evaluate the susceptibility of different
fish species or strains.
Effect on host
Mortality
Mortalities associated with epitheliocystis that have
been reported ranged from 4% to 100% and
occurred only in cultured fish (Table 3). However,
even in cultured fish, the condition was usually
non-proliferative (not causing proliferative tissue
reaction in host) and benign (with no apparent
effect on the host). Most of the mortalities were
associated with early life stages, with the exception
of a few species (Table 5). Epitheliocystis associated
mortalities have been reported in Atlantic salmon
cultured in Norway (Nylund et al. 1998; Brun,
Poppe, Skrudland & Jarp 2003). However, no
mortalities related to this condition were reported
in Atlantic salmon cultured in Tasmania despite
epitheliocystis being present in up to 20% of the
fish (Nowak & Clark 1999). The infection had very
low intensity in salmon from Tasmania with only
one or two cysts present in a section of a gill arch. In
contrast, in an outbreak of epitheliocystis in
Atlantic salmon in Ireland and Norway, the cysts
affected 60–80% of gill lamellae on each gill arch
(Draghi et al. 2004). While mortality in older fish
often occurs as a result of proliferative epithelio-
cystis, no proliferative host response is usually
associated with epitheliocystis-related mortalities in
early life stages of fish (Table 5).
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B F Nowak and S E LaPatra Epitheliocystis in fish
True prevalence
(at 95% confidence) Reference
29 Kent et al. (1998)
19 Kent et al. (1998)
25 Kent et al. (1998)
20 Kent et al. (1998)
25 Kent et al. (1998)
27 Kent et al. (1998)
29 Kent et al. (1998)
27 Kent et al. (1998)
27 Kent et al. (1998)
3 Nowak et al. (2004)
4 Nowak et al. (2004)
8 Lewis et al. (1992)
Pathology
Epitheliocystis affects mostly the gills and skin of
fish. It is characterized by the presence of hyper-
trophied cells containing fine basophilic granular
inclusions (Paperna 1977). The bacteria are present
in one large inclusion, delineated by a bilaminated
membrane and displacing cytoplasm of the host cell
(Paperna et al. 1981). The infected cells are usually
reported to be epithelial cells, for example in
Atlantic salmon (Nowak & Clark 1999; Draghi
et al. 2004), leafy sea-dragon (Langdon et al.
1991), barramundi (Anderson & Prior 1992),
brown bullhead, Ameiurus nebulosus (Lesueur)
(Desser, Paterson & Steinhagen 1988), Connecticut
striped bass, Morone saxatilis (Walbaum), and white
perch, Morone americana (Gmelin) (Wolke, Wyand
& Khairallah 1970). In other fish species other
types of cells were reported to be infected, including
chloride cells in amberjack, Seriola dumerili (Risso)
(Crespo et al. 1990; Grau & Crespo 1991), and
Atlantic salmon (Nylund et al. 1998), mucous cells
in carp (Paperna & Alves de Matos 1984),
macrophages in brown bullhead (Desser et al.
1988) and pillar cells in red sea bream, Pagrus
major (Temminck & Schlegel) and tiger puffer,
Takifugu rubripes (Temminck & Schlegel) (Miya-
zaki, Fujimaki & Katai 1986). It has also been
suggested that an epithelial cell may adapt to the
infection by increasing the number of mitochon-
dria, which may then result in an incorrect
identification of this cell as a chloride cell (Morrison
& Shum 1983). Similar changes to host cells were
reported in microsporidian infections (Morrison &
 Journal of Fish Diseases 2006, 29, 573–588 B F Nowak and S E LaPatra Epitheliocystis in fish

Sprague 1981). Chlamydiae lack the ability to

J.S. Langdon & S.C. Battaglene, unpublished data

C. Johnston & P. Phillips, personal communication
synthesize high-energy compounds necessary for

Crespo et al. (1990); Grau & Crespo (1991)

metabolism and respiration, which must be pro-
vided by the host cell. This could lead to

Miyaki, Mizuta, Yamamoto, Yoshikoshi,

preferential infection of chloride cells or to
adaptation of epithelial cells to the infection by
increasing the number of mitochondria.

