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WHIRLING DISEASE OF SALMONID FISH: LIFE CYCLE, BIOLOGY, AND

DISEASE
Author(s): Michael A. Gilbert and Willard O. Granath Jr.
Source: Journal of Parasitology, 89(4):658-667. 2003.
Published By: American Society of Parasitologists
DOI: http://dx.doi.org/10.1645/GE-82R
URL: http://www.bioone.org/doi/full/10.1645/GE-82R

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J. Parasitol., 89(4), 2003, pp. 658667


q American Society of Parasitologists 2003

WHIRLING DISEASE OF SALMONID FISH: LIFE CYCLE, BIOLOGY, AND DISEASE


Michael A. Gilbert and Willard O. Granath, Jr.
Division of Biological Sciences, University of Montana, Missoula, Montana 59812. e-mail: snail@selway.umt.edu
ABSTRACT: Myxobolus cerebralis is the myxozoan parasite responsible for causing whirling disease in salmonid fish. Although
the parasite was first described nearly 100 yr ago, it received relatively little attention until the discovery of its 2-host life cycle
in the mid 1980s. This was the first, complete, myxozoan life cycle to be described, and it was greeted with some skepticism
because it united 2 stages of M. cerebralis that were previously classified in 2 separate taxa. In the last decade, there has been
a renewed interest in this parasite because whirling disease has been implicated in the decline of wild trout populations in several
western states in the United States. Subsequent research efforts have dramatically increased the understanding of the biology of
M. cerebralis and the numerous factors that affect the severity of whirling disease in salmonid hosts. These efforts also have
provided a great deal of new information concerning interactions between M. cerebralis and its aquatic oligochaete host Tubifex
tubifex. This review examines the current state of M. cerebralis in relation to 3 categories: the life cycle, the salmonid hosts, and
the oligochaete host.

from Europe (Hoffman, 1990). The hypothesis of this relatively


recent introduction is supported by sequence analysis of 18S
ribosomal genes of M. cerebralis from Europe and the United
States (Andree et al., 1999). Presently, whirling disease has
been reported from a total of 22 states and 26 different countries
(Hoffman, 1990; Bartholomew and Reno, 2002).
Unfortunately, knowledge of the biology of M. cerebralis has
not kept pace with its spread. Despite the fact that the parasite
was described 100-yr ago (Hofer, 1903), it was not until 1986
that researchers finally elucidated its 2-host life cycle (Wolf et
al., 1986). Before this time, the actinosporean and myxosporean
stages (Fig. 1) of the parasite were believed to be 2 separate
organisms, each having a direct, 1-host life cycle. Thus, when
a life cycle was proposed that united 2 organisms previously
classified as 2 separate taxa, it was greeted with some skepticism (Hamilton and Canning, 1987; Lom, 1987). The unified
life cycle was not accepted widely until the experiments of Wolf
and Markiw (1984) were repeated by other researchers (El-Matbouli and Hoffmann, 1989) and molecular studies were able to
provide additional evidence to unite the myxospore and actinospore stages of the parasite (Markiw, 1989; Andree et al.,
1997). Similar 2-host life cycles have since been confirmed for
other myxozoan species (El-Matbouli and Hoffmann, 1989,
1993; Burtle et al., 1991; Ruidisch et al., 1991; Yokoyama et
al., 1991, 1995a; El-Matbouli et al., 1992a; Kent et al., 1993;
Uspenskaya, 1995; Szekely et al., 2001). When the life cycle
of M. cerebralis was confirmed, an entire class within the phylum Myxozoa (class Actinosporea) was abolished. This created
numerous taxonomic problems within the phylum as attempts
were made to match actinosporeans to their corresponding
myxospore stage (Corliss, 1985; Kent et al., 1994). Although
the life-cycle debate was put to rest, the taxonomic situation
became further complicated by studies showing that myxozoans
are not protists. Oddly, myxozoans were classified traditionally
as protozoans despite being multicellular. However, in the past
several years, 18S ribosomal deoxyribonucleic acid (rDNA)
analysis confirmed the suspicions of many investigators that the
Myxozoa are, in fact, Metazoa, although their nearest metazoan
relative remains an issue of debate (Smothers et al., 1994; Siddall et al., 1995; Schlegel et al., 1996). Officially, the phylum
Myxozoa is still grouped with Protozoa, but the situation is
being reviewed, and it is likely that this classification will be
revised (Hedrick et al., 1998).

Salmonid whirling disease continues to be a significant health


problem in hatcheries and wild populations of trout. The myxozoan parasite Myxobolus cerebralis is the causative agent of
this disease, and it can induce a range of pathologies from a
whirling behavior in fish, for which the disease is named, to
blackened caudal regions and severe skeletal deformities.
Heavy infections in young fish often result in death. Rainbow
trout (Oncorhynchus mykiss) are most susceptible to disease,
although the parasite is able to infect numerous species of salmonid fishes, including sockeye salmon (O. nerka), golden trout
(O. aguabonita), cutthroat trout (O. clarki), brook trout (Salvelinus fontinalis), bull trout (S. confluentus), steelhead (O. mykiss), chinook salmon (O. tshawytscha), Atlantic salmon (Salmo
salar), and brown trout (S. trutta) (OGrodnick, 1979; Hoffman,
1990; Hedrick, McDowell, Gay et al., 1999; Hedrick, McDowell, Mukkatira et al., 1999).
Until the 1990s, M. cerebralis was primarily a problem in
hatchery settings. However, during the past decade, whirling
disease has been implicated in the decline of wild trout populations in several western states in the United States (Nehring
and Walker, 1996; Vincent, 1996; Hedrick et al., 1998). For
example, whirling disease was blamed for the 90% decrease in
wild rainbow trout inhabiting a 93-km stretch of the Madison
River in Montana that occurred between 1991 and 1995 (Rognlie and Knapp, 1998). Thus, the parasite has the potential to
cause severe economic damage in the United States, especially
in western states that depend on sport fishing to provide tourism
revenues, e.g., in its 1999 report, the Montana Whirling Disease
Task Force estimated that trout fishing generated US
$300,000,000 in recreational expenditures in Montana alone
(http://www.whirlingdisease.org). In addition, the ability of the
parasite to infect a wide variety of salmonid hosts makes it a
potential hazard to several threatened and endangered species,
e.g., bull trout, cutthroat trout, and steelhead.
Myxobolus cerebralis was first described in Germany (Hofer,
1903) after signs of disease were observed in rainbow trout
imported from North America. It is believed that M. cerebralis
evolved in Europe as a parasite of the brown trout (Hoffman,
1970), where infections are typically asymptomatic. Thus, it
was the introduction of the nonnative rainbow trout into Europe
that led to the discovery of the parasite. Myxobolus cerebralis
was first detected in the United States in 1958 at the Benner
Springs fish hatchery in Pennsylvania (Hoffman et al., 1962).
Many researchers believe that the parasite was introduced into
this country through the importation of frozen rainbow trout
658

GILBERT AND GRANATHWHIRLING DISEASE

659

FIGURE 1. Life cycle of Myxobolus cerebralis. The life cycle starts when myxospores of M. cerebralis are released from the cartilage of an
infected fish. A. Photograph of a juvenile rainbow trout infected with M. cerebralis. Bottom scale is in millimeters. B. Photomicrograph of a
histological section through the cartilage of a trout heavily infected with M. cerebralis showing numerous myxospores. Bar 5 20 mm. C.
Photomicrograph of a free myxospore of M. cerebralis. Bar 5 10 mm. For the life cycle to continue, these myxospores must be ingested by the
aquatic oligochaete Tubifex tubifex. D. Photograph of a single T. tubifex. Scale is in millimeters. Inside the lumen of the worms intestine the
myxospore will extrude its polar filaments and attach to the intestinal mucosa. The infective germ cell contained within the myxospore will then
migrate to the intercellular space of the intestinal epithelium, where it will undergo reproduction and development to the triactinomyxon (TAM)
stage during a 70- to 120-day period. Next, TAMs are released in the feces of T. tubifex and gain access to the water column, where they can
infect a fish host. E. Photomicrograph of a fecal packet of T. tubifex containing M. cerebralis TAMs. Some of the TAMs have escaped from the
packet. Bar 5 150 mm. F. Micrograph of a free M. cerebralis TAM. Bar 5 150 mm. Susceptible salmonids become infected when they come
into contact with the TAMs, which attach themselves to the fishs skin using their polar filaments. After attachment, the sporoplasm will actively
penetrate the fish host as an intact unit. After reproduction in the epidermis, the parasites migrate through the CNS to the associated cartilage,
where they mature into large plasmodia containing vegetative nuclei as well as generative cells that actively feed on the cartilage. Approximately
80 days after exposure to TAMs, the generative cells initiate sporogenesis, which will ultimately give rise to the myxospore stage, completing
the life cycle.

THE LIFE CYCLE


The Myxozoa contains approximately 1,350 species, all of
which are parasitic (Lom, 1987; Lom and Dykova, 1992). In
nearly all of the known myxozoan life cycles, the parasites alternate between a teleost fish host and an aquatic oligochaete
(Kent et al., 1994). During its life cycle, M. cerebralis parasites
alternate between its obligate, oligochaete host Tubifex tubifex
and any of the numerous species of salmonid hosts. The life
cycle (Fig. 1) starts when the myxospores of the parasite are
released from the cartilage of an infected fish (Fig. 1A, B). This
occurs when the fish dies and decomposes or when a predator
ingests the fish and subsequently releases the myxospores (Fig.
1B, C) in its feces. For the life cycle to continue, these myxospores must be ingested by T. tubifex (Figs. 1D, 2). Once inside

the lumen of the oligochaetes intestine, the spores extrude their


polar filaments and attach themselves to the intestinal mucosa.
The stimulus that triggers the extrusion of the polar filaments
is unknown. After attaching to the intestinal epithelium, the
binucleate, infective germ cell contained within the myxospore
migrates into the intercellular space, where it undergoes reproduction and development (El-Matbouli and Hoffmann, 1998).
The parasite can be detected as early as 24 hr after exposure to
myxospores, and the initial stages of replication can be detected
by in situ hybridization at 5 days postexposure (PE) (Antonio
et al., 1999). Although the parasite has long been known to
reproduce asexually in the intestine of T. tubifex, evidence suggests that the asexual, schizogenic phase is followed by a sexual
phase (gametogony), with each schizont resulting in the for-

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THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 4, AUGUST 2003

FIGURE 2. Photomicrographs of the intestine of Tubifex tubifex. A


B. Micrographs of intestines of T. tubifex that were actively shedding
triactinomyxons (TAMs) of Myxobolus cerebralis. Note the numerous
circular areas of discoloration that give the intestines a grainy appearance in the segments indicated by the arrows. CD. Micrographs of
intestines of uninfected T. tubifex. Note that these intestines lack the
grainy appearance seen in worms actively shedding TAMs. Bar 5 1
mm.

mation of 16 (8a and 8b) haploid gametocytes (El-Matbouli


and Hoffmann, 1998; El-Matbouli et al., 1998). The discovery
of sexual reproduction in T. tubifex contradicted the unsubstantiated dogma that the salmonid host was the definitive host of
M. cerebralis. The final stage of development in the worm (sporogony) is initiated by the fusion of a and b gametocytes to
form diploid zygotes. Each zygote, after subsequent divisions
and cell differentiation, gives rise to a single triactinomyxon
(TAM) stage (El-Matbouli and Hoffmann, 1998). The development of the parasite within T. tubifex to the TAM stage appears to be dependent on temperature (El-Matbouli, McDowell
et al., 1999). The TAM stage is released from the worm into
the water column, where it can infect the next fish host (Fig.
1E, F). It has been suggested that the TAMs enter the water
column either by egestion or after the death of the tubificids
(El-Matbouli et al., 1992b). However, recent evidence revealed
that TAMs are released in the feces of the worms (Gilbert and
Granath, 2001). TAMs have been detected in laboratory experiments anywhere from 74 to 120 days after the exposure of
worms to myxospores (Markiw, 1986; El-Matbouli and Hoffmann, 1998; El-Matbouli, McDowell et al., 1999; Gilbert and
Granath, 2001; Stevens et al., 2001). However, there is no apparent correlation between the number of myxospores available
to a worm and an optimum production of TAMs (Markiw,
1986; Stevens et al., 2001).
Susceptible salmonid fish become infected with M. cerebralis
when they come into contact with TAMs, which attach themselves to the fishs skin using their polar filaments. After attachment, a group of infective germ cells enclosed in an envelope cell (collectively referred to as a sporoplasm) actively
penetrate the fish host as an intact unit. Electron microscopy
has shown that penetration occurs through the secretory openings of mucous cells, most often on the epidermis (near the
fins), on the respiratory epithelium, and in the buccal cavity
(El-Matbouli, Hoffmann et al., 1999), and appears to be facilitated by serine and cystiene proteases released from the spo-

roplasm (Hedrick and El-Matbouli, 2002). Penetration of the


epidermis can be completed 1 min after exposure to TAMs (ElMatbouli, Hoffmann et al., 1999). Unlike development in T.
tubifex, both intercellular and intracellular stages of the parasite
occur in the salmonid host. In addition, reproduction in salmonids appears to be asexual (El-Matbouli et al., 1995), despite
earlier reports to the contrary (Lom, 1987). The first phase of
reproduction in the fish occurs in the epidermis, where intercellular and then intracellular stages of M. cerebralis were detected for up to 2 days after exposure of rainbow trout to TAMs
(El-Matbouli et al., 1995). After 2 days, parasites were no longer detected in the epidermis but were observed in the intercellular space of the subcutis, and all the parasites had migrated
to the nerve tissue (El-Matbouli et al., 1995) by 4 days PE.
Some developmental stages were seen in peripheral nerves of
the trout; however, the majority of the parasites were located in
the central nervous system (CNS) and in the meninges (ElMatbouli et al., 1995). The parasites then migrated through the
CNS to the associated cartilage where they first appeared at 20
days PE (El-Matbouli et al., 1995). The parasites then matured
into large plasmodia containing both vegetative nuclei and generative cells, which fed on the cartilage (El-Matbouli et al.,
1995). Approximately 80 days after exposure to M. cerebralis
TAMs, the fusion of generative cells initiated sporogenesis,
which ultimately gave rise to the myxospore stage of the parasite, completing the life cycle (El-Matbouli et al., 1995).
The spread of M. cerebralis has been attributed largely to
the resistance of the myxospore stage to harsh environmental
conditions. So, numerous studies have examined their resistance
to freezing, drying, and various disinfectants (Hoffman and
Putz, 1969, 1971; G. L. Hoffman and G. L. Hoffman Jr., 1972;
Hoffman and OGrodnick, 1977; Wolf and Markiw, 1982).
However, all these studies were done before the determination
of the parasites 2-host life cycle, so these data must be reinterpreted keeping the 2-host cycle in mind. Unfortunately, such
reevaluations are often inconclusive when considering the disinfection of myxospores. More recent studies have shown that
myxospores can survive freezing at 220 C for at least 3 mo as
well as passage through the alimentary tract of predators without losing infectivity (El-Matbouli and Hoffmann, 1991b).
In contrast to the myxospores, the waterborne TAM stage of
M. cerebralis is fragile and relatively short lived. The length of
time a TAM is infective to the fish host appears to be dependent
on temperature. One study reported that TAMs were infective
for 2 days at 23 C and 5 days at 7 C (Markiw, 1992b), whereas
another study reported that TAMs were infective for periods
longer than 15 days at temperatures up to 20 C (El-Matbouli,
McDowell et al., 1999). In light of these conflicting reports, it
seems likely that factors other than temperature are also involved, e.g., the fluid-filled processes of the TAMs may make
them extremely sensitive to changes in osmotic potential. The
length of time that TAMs are viable is an important aspect of
the epizootiology of whirling disease that warrants further investigation. For example, TAMs are neutrally buoyant and will
drift with the current of a river or stream. Therefore, the length
of time that TAMs remain viable will ultimately determine how
far they can travel from their infected T. tubifex host while
remaining infective for the salmonid host.
Another important epizootiological factor influencing the severity of whirling disease in a watershed is the number of M.

