Comparative Susceptibility of Deschutes River,Oregon, Tubifex tubifex Populations to Myxobolus cerebralis

Abstract

Dams along the Deschutes River (DR) in central Oregon have blocked fish migration for over 40 years. Reestablishment of anadromous fish runs above the dams as part of a fish passage plan may introduce fish pathogens, such as Myxobolus cerebralis, the myxozoan parasite that causes salmonid whirling disease. This parasite is carried by adult salmon that stray into the DR system during their return to enzootic areas of the upper Columbia River basin, and it is now known to be established in at least one lower DR tributary. The life cycle of M. cerebralis involves two obligate hosts: a salmonid and the oligochaete worm Tubifex tubifex. To determine the likelihood of parasite establishment above the DR dams, we conducted benthic sediment surveys between 1999 and 2007 and found that T. tubifex had a patchy distribution and low relative abundance. Mitochondrial 16S ribosomal DNA gene analysis indicated that two lineages of T. tubifex (III and VI) were present both above and below the dams. Laboratory susceptibility studies to characterize differences in infection prevalence and parasite production between nine T. tubifex populations revealed that production varied considerably among exposed groups and was proportional to the number of lineage III worms present. Our results suggest that M. cerebralis could become established above the dams if infected fish are allowed passage into the upper DR system, but not all areas of the DR basin can be classified as having the same likelihood for parasite establishment, and the potential impact will be location dependent.

Received July 2, 2010; accepted October 24, 2010     

Reduced Myxobolus cerebralis Actinospore Production in a Colorado Reservoir May Be Linked to Changes in Tubifex tubifex Population Structure

Abstract

Elucidating the dynamics of a parasitic infection requiring two hosts in a natural ecosystem can be a daunting task. Myxobolus cerebralis (Mc), the myxozoan parasite that causes whirling disease in some salmonids, was detected in the Colorado River upstream of Windy Gap Reservoir (WGR) in 1988. Subsequently, whirling disease was implicated in the decline of wild Rainbow Trout Oncorhynchus mykiss in the river when WGR was identified as a point source of Mc triactinomyxons (TAMs). Between 1997 and 2004, numerous investigations began to elucidate the etiology of Mc in WGR. During this period, Mc TAM production in WGR declined more than 90%. Explanations for the decline have included differences in stream discharge between years, changes in the thermal regime of the lake, severe drought, changes in the fish population structure in WGR, and reductions in the prevalence and severity of Mc infection in salmonids in the Colorado and Fraser rivers upstream of WGR. All of these have been discredited as explanations for the reduced TAM production. In 2005, a new study was conducted to replicate the studies completed in 1998. In this paper, the results of a new real-time polymerase chain reaction assay utilized to quantify the mitochondrial 16S rDNA specific to each of four lineages of Tubifex tubifex in pooled samples of 50 oligochaetes are presented. These results suggest that compared with 1998, the densities of aquatic oligochaetes and T. tubifex have increased, TAM production has been greatly reduced, and the decline is congruent with the dominance of lineages I, V, and VI of T. tubifex—three lineages that are refractory or highly resistant to Mc infection—in the oligochaete population. While it is possible that the resistant lineages function as biofilters that deactivate Mc myxospores, the reason for the decline in TAM production in WGR remains an enigma.

Received February 15, 2012; accepted March 17, 2013     

Whirling disease dynamics: An analysis of intervention strategies

Whirling disease (WD), a severe and widespread disease of salmonids, is caused by the myxosporean parasite Myxobolus cerebralis. It is further characterized by a unique two-host life cycle, utilizing the oligochaete Tubifex tubifex as an intermediate host. M. cerebralis is an invasive species that has been affecting populations in the United States including epidemics that killed in excess of 90% of populations in Colorado and Montana streams within the past 20 years. Currently, there is no known cure for WD, and the accepted method of control is removal of infected fish from the population. We have created a compartmental model of the WD system in order to assess more efficient means of control and management of the disease. Using data gathered from the literature, we used Bayesian model fitting to estimate model parameters and estimated that R₀ ≈ 1.51 (95% CI: 1.39, 1.72), a value which implies that WD can be controlled using available strategies. To this end, we posit several parameters that we expect to be most influential to WD propagation, namely: release of triactinomyxons by T. tubifex, release of spores by salmonids, and infectious particle loads in each respective host. Based on currently available control strategies, approaches targeting the infectious particles and the oligochaete host appear the most effective alternative strategies for management and control of WD.     