Venizelos & Benetti (1996)

The direct link between epitheliocystis and mortality has not been confirmed, except that severe epitheliocystis occurred in the mortalities and not in surviving fish or fish from other populations.
The intensity of infection can vary (Table 6).

Kanai & Tabeta (1998)

Szakolczai et al. (1999)
Goodwin et al. (2005)

Miyazaki et al. (1986)

Bradley et al. (1988)
Only one filament on the gill arch was affected and

Nylund et al. (1998)

Groff et al. (1996)

in most cases only one cyst was present in Atlantic

Paperna (1977)
salmon in Tasmania (Nowak & Clark 1999). This
Reference

is consistent with a very low infection level. A much

greater percentage of affected filaments were
reported for Atlantic salmon in Norway (Nylund
et al. 1998; Draghi et al. 2004), leafy sea dragon
Environment

Fresh water
Fresh water

Fresh water
Fresh water

(Langdon et al. 1991) and silver perch (Frances

et al. 1997). While non-proliferative hyperinfection
Marine
Marine
Marine
Marine

Marine
Marine
Marine
Marine
was reported in sea bass, Dicentrarchus labrax (L.),
epitheliocystis hyperinfections in sea bream or the
amberjack result in significant host response (Grau
mild hyperplasia
Proliferative host

No response or

& Crespo 1991; Crespo et al. 1999; Crespo, Zarza

& Padros 2001). For sea bream or mugilids no
response

relationship was found between state and level of

Table 5 Mortalities reported in cultured fish in association with epitheliocystis (as confirmed by histopathology)

Yes

Yes
Yes
Yes
Yes

Yes
Yes

infection or cyst size and the stage of development

No
No

No
No

of the pathogen (Paperna et al. 1981).

Epitheliocystis can cause a significant prolifera-
Larva, early juvenile

Larva, early juvenile

tive reaction (Table 6). In cultured sea bream, cysts

Adult (1–1.5 kg)
7 months old

were present in the majority of interlamellar spaces

6 month old
600–650 g
1 year old

and were associated with epithelial proliferation

0+, 1+

Juvenile

Juvenile

Smolt

forming concentric layers around the cysts (Paperna

Age

0+

1977). In contrast, wild sea bream developed

epitheliocystis in captivity, but it was never associ-
Mortality (%)

ated with host response except for the enclosure of

26/10 days

the cyst surface by one or more layers of epithelium

30–100
98–100

Severe
Severe
30–40
1/day
4–8

(Paperna 1977). Atlantic salmon in Tasmania with

95

20
85

10

epitheliocystis had about one-third of the cysts

associated with some proliferative cellular response
Australian bass, Macquaria novemaculatea (Steindachner)

and inflammation (Nowak & Clark 1999). Pre-

White sturgeon, Acipenser transmontanus (Richardson)

Red sea bream, Pagrus major (Temminck & Schlegel)

Largemouth bass, Micropterus salmoides (Lacepède)

vious observations of epitheliocystis in salmonids

have suggested little or no reaction in the
Lake trout, Salvelinus namaycush (Walbaum)

Pacu, Piaractus mesopotamicus (Holmberg)

surrounding cells (Turnbull 1993). However,

Bartail flathead, Platycephalus sp. (Bloch)

samples submitted from farmed Atlantic salmon

Yellowtail, Seriola lalandi (Valenciennes)

Kingfish Seriola lalandi (Valenciennes)

from Norway in 2000 and Ireland in 1999 showed

Amberjack, Seriola dumerili (Risso)

proliferative lesions associated with epitheliocystis

Atlantic salmon, Salmo salar L.
Sea bream, Sparus aurata L.

affecting 60–80% of lamellae (Draghi et al. 2004).