GILBERT AND GRANATHWHIRLING DISEASE

cerebralis TAMs necessary to establish an infection in salmonid fish. One major consideration is the age of the fish at the
time of exposure. For example, 2-day-old sac fry of rainbow
trout became heavily infected when exposed to 10 TAMs per
fish (Markiw, 1991), whereas no infection was established in
2-mo-old rainbow trout fry exposed to the same dosage (Markiw, 1992a). One hundred TAMs or more were required to infect
and produce disease in the 2-mo-old fry (Markiw, 1992a). Although M. cerebralis TAMs are able to infect trout as young
as 2 days old, the eggs of salmonid fish do not appear to be
susceptible to infection (Markiw, 1991). As with myxospores,
the stimulus that triggers extrusion of TAM polar filaments remains unknown. The latest hypothesis suggests that extrusion
results from a combination of mechanostimulations, such as the
swimming motions of a fish, and a chemoreceptor that is specific for a molecule on the body of the fish (El-Matbouli, Hoffmann et al., 1999). However, it is not known if these stimuli
are species specific. For example, the actinosporean stage of M.
cultus reacted nonspecifically with numerous species of fish
(Yokoyama et al., 1995b), whereas M. cerebralis TAMs were
reported to exhibit some degree of host specificity (El-Matbouli,
Hoffmann et al., 1999).
THE SALMONID HOSTS
Although nearly all the 1,350 known species of myxozoans
are parasites of fish at some point in their life cycle, the vast
majority of these species are relatively nonpathogenic (Lom,
1987; Lom and Dykova, 1992). In comparison, M. cerebralis
is 1 of the most pathogenic myxozoans known for fish (Hedrick
et al., 1998) and is a likely reason why whirling disease was
the first myxosporean disease described (Lom, 1987). It was
once thought that the whirling behavior exhibited by some infected fish was caused by a damaged auditoryvestibular apparatus, resulting in loss of equilibrium (Hoffman et al., 1962).
However, more recent studies have shown that damage to the
auditoryvestibular apparatus in fish results in an impaired ability to maintain an upright orientation of the body (Platt, 1983).
This translates to a corkscrew swimming motion in fish, not the
circular swimming associated with whirling disease (Rose et
al., 2000). The whirling behavior is more likely due to constrictions in areas of the spinal cord and lower brain stem caused
by an inflammatory response that is triggered by the parasite
feeding on nearby cartilage (Rose et al., 2000). Inflammation
in the posterior region of the spine may also cause damage to
the caudal nerves responsible for controlling pigment deposition resulting in the black tails that are sometimes observed in
this disease (Halliday, 1976). This hypothesis seems plausible
given the fact that host immune reactions are responsible for
clinical signs of disease caused by other myxozoan parasites,
such as Myxidium lieberkuhni and Sphaerospora renicola (Hedrick et al., 1998). Sphaerospora renicola, like M. cerebralis, is
especially lethal to young fish (Lom, 1987).
Myxobolus cerebralis parasites are able to infect numerous
species of salmonids (OGrodnick, 1979; Hoffman, 1990; Hedrick, McDowell, Mukkatira et al., 1999). However, not all fish
that are infected with M. cerebralis present clinical signs of
whirling disease. Among species that exhibit signs of disease,
rainbow trout suffer the worst pathology (OGrodnick, 1979;
Hedrick, McDowell, Mukkatira et al., 1999), although the se-

661

verity of disease in members of this species is dependent on


the size and age at the time of exposure, the strain of rainbow
trout infected, and the dose of TAMs the fish received (Halliday, 1973; Hoffman and Byrne, 1974; Markiw, 1991, 1992a;
Hedrick, McDowell, Mukkatira et al., 1999). One-year-old rainbow trout exposed to approximately 20,000 TAMs/fish produced 10 times as many myxospores as 3-yr-old fish exposed
to approximately 100,000 TAMs/fish. As a general rule, heavy
infections in young fish often cause severe clinical signs of
disease and usually results in death. This is in contrast to infections in older fish that have less severe symptoms and are
often asymptomatic. This is consistent with the presence of
more highly ossified skeletons and, therefore, less cartilage on
which the parasite can feed in older fish.
Other species of salmonids appear to be more resistant to
parasite infection. Infections in brown trout are usually asymptomatic. However, brown trout exposed to more than 1,000
TAMs/fish in the laboratory exhibited blackened caudal regions
(Hedrick, McDowell, Gay et al., 1999), and naturally infected
brown trout can exhibit this clinical sign (Nehring and Walker,
1996). Interestingly, there is no report of brown trout exhibiting
whirling behavior. It has been suggested that the resistance to
disease exhibited by brown trout is evidence that M. cerebralis
species coevolved with the brown trout in Europe (Hoffman,
1970). However, Hedrick et al. (1998) noted that this hypothesis fails to consider the high susceptibility of the European
Danube salmon (Hucho hucho) (El-Matbouli et al., 1992b), or
the resistance of coho salmon (OGrodnick, 1979), native to
western North America, to whirling disease. The mechanism
that conveys the varying levels of resistance observed in salmonid species remains unknown and it appears that different
species may limit the disease by different mechanisms. For example, bull trout are susceptible to infection, and the parasite
develops normally in the skin and nerves, but lesions in the
cartilage and parasite numbers are resolved eventually over
time (Hedrick, McDowell, Mukkatira et al., 1999). This species
rarely exhibits clinical signs of the disease. Arctic grayling
(Thymallus arcticus), another salmonid, appears to be completely refractory to infection. In laboratory exposures of arctic grayling to M. cerebralis TAMs, no developmental stage of the
parasite was observed in the nerves or skin at 35 days PE,
indicating that this species may be able to inhibit the development of the early stages of the parasite (Hedrick, McDowell,
Mukkatira et al., 1999). No strain of rainbow trout that is naturally resistant to infection with M. cerebralis has yet been
identified, although there are strains of rainbow trout that are
naturally resistant to Ceratomyxa shasta, another myxozoan
parasite of salmonid fish (Bartholomew, 1998). In the laboratory, C. shastaresistant strains of rainbow trout were highly
susceptible to M. cerebralis, suggesting different mechanisms
of resistance for the 2 parasites (Hedrick et al., 1998).
During the development of M. cerebralis in the epidermis of
rainbow trout, it appears as though some of the parasites are
killed (El-Matbouli et al., 1995), possibly by a humoral response in the fishs skin (Hedrick et al., 1998). In contrast, these
parasites are sheltered from host immune reactions while migrating through the CNS (El-Matbouli et al., 1995). In a histological study of infected rainbow trout, the first clear association between the parasite and host immune cells did not occur
until after the parasites had invaded and begun to phagocytize

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THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 4, AUGUST 2003

cartilaginous tissues at 20 days PE (El-Matbouli et al., 1995).