Experimentally Induced Whirling Disease I. Dose Response of Fry and Adults of Rainbow Trout Exposed to the Triactinomyxon Stage of Myxobolus cerebralis

Abstract

The intensity and prevalence of whirling disease was tested by exposure of 2-monthold fry and 1-, 2-, and 3.5-year-old adults of rainbow trout Oncorhynchus mykiss to a known number of laboratory-produced Myxobolus cerebralis at the actinosporean triactinomyxon stage. Fry exposed to graded concentrations of infectivity (triactinomyxons) for 3 h were individually examined for spores of Myxobolus cerebralis 5 and 6 months later. Exposure of fish to the lowest doses, 1 and 10 triactinomyxons per fish, did not result in detectable myxosporean spores. Fish that became lightly infected by a dose of 100 triactinomyxons per fish experienced a decrease in the incidence of infection between 5 and 6 months after exposure. A linear relationship was found between the numbers of recovered myxosporean spores and doses of 100–10,000 triactinomyxons per fish, and the spore burden appeared to plateau at doses of 10,000–100,000 triactinomyxons per fish. Adult fish continuously exposed to the highest dose of triactinomyxons for 3.5 months were infected and asymptomatic, however, the severity of myxosporean infection decreased with increased age of fish. This information may help in controlling whirling disease in salmonids.     

Experimentally Induced Whirling Disease II. Determination of Longevity of the Infective Triactinomyxon Stage of Myxobolus cerebralis by Vital Staining

Abstract

Effects of temperature and aging on viability and infectivity of laboratory-produced actinosporean triactinomyxon spores (infective stage of the organism causing whirling disease) were studied. In vitro staining of triactinomyxon spores with vital fluorescein diacetate correlated with the ability of the spores to infect fry of rainbow trout Oncorhynchus mykiss. The experimentally produced actinosporean stage of Myxobolus cerebralis was short-lived, persisting for only 3–4 d at 12.5°C and for less time at warmer temperatures. The vital staining method has potential for screening therapeutants intended to control myxosporean infection of fish.     

Three Myxosporeans Found in the Cranial and Branchial Tissues of Rainbow Trout in California

Abstract

Three myxosporeans were encountered in the cranial tissues of a California population of rainbow trout Oncorhynchus mykiss examined for the presence of Myxobolus cerebralis, the causative agent of whirling disease. Typical spores of M. cerebralis and a previously undescribed species of Myxobolus were found in the cranial tissues prepared by the pepsin HCl-trypsin digestion method. Henneguya zschokkei was also detected in digest preparations of cranial tissues, but was more numerous when branchial cartilage was included in the preparations. Microscopic examinations of tissues of individual rainbow trout showed occasional infections with both myxobolid species. Myxobolus cerebralis trophozoites and spores were found in the cranial and gill cartilage, and Myxobolus sp. was found in the brain and spinal cord. Henneguya zschokkei was also found within granulomas in the connective tissues below the gill arch. Both M. cerebralis and H. zschokkei were associated with a chronic inflammatory response in their respective tissues. In contrast, the Myxobolus sp. spores were found in pockets within the nervous tissues with no detectable host response. The spore measurements, calculated from fresh digests of infected tissues for the three myxosporeans (N = 20), for length × width × thickness in micrometers (SD) were 11.7 (0.6) without tails and 42.6 (5.2) with tails × 7.7 (0.8) × 7.0 (0.1) for H. zschokkei, 9.9 (0.4) × 8.4 (0.1) × 6.5 (0.3) for M. cerebralis, and 12.7 (0.7) × 10.5 (1.0) × 9.5 (0.8) for Myxobolus sp. Examined under scanning electron microscopy, the latter two species were morphologically similar although distinctive in size.     

Detection of Myxobolus cerebralis in Rainbow Trout and Oligochaete Tissues by Using a Nonradioactive in Situ Hybridization (ISH) Protocol

Abstract

A nonradioactive in situ hybridization (ISH) protocol was developed to detect Myxobolus cerebralis, the causative organism of whirling disease, in its primary host, rainbow trout Oncorhynchus mykiss, and in its alternate oligochaete host, Tubifex tubifex. A cocktail of three oligonucleotide primers (derived from the small subunit ribosomal DNA sequence) directed at target sequences of the parasite DNA was tailed at the 3′ end with digoxigenin-labeled deoxyuridine triphosphate (DIG-dUTP). Labeled probes were hybridized to parasite DNA present in deparaffinized tissue sections from infected trout and oligochaetes. The bound probes were visualized after modifications of existing ISH protocols. By using the new ISH procedure, the parasite was found in target tissues of subclinically and clinically infected fish and tubificid oligochaetes after exposures of these hosts to triactinomyxons and mature spores, respectively. The probe did not bind with salmonid tissues infected with two other myxosporean parasites, Ceratomyxa shasta or the PKX organism, or to a Myxobolus sp. infecting the cartilage of plain sculpin Myoxocephalus jaok. These initial results indicate that ISH is an effective and specific test for detecting Myxobolus cerebralis in its fish and oligochaete hosts.     

Comparison of Susceptibility of Five Cutthroat Trout Strains to Myxobolus cerebralis Infection

Abstract

Susceptibility to infection by the myxosporean parasite Myxobolus cerebralis was compared among strains of cutthroat trout Oncorhynchus clarki in two separate exposure tests in the laboratory. In both tests, each strain was exposed to 1,000 triactinomyxons/fish for 2 h in 8.0 L of water. In the first test, three strains of 10-week-old cutthroat trout were compared: two strains of Bonneville cutthroat trout O. c. utah (Bear Lake and southern Bonneville strains) and Yellowstone cutthroat trout O. c. bouvieri. In the second test, these strains plus Snake River fine-spotted cutthroat trout O. c. subsp. and Colorado River cutthroat trout O. c. pleuriticus were exposed at either 5 or 10 weeks of age. The prevalence of the M. cerebralis infection was determined by single-round polymerase chain reaction (PCR) assay 5 weeks after exposure. In the first test, the prevalence was significantly lower in the Bear Lake strain of Bonneville cutthroat trout (78.5%) than in the Yellowstone (97.8%) or southern Bonneville (100%) strains when exposed at 10 weeks of age. In the second test, the Bear Lake strain also had significantly lower infection rates after exposure at 5 (54%) or 10 weeks (82%) of age than the other four strains, which did not differ from each other (94–100%). The severity of the infection was also significantly reduced in Bear Lake Bonneville cutthroat trout, as suggested by the strength of the product of the single-round PCR assay. These results suggest that intraspecific differences in susceptibility to M. cerebralis infection exist, further supporting the need to maintain the genetic diversity among subspecies and geographic variants of cutthroat trout.     

Susceptibility of Juvenile and Yearling Bull Trout to Myxobolus cerebralis and Effects of Sustained Parasite Challenges

Abstract

Evaluation of the susceptibility of bull trout Salvelinus confluentus fry to Myxobolus cerebralis infection using two different laboratory challenge models demonstrated that the method of administering the exposure dose affected infection prevalence. Administration of a low parasite dose (500 per fish) in a single exposure did not establish infection, but when the same cumulative dose was administered over 21 d the prevalence of infection was 45%. The results of challenges at a high exposure dose (5,000 per fish) were similar, infections being detected in 24% of the fish receiving a single dose and 40% of those administered the same dose in multiple exposures. Clinical disease was not detected in fish exposed via either challenge method. The susceptibility of yearling bull trout was tested by means of a single high dose of 10,000 per fish, and infection was detected in only 5% of those fish.     

Spatial and Temporal Variation of Whirling Disease Risk in Montana Spring Creeks and Rivers

Abstract

Spring creeks are important spawning and rearing areas for wild trout, but the stable flows, cool temperatures, and high nutrient levels that characterize these unique habitats may also make them highly susceptible to establishment and proliferation of the whirling disease pathogen Myxobolus cerebralis. We evaluated the spatial and temporal dynamics in whirling disease risk by using sentinel rainbow trout Oncorhynchus mykiss fry in nine different spring creeks and their conjoining rivers or reservoirs in Montana over a 20-month period. Whirling disease risk was high in five of the seven pathogen-positive spring creek study sites; at these sites, prevalence levels exceeded 90% and over 50% of sentinel fry had moderate to high infection severity scores. Spring creeks generally had higher disease prevalence and severity than paired river or reservoir sites. Fine sediment levels varied widely among springs creeks with high and low whirling disease risk, and we found no significant association between fine sediment level and infection severity. The low risk measured for some spring creeks was likely attributable to the pathogen invasion being in its early stages rather than to environmental characteristics limiting the severity of infection. High whirling disease risk occurred over a wide range of temperatures at spring creek sites (4.5–13°C) and river sites (1.7–12.5°C). There was an unusual seasonal cycle of infection in spring creeks, with peak infection levels occurring from late fall to early spring and declining to near zero in late spring to early fall. The low infection risk during spring suggests that spring-spawning trout would be at a low risk of infection, even in spring creeks with otherwise high disease severity. In contrast, fry of fall-spawning trout may be much more susceptible to infection in spring creek environments.

Received November 22, 2011; accepted May 7, 2012     

Susceptibility of Selected Inland Salmonids to Experimentally Induced Infections with Myxobolus cerebralis,the Causative Agent of Whirling Disease

Abstract

Laboratory exposures to the infectious stages (triactinomyxons) of Myxobolus cerebralis demonstrated a range of susceptibility to whirling disease among four species of inland salmonids. Replicate groups of each species were exposed to two concentrations of triactinomyxons, a low dose (100–200 per fish) and a high dose (1,000–2,000 per fish). Exposed fish were evaluated for clinical signs, for severity of microscopic lesions at 35 d, 2 and 5 months, and for spore concentrations in the head cartilage at 5 months. A standard strain of rainbow trout Oncorhynchus mykiss matched for age served as a susceptible species control. Rainbow trout, westslope cutthroat trout O. clarki lewisi, Yellowstone cutthroat trout O. clarki bouvieri, and bull trout Salvelinus confluentus were susceptible to M. cerebralis infections. Clinical signs, including radical swimming (“whirling”) and black tails, were observed at 7 weeks postexposure among rainbow and cutthroat trout challenged at 3 weeks of age. Clinical signs were rare among bull trout exposed at an age of 4 weeks and absent among rainbow and cutthroat trout exposed at 3 months posthatch. Most rainbow, cutthroat, and bull trout were found to be infected when examined at 5 months postexposure. The most severe microscopic lesions among infected fish at 5 months postexposure were found among rainbow trout. Cutthroat trout had less severe lesions, bull trout had mild infections, and no evidence of infection was found among Arctic grayling Thymallus arcticus. Mean spore concentrations among infected fish correlated with the severity of microscopic lesion scores. Rainbow trout had mean concentrations of spores in head cartilage reaching 10⁶, whereas more resistant species such as bull trout had 10⁴ spores; no spores were found among Arctic grayling at 5 months postexposure.     

Detection of Myxobolus (Myxosoma) cerebralis in Salmonid Fishes in Oregon

Abstract

Myxobolus (Myxosoma) cerebralis, the etiological agent of whirling disease, was detected in salmonid fish populations in northeastern Oregon. This is the first record of M. cerebralis in the Pacific Northwest of the USA. During an epizootiological survey for the parasite, two methods for spore detection were compared, and an efficient procedure for determining M. cerebralis infection in adult fish was developed. The enzyme digest method was more efficient than the plankton centrifuge procedure for examination of numerous individual lots of fish processed during the survey. Sampling only the area around the otoliths was at least as effective as sampling entire heads for detection of spores in infected fish.     

The Effects of Myxobolus cerebralis on the Physiological Performance of Whirling Disease Resistant and Susceptible Strains of Rainbow Trout

Abstract

The development of rainbow trout Oncorhynchus mykiss strains that are resistant to whirling disease has shown promise as a management tool for populations in areas where Myxobolus cerebralis is present. However, the physiological effects of the disease on characteristics necessary for fish survival in natural river conditions have not been tested in many of these strains. Five rainbow trout strains were evaluated for their swimming ability and growth characteristics in relation to M. cerebralis exposure: the resistant German rainbow trout (GR) strain (Hofer strain), the susceptible Colorado River rainbow trout (CRR) strain, and three intermediate (hybrid) strains (F₁ = GR × CRR; F₂ = F₁ × F₁; B₂ = backcross of F₁ × CRR). Three broad response patterns among strain and exposure were evident in our study. First, exposure metrics, growth performance, and swimming ability differed among strains. Second, exposure to the parasite did not necessarily produce differences in growth or swimming ability. Exposure to M. cerebralis did not affect batch weight for any strain, and critical swimming velocity did not differ between exposed and unexposed families. Third, although exposure did not necessarily affect growth or swimming ability, individuals that exhibited clinical deformities did show reduced growth and swimming performance; fish with clinical deformities were significantly smaller and had lower critical swimming velocities than exposed fish without clinical deformities. Research and management have focused on GR × CRR hybrid strains; however, given the performance of the GR strain in our study, it should not be discounted as a potential broodstock. Additional field trials comparing the GR and F₁ strains should be conducted before wholesale adoption of the GR strain to reestablish rainbow trout populations in Colorado.

Received September 9, 2010; accepted May 27, 2011     

A Packed-Bed Filtration System for Collection of Myxobolus cerebralis Triactinomyxons

Abstract

Information on the distribution and abundance of Myxobolus cerebralis triactinomyxons in natural systems is limited because direct and accurate sampling methods for this life stage have not been developed. Existing methods are based on indirect measures of triactinomyxon densities and are therefore confounded. Direct estimation of triactinomyxon concentrations would more exactly pinpoint the ambient infection risk to wild fish and allow evaluation of management strategies designed to mitigate the effects of the disease. We developed a mobile packed-bed filtration system that quickly, accurately, and precisely collects, concentrates, and quantifies triactinomyxons. The system includes pumping, prefiltration, two rounds of packed-bed filtration, and centrifugation. Laboratory tests of the completed system using known quantities of triactinomyxons resulted in a mean recovery rate of 91% with a minimum detectable concentration of triactinomyxons of 0.04/L. We subsequently field-tested the system at a site known to be positive for the parasite and recovered triactinomyxons at densities of 0.7–1.4/L. The packed-bed filtration system has the potential to quickly determine the temporal and spatial variation in infection risk and to test the efficacy of various management strategies.     

Comparison of Hatchery and Field Performance between a Whirling-Disease-Resistant Strain and the Ten Sleep Strain of Rainbow Trout

Abstract

A whirling-disease-resistant strain of rainbow trout Oncorhynchus mykiss (GRHL strain) derived from a backcross of an F1 hybrid of two strains (German strain × Harrison Lake strain) with German strain females, was compared with the Ten Sleep (TS) strain of rainbow trout. The GRHL strain had consistently superior growth and feed conversion in two consecutive hatchery trials. Hatching and mortality rates were similar between strains. Both strains were stocked into two Utah reservoirs (Hyrum, Porcupine), and a third, Causey Reservoir, was monitored as a control for seasonal variation in prevalence of Myxobolus cerebralis. A total of 1,323 salmonids captured by gill net in spring and fall sampling between 2006 and 2008 were tested for M. cerebralis via pepsin-trypsin digest methods. Only eight of these (<1% per species) had clinical signs consistent with whirling disease. In both reservoirs, GRHL survived better than the TS and had higher growth rates. The prevalence of M. cerebralis was significantly lower for GRHL (18.1%) than TS (50.0%) in Porcupine Reservoir. In Hyrum Reservoir the trend was similar, but prevalence was lower and did not significantly differ between GRHL (9.6%) and TS (23.1%). For infected fish, no significant differences were observed between strains in myxospore counts in either Hyrum (GRHL = 911–28,244 spores/fish [spf], TS = 1,822–155,800 spf) or Porcupine (GRHL = 333–426,667spf, TS = 333–230,511 spf) reservoirs. Unmarked rainbow trout in both reservoirs had significantly higher myxospore counts than stocked fish of either strain. There were significant differences in M. cerebralis prevalence and myxospore loads among other naturally reproducing salmonids in the reservoirs. The trend in susceptibility was cutthroat trout Oncorhynchus clarkii > kokanee Oncorhynchus nerka > brown trout Salmo trutta. The GRHL performed well in both hatchery and field settings and is recommended for stocking programs.

Received December 28, 2011; accepted February 2, 2012     

The Effects of Whirling Disease on Growth and Survival of Snake River Cutthroat and Colorado River Rainbow Trout Fingerlings

Abstract

Two sizes of fingerling Snake River cutthroat trout Oncorhynchus clarkii behnkei and Colorado River rainbow trout O. mykiss were raised at hatcheries testing negative for Myxobolus cerebralis and stocked into the Dolores and Cache la Poudre rivers from 1999 to 2001. Populations were resampled over a 2-year period to determine which species and size combination had the highest growth and survival rates. Fish were tested for M. cerebralis via polymerase chain reaction and pepsin?trypsin digest analyses. Growth and survival rates between the species and size groups were not significantly different in either river. In the Dolores River, annual survival for both species and sizes of fish combined ranged from 0.063 to 0.12. In the Cache la Poudre River, survival for both sizes of rainbow trout was 0.004; survival for cutthroat trout ranged from 0.182 to 0.53. Larger fish had higher growth rates than smaller fish, and cutthroat trout had higher rates than similar sizes of rainbow trout. In both rivers, a higher percentage of the rainbow trout sample was infected than in the cutthroat trout sample. Rainbow trout also had a higher mean number of spores per head than cutthroat trout, and small rainbow trout had higher spore counts than large rainbow trout. Survival rates for cutthroat trout in the Cache la Poudre River were the highest of any of the groups, suggesting a difference that is biologically significant. Raising fingerlings to sizes greater than 100 mm can improve poststocking survival. If rainbow trout are stocked into contaminated waters, raising fingerlings to a larger size does not appear to improve growth or survival rates. Stocking rainbow trout in the spring could maximize growth rates but will expose fish to greater triactinomyxon densities, resulting in higher intensities of infection.     

Assessment of the Long-Term Viability of the Myxospores of Myxobolus cerebralis as Determined by Production of the Actinospores by Tubifex tubifex

While whirling disease was first observed in Rainbow Trout Oncorhynchus mykiss in 1893, the complete life cycle of Myxobolus cerebralis (Mc), the causative agent of the disease, was not understood until 1984, when it was shown to involve two obligate hosts, a salmonid fish and the aquatic oligochaete Tubifex tubifex (Tt). The viability of the triactinomyxon (TAM) actinospores produced by Tt has been well studied, and is known to be temperature dependent and measured in days and weeks. Assertions that Mc myxospores produced by infected fish remain viable for years or even decades were made during the mid-20th century, decades before the Mc life cycle was described. Moreover, the duration of myxospore viability has not been well studied since the life cycle was elucidated. In a series of time-delay treatments, we assessed the long-term viability of Mc myxospores by exposure to Mc-susceptible Tt oligochaetes and quantified TAM production. As the time delay between inoculation and incubation of Mc myxospores in sand and water and exposure to Tt oligochaetes increased, TAM production decreased exponentially. Production among the 15-d time-delay replicates was reduced 74.7% compared with the 0-d treatment. Likewise, total TAM production was reduced 94.5, 99.4, and 99.9%, respectively, in the 90-, 120-, and 180-d time-delay treatments. Linear regression analysis of our data and the absence of TAM production among replicates of Mc myxospores held at 5°C for 365 d prior to exposure to Mc-susceptible Tt oligochaetes indicate that the long-term viability of Mc myxospores is less than 1 year under the conditions of this study.

Received May 14, 2014; accepted September 27, 2014     

A Simple Technique Used to Filter and Quantify the Actinospore of Myxobolus cerebralis and Determine Its Seasonal Abundance in the Colorado River

Abstract

We used a simple technique for filtering the actinospores of Myxobolus cerebralis from natural waters to observe seasonal periodicity at eight sites in the upper Colorado River drainage. We used a tub lined with 20-μm-mesh Pecap screen to concentrate actinospores from 1,900-L samples and estimate density by microscope count. Identity of the observed actinospores as those of M. cerebralis was confirmed in 86 samples by the use of a polymerase chain reaction test. The 42-ha Windy Gap Reservoir appeared to be a point source of actinospores; the highest densities observed were consistently from samples taken at sites just below the reservoir. Both densities and the frequency of detection were much lower 26 km below the reservoir. The actinospores first appeared in abundance after the runoff in both years of the study. Actinospore densities tended to be greatest during summer and early fall and lowest during spring. In August 1997, a series of significant flow fluctuations and attendant water temperature swings appeared to alternately inhibit and stimulate the release of actinospores. Populations of rainbow trout Oncorhynchus mykiss continue to suffer recruitment failures throughout the study reach, apparently because of the effects of whirling disease in age-0 fish. This suggests that the detection of low numbers of actinospores by this technique at some sampling locations may indicate a level of infectivity that is destructive for the susceptible rainbow trout.     

Myxobolus cyprini Doflein, 1898, in Peamouth: First Report of This Myxosporean in the Western Hemisphere

Abstract

A survey of 3–8-year-old peamouths Mylocheilus caurinus collected from the lower Fraser River, British Columbia, revealed 100%, infection by a myxosporean identified as Myxobolus cyprini based on spore morphology, site of infection, and associated pathological changes. Myxobolus cyprini is a common myxosporean parasite of several species of cyprinid fishes (family Cyprinidae) in Europe, the former USSR, and China, but this is the first report of this myxosporean in the Western Hemisphere. The source of the infection in peamouths—whether the parasite is endemic to North America or has been imported with exotic cyprinids from Europe or Asia—is unknown. It is also possible that the myxosporean from peamouths is very closely related to, but a distinct species from, M. cyprini of Eurasia. As reported from European cyprinid fishes, the myxosporean in peamouths developed in the skeletal muscle, and disseminated spores were found associated with chronic inflammation in the visceral organs.     

Distribution of Myxobolus cerebralis in Salmonid Fishes in Montana

Abstract

Over an approximately 2-year period, 20,974 fish (trout and other salmonid species) from 230 separate waters (creeks, rivers, lakes, reservoirs, ponds, hatcheries, and irrigation ditches) within 21 of the 22 major drainages in Montana were examined for Myxobolus cerebralis. Nine of the major river drainages have waters containing infected fish: Beaverhead, Big Hole, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the south fork of the Flathead River, Jefferson, Madison, Missouri above the Marias River, and Yellowstone above the Bighorn River. The Beaverhead, Clark Fork above the Bitterroot River, Jefferson, Madison, and Missouri above the Marias River have the greatest number of waters containing fish infected with M. cerebralis. Comparisons of infection levels (number of pooled samples that contain fish infected with M. cerebralis) between species among these drainages show significantly lower levels of infection in brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss in the Missouri above the Marias River drainage and significantly higher levels of infection of rainbow trout in the Jefferson. Comparisons of differences in infection levels between drainages among species show that, in the Beaverhead, Clark Fork above the Bitterroot River, and Madison, infection levels in brown trout are significantly higher than in rainbow trout. This is partially attributed to losses of juvenile rainbow trout because of M. cerebralis infection, leading to biased samples. Histopathologic studies showed lesions were consistently less severe in brown trout than other species and occurred in a different location (gill arches versus ventral calvarium). In six of the nine affected drainages (Beaverhead, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the South Fork, Jefferson, and Madison), infected fish were found at or near the time that intensive sampling was initiated in the spring of 1995. In the three remaining affected drainages (Missouri above the Marias River, Yellowstone above the Bighorn River, and the Big Hole), infected fish were not identified until at least 15 months after the initiation of widespread testing. This indicates that in the first six drainages listed above, the infection was well established prior to 1995 but spread into the last three drainages in the ensuing months. Methods of transmission and the sources of infection are unknown, although the absence of infected fish in state, private, and federal hatcheries in Montana indicates hatchery fish from these sources are not likely to be responsible.