In contrast, Atlantic salmon gills samples in Ireland
in 1995 had no proliferative changes (Draghi et al.
2004). This suggests that a proliferative response is
not related to host species or geographical location.
Granular and amorphous cyst contents were
Species

described in sea bream (Crespo et al. 1999). If a

cyst appeared granular it did not result in a

 2006
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 2006
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582
Table 6 Examples of pathology and host response to epitheliocystis

Fish species Intensity of infection Cyst size (lm) Host reaction Reference

American plaice, Hippoglosoides Highly variable 25 Cyst surrounded by squamous host Morrison & Shum (1983)
platessoides (Fabricius) cells
White sturgeon, Acipenser Highly variable 11–20 · 15–28 Absent or limited to a mild epithelial Groff et al. (1996)
transmontanus (Richardson) hyperplasia
Leafy seadragon, Phycodurus NA 20–90 No reaction Langdon et al. (1991)
eques (Günther)
Atlantic salmon Tasmania, Low – maximum three 18–71 Variable, including: lamellar fusion Nowak & Clark (1999)
Salmo salar L. cysts/gill arch inflammation
Journal of Fish Diseases 2006, 29, 573–588

Atlantic salmon Norway, 2000 samples – 60–80% of NA Proliferative lesions, multifocal to Nylund et al. (1998);
Salmo salar L. lamellae affected in 2000 diffuse hyperplasia, focal necrosis of Draghi et al. (2004)
1996 samples – <20% epithelial cells, macrophage
lamellae fused infiltrates
Atlantic salmon Ireland, 1995 samples – few cysts NA 1995 samples – no proliferative Draghi et al. (2004)
Salmo salar L. 1999 samples – 60–80% changes
of lamellae 1999 samples – proliferative lesions,
multifocal to diffuse hyperplasia,
focal necrosis of epithelial cells,
macrophage infiltrates
Silver perch, Bidyanus bidyanus 2–96% filaments 10–87 Epithelial hyperplasia Frances et al. (1997)
(Mitchell)
Sea bream, Sparus aurata L. 100% filaments 100 · 55 Hyperplastic epithelial cells Paperna (1977)
aggregated in concentric layers
around the cysts, infiltration with
macrophages and eosinophils
Amberjack, Seriola dumerili (Risso) NA 250 · 210 maximum Proliferative reaction Crespo et al. (1990)
Carp Cyprinus carpio L. NA 1–10 · 3–5 Non-proliferative and proliferative Paperna & Alves de Matos (1984)
Pacu Piaractus mesopotamicus (Holmberg) Majority of filaments 10–20 (homogeneous content) Proliferative, well-defined wall around Szakolczai et al. (1999)
30–40 (granular content) the cyst, epithelial hyperplasia,
hyperaemia and oedema but no
inflammation except around the
larger cysts
Channel catfish, Ictalurus punctatus NA 9–19 · 16–22 Non-proliferative, no host reaction, Zimmer et al. (1984)
(Rafinesque) refractile pink wall

NA, no information available.