At this time, macrophages could be seen at the periphery of
lesions in the cartilage (El-Matbouli et al., 1995). Unlike rainbow trout, the lesions in brown trout are found more often in
the fin rays, ribs, and gill arches than in the cranium, and eosinophilic granular leukocytes are common in the cranial nerve
ganglia and nerve roots of brown trout but not of rainbow trout
(Hedrick, McDowell, Gay et al., 1999).
The interactions between M. cerebralis and cells of the salmonid immune system have not been studied in detail. Recently, Shum et al. (2001) examined differences in the major
histocompatibility complex class I and class II genes from rainbow and brown trout in hopes of identifying alleles that may
confer resistance to M. cerebralis. Thus far, these researchers
have isolated a total of 15 class I and 14 class II alleles from
rainbow trout as well as 10 class I and 12 class II alleles from
brown trout. They also have found that only a single class I
allele and no class II allele was shared between these fish species (Shum et al., 2001).
Fumagillin is the only drug that has been studied for the
prevention or treatment of whirling disease in salmonids (ElMatbouli and Hoffmann, 1991a; Molnar, 1993; Staton et al.,
2002). This antibiotic was first used to treat Nosema apis, a
microsporean parasite causing Nosema disease in honey bees
(Katznelson and Jamieson, 1952). Later, it was shown to be
effective in the treatment of 2 myxozoan-related diseases, renal
sphaerosporosis in carp (Molnar et al., 1987) and proliferative
kidney disease in salmonids (Hedrick et al., 1988). In 1991, ElMatbouli and Hoffmann (1991a) showed that orally administered fumagillin reduced both the prevalence and severity of M.
cerebralis infection in rainbow trout that had been exposed to
TAMs in a laboratory setting. However, in a recent study following a pivotal efficacy trial protocol, i.e., protocols that
meet the U.S. Food and Drug Administration requirements for
new animal drug approval, Staton et al. (2002) found that fumagillin was not efficacious in the prevention or control of
whirling disease. Thus, there is currently no known effective
drug for the control of whirling disease in salmonid fish.
THE OLIGOCHAETE HOST
In comparison with the interactions between M. cerebralis
and its salmonid hosts, very little is known about the interactions between the parasite and its oligochaete host T. tubifex.
Tubifex tubifex was first described in 1774; yet, debate continues over the identification and taxonomy of this species and the
possibility of subspecies (reviewed by Brinkhurst, 1986). Much
of this debate stems from the fact that positive identification of
T. tubifex is based on external morphology, e.g., chaetae, and
morphology of the reproductive structures, e.g., penis sheath.
Further complicating identification of these worms is that after
breeding they resorb their reproductive organs (Poddubnaya,
1984), and chaetae of T. tubifex can change forms depending
on environmental conditions, such as salinity (Chapman and
Brinkhurst, 1987). The results of recent molecular studies (Anlauf, 1994, 1997; Anlauf and Neumann, 1997; Sturmbauer et
al., 1999; Beauchamp et al., 2001, 2002) suggest the existence
of cryptic species of Tubifex that vary in a variety of physiological characteristics, including their susceptibility to M. cerebralis infection.

Tubifex tubifex is the only species of aquatic oligochaete


known to be a suitable host for M. cerebralis (Gilbert and Granath, 2002). For example, Limnodrilus hoffmeisteri, Ilyodrilus
templetoni, Quistadrilus multisetosus (Wolf et al., 1986), Dero
spp., Stylaria spp., Aeolomsoma spp. (Markiw and Wolf, 1983),
and Tubifex ignotus (El-Matbouli and Hoffman, 1989) are
known to be unsuitable hosts for M. cerebralis. In addition, our
laboratory, as well as several others, have tested additional species of aquatic oligochaetes and found that only T. tubifex
worms are able to support the development and transmission of
M. cerebralis. Although no worldwide survey for T. tubifex has
been conducted, it is widely believed that these worms are cosmopolitan, and the distribution of M. cerebralis over several
continents (Hoffman, 1970, 1990; Bartholomew and Reno,
2002) supports this view.
Tubifex tubifex worms inhabit sediments at the bottom of
lakes and streams, where they feed by ingesting whole sediments and selectively digesting some of the bacterial species
present (Wavre and Brinkhurst, 1971). The worms play an important role in the ecosystem by reworking sediments and providing a valuable food source for leeches, crustaceans, fishes,
and numerous species of insects. Unfortunately, T. tubifex species are probably best known for their ability to survive eutrophic conditions and have thus earned the nickname sludge
worms. They are quite capable of surviving in pristine conditions, but in most cases the numbers of T. tubifex are kept
low by competition, predation, or both. Therefore, under such
conditions, they are difficult to detect without extensive sampling efforts.
Within a watershed, Tubifex distribution is influenced by the
composition and organic content of the substratum, although
this relationship is indirect. That is, the microflora associated
with different substrata is the primary factor in substratum selection by the worms and not the substratum itself (McMurtry
et al., 1983; Lazim and Learner, 1987; Lestochova, 1994). Tubifex tubifex abundance also is positively correlated with both
sedimentation rates and organic carbon flux (Robbins et al.,
1989). These worms are often found in close association with
other oligochaete species such as L. hoffmeisteri. These characteristic species associations are believed to be caused by mutualistic relationships, in which 1 species feeds on the bacteria
associated with the fecal pellets of the other species and vice
versa (Brinkhurst, 1971; Brinkhurst, 1974; Milbrink, 1993).
In addition to feeding on bacteria, T. tubifex worms are able
to absorb small, dissolved, organic molecules directly through
their body wall. In areas with large concentrations of dissolved
organic materials, it is estimated that the worms can obtain up
to 40% of their oxidative requirements by absorbing molecules
directly from the water (Hoffmann et al., 1987). Tubifex tubifex
worms are capable of anaerobic respiration and have survived
16 wk in the laboratory under anoxic conditions (Reynoldson,
1987). The end products of glucose breakdown in the absence
of oxygen are acetate and propionate, and these products can
be reabsorbed through the body wall on return to aerobic conditions (Hoffmann et al., 1987). This adaptation is 1 of the
reasons why T. tubifex worms are able to survive in eutrophic
environments, which often result from large amounts of organic
pollution. In extreme cases, T. tubifex and L. hoffmeisteri may
be the only benthic macroinvertebrates present in such environments (Brinkhurst, 1996). Tubifex tubifex also have a unique

GILBERT AND GRANATHWHIRLING DISEASE

ability to survive drought and food shortages by secreting a


protective cyst and lowering its metabolic rate. Cysts containing
live T. tubifex have been recovered from a cattle pond after 5
mo of drought, and worms in the laboratory survived 6 mo of
starvation in cysts (Anlauf, 1990). Food shortage, which often
occurs as a result of drought, is believed to be the main factor
in cyst formation because the addition of food is the only means
by which worms exit the cysts (Anlauf, 1990). Some researchers have speculated that T. tubifex can be dispersed to new
locations in these cysts. If worms infected with M. cerebralis
could use this type of dispersal mechanism, it could have significant impacts on efforts to control the spread of whirling
disease.
Like other oligochaetes, T. tubifex is hermaphroditic. They
usually breed sexually but have the ability to reproduce asexually by parthenogenesis (Poddubnaya, 1984). Fertilized eggs
are deposited in a cocoon, and young worms hatch directly from
the cocoon 726 days later, depending on the temperature. The
newly hatched worms will become sexually mature within 13
mo, and after breeding, the worms can undergo several cycles
of resorption and regeneration of their sexual organs (Poddubnaya, 1984). Reports concerning the lifespan of T. tubifex are
highly variable. Poddubnaya (1984) reported that the lifespan
of reproducing parthenogenetic individuals varies from 70 to
530 days. Matsumoto and Jammoto (1966) indicated that T.
hattai (a closely related species) lives from 4 to 6 yr. Tubifex
tubifex specimens, in our laboratory, have been kept alive for
more than 3 yr, although their age at the time of acquisition
was unknown. Although the exact numbers vary, it is clear that
these worms live long enough to span several seasons.
Individual T. tubifex can remain persistently infected with M.
cerebralis throughout their lifespan. Gilbert and Granath (2001)
determined the periodicity and length of the release of the TAM
stage of M. cerebralis by T. tubifex. In this study, individual T.
tubifex worms were examined daily for the release of M. cerebralis TAMs, and the number of waterborne TAMs released
by each worm was quantified. The duration of the infection in
these worms was also monitored using a polymerase chain reaction (PCR) diagnostic test (Andree et al., 1998). TAMs were
first released 74 days PE and continued to be released until 132
days PE. During this period, each worm released, on average,
1.5 3 103 waterborne TAMs 12 times; however, no pattern or
periodicity was noted. The results of the PCR diagnostic tests
conducted at 5, 7, 9, and 15 mo PE were positive, and the
persistent infection was confirmed at 606 days PE, when the
remaining worms began releasing TAMs again. Similar results
were observed in naturally infected T. tubifex, indicating that
these worms remain infected for the duration of their normal
lifespan and are capable of shedding viable TAMs in temporally
separate periods.
El-Matbouli, McDowell et al. (1999) examined the effect of
water temperature on the development, release, and survival of
TAMs in T. tubifex. They examined infected worms held at 5,
10, 15, 20, 25, and 30 C and found that the highest level of
TAM production occurred at 10 and 15 C; at higher or lower
temperatures, TAM release was minimal. This finding probably
explains why researchers detect the most severe cases of whirling disease in trout in the 1015 C temperature range. For example, Baldwin et al. (2000), using sentinel cage methods, observed a direct correlation between both the prevalence and the

663

severity of infection of M. cerebralis in rainbow trout with temperature. The highest prevalence of infection and the most severe lesions were found in trout that were held in an enzootic
area when water temperatures were between 10 and 12 C. Thus,
if young trout, which are the most susceptible to the disease,
are emerging from their redds at these temperatures, it could
severely affect the fish population. Additional studies conducted
by the Montana Department of Fish, Wildlife and Parks
(MDFWP) also show a correlation between water temperature
and infection rates in sentinel fish. MDFWP observed that peak
infection rates occurred in fish during the spring warming and
fall cooling periods, when average daily water temperatures
were between 11 and 14 C (E. R. Vincent, pers. comm.). This
information, coupled with the fact that T. tubifex worms can
remain persistently infected with the parasite, release TAMs
more than once during its life, and live for several years (Gilbert and Granath, 2001), presents the possibility of a seasonal
periodicity in TAM release. A seasonal periodicity would explain the peaks in infection rates of sentinel fish seen by
MDFWP and the correlation between water temperature and
severity of M. cerebralis infections in rainbow trout (Baldwin
et al., 2000).
There is a lack of publications directly addressing pathology
in T. tubifex caused by M. cerebralis. However, alterations in
the coloration as well as distortions of the intestine can be observed in infected T. tubifex that are actively shedding TAMs
(Fig. 2). These tissue changes are likely caused by large clusters
of M. cerebralis cells and a resulting hypotrophy of intestinal
epithelial cells that is likely to decrease the absorptive surface
of the intestine (El-Matbouli and Hoffman, 1998; Hedrick and
El-Matbouli, 2002).
Recent data also indicate that the parasite may differentially
affect worm populations geographically. Stevens et al. (2001)
examined the effects of various doses of M. cerebralis myxospores on TAM production and the biology of T. tubifex from
2 different regions where whirling disease is enzootic (California and Montana). They exposed these worms to 50, 500, or
1,000 myxospores and determined TAM production and the effect of the parasite on biomass, abundance, and individual
weight of the oligochaetes. Results indicated that the California
worms produced significantly more TAMs than the Montana T.
tubifex, but within each population, TAM production did not
vary with spore dose. Furthermore, when compared with uninfected controls, total worm biomass, abundance, and individual weight were decreased in both geographic populations.
Thus, this study shows that T. tubifex populations vary in their
ability to produce TAMs and that the parasite appears to have
a deleterious effect on both worm populations. Although no
developmental stage of M. cerebralis was seen in the gonads
of worms during histological studies of infected T. tubifex (ElMatbouli and Hoffmann, 1998), the parasite may have an indirect effect on worm reproduction. Whether the effects on
worm physiology are a result of parasitic castration remains to
be determined.
The wide distribution of T. tubifex and the ability of this
organism to occupy a variety of habitat types led some researchers to suggest that this species may consist of different
ecological races (Milbrink, 1973; Poddubnaya, 1980) or even
cryptic species. To begin addressing such ecological questions,
Anlauf (1994, 1997) used horizontal starch gel electrophoresis

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THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 4, AUGUST 2003

to examine polymorphisms of 2 different enzymes from T. tubifex, i.e., isocitrate dehydrogenase and phosphoglucose isomerase. This study concluded that T. tubifex populations from
Germany consisted primarily of 3 dominant genotypes and that
these genotypes differed in their adult weights, reproduction,
and relative mortality in response to temperature shifts. Anlauf
and Neumann (1997) also used these techniques to address the
question of possible genetic variability of T. tubifex in relation
to habitat type. For this study, they examined the polymorphisms of 6 different enzymes from 20 different populations of
worms that were collected from 4 habitat types. These habitats
were classified as type I, lakes without an anoxic period in the
hypolimnion; type II, lakes with an extended anoxic period in
the hypolimnion; type III, creeks with a permanent flow of water; and type IV, pools without a permanent flow of water and
seasonal drying or freezing. Their results indicated a good correlation between habitat type and the frequency of some alleles.
Some alleles were observed mainly from shallow-water habitats
(types III and IV), whereas others were observed mainly from
deep-water habitats (types I and II). In addition, other individuals were found at almost every shallow-water site tested but
were only found at 2 deep-water sites. Most of the alleles that
were restricted to shallow-water habitats were seen in these individuals as well. Although the results of these studies support
the concept of different ecological races as postulated by Milbrink (1973) and Poddubnaya (1980), they must be interpreted
cautiously. For example, Kinne (1962) looked at the effects of
osmotic shock and rearing temperature on developmental stages
of fish. This study found that the variation seen in some physiological characters becomes fixed during the course of ontogeny under different environmental conditions. Therefore, if only
individuals that are adults when collected are tested using protein-based molecular markers, irreversible, nongenetic adaptations might be interpreted as ecological or geographical
variation (which remain stable in the laboratory) but are, in
reality, manifestations of the environment during ontogeny. Although the effects of adaptation versus ecological or geographical variation on the development of T. tubifex has not been
examined, until more is known about the developmental biology of tubificid worms, it should be noted that Anlauf and Neumann (1997) used only mature T. tubifex.
Avoiding the problems associated with protein-based molecular markers, Sturmbauer et al. (1999) used a segment of the
mitochondrial 16S rDNA to examine the phylogeny of T. tubifex collected from locations throughout Europe. This study
found that T. tubifex species consisted of 5 morphologically
indistinguishable lineages (designated I through V) that differed
significantly in their resistance to cadmium. In addition, the
genetic distances observed between some of these lineages
were large enough to suggest the existence of at least 2 cryptic
species. Beauchamp et al. (2001) used this technique to examine the phylogeny of T. tubifex and other oligochaetes collected from a variety of locations in both North America and
Europe. The results of this study were similar to those seen by
Sturmbauer et al. (1999). For example, the T. tubifex species
analyzed by Beauchamp et al. (2001) formed several distinct
lineages, and the genetic distances between some of these lineages were similar to those for well defined species within Limnodrilus, suggesting the existence of cryptic species within the
Tubifex. Interestingly, 2 of the European lineages observed by

Sturmbauer et al. (1999), lineages II and IV, were absent in the


samples analyzed by Beauchamp et al. (2001). Furthermore, 1
North American lineage observed by Beauchamp et al. (2001),
lineage VI, was absent from the European samples analyzed by
Sturmbauer et al. (1999). These results are the first to suggest
distinct genetic differences among T. tubifex populations from
North America and Europe. Beauchamp et al. (2002) then proceeded to examine the genetic differences and the susceptibility
to M. cerebralis infection of 2 different populations of T. tubifex collected from the Upper Colorado River. Samples were
collected from Windy Gap Reservoir, a locality where wild
rainbow trout exhibit high levels of M. cerebralis infection, and
Breeze Bridge, a downstream location where infections in wild
trout are much less severe. The results of their study revealed
that lineages I and III, which contain the most oligochaetes
susceptible to infection with M. cerebralis, were the most prominent lineages at Windy Gap Reservoir (Beauchamp et al.,
2002). In addition, their study found that lineages V and VI,
which are refractory to M. cerebralis infections, were more
prominent at Breeze Bridge, the location where infections in
wild trout are much less severe. These results suggest that genetic differences among populations of T. tubifex may influence
the severity of whirling disease in trout inhabiting the same
region. This is epizootiologically interesting for several reasons.
First, habitat conditions at Windy Gap Reservoir and Breeze
Bridge are quite different from each other. This fact combined
with the results observed by Anlauf and Neumann (1997) suggests that environmental factors present in different habitat
types may select for different genotypes of T. tubifex. Therefore, if genetic differences among populations of T. tubifex do
indeed influence the severity of whirling disease in trout, then
habitat conditions may ultimately be responsible for the severity
of whirling disease in a given region. For example, Anlauf and
Neumann (1997) found that shallow-water populations exhibited a lower level of heterozygosis than expected, based on
HardyWeinberg calculations, which would result from a high
degree of inbreeding or parthenogenic reproduction. Thus, if
habitat conditions were to significantly lower a population and
select for an ecological race or genotype of T. tubifex that was
susceptible to infection, or if a susceptible genotype were to
colonize a new watershed, rapid reproduction by parthenogenesis and subsequent inbreeding would create a large population
of susceptible worms. This, in turn, could exacerbate an existing whirling disease problem or put a watershed at greater risk
of a whirling disease epizootic. Conversely, if a nonsusceptible
race were selected for, or if it colonized a new area, then T.
tubifex populations could increase with little or no effect on the
transmission of M. cerebralis. Therefore, if the environmental
factors responsible for this selection were to be identified, then
habitat restoration or manipulation may be an effective means
to control whirling disease in many situations.
SUMMARY
Whirling disease continues to be a significant health problem
in some hatcheries and many wild populations of trout. In addition, the ability of M. cerebralis to infect a wide variety of
salmonid hosts makes this parasite a potential threat to several
threatened and endangered fish species. Significant work remains to be done, in a wide variety of fields, before the epi-

