Susceptibility-related differences in the quantity of developmental stages of Myxobolus spp. (Myxozoa) in fish blood.

Here, we investigated the early development of two closely related myxozoan parasites, the highly pathogenic Myxobolus cerebralis, the causative agent of the whirling disease in salmonids, and Myxobolus pseudodispar, a common, non-pathogenic parasite of cyprinids. The aim of our study was to examine under in vivo laboratory conditions whether fish blood is involved in the intrapiscine development of the two parasite species and investigate if there is dissimilarity between the parasite infection intensity in blood and if it varies in terms of host susceptibility and parasite pathogenicity. Highly susceptible, less susceptible and non-susceptible hosts were involved. Blood samples were taken 1 day, 1 week and 1 month post exposure to M. cerebralis and M. pseudodispar, respectively. The prevalence and infection intensity was estimated by parasite-specific quantitative real-time PCR. Although previous findings assumed that M. cerebralis might escape from host immune system by migrating via peripheral nerves, our experimental results demonstrated that M. cerebralis is present in blood during the early stage of intrapiscine development. For the non-pathogenic M. pseudodispar, the highest infection prevalence was found in the original host, common roach Rutilus rutilus, whereas the highest infection intensity was detected in rudd Scardinius erythrophthalmus, a "dead-end" host of the parasite. The presence of M. pseudodispar developmental stages in the blood of both susceptible and non-susceptible cyprinids suggests that the susceptibility differences remain hidden during the early stage of infection. Our findings supply further evidence that host specificity is not determined during the early, intrapiscine development involving the vascular system. Furthermore, we found remarkable differences in the infection dynamics of the two parasite species examined, possibly due to their distinct pathogenicity or variations in adaptive capabilities to immune components in host blood.

Counter-insurgents of the blue revolution? Parasites and diseases affecting aquaculture and science.

Aquaculture is the fastest-growing segment of food production and is expected to supply a growing portion of animal protein for consumption by humans. Because industrial aquaculture developed only recently compared to industrial agriculture, its development occurred within the context of a growing environmental awareness and acknowledgment of environmental issues associated with industrial farming. As such, parasites and diseases have become central criticisms of commercial aquaculture. This focus on parasites and diseases, however, has created a nexus of opportunities for research that has facilitated considerable scientific advances in the fields of parasitology and aquaculture. This paper reviews Myxobolus cerebralis , Lepeophtheirus salmonis , white spot syndrome virus, and assorted flatworms as select marquee aquaculture pathogens, summarizes the status of the diseases caused by each and their impacts on aquaculture, and highlights some of the significant contributions these pathogens have made to the science of parasitology and aquaculture.

Life cycles of three Myxobolus spp. from cyprinid fishes of Lake Balaton, Hungary involve triactinomyxon-type actinospores.

A study on the actinosporean fauna of oligochaetes of Lake Balaton was carried out from 2009 to 2011. The morphology of actinosporean stages of myxosporeans obtained from oligochaetes was studied, and their 18S rDNA structure was analyzed by molecular biological methods. Three triactinomyxon types were released from the oligochaete Isochaetides michaelseni (Tubificidae). The sequences of Triactinomyxon type 1 proved to be identical with those of Myxobolus fundamentalis. The sequences of Triactinomyxon type 2 showed 99.9% similarity to Myxobolus eryhtrophthalmi, while the sequences of Triactinomyxon type 3 showed a 99.9% similarity to those of Myxobolus shaharomae. The life cycles of the above species, just like those of other species with a known life cycle, suggest that most Myxobolus spp. develop through triactinomyxon-type actinosporean stages.

Effects of habitat alteration on the epizootiology of Myxobolus cerebralis, the causative agent of salmonid whirling disease.

Whirling disease, caused by the myxozoan parasite Myxobolus cerebralis , is a serious health threat to salmonid fish and its control remains problematic. The parasite has a 2-host life cycle involving a salmonid and the aquatic oligochaete Tubifex tubifex . A commonly used strategy to control parasites that requires an obligatory invertebrate host is to eliminate or reduce the host population size to a point where parasite transmission can no longer occur. Large numbers of T. tubifex are frequently found in degraded habitats that are characterized by an abundance of fine sediments, organic matter, and a lack of aquatic invertebrate diversity. If such environments are rehabilitated, then the normal flora and fauna should re-establish and the numbers of T. tubifex should decline due to their inability to compete with the re-established invertebrates. During an epizootiological study on Rock Creek, located in west-central Montana, 2 opportunities were available to examine the effects of habitat restoration on the transmission of M. cerebralis . The Puyear Ranch re-establishment project was a major endeavor conducted on the main channel of Rock Creek, a little more than midway upstream. Another significant restoration was conducted on Upper Willow Creek, a tributary of Rock Creek, located closer to the headwaters. Sentinel trout studies, along with examining T. tubifex for the parasite and measuring various water-quality parameters, revealed that the restoration of the Puyear Ranch locality had no significant effect on reducing the intensity of M. cerebralis in trout. This was likely due to the restored area being located mid-river, just downstream from a "hot spot" of infected T. tubifex . In comparison, there was a significant reduction in the intensity of M. cerebralis in sentinel fish after the Upper Willow Creek restoration project was completed. Unlike the Puyear Ranch locality, there was no hot spot of infected T. tubifex above the area rehabilitated on Upper Willow Creek. Further, the relative abundance of T. tubifex and M. cerebralis -infected worms was reduced. Although further study is needed, it appears that habitat rehabilitation can reduce the transmission of M. cerebralis . Since the triactinomyxon stage of the parasite released from T. tubifex (which infects trout) can float for many kilometers, the rehabilitation of a hot spot may reduce the infection of trout downstream where they inhabit a healthy environment with no M. cerebralis -infected T. tubifex in the vicinity. Thus, rehabilitation of a relatively small area could significantly affect the drainage for many kilometers beyond the improved habitat.

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.

The susceptibility of diverse species of cultured oligochaetes to the fish parasite Myxobolus pseudodispar Gorbunova (Myxozoa).

This study provides detailed information on the invertebrate hosts of Myxobolus pseudodispar (Myxozoa) and explores the susceptibility range of several species and analyses the relevance of the species composition of an oligochaete population. Our findings demonstrate that the oligochaete host range of M. pseudodispar is similarly wide as the number of vertebrate host species. Besides Tubifex tubifex and Limnodrilus hoffmeisteri, Psammoryctides barbatus and Psammoryctides moravicus were also found to be susceptible invertebrate hosts. The genetic characterization of the mitochondrial 16S rDNA of T. tubifex sensu lato revealed that lineages I, II and III are susceptible to M. pseudodispar, whereas T. tubifex lineage VI seems to be non-susceptible. T. tubifex lineage V and L. hoffmeisteri specimens were positive in a M. pseudodispar-specific PCR, but in most cases, the release of mature actinospores could not be detected. Hence, these non-susceptible oligochaetes likely serve as `biological filters` as they remove myxospores from the sediment without producing actinospores. Together with the phylogenetic analysis of the susceptible and non-susceptible oligochaete hosts on the basis of mt 16S rDNA sequences, the route of the development of M. pseudodispar in the oligochaete hosts was tracked by in situ hybridization. According to our findings, the gut epithelia seem to be a portal of entry of the sporoplasms, where the development of the parasite also takes place. The basal lamina seems to be involved in the migration of the parasite, and the worm's cellular immune response is activated by the infection.

The development of Myxobolus pavlovskii (Myxozoa: Myxobolidae) includes an echinactinomyxon-type actinospore.

Echinactinomyxon-type actinospores were found in a mixed-species oligochaete culture originating from the Temperate Water Fish Hatchery near Budapest, Hungary. On the basis of DNA sequence analysis, the actinospores were identified as Myxobolus pavlovskii (Akhmerov, 1954), the 18S rDNA sequence from myxospores of which is available in GenBank. Silver carp Hypophthalmichthys molitrix (Valenciennes) fry specimens were successfully infected by cohabitation with the echinactinomyxon-releasing oligochaetes, which confirmed the molecular data congruence. The echinactinomyxons and the myxospores that developed in the gills of exposed fish fry were analysed morphologically and on DNA basis. The infected gill tissue was examined histologically. As typical characters of M. pavlovskii, numerous small plasmodia were observed in the epithelia of gill lamellae. Plasmodia contained thousands of myxospores with polar capsules unequal in size and witl; large intercapsular processes. The 18S rDNA sequence from actinospores and those from myxospores originating from the experimentally infected fish were identical. The oligochaete species releasing actinospores was morphologically determined as Limnodrilus sp. This is the first record of an echinactinomyxon as an alternate stage within the genus Myxobolus.

Complete life cycle of Myxobolus rotundus (Myxosporea: Myxobolidae), a gill myxozoan of common bream Abramis brama.

The life cycle of Myxobolus rotundus Nemeczek, 1911, a myxosporean parasite of the gills of common bream Abramis brama L., was studied under laboratory conditions. Mature Myxobolus spp. spores from plasmodia in the gills of wild bream were used to infect naïve oligochaete worms in a flow-through system of aquaria. Triactinomyxon-type actinospores were released from the oligochaetes 1 yr later and allowed to continually flow into a tank containing uninfected bream fry. The gills of the fry were checked for development of plasmodia in squash preparations 3 d postexposure, and then at weekly intervals for 8 wk. Tissue samples were fixed at each time point. Developing plasmodia were first observed 17 d post-exposure (Day 17). Mature spores were collected from plasmodia on Day 56 and were added to plastic dishes containing parasite-free Tubifex tubifex oligochaetes. Second-generation actinospores were released from these worms 8 mo post-exposure, and were morphologically identical to first-generation spores. Myxospores obtained from the bream fry were morphologically identical to those identified in wild bream as M. rotundus. Small subunit ribosomal RNA gene sequences obtained from first- and second-generation actinospores and the bream fry myxospores were 100% similar to M. rotundus spores from the original wild fish.

The life cycle of Myxobolus lentisuturalis (Myxozoa: Myxobolidae), from goldfish (Carassius auratus auratus), involves a Raabeia-type actinospore.

We studied a natural infection of the oligochaete Branchiura sowerbyi Beddard, 1892 with the Raabeia-type actinosporean stage of Myxobolus lentisuturalis Dyková, Fiala et Nie, 2002 which infected goldfish Carassius auratus auratus (L.) in Italy, using molecular analysis of the SSU rRNA gene. The existence of intraoligochaete development shows that this parasite follows the life-cycle pattern described by Wolf and Markiw (1984) for Myxobolus cerebralis. Histological examinations of the goldfish infected by M. lentisuturalis showed at low magnification the presence of two bilateral crescent-shaped masses in the dorsal epaxial muscle. These lesions were not circumscribed, presented irregular edges and infiltrated the underlying bundles of skeletal muscle and interstitial tissue. At higher magnification, disappearance of muscle fibres and substitution of the muscle tissue with Myxobolus spores and plasmodia were observed.

Differentially expressed parasite genes involved in host recognition and invasion of the triactinomyxon stage of Myxobolus cerebralis (Myxozoa).

The host recognition and invasion process of Myxobolus cerebralis actinospores (triactinomyxon, TAM) was studied on a genetic level. A small-scale in vitro assay was developed to activate a large number of TAMs simultaneously, and to monitor the host invasion in the absence of live fish. The transcriptomes of non-activated and in vitro-activated TAMs were compared by suppressive subtractive hybridization (SSH) to identify parasite genes involved in the host invasion process. Differential screening and a subsequent BLAST search revealed 15 of 452 SSH-library clones expressed differently in activated TAMs. None of the 15 transcripts obtained has previously been identified from M. cerebralis. Quantitative real-time PCR was used to examine the relative expression profile of 8 selected transcripts upon TAM activation and after penetration of the host. Four of these were found to be up-regulated in activated TAMs, while expression was relatively low in non-activated TAMs and in infected fish tissue, indicating that they are relevant genes during host recognition or subsequent host invasion of M. cerebralis TAMs.