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1.
Abstract

Water temperature, a pivotal factor influencing interactions between teleosts and pathogens, was examined to determine its effects on the kinetics of xenoma formation and dissolution subsequent to experimental exposure of rainbow trout Oncorhynchus mykiss to the microsporidian gill pathogen Loma salmonae. The permissive water temperature range in which xenomas developed was between 9° and 20°C. Parasite development was arrested at temperatures outside this range, as indicated by the absence of visible xenomas among exposed fish. In addition, when these trout were subsequently moved to temperatures within the permissive range, xenomas failed to develop. Water temperature, within the permissive range, had no significant effect on either the number of xenomas that formed or the proportion of fish that developed xenomas following gastric intubation with a standard dose of spores. The relationship between water temperature and xenoma onset-time was best described (R 2 = 88.3%) by polynomial regression analysis: onset = 320 ? 33.4T + 0.9547T 2, where T is temperature (°C). Xenoma onset rate was also described through a modified degree-days model, yielding a predictive equation appropriate for use under conditions of fluctuating temperature. The thermal units, expressed as days × (°C above 7°C) necessary for xenoma onset were 298.6 on average. Xenoma dissolution rates, from the time of onset, also appeared to have a trend; more rapid dissolution occurred as temperatures increased. However, this trend correlated minimally with regression models.  相似文献   

2.
The intracellular microsporidian parasite Loma salmonae affects salmonids of the genus Oncorhynchus and is a significant cause of economic losses in pen-reared Chinook salmon (O. tshawytscha) in British Columbia. Loma salmonae infection is easily recognized by the xenomas that form in the gills, but early stages of infection are difficult to detect in histologic sections. In situ hybridization (ISH), using an L. salmonae-specific digoxigenin-labeled single-stranded DNA probe, was used to detect the parasite during the early stages of infection. Loma salmonae was detected in the gut mucosal epithelium as early as 24 hours postexposure (PE), and it localized in the lamina propria of the intestine within 24 hours of infection. After the parasite was detected in the lamina propria, dividing merogonic stages in infected cells in the heart were detected by ISH as early as 2 days PE, providing the first evidence of parasitaemia and hematogenous distribution of this parasite in infected blood cells. The parasites inside the infected cells appeared to be undergoing merogony as they passed through the heart, indicating that proliferation may start at the site of infection, before the parasite arrives to the gills for their final developmental phase. This is the first time that L. salmonae passage through the intestinal wall and migration to the heart has been visualized; however, the identity of the cells harboring the parasite has yet to be determined.  相似文献   

3.
Loma salmonae, a microsporidian parasite of salmonids of the genus Oncorhynchus, is a significant cause of economic loss in pen-reared chinook salmon (O. tschawytscha). Final stages of L. salmonae infections are easily recognized by the xenomas that form in the gills during sporogony. However, early prexenoma stages of infection (3 weeks or less after infection) are difficult to detect on histologic slides. An L. salmonae-specific single-stranded DNA probe labeled with digoxigenin was used to detect these prexenoma stages of L salmonae by in situ hybridization in experimentally infected rainbow trout. This method allows detection of the parasite in the gills only 2 weeks after infection, providing a sensitive and specific way of detecting L. salmonae during the early stages of infection.  相似文献   

4.
Specimens of western pygmy perch, Edelia vittata Castelnau 1873, were submitted for examination because of the presence of black masses in the body wall in the abdominal area. The owner reported that approximately 10% of mature fish in one dam were affected. The masses had no apparent effect on the health of affected fish, but they were unsuitable for sale for aesthetic reasons. After necropsy and histological examination, the masses were found to be microsporidial xenomas within the abdominal cavity. Transmission electron microscopy results suggest that this microsporidian is a previously unreported species, because the melanised xenoma wall and internal structure of the spores differ from previously described species. The epidemiology of the parasite remains largely unknown and highlights the need for further surveys of the disease status and parasites of Australian native fish, especially those of interest to aquaculturists.  相似文献   

5.
Abstract

A series of dams on the Deschutes River, Oregon, act as migration barriers that segregate the river system into upper and lower basins. Proposed fish passage between basins would reunite populations of native potamodromous fish and allow anadromous fish of Deschutes River origin access to the upper basin. We assessed the potential redistribution of host-species-specific genotypes (O, I, II, III) of the myxozoan parasite Ceratomyxa shasta that could occur with fish passage and examined the influence of nonnative fish on genotype composition. To determine the present distribution of the parasite genotypes, we exposed eight salmonid species—three native and five stocked for sport fishing—in present and predicted anadromous salmonid habitats. We monitored fish for infection by C. shasta and sequenced a section of the parasite ribosomal DNA gene from fish and water samples to determine parasite genotype. Genotype O was present in both upper and lower basins and detected only in steelhead Oncorhynchus mykiss. Genotype I was spatially limited to the lower basin, isolated predominately from Chinook salmon O. tshawytscha, and lethal for this species only. Genotype II was detected in both basins and in multiple species, but only as a minor component of the infection. Genotype III was also present in both basins, had a wide host range, and caused mortality in native steelhead and multiple nonnative species. Atlantic salmon Salmo salar and kokanee O. nerka were the least susceptible to infection by any genotype of C. shasta. Our findings confirmed the host-specific patterns of C. shasta infections and indicated that passage of Chinook salmon would probably spread genotype I into the upper Deschutes River basin, but with little risk to native salmonid populations.

Received April 20, 2012; accepted July 19, 2012  相似文献   

6.
Abstract

We determined the sensitivity and specificity of a nested polymerase chain reaction (PCR) for detection of the microsporidian parasite Nucleospora salmonis in kidney tissue of rainbow trout Oncorhynchus mykiss. Kidney tissues were sampled on three dates from 162 juvenile rainbow trout obtained from a California State fish hatchery where the organism was endemic. Kidney tissues were used to prepare imprints stained with May–Grünwald Giemsa and for extraction of genomic DNA for a nested PCR test for N. salmonis. Positive PCR results for N. salmonis were obtained from 1 of 100, 2 of 32, and 27 of 30 kidneys collected on the first, second, and third sample dates, respectively. Kidney tissues from 3 of 27 trout in the third sample that tested positively by PCR also had microscopic evidence of parasites in stained kidney imprints. No parasites were detected in the remaining 159 kidney samples examined microscopically. Sensitivity and specificity of the PCR assay were estimated by using maximum likelihood estimation based on cross-classified test results. This method yielded estimates of sensitivity of 99.99% and specificity of 99.87%. This field evaluation supports experimental evidence that the nested PCR test will be a valuable diagnostic tool for prevention and control of N. salmonis as well as for risk assessment associated with fish movements.  相似文献   

7.
Abstract

Bacterial kidney disease (BKD) is an important contributor to mortality of salmonids in hatcheries in the Columbia River basin. However, the impact of BKD on the survival of downstream migrants is difficult to determine because there is little information on the disease-related mortality among these fish. In this study, the impact of BKD on juvenile salmonids was examined by determining the percentage of downriver migrants infected with Renibacterium salmoninarum (the causative agent of BKD) and evaluating the effects of salt water on the progress of the disease. During the 2 years of this study, approximately 20% of the three species of migrating hatchery and wild salmonids (Oncorhynchus spp.) collected were infected with R. salmoninarum. Mortality caused by BKD increased when fish were held in salt water.  相似文献   

8.
Abstract

Two copepods and three isopods, representing five families of Crustacea, are noted from ten hosts, eight of these representing new host records. Cardiodectes rubosus (Copepoda: Pennellidae) is reported from the Atlantic Ocean for the first time. Excorallana tricornis (Isopoda: Corallanidae) may be a more important fish parasite than previously thought.  相似文献   

9.
Early-rearing salmonids in Ontario Ministry of Natural Resources (OMNR) fish hatcheries have been consistently affected by bacterial gill disease (BGD) (causative agent: Flavobacterium branchiophilum) for many years. Separate retrospective epidemiological investigations of BGD treatments at two OMNR fish hatcheries (Hatcheries A and B) for the 1999 production year were conducted using on-site hatchery records. Both investigations were carried out at the rearing unit-level, with early-rearing (<9 months of age) “tank-lot” as the unit of analysis to identify unique fish populations over time. Multivariable repeated measures logistic regression models were created for both hatchery datasets, controlling for lot-level and species effects. For Hatchery A, the species brook trout (Salvelinus fontinalis) and brown trout (Salmo trutta) were significantly associated with BGD treatment, as well as lower water exchange rate, and higher feeding and mortality percentages during the 2 weeks previous to BGD treatment. At Hatchery B, the species brook trout (S. fontinalis) and splake (Salvelinus namaycush × S. fontinalis) were significantly associated with BGD treatment, as well as lower individual fish weights and treatment for BGD during the previous week. These results emphasize the importance of water quality, feeding rate, fish size and prior mortality on the development of BGD. Significant hatchery and species effects were evident, and future observational research on BGD must account for these factors in their design and analysis.  相似文献   

10.
Abstract

The biflagellated, single-celled parasite Ichthyobodo necator can cause significant losses among fish populations, particularly those cultured in tanks. Treatments of KMnO4 and CuSO4 were evaluated against a naturally occurring I. necator infestation on sunshine bass (female white bass Morone chrysops × male striped bass M. saxatilis) raised in tanks. Four-hour static treatments with 3 mg of KMnO4/L of water (2.5 mg/L above the determined KMnO4 demand) or 2 mg of CuSO4/L of water (total alkalinity = 207 mg/L; total hardness = 95 mg/L) were randomly applied to 4 tanks/treatment (23 fish/tank); the same treatments were reapplied 2 d later. Four tanks were used as positive controls. By 2 d posttreatment (after the second treatment), only 17.4% of the untreated control fish survived, and a sample of the remaining fish was heavily infested with I. necator. All remaining control fish were dead by 5 d posttreatment. The KMnO4 treatment significantly curtailed the initial mortality (survival = 92.4%) and slightly reduced the high parasite loads at 2 d posttreatment. However, fish mortalities increased dramatically over the next 3 d (survival at 5 d posttreatment = 37.5%), and parasite loads from sampled fish remained high. The CuSO4 treatment was effective in significantly lowering the parasite load (almost eliminating I. necator) and maintaining a high fish survival (87.5%) by 5 d posttreatment. The findings in this study clearly demonstrate that CuSO4 is a viable treatment for ichthyobodosis in tanks.  相似文献   

11.
Abstract

Specific-pathogen-free channel catfish Ictalurus punctatus were exposed to sediment and mud from a pond containing channel catfish with proliferative gill disease. In one experiment, fish were to exposed to mud and sediment for 2 months in water maintained at 19°C. Fish were necropsied weekly, and certain tissues were examined histologically and ultrastructurally. Four trials were conducted with sediment samples from different epizootics of proliferative gill disease. In a second experiment, fish were exposed to sediment for 7 d in water maintained at 16, 19, or 26°C; the fish were then moved to clean water held at 16, 19, or 26°C. Fish were necropsied before transfer to clean water and weekly thereafter for 2 months. Channel catfish held at 19°C developed proliferative gill disease within 2 d of exposure to sediment. Primary cells of a uninucleate myxosporean parasite were present in the gills at the base of lamellae. These developed into plasmodia with numerous secondary cells, and some primary cells disintegrated, releasing their internal secondary cells. Similar development was observed in internal organs 1 week after appearance of the parasite in gills. Complete sporogony did not occur over the 2 months of this study. Plasmodia became necrotic and were not detected after 60 d. In fish exposed to sediment for 7 d at 16, 19, and 21°C, similar organisms were detected, but clinical disease occurred only at 19 and 26°C. Proliferative gill disease may be attributed to extrasporogonic stages of a myxosporean resembling Sphaerospora spp.  相似文献   

12.
Abstract

Nucleospora salmonis is an intranuclear microsporidian that primarily infects lymphoblast cells and contributes to chronic lymphoblastosis and a leukemia-like condition in a range of salmonid species. The primary goal of this study was to evaluate the prevalence of N. salmonis in out-migrating juvenile hatchery and wild Chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss from the Snake River in the U.S. Pacific Northwest. To achieve this goal, we first addressed the following concerns about current molecular diagnostic tests for N. salmonis: (1) nonspecific amplification patterns by the published nested polymerase chain reaction (nPCR) test, (2) incomplete validation of the published quantitative PCR (qPCR) test, and (3) whether N. salmonis can be detected reliably from nonlethal samples. Here, we present an optimized nPCR protocol that eliminates nonspecific amplification. During validation of the published qPCR test, our laboratory developed a second qPCR test that targeted a different gene sequence and used different probe chemistry for comparison purposes. We simultaneously evaluated the two different qPCR tests for N. salmonis and found that both assays were highly specific, sensitive, and repeatable. The nPCR and qPCR tests had good overall concordance when DNA samples derived from both apparently healthy and clinically diseased hatchery rainbow trout were tested. Finally, we demonstrated that gill snips were a suitable tissue for nonlethal detection of N. salmonis DNA in juvenile salmonids. Monitoring of juvenile salmonid fish in the Snake River over a 3-year period revealed low prevalence of N. salmonis in hatchery and wild Chinook salmon and wild steelhead but significantly higher prevalence in hatchery-derived steelhead. Routine monitoring of N. salmonis is not performed for all hatchery steelhead populations. At present, the possible contribution of this pathogen to delayed mortality of steelhead has not been determined.

Received September 14, 2010; accepted November 14, 2010  相似文献   

13.
Abstract

After an epizootic of amyloodiniosis (caused by the protozoan Amyloodinium ocellatum) in a commercial aquaculture facility for hybrid striped bass (female striped bass Morone saxatilis × male white bass M. chrysops), sera from these fish, as well as from others that had been experimentally immunized with the parasite, were evaluated by an enzyme-linked immunosorbent assay for antibody specific for the parasite. Titers were similar between the fish infested in culture and the experimentally immunized fish, and were significantly higher in both the cultured and artificially exposed fish than in unexposed fish. These results suggest that an infestation of A. ocellatum can stimulate the production of humoral antibodies to the parasite, providing further evidence that natural infestations of the parasite may confer protective resistance in fish that survive the initial parasitic infestation.  相似文献   

14.
Abstract

The efficacy of hydrogen peroxide to control external parasitic infestations on juvenile (10–33-g) rainbow trout Oncorhynchus mykiss was evaluated in three clinical field trials. Fish were exposed to hydrogen peroxide concentrations ranging from 0 to 560 mg/L for 30 min once every other day for a total of three treatments. Pre- and posttreatment skin scrapes and gill wet mounts of test fish were microscopically examined to identify and enumerate external parasites. Infestation severity was classified as nonexistent (0 organisms), low (1–10 organisms), moderate (11–20 organisms), or high (?21 organisms). In trial 1, pretreatment skin examinations revealed a severe infestation of the protozoan Ambiphrya on all fish examined. Posttreatment skin examinations conducted within 24 h of the last treatment indicated that all hydrogen peroxide treatments eliminated Ambiphrya, whereas control fish remained severely infested with the protozoan. In trial 2, pretreatment examinations of skin and gill samples indicated a high infestation of the trematode Gyrodactylus (skin) and the protozoan Trichodina (gills) on all fish. Posttreatment examinations conducted within 24 h of the last treatment indicated that Gyrodactylus was eliminated from the skin of all treated fish; however, the high infestation of Trichodina remained on the gills of the test fish. All control fish had high infestation levels of both parasites. A high infestation of Ambiphrya was found on the skin of test fish before treatment (trial 3). Posttreatment examinations conducted 14 d after the last treatment revealed that 56% of the fish were parasite free, whereas the remaining test fish had low infestation levels. Control fish remained severely infested with the parasite. Based on the efficacy data, all hydrogen peroxide treatment regimens were efficacious in the control of Ambiphrya and Gyrodactylus.  相似文献   

15.
Abstract

Proliferative gill disease (PGD) was first reported in channel catfish Ictalurus punctatus at commercial farms in 1981 and is caused by the myxozoan parasite Henneguya ictaluri. The disease affects the gills and is characterized by severe branchial inflammation, epithelial hyperplasia, lamellar fusion, and lysis of chondrocytes. Presumptive diagnosis is based on the presence of lytic areas in the cartilage of the primary lamellae on microscopic examination and is confirmed histologically by the presence of the organism. In these trials, PGD was induced by exposing channel catfish fingerlings to fresh or aged infectious water collected from a pond containing fish diagnosed with severe PGD. The severity of disease was graded by histological scoring and microscopic examination of wet mounts to determine the percentage of gill filaments containing chondrolytic lesions. Exposure of fish to infectious pond water was shown to produce pathological lesions consistent with PGD, and the percentage of gill filaments containing chondrolytic lesions was positively correlated with histological scoring of gill pathology. The number of trophozoite stages in the gills was shown to increase with the severity of the disease. In most cases, however, parasitic cells were not observed in tissue samples with chondrolytic lesions during the early stages of infection. These observations indicate that pathology and lysis of chondrocytes can occur prior to detection of the organism by histopathology. Exposing fish to infectious pond water that was aged for 1 d produced negligible gill pathology and implies that the infectivity of the H. ictaluri actinospore stage is short lived. Removing fish from the source of infection promoted repair of damaged gill tissue; within 14 d of fish transfer to clean water, gill pathology associated with the acute infection was negligible.  相似文献   

16.
Abstract

Length of survival of the monogenetic trematode Gyrodactylus stellatus in serum and mucus collected from English soles Pleuronectes vetulus at different stages of a laboratory epizootic suggests that both the mucus and serum may be involved in resistance to the parasite. In general, trematode survival was shorter in the serum and mucus samples collected from English soles at the later, recovering stages of infection. A rabbit antiserum against English sole whole serum was used in a gel diffusion immunoassay to show that mucus from infested English soles contained proteins antigenically similar to English sole serum proteins. Precipitation reactions appeared strongest in mucus collected during later stages of infection. No precipitation reactions were detected in the mucus of uninfested fish, indicating that the precipitation bands that were observed were associated with G. stellatus infections.  相似文献   

17.
Abstract

We examined the gills of wild fish collected from central Texas for Centrocestus formosanus metacercariae to determine whether this temperature-restricted parasite had invaded the thermally dynamic Guadalupe River via an introduced population in its thermally stable tributary, the Comal River. We collected fish from three sites in the Guadalupe River near its confluence with the Comal River (upstream, at, and downstream) and one site in the Comal River. Centrocestus formosanus infected 14 of the 25 species examined (56.0%) and 171 of the individual fish (27.1%). Several of the infected fish represent new host records for the parasite, and two are listed as species of special concern by the state of Texas. Mean metacercarial intensities varied from 8 to 616 among species, and the highest recorded intensity was greater than 800 in two Guadalupe roundnose minnow Dionda nigrotaeniata. Among the 24 species examined from the Guadalupe River, 11 (45.8%) were infected with C. formosanus. Thorough surveys at the study sites yielded no living specimens of the first obligate intermediate snail host (red-rim melania Melanoides tuberculatus), which must be present to perpetuate the parasite. Thus, the infections were probably due to drifting cercariae that had been shed into the water column upstream of the study area in the Comal River. We therefore investigated spatial patterns in cercarial acquisition using caged fish to determine whether drifting cercariae were present in the water column at the study sites. Of 57 uninfected blacktail shiners Cyprinella venusta exposed to Guadalupe River water downstream from and at the confluence, 52 (91.2%) became infected with C. formosanus metacercariae at a mean rate of 4 metacercariae/d. This finding extends the known geographic range of this invasive exotic parasite and is the first report of the life cycle being advanced in the fish assemblage of a thermally variable temperate stream in the USA.

Received October 18, 2010; accepted April 7, 2011  相似文献   

18.
The recent identification of multiple genotypes of the salmonid parasite Ceratonova shasta with different virulence levels in Chinook Salmon Oncorhynchus tshawytscha suggests that it is possible to immunize fish against subsequent infection and disease. We hypothesized that exposure of Chinook Salmon to the less-virulent parasite genotype (II) prior to the more virulent parasite genotype (I) would decrease disease and/or result in fewer mature parasites compared with fish only infected with the more virulent genotype. To test this hypothesis, fish were challenged in a combination of field and laboratory exposures, and we measured infection prevalence, percent morbidity, and mature parasite production. Neither mortality nor mature parasite production were reduced when fish were exposed to genotype II prior to genotype I compared with fish exposed only to genotype I, suggesting that protection against C. shasta using a less-virulent genotype of the parasite does not occur.

Received June 9, 2014; accepted September 19, 2014  相似文献   


19.
An epidemiological cohort study of Cryptosporidium scophthalmi in cultured turbot Psetta maxima L. of Northwestern Spain was conducted along a four-year period. Four different ongrowing cohorts were monitored monthly from introduction into the ongrowing tanks (10–50 g) until reaching market size (400–1400 g). The association of host and environmental factors with five categories of parasite abundance was assessed using a multivariable regression framework. Epidemiological factors assessed here were water temperature, weight, length, month of collection, season, age, origin, condition factor, water filtration, and status to the myxozoan Enteromyxum scophthalmi infection. E. scophthalmi was included into the analysis because it targets the same organ than C. scophthalmi and it was prevalent in the studied population. The multivariable analysis demonstrated the statistically significant association between several factors and parasite abundance. C. scophthalmi abundance was associated (P < 0.05) with age, condition factor, season, and status to E. scophthalmi infection. Young animals, with poor condition factor, during spring or summer, and not infected with the myxozoan were most likely to be highly infected by C. scophthalmi. Inclusion of these four variables significantly (P < 0.05) improved the model, compared to the model that did not include any of these epidemiological factors. Increasing levels of C. scophthalmi abundance were associated (P < 0.01) with higher severity of C. scophthalmi-compatible lesions. The frequency of distribution of C. scophthalmi abundance was clearly right-skewed and fitted a negative binomial distribution, whereas the intensity of infection fitted a Poisson distribution. The quantification of the variance-to-mean ratio stratified by age demonstrated overdispersion for 8–16 months old fish, although this bivariate association is likely affected by several other factors, as suggested by the results of the multivariable analysis. The negative relation between C. scophthalmi abundance and status to E. scophthalmi infection suggests differences in the transmission, onset, and course of both infections. The coarse filtration used in some cohorts did not significantly affect the levels of infection. C. scophthalmi was probably introduced into the ongrowing tanks mainly with carrier fish, though the involvement of infective oocysts from the water supply cannot be disregarded. Infection prevalence and mean intensity decreased with fish age and a seasonal distribution was found. Results presented here will help to understand the epidemiology of C. scophthalmi in turbot, to estimate the expected levels of infection associated with presence or absence of epidemiological factors, and to quantify the impact that the disease may have on susceptible turbot populations. The multivariable model used here is more powerful than the visual inspection of graphics for exploring associations in cooperative processes and can be easily extended to the assessment of epidemiological associations in other population and parasitic diseases.  相似文献   

20.
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.  相似文献   

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