GILBERT AND GRANATHWHIRLING DISEASE

zootiology of this disease is understood well enough to manage


threatened populations of salmonids. However, there are numerous field and laboratory studies currently underway that are
providing the necessary information to better manage fish populations. If the scientific advances seen in recent years are any
indication of what is to come, fish populations currently suffering from whirling disease have a promising future.
ACKNOWLEDGMENTS
The authors thank the Whirling Disease Foundation (Bozeman, Montana; http://www.whirling-disease.org) for their generous financial support used in the preparation of the manuscript. We also gratefully thank
Ms. Elizabeth MacConnell (U.S. Fish and Wildlife Service, Bozeman,
Montana) for providing the photographs of the fish and myxospores
used in Figure 1A, B and the National Partnership on the Management
of Wild and Native Coldwater Fisheries, who supported some of the
work.

LITERATURE CITED
ANDREE, K. B., M. EL-MATBOULI, R. W. HOFFMANN, AND R. P. HEDRICK.
1999. Comparison of 18S and ITS-1 rDNA sequences of selected
geographic isolates of Myxobolus cerebralis. International Journal
for Parasitology 29: 771775.
, S. J. GRESOVIAC, AND R. P. HEDRICK. 1997. Small subunit RNA
sequences unite alternate actinosporean and myxosporean stages of
Myxobolus cerebralis the causative agent of whirling disease in
salmonid fish. Journal of Eukaryotic Microbiology 44: 208215.
, E. MACCONNELL, AND R. P. HEDRICK. 1998. A nested polymerase chain reaction for the detection of genomic DNA of Myxobolus
cerebralis in rainbow trout (Oncorhynchus mykiss). Diseases of
Aquatic Organisms 34: 145154.
ANLAUF, A. 1990. Cyst formation in Tubifex tubifex (Muller)An adaptation to survive food deficiency and drought. Hydrobiologia
190: 7982.
. 1994. Some characteristics of genetic variants of Tubifex tubifex (Muller 1774) (Oligochaeta: Tubificidae) in laboratory cultures. Hydrobiologia 278: 16.
. 1997. Enzyme variability of Tubifex tubifex (Muller) (Oligochaeta, Tubificidae) and seven other tubificid species. Archiv fur
Hydrobiologie 139: 83100.
, AND D. NEUMANN. 1997. The genetic variability of Tubifex
tubifex (Muller) in 20 populations and its relation to habitat type.
Archiv fur Hydrobiologie 139: 145162.
ANTONIO, D. B., M. EL-MATBOULI, AND R. P. HEDRICK. 1999. Detection
of early developmental stages of Myxobolus cerebralis in fish and
tubificid oligochaete hosts by in situ hybridization. Parasitology
Research 85: 942944.
BALDWIN, T. J., E. R. VINCENT, R. M. SILFLOW, AND D. STANEK. 2000.
Myxobolus cerebralis infection in rainbow trout (Oncorhynchus
mykiss) and brown trout (Salmo trutta) exposed under natural
stream conditions. Journal of Veterinary Diagnostic Investigation
12: 312321.
BARTHOLOMEW, J. L. 1998. Host resistance to infection by the myxosporean parasite Ceratomyxa shasta: A review. Journal of Aquatic
Animal Health 10: 112120.
, AND P. RENO. 2002. The history and dissemination of whirling
disease. In Whirling disease: Reviews and current topics, J. L. Bartholomew and J. C. Wilson (eds.). American Fisheries Society
Symposium 29, Bethesda, Maryland, p. 324.
BEAUCHAMP, K. A., M. GAY, G. O. KELLEY, M. EL-MATBOULI, R. D.
KATHMAN, R. B. NEHRING, AND R. P. HEDRICK. 2002. Prevalence
and susceptibility of infection to Myxobolus cerebralis, and genetic
differences among populations of Tubifex tubifex. Diseases of
Aquatic Organisms 51: 113121.
, R. D. KATHMAN, T. S. MCDOWELL, AND R. P. HEDRICK. 2001.
Molecular phylogeny of tubificid oligochaetes with special emphasis on Tubifex tubifex (Tubificidae). Molecular Phylogenetics and
Evolution 19: 216224.
BRINKHURST, R. O. 1971. Interspecific interactions and selective feeding

665

by tubificid oligochaetes. Limnology and Oceanography 17: 122


133.
. 1974. Factors mediating interspecific aggregation of tubificid
oligochaetes. Journal of the Fisheries Research Board of Canada
31: 460462.
. 1986. Guide to the freshwater aquatic microdrile oligochaetes
of North America, publication number 84. Department of Fisheries
and Oceans, Scientific Information and Publications Branch, Ottawa, Canada, 259 p.
. 1996. On the role of tubificid oligochaetes in relation to fish
disease with special reference to the Myxozoa. Annual Review of
Fish Diseases 6: 2940.
BURTLE, G. J., L. R. HARRISON, AND E. L. STYER. 1991. Detection of a
triactinomyxid Myxozoan in an oligochaete from ponds with proliferative gill disease of channel catfish. Journal of Aquatic Animal
Health 3: 281287.
CHAPMAN, P. M., AND R. O. BRINKHURST. 1987. Hair today gone tomorrow: Induced chaetal changes in tubificid oligochaetes. Hydrobiologia 155: 4555.
CORLISS, J. O. 1985. Consideration of taxonomic-nomenclatural problems posed by report of myxosporidians with a two-host life cycle.
Journal of Protozoology 32: 589591.
EL-MATBOULI, M., T. FISCHER-SCHERL, AND R. W. HOFFMANN. 1992a.
Transmission of Hoferellus carassi (Achmerov 1960) to goldfish
via an aquatic oligochaete. Bulletin of the European Association of
Fish Pathology 12: 5456.
, , AND . 1992b. Present knowledge on the life
cycle, taxonomy, pathology and therapy of some Myxosporea spp.
important for freshwater fish. Annual Review of Fish Diseases 3:
367402.
, AND R. W. HOFFMANN. 1989. Experimental transmission of two
Myxobolus spp. developing bisporogeny via tubificid worms. Parasitology Research 75: 461464.
, AND . 1991a. Prevention of experimentally induced
whirling disease in rainbow trout (Oncorhynchus mykiss) by fumagillin. Diseases of Aquatic Organisms 10: 109113.
, AND . 1991b. Effects of freezing, aging and passage
through the alimentary canal of predatory animals on the viability
of Myxobolus cerebralis spores. Journal of Aquatic Animal Health
3: 260262.
, AND . 1993. Myxobolus carassii (Klokaceva, 1914) also
requires an aquatic oligochaete, Tubifex tubifex, as an intermediate
host in its life cycle. Bulletin of the European Association of Fish
Pathology 13: 189192.
, AND . 1998. Light and electron microscopic studies on
the chronological development of Myxobolus cerebralis to the actinosporean stage in Tubifex tubifex. International Journal for Parasitology 28: 195217.
, , AND C. MANDOK. 1995. Light and electron microscopic
observations on the route of the triactinomyxon-sporoplasm of Myxobolus cerebralis from epidermis to rainbow trout cartilage. Journal of Fish Biology 46: 919935.
, , H. SCHOEL, T. S. MCDOWELL, AND R. P. HEDRICK. 1999.
Whirling disease: Host specificity and interactions between the actinosporean stage of Myxobolus cerebralis and rainbow trout (Oncorhynchus mykiss). Diseases of Aquatic Organisms 35: 112.
, T. W. HOLSTEIN, AND R. W. HOFFMANN. 1998. Determination of
nuclear DNA concentration in cells of Myxobolus cerebralis and
triactinomyxon spores, the causative agent of whirling disease. Parasitology Research 84: 694699.
, T. S. MCDOWELL, D. B. ANTONIO, K. B. ANDREE, AND R. P.
HEDRICK. 1999. Effect of water temperature on the development,
release and survival of the triactinomyxon stage of Myxobolus cerebralis in its oligochaete host. International Journal for Parasitology
29: 627641.
GILBERT, M. A., AND W. O. GRANATH JR. 2001. Persistent infection of
Myxobolus cerebralis, the causative agent of salmonid whirling disease, in Tubifex tubifex. Journal of Parasitology 87: 101107.
, AND . 2002. The role Tubifex tubifex (Annelida: Oligochaeta: Tubificidae) in the transmission of Myxobolus cerebralis
(Myxozoa: Myxosporea: Myxobolidae). In Whirling disease: Reviews and current topics, J. L. Bartholomew and J. C. Wilson

666

THE JOURNAL OF PARASITOLOGY, VOL. 89, NO. 4, AUGUST 2003

(eds.). American Fisheries Society Symposium 29, Bethesda,


Maryland, p. 7985.
HALLIDAY, M. M. 1973. Studies on Myxosoma cerebralis; a parasite of
salmonids II. The development and pathology of Myxosoma cerebralis in experimentally infected rainbow trout (Salmo gairdneri)
fry reared at different water temperatures. Nordic Veterinary Medicine 25: 349358.
. 1976. The biology of Myxosoma cerebralis, the causative organism of whirling disease of salmonids. Journal of Fish Biology
9: 339357.
HAMILTON, A. J., AND E. U. CANNING. 1987. Studies on the proposed
role of Tubifex tubifex (Muller) as an intermediate host in the life
cycle of Myxosoma cerebralis. Journal of Fish Diseases 10: 145
151.
HEDRICK, R. P., AND M. EL-MATBOULI. 2002. Recent advances with taxonomy, life cycle and development of Myxobolus cerebralis in the
fish and oligochaete hosts. In Whirling disease: Reviews and current topics, J. L. Bartholomew and J. C. Wilson (eds.). American
Fisheries Society Symposium 29, Bethesda, Maryland, p. 4553.
, , M. A. ADKINSON, AND E. MACCONNELL. 1998. Whirling disease: Re-emergence among wild trout. Immunological Reviews 166: 365376.
, J. M. GROFF, P. FOLEY, AND T. S. MCDOWELL. 1988. Oral administration of fumagillin DHC protects chinook salmon (Oncorhynchus tshawytscha) from experimentally induced proliferative
kidney disease. Diseases of Aquatic Organisms 4: 165168.
, T. S. MCDOWELL, M. GAY, G. D. MARTY, M. P. GEORGIADIS,
AND E. MACCONNELL. 1999. Comparative susceptibility of rainbow
trout Oncorhynchus mykiss and brown trout Salmo trutta to Myxobolus cerebralis, the cause of salmonid whirling disease. Diseases of Aquatic Organisms 37: 173183.
, , K. MUKKATIRA, M. P. GEORGIADIS, AND E. MACCONNELL. 1999. Susceptibility of selected inland salmonids to infections with Myxobolus cerebralis, the causative agent of whirling
disease. Journal of Aquatic Animal Health 11: 330339.
HOFER, B. 1903. Ueber die drehkrankheit der regenbogenforelle. Allgemeine Fischerei Zeitschrift 28: 78.
HOFFMAN, G. L. 1970. Intercontinental and transcontinental dissemination and transfaunation of fish parasites with emphasis on whirling
disease (Myxobolus cerebralis) and its effects on fish. In Symposium on disease of fishes and shellfishes, S. F. Snieszko (ed.). Special publication 5. American Fisheries Society, Bethesda, Maryland, p. 6981.
. 1990. Myxobolus cerebralis, a worldwide cause of salmonid
whirling disease. Journal of Aquatic Animal Health 2: 3037.
, AND C. J. BYRNE. 1974. Fish age as related to susceptibility to
Myxosoma cerebralis, cause of whirling disease. Progressive Fish
Culturalist 36: 151.
, C. E. DUNBAR, AND A. BRADFORD. 1962. Whirling disease of
trout caused by Myxosoma cerebralis in the United States. U.S.
Fish and Wildlife Service Special Scientific Report Fisheries 427:
114.
, AND G. L. HOFFMAN JR. 1972. Studies on the control of whirling
disease (Myxosoma cerebralis): The effects of chemicals on spores
in vitro, and of calcium-oxide as a disinfectant in simulated ponds.
Journal of Wildlife Diseases 8: 4953.
, AND J. J. OGRODNICK. 1977. Control of whirling disease (Myxosoma cerebralis) effects of drying and disinfection with hydrated
lime or chlorine. Journal of Fish Biology 10: 175179.
, AND R. E. PUTZ. 1969. Host susceptibility and the effect of
aging, freezing, heat and chemicals on the spores of Myxosoma
cerebralis. Progressive Fish Culturalist 31: 3537.
, AND . 1971. Effect of freezing and aging on the spores
of Myxosoma cerebralis, the causative agent of salmonid whirling
disease. Progressive Fish Culturalist 33: 9598.
HOFFMANN, K. H., E. HIPP, AND U. SEDLMEIER. 1987. Aerobic and anaerobic metabolism of the freshwater oligochaete Tubifex sp. Hydrobiologia 155: 157158.
KATZNELSON, H., AND C. A. JAIMIESON. 1952. Control of Nosema of
honey bees with fumagillin. Science 115: 7071.
KENT, M. L., L. MARGOLIS, AND J. O. CORLISS. 1994. The demise of a
class of protists: Taxonomic and nomenclatural revisions proposed

for the protist phylum Myxozoa Grasse, 1970. Canadian Journal of


Zoology 72: 932937.
, D. J. WHITAKER, AND L. MARGOLIS. 1993. Transmission of Myxobolus arcticus (Pugachev and Khokholov, 1979), a myxosporean
parasite of pacific salmon, via a triactinomyxon from the aquatic
oligochaete Stylodrilus heringianus (Lumbriculidae). Canadian
Journal of Zoology 71: 12071211.
KINNE, O. 1962. Irreversible non-genetic adaptation. Comparative Biochemistry and Physiology 5: 265282.
LAZIM, M. N., AND M. A. LEARNER. 1987. The influence of sediment
composition and leaf litter on the distribution of tubificid worms
(Oligochaeta). Oecologia 72: 131136.
LESTOCHOVA, E. I. 1994. Influence of small river conditions on the abundance of tubificidae. Hydrobiologia 278: 129131.
LOM, J. 1987. Myxosporea: A new look at long-known parasites of fish.
Parasitology Today 3: 327332.
, AND I. DYKOVA. 1992. Protozoan parasites of fishes. Elsevier
Publishing Company, Amsterdam, The Netherlands, 314 p.
MARKIW, M. 1986. Salmonid whirling disease: Dynamics of experimental production of the infective stage-the triactinomyxon spore.
Canadian Journal of Fisheries and Aquatic Science 43: 521526.
. 1989. Salmonid whirling disease: Myxosporean and actinosporean stages cross-react in direct fluorescent antibody test. Journal of Fish Diseases 12: 137141.
. 1991. Whirling disease: Earliest susceptible age of rainbow
trout to the triactinomyxid of Myxobolus cerebralis. Aquaculture
92: 16.
. 1992a. Experimentally induced whirling disease: Dose response of fry and adults of rainbow trout exposed to the triactinomyxon stage of Myxobolus cerebralis. Journal of Aquatic Animal Health 4: 4043.
. 1992b. Experimentally induced whirling disease: Determination of longevity of the infective triactinomyxon stage of Myxobolus cerebralis by vital staining. Journal of Aquatic Animal
Health 4: 4447.
, AND K. WOLF. 1983. Myxosoma cerebralis (Myxozoa: Myxosporea) etiologic agent of salmonid whirling disease requires tubificid worm (Annelida: Oligochaeta) in its life cycle. Journal of
Protozoology 30: 561564.
MATSUMOTO, M., AND G. JAMMOTO. 1966. On the seasonal rhythmicity
of oviposition in the aquatic oligochaete Tubifex hattai. Japanese
Journal of Ecology 16: 134139.
MCMURTRY, M. J., D. J. RAPPORT, AND K. E. CHUA. 1983. Substrate
selection by tubificid oligochaetes. Canadian Journal of Fisheries
and Aquatic Science 40: 16391646.
MILBRINK, G. 1973. On the use of indicator communities of Tubificidae
and some Lumbriculidae in the assessment of water pollution in
Swedish lakes. Zoon 1: 125139.
. 1993. Evidence for mutualistic interactions in freshwater oligochaete communities. Oikos 68: 317322.
MOLNAR, K. 1993. Recent achievements in the chemotherapy of myxosporean infections of fish. Acta Veterinaria Hungarica 4: 5158.
, F. BASKA, AND C. SZEKELY. 1987. Fumagillin, an efficacious
drug against renal sphaerosporosis of the common carp (Cyprinus
carpio). Diseases of Aquatic Organisms 2: 187190.
NEHRING, R. B., AND P. G. WALKER. 1996. Whirling disease in the
wild: The new reality in the intermountain west. Fisheries 21:
2830.
OGRODNICK, J. J. 1979. Susceptibility of various salmonids to whirling
disease (Myxosoma cerebralis). Transactions of the American Fisheries Society 108: 187190.
PLATT, C. 1983. The peripheral vestibular system of fishes. In Fish
neurobiology, R. G. Northcutt and R. E. Davis (eds.). University
of Michigan Press, Ann Arbor, Michigan, p. 89123.
PODDUBNAYA, T. L. 1980. Life cycles of mass species of Tubificidae. In
Aquatic oligochaete biology, R. O. Brinkhurst and D. G. Cook
(eds.). Plenum, New York, p. 175184.
. 1984. Parthenogenesis in tubificidae. Hydrobiologia 115: 97
99.
REYNOLDSON, T. B. 1987. The role of environmental factors in the ecology of tubificid oligochaetesAn experimental study. Holarctic
Ecology 10: 241248.
ROBBINS, J. A., T. KEILTY, D. S. WHITE, AND D. N. EDGINGTON. 1989.

GILBERT AND GRANATHWHIRLING DISEASE

Relationships among tubificid abundances, sediment composition,


and accumulation rates in Lake Erie. Canadian Journal of Fisheries
and Aquatic Science 46: 223231.
ROGNLIE, M. C., AND S. E. KNAPP. 1998. Myxobolus cerebralis in Tubifex tubifex from a whirling disease epizootic in Montana. Journal
of Parasitology 84: 711713.
ROSE, J. D., G. S. MARRS, C. LEWIS, AND G. SCHISLER. 2000. Whirling
disease behavior and its relation to pathology of brain stem and
spinal cord in rainbow trout. Journal of Aquatic Animal Health 12:
107118.
RUIDISCH, S., M. EL-MATBOULI, AND R. W. HOFFMANN. 1991. The role
of tubificid worms as an intermediate host in the life cycle of Myxobolus pavlovskii (Akhmerov, 1954). Parasitology Research 77:
663667.
SCHLEGEL, M., J. LOM, A. STECHMANN, D. BERNHARD, D. LEIFE, I. DY , AND M. SOGIN. 1996. Phylogenetic analysis of complete
KOVA
small subunit ribosomal RNA coding region of Myxidium lieberkuehni: Evidence that the Myxozoa are Metazoa and related to the
Bilateria. Archiv fur Protistenkunde 147: 19.
SHUM, B. P., L. GUETHLEIN, L. R. FLODIN, M. A. ADKISON, R. P. HEDRICK,
R. B. NEHRING, R. J. M. STET, C. SECOMBES, AND P. PARHAM. 2001.
Modes of salmonid MHC class I and II evolution differ from the
primate paradigm. Journal of Immunology 166: 32973308.
SIDALL, M. E., D. S. MARTIN, D. BRIDGE, S. S. DESSER, AND D. K. CONE.
1995. The demise of a phylum of protists: Phylogeny of Myxozoa
and other parasitic cnidarians. Journal of Parasitology 81: 961967.
SMOTHERS, J. F., C. D. VON DOHLEN, L. H. SMITH JR., AND R. D. SPALL.
1994. Molecular evidence that the myxozoan protists are metazoans. Science 265: 17191721.
STATON, L., D. ERDAHL, AND M. EL-MATBOULI. 2002. Efficacy of Fumagillin and TNP-470 to prevent experimentally induced whirling
disease in rainbow trout Oncorhynchus mykiss. In Whirling disease:
Reviews and current topics, J. L. Bartholomew and J. C. Wilson
(eds.). American Fisheries Society Symposium 29, Bethesda,
Maryland, p. 239247.
STEVENS, R., B. L. KERANS, J. C. LEMMON, AND C. RASMUSSEN. 2001.
The effects of Myxobolus cerebralis myxospore dose on triactino-

667

myxon production, and biology of Tubifex tubifex from two geographic regions. Journal of Parasitology 87: 315321.
STURMBAUER, C., G. OPADIYA, H. NIEDERSTATTER, A. RIEDMANN, AND R.
DALLINGER. 1999. Mithochondrial DNA reveals cryptic oligochaete
species differing in cadmium resistance. Molecular Biology and
Evolution 16: 967974.
SZEKELY, CS., K. MOLNAR, AND O. RACZ. 2001. Complete developmental cycle of Myxobolus pseudodispar (Gorbunova) (Myxosporea:
Myxobolidae). Journal of Fish Diseases 24: 461468.
USPENSKAYA, A. V. 1995. Alteration of actinosporean and myxosporean
phases in the life cycle of Zschokkella nova (Myxozoa). Journal of
Eukaryotic Microbiology 42: 665668.
VINCENT, E. R. 1996. Whirling disease and wild trout: The Montana
experience. Fisheries 21: 3233.
WAVRE, M., AND R. O. BRINKHURST. 1971. Interactions between some
tubificid oligochaetes and the bacteria found in the sediments of
Toronto Harbour, Ontario. Journal of the Fisheries Research Board
of Canada 28: 335341.
WOLF, K., AND M. MARKIW. 1982. Myxosoma cerebralis: Inactivation
of spores by hot smoking of infected trout. Canadian Journal of
Fisheries and Aquatic Science 39: 926928.
, AND . 1984. Biology contravenes taxonomy in the Myxozoa: New discoveries show alternation of invertebrate and vertebrate hosts. Science 225: 14491452.
, , AND J. K. HILTUNEN. 1986. Salmonid whirling disease:
Tubifex tubifex identified as the essential oligochaete in the protozoan life cycle. Journal of Fish Diseases 9: 8385.
YOKOYAMA, H., K. OGAWA, AND H. WAKABAYASHI. 1991. A new collection method of actinosporeansA probable infective stage of
myxosporeans to fishesfrom tubificids and experimental infection
of goldfish with the actinosporean Raabeia sp. Fish Pathology 26:
133138.
, , AND . 1995a. Myxobolus cultus n. sp. (Myxosporea: Myxobolidae) in the goldfish (Carassius auratus) transformed from the actinosporean stage in the oligochaete Branchiura
sowerbyi. Journal of Parasitology 81: 446451.
, , AND . 1995b. Chemoresponse of actinosporean
spores of Myxobolus cultus to skin mucous of goldfish (Carassius
auratus). Diseases of Aquatic Organisms 21: 711.