Description of two new actinosporean types from a brook of Fuji Mountain,Honshu, and from Chitose River,Hokkaido, Japan

Actinospore infection of oligochaetes living in the mud of 3 freshwater biotopes in Japan was studied. Using the cell-well plate method, a new aurantiactinomyxon type was found in 0.77% of the examined Tubifex tubifex oligochaete specimens from a brook near Yamanashi Prefectural Fisheries Experimental Station on Fuji Mountain. In 0.14% of Lumbriculus variagetus collected from Chitose River, near Chitose Salmon Hatchery, a new siedleckiella type was found, while at the same time 8.1% of the Lumbriculus spp. oligochaetes released triactinomyxons of Myxobolus arcticus. Of the examined Rhyacodrilus komarovi oligochaetes collected from the Mena River system, Hokkaido, 0.2, 0.6, 0.5 and 0.8% were infected with echinactinomyxon, neoactinomyxum and 2 types of triactinomyxon spores, respectively, and described in our previous paper. The oligochaetes released actinospores for several weeks. Actinospore infection showed high intensity in positive oligochaetes in the case of all the actinosporean types. Two of the actinospore types (aurantiactinomyxon and siedleckiella) presented here have not been previously described.     

Actinosporean infections in oligochaetes in a river system in southwest Ireland with descriptions of three new forms

Sediment samples were collected in April, June and August 1998 from a river in southwest Ireland. A total of 1,836 oligochaete specimens were collected from these sediments, and examined for actinosporean infections. Actinosporean-infected oligochaetes have been determined, symptoms and course of infection have been considered and the spores released from them have been described. Spore characteristics for each actinosporean were defined and the prevalence of infection in each oligochaete species was also determined. The overall prevalence of infection with actinosporeans within the oligochaete sample was approximately 2%. Seven types were distinguished, belonging to echinactinomyxon, aurantiactinomyxon and neoactinomyxum forms, from Tubifex ignotus, Lumbriculus variegatus and Limnodrilus hoffmeisteri, and compared with actinosporean types previously reported in the literature. Three of these forms have not been previously described, while the other 4 are morphologically and morphometrically similar to known forms. These actinosporeans may be stages in myxozoan life cycles.     

Description of a new actinosporean type from South African freshwaters

Actinospore infection of oligochaetes collected from the mud of 2 freshwater biotopes in South Africa was studied. Using the 'cell-well plate method', a new aurantiactinomyxon type was found in 1.1% of the examined Branchiura sowerbyi oligochaete specimens from the Rietvlei River, north of Johannesburg, Gauteng, South Africa. In 1.5% of B. sowerbyi collected in a pond (Padda Dam), near the Rand Afrikaans University, Johannesburg, the same aurantiactinomyxon type was found. Infected oligochaetes were found only after collection and no actinosporean release was recorded in Branchiura specimens kept alive for several weeks. Actinospore infection showed a high intensity in oligochaetes in both positive cases. Until now, no actinosporean stages of myxosporeans have been described from South Africa. The aurantiactinospore type presented in this communication differs from the known types already described in the literature.     

Actinosporean stages of Myxozoan parasites of oligochaetes from Nera River basin, Central Italy

The aim of this study was to check the occurrence of actinosporean stages of Myxozoan parasites of oligochaetes in the Nera River basin (Central Italy). The investigation was conducted near twelve trout farms (rainbow and brown trout) along the Nera River and in the littoral of the Piediluco Lake. Four actinosporean types, belonging to the collective groups aurantiactinomyxon, triactinomyxon, raabeia and echinactinomyxon, were detected in some of the biotopes investigated (Piediluco Lake and Sordo River). Differential diagnosis, carried out on a sub-sample of spores, showed a complete fitting with forms previously described in literature, with the exception of the triactinomyxon type.     

Description of a new synactinomyxon type from the River Sousa, Portugal

Actinospore infection of oligochaetes collected from the mud of 2 freshwater biotopes in Portugal was studied. Using the 'cell-well plate method', a new synactinomyxon type was found in 2 specimens (1.3%) of the examined Tubifex tubifex oligochaetes from the River Sousa north of Porto, Portugal. In Criodrilus lacuum and Dero digitata specimens collected from the same river, no actinosporeans were released during the 12 wk observation period. Infected oligochaetes were only found immediately post-collection, and no further actinosporean release was recorded in Tubifex specimens kept alive for several weeks. Actinospore infection showed high intensity in oligochaetes in both positive cases. No actinosporean stages of myxosporeans have as yet been described from Portugal. On the basis of spore morphology and 18S rDNA sequence data, the synactinomyxon type presented in this paper differs from those already known and described in the literature.     

Development of Myxobolus portucalensis Saraiva & Molnár, 1990 (Myxosporea: Myxobolidae) in the oligochaete Tubifex tubifex (Müller)

The extrapiscine development of Myxobolus portucalensis, a myxosporean parasite of the subcutaneous connective tissue of eel Anguilla anguilla L. was studied in the experimentally infected oligochaete Tubifex tubifex. After infecting parasite-free tubificids with mature spores of M. portucalensis, development of actinosporean stages was first observed with a light microscope 26 days after infection. Triactinospores of M. portucalensis emerged from the worms after 160 days of intra-oligochaete development. It was observed in histological sections that early pansporocysts were located in the gut epithelium of the experimental oligochaetes. Mature pansporocysts, each containing eight triactinospores, appeared 149 days after infection. After the rupture of pansporocysts, free actinospores were found in the lumen of the oligochaete gut. Released actinospores were floating in the water and showed a typical triactinomyxon form. Each triactinospore had three pyriform polar capsules, a sporoplasm with 32 secondary cells, a moderately long style and three slightly curved, sharply pointed caudal processes. The prevalence of infection in T. tubifex proved to be 52.5% (n = 40). No infection was found in Limnodrilus hoffmeisteri or in control Tubifex specimens     

Molecular relationships and phylogeny in a community of myxosporeans and actinosporeans based on their 18S rDNA sequences

The community of myxosporeans and actinosporeans inhabiting a typical Scottish highland stream and the outflow area of an adjacent salmon hatchery was analysed on the basis of their 18S rDNA sequences. Nine myxosporeans belonging to the genera Sphaerospora, Chloromyxum, Zschokkella, Myxidium, Hoferellus and Myxobilatus were identified from mature spores in different organs of the fish species present. Twelve actinosporean types belonging to the collective groups of neoactinomyxum, aurantiactinomyxon, raabeia, echinactinomyxon and synactinomyxon were found to be released from oligochaete worms collected from sediments. Twenty of the 21 sequences obtained from these myxozoans are new entries to the myxozoan database, and the genera Chloromyxum, Hoferellus and Myxobilatus were entered for the first time. Study of the molecular relationships between the different taxa and with other myxozoan sequences available showed that the myxosporeans inhabiting the urinary system clearly cluster together, independently of host species or spore morphology. However, the sequences of the two Sphaerospora species encountered show considerable differences from other members of this group and all other freshwater myxosporeans, and they were found to occupy an ancestral marine position. Three actinosporeans, i.e. Neoactinomyxum eiseniellae, Aurantiactinomyxon pavinsis and Raabeia 'type 3' were found to represent alternate life cycle stages of Chloromyxum sp., Chloromyxum truttae and Myxidium truttae, respectively (approximately 1400 identical base pairs each). Three other actinosporeans encountered (two echinactinomyxon and one raabeia type) showed over 92% sequence identity with myxosporeans from GenBank, whereas all other actinosporeans formed a closely related group devoid of any known myxosporeans.     

Environmental factors affecting the distribution and abundance of cyst-forming Myxobolus spp. and their cyprinid hosts in 3 lakes in Algonquin Park,Ontario

In 1999, 4 species of cyprinid were surveyed for myxozoan parasites in a watershed in Algonquin Park, Canada, Kathlyn Lake. Broadwing Lake, and Lake Sasajewun were included. Eight species of Myxobolus were found that differed in their prevalence and distribution among the 3 lakes. The oligochaetes and environmental parameters, including sediment types and aquatic plants, of these 3 lakes were surveyed the following year. Oligochaetes belonging to 17 species were collected from the 3 lakes. The distribution patterns of the oligochaete fauna, with respect to the environmental variables, were examined using canonical correspondence analysis. Naidids were predominant in all 3 lakes, particularly in the pebbly and sandy sediment of Lake Sasajewun. The highest percentage of tubificids occurred in the detritus and muddy substrate of Broadwing Lake. These findings indicate that the prevalence of certain oligochaetes is congruent with the absence or presence of particular myxozoan species and that substrates and aquatic plants influence the distribution of certain oligochaete species.     

Biodiversity of groundwater oligochaetes from a karst unit in northern Iberian Peninsula: ranking subterranean sites for conservation management

The present study suggests that the Santa Eufemia-Ereñozar karst unