Beruflich Dokumente
Kultur Dokumente
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
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and
environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published
by nonprofit societies, associations, museums, institutions, and presses.
Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of
BioOnes Terms of Use, available at www.bioone.org/page/terms_of_use.
Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries
or rights and permissions requests should be directed to the individual publisher as copyright holder.
BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research
libraries, and research funders in the common goal of maximizing access to critical research.
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.
660
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
662
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
664
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
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
666
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.