B F Nowak and S E LaPatra Epitheliocystis in fish
 Journal of Fish Diseases 2006, 29, 573–588
proliferative reaction. The morphology of the
organisms within the cysts and their staining
properties (in particular with Azan and Macchiav-
ello), as well as the morphology of their capsules
and their location in fish tissue, were different
(Crespo et al. 1999). This was related to two
distinct developmental cycles of the pathogen. Cysts
in some other fish species were reported to be
surrounded by a layer of squamous epithelial cells
(Morrison & Shum 1983; Crespo et al. 1990).
The cellular response to the infection is often
variable, even in the same host (Table 6). For
example, in Atlantic salmon cultured in Tasmania
56% of cysts resulted in almost no reaction except
for encapsulation of the cyst with a thin layer of
squamous epithelial cells and lamellar fusion due to
the size of the cyst. However, other cysts resulted in
a dramatic cellular response. About 53% of the cysts
exhibited lamellar fusion. Inflammation was associ-
ated with 36% of the cysts, epithelial hyperplasia
with 34% and proliferation of mucous cells with
21%. Epithelial lifting was seen in 8% of cysts and
necrotic cells were associated with 6% of the cysts.
Other hosts show consistently the same reaction, for
example epitheliocystis was reported to cause a
proliferative reaction in silver perch (Frances et al.
1997; Meijer et al. 2006) and red sea bream
(Miyazaki et al. 1986; Ototake & Matsusato 1987).
In Atlantic salmon, cysts were present mostly in
the middle of the filament (44%), followed by the
base of the filament (32%) and the tip of the
filament (24%) (Nowak & Clark 1999). Most of
the cysts were positioned in the filamentous
epithelium. However, 22% of the cysts were present
at the tip of a lamella. The infected epithelial cells
were mostly present towards the tip of the lamellae
in epitheliocystis infection in Atlantic salmon from
Norway (Nylund et al. 1998). The position of the
cyst did not seem to have an effect on the cellular
response (Nowak & Clark 1999).
In Atlantic salmon the diameter of the cysts ranged
from 18 to 73.5 lm (mean 43.6 lm, SE ¼
1.7 lm). There was an increase in the mean size of
the cyst dependent on when in the first year of sea life
the sample was obtained, so the mean size of the cyst
was larger the longer fish were in the cages. However,
this relationship was not present in the second year of
sampling (Nowak & Clark 1999). There was no
relationship between prevalence or water tempera-
ture and size of the cysts. The cellular response was
not related to the size of the cyst (Nowak & Clark
1999). The average diameter of cysts was within the
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B F Nowak and S E LaPatra Epitheliocystis in fish
range reported previously in other fish species
(Miyazaki et al. 1986; Noga 1996; Frances et al.
1997) and approaches the lower range of the size
reported for mature cysts in salmonids (Bruno &
Poppe 1996).
Treatment and control
Oxytetracycline (25 ppm active ingredient, twice a
day for 3 days) was successfully used to treat
epitheliocystis in farmed largemouth bass, Micro-
pterus salmoides (Lacepède), resulting in a rapid
decline in mortalities to 0 in 3 days post-treatment
and no epitheliocystis was detected histologically
after 2 weeks (Goodwin, Park & Nowak 2005). In
the past, chloramphenicol mixed with ground
chicken livers (1:100) and fed 100 g a day for
3 days was reported to be successful in treating
epitheliocystis in cultured sea bream (Paperna
1977) with mortalities stopping after 1 week.
However, this antibiotic is not now suitable for
use in fish destined for human consumption.
Sterilization of rearing water using ultra violet light
was reported to control outbreaks of epitheliocystis
in amberjack, Seriola dumerili (Risso), and Plec-
tropomus leopardus (Lacepède) (Miyaki, Mizuta,
Yamamoto, Yoshikoshi, Kanai & Tabeta 1998).
Risk factors
Cultured fish appear to have a greater prevalence of
epitheliocystis than wild fish. Furthermore, prolif-
erative hyperinfection has been so far reported only in
farmed fish (Paperna 1977; Grau & Crespo 1991).
This suggests that culture conditions may exacerbate
epitheliocystis, perhaps due to increased fish densi-
ties, the presence of nutrients and/or stress. No
epitheliocystis was found in sea bream collected for
stocking, however, within 1 month mild epithelio-
cystis was found in up to 70% of the fish (Paperna
1977). Wild sea bream showed only mild and
sporadic epitheliocystis, whereas hatchery-reared sea
bream were affected by hyperinfection resulting in
26% mortality (Paperna 1977). While epitheliocystis
was present in wild kingfish, the prevalence and
intensity of infection was greater in cultured yellow-
tail kingfish (B.F. Nowak, unpublished data).
Aquaculture may also affect the prevalence of
epitheliocystis in wild fish. Mean prevalence of
epitheliocystis was greater in mackerel, Trachurus
declivis (Jenyns), collected from salmon farms (from
salmon cages or next to the cages) than in mackerel
 Journal of Fish Diseases 2006, 29, 573–588
from other locations, however, this difference was
not statistically significant (Nowak, Dawson, Bas-
son, Deveney & Powell 2004). This may be related
to increased concentrations of nutrients in some
aquaculture areas. Higher concentrations of nutri-
ents are also reported from areas close to sewage
outfalls. Mean prevalence of epitheliocystis was
greater in red morwong, Cheilodactylus fuscus
(Castelnau), and rock cale, Crinodus lophodon
(Günther), collected near Sydney, Australia sewage
outfalls than in fish of the same species collected
further from the outfalls, however, this difference
was not statistically significant (Nowak 1996).
Juvenile silver perch experimentally exposed to
metabolic ammonia (unionized ammonia –
0.02 mg L)1) had a lower percentage of filaments
affected by epitheliocystis than control fish. Epi-
theliocystis was observed in 75% of all experimental
fish including controls and fish exposed up to
0.36 mg L)1 unionized ammonia for 39 days
(Frances, Nowak & Allan 2000).
Season has been suggested to be a risk factor for
epitheliocystis. For Atlantic salmon in Tasmania,
month had a statistically significant effect on the
prevalence of epitheliocystis (Nowak & Clark
1999). Additionally, the prevalence of epitheliocys-
tis appeared greater during summer months, which
may be a result of the increased water temperature.
There was also a trend of a greater prevalence in the
second year of sampling, although it was not
statistically significant. Interestingly, the second
sampling year was warmer than the first, suggesting
that the prevalence of epitheliocystis in Atlantic
salmon in Tasmania was positively correlated with
temperature. Furthermore, all the Atlantic salmon
which had more than one cyst in the gill arch were
collected during summer, mostly in the second
(warmer) sampling season. However, the mean size
of the cyst was not affected by the temperature
difference (Nowak & Clark 1999).
In contrast, reports for other species indicated an
association of epitheliocystis with low temperature,
in particular the progression from a chronic to
proliferative form of the disease (Crespo et al. 1990;
Turnbull 1993). The difference is most probably
due to host-specific and pathogen-specific differ-
ences, as well as the fact that Atlantic salmon in
Tasmania are cultured at high temperatures relative
to Atlantic salmon cultured elsewhere. The lack of
statistically significant differences between farms
culturing salmon at different salinities (brackish
water – fully marine) suggests that salinity does not
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B F Nowak and S E LaPatra Epitheliocystis in fish
affect the prevalence of epitheliocystis in Atlantic
salmon in Tasmania (Nowak & Clark 1999).
Age-dependent morphology of epitheliocystis
cysts was reported in sea bream, with granular cysts
in 0.7–2 g fingerlings, granular and amorphous
cysts in juveniles and only amorphous cysts in
400 g bream (Crespo et al. 1999). It was suggested
that age of fish may play a role in switching between
different developmental cycles.
Prevalence of epitheliocystis has been suggested
to be positively correlated with age in wild winter
flounder, Pleuronectes americanus (Walbaum) (Mac-
Lean 1993). This is in contrast to the results for
Atlantic salmon in Tasmania (Nowak & Clark
1999). The prevalence of epitheliocystis did not
seem to be related to the age of the salmon, if
anything it decreased with the age of the fish. This
discrepancy may be either due to species-specific
differences or the fact that the winter flounder were
collected only on two occasions, in June and August
and the temperatures were greater in August when
the fish were older (MacLean 1993). In this case,
temperature was confounded with age and it is
impossible to separate between the effects of these
two factors. Thus, temperature and not age could
have influenced epitheliocystis prevalence in floun-
der.
As some human bacterial pathogens infect
amoebae and Chlamydia-like bacteria are endosym-
bionts of amoebae, gene sequences suggest some
similarity between one of the epitheliocystis agents
and Acanthoamoeba sp. endosymbiont (Draghi
et al. 2004), it has been suggested that amoebae
may be carriers of the epitheliocystis agent (Corsaro
& Venditti 2004; Draghi et al. 2004). However, it
is unlikely that Neoparamoeba sp., the causative
agent of amoebic gill disease (AGD) in Atlantic
salmon is involved. Epitheliocystis is rare in AGD-
affected fish and if it occurs it has a low prevalence
and very low intensity of infection (Nowak & Clark
1999). Furthermore, 16S ribosomal RNA gene
analysis used to assess the bacterial community of
AGD-affected Atlantic salmon gills did not detect
any chlamydial sequences (Bowman & Nowak
2004). While this does not preclude the role of
amoebae as carriers of epitheliocystis, it suggests
that neoparamoebae are not involved.
Co-infections
Paramyxovirus, in addition to the presence of
bacteria, has been associated with some cases of
 Journal of Fish Diseases 2006, 29, 573–588
epitheliocystis and associated mortalities (Hoffman
et al. 1969; Bradley et al. 1988; Nylund et al. 1998;
Kvellestad, Dannevig & Falk 2003; Fridell, Devold
& Nylund 2004; Kvellestad, Falk, Nygaard, Flesja
& Holm 2005). Atlantic salmon paramyxovirus
(ASPV) was present in the gills of some fish infected
by epitheliocystis (Kvellestad et al. 2003, 2005;
Fridell et al. 2004). However, challenge experi-
ments with the virus isolated from Atlantic salmon
with epitheliocystis did not result in any mortality
of experimental salmon (Fridell et al. 2004). Fur-
thermore, the association between ASPV and
epitheliocystis was not consistent (A. Kvellestad,
personal communication). Co-infection of epithe-
liocystis and lymphocystis was reported in barram-
undi (Meijer et al. 2006). Interestingly, some of the
lymphocystis cysts in the skin reacted weakly with
Chlamydiales 16S rRNA-specific oligoprobe and
antibodies against Chlamydiaceae LPS (Meijer et al.
2006), suggesting that the lymphocystis cysts
contained some bacteria. Mixed infection of differ-
ent Chlamydiaceae strains and a porcine epidemic
diarrhoea virus were reported in an in vitro study,
which showed that inclusions of all chlamydial
strains were larger in the dually infected cells than
in monoinfected ones (Stuedli, Grest, Schiller &
Pospischil 2005).
Virus-like particles and Trichodina sp. were
present in gills of Atlantic salmon affected by
epitheliocystis (Nylund et al. 1998). Epitheliocystis
and Trichodina sp. co-infection was reported in
cultured red sea bream (Syasina, Park & Kim
2004). Co-infection of epitheliocystis with Tricho-
dina sp. and Gyrodactylus sp. was described in
cultured pacu, Piaractus mesopotamicus (Holmberg)
(Szakolczai et al. 1999). Epitheliocystis has been
detected in gills of fish infected by other monogen-
eans, for example in amberjack infected with
Zeuxapta seriolae (Montero, Crespo, Padrós, De la
Gándara, Garcı́a & Raga 2004) and in kingfish
infected with the same monogenean (Mansell et al.
2005). An association was seen between the severity
of epitheliocystis and presence of gill monogeneans
(Bivagina pagrosomi and Lamellodiscus pagrosomi) in
snapper, Pagrus major (Temminck & Schlegel), and
when the fluke problem was overcome there was a
reduction in epitheliocystis (B. Diggles, personal
communication), suggesting that there was either a
link between these two infections or the same risk
factors were involved. Co-infections by the mono-
geneans Microcotyle sp., Lamellodiscus sp. and
epitheliocystis (Padros & Crespo 1995) and the
 2006
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B F Nowak and S E LaPatra Epitheliocystis in fish
monogeneans Microcotyle sp., Lamellodiscus ignora-
tus, the ciliate Trichodina sp., two species of Vibrio
and epitheliocystis (Cruz E Silva, Freitas & Orge
1997) were reported in cultured sea bream.
Further research
Our knowledge of epitheliocystis has greatly
increased in the last few years, mostly due to the
application of molecular techniques. However,
more information is required to fully understand
this condition and its significance to cultured and
wild fish. Development of real-time PCR for
epitheliocystis agents in Atlantic salmon will allow
epidemiological study for this host species. Addi-
tional sequence data from epitheliocystis agents are
required to determine the relationship between
bacteria from different hosts or even bacteria from
the same host, and also from different locations and
times of outbreaks, and to design effective diagnos-
tic and research tools. There is a need to develop
in vitro culture techniques and a challenge model
for this disease to assess its real significance for
aquaculture and fisheries.
Acknowledgements
We would like to thank Dr Duncan Colquhoun
for contributing ideas and information. We are
grateful to Dr Joseph Groff for his invaluable
study of the immunostaining of epitheliocystis
specimens. We would like to thank Drs R.
Jimenez, Silvia Crespo, Gary Marty, Alicia Noble,
Bev Dixon, Rod Getchell, Mr John Morrison,
Ms Beth MacConnell and Mr Terry Patterson for
supplying samples. We are grateful to Dr Brian
Jones, Dr Colin Johnston, Dr Agnar Kvellestad,
Mr Terje Steinum, Dr Peter Phillips and Dr Ben
Diggles for providing information on epitheliocys-
tis outbreaks and development of new diagnostic
methods. We thank Dr Andy Goodwin for
providing photographs of wet preparations and
histopathology of epitheliocystis.
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Received: 23 January 2006
Revision received: 1 June 2006
Accepted: 1 June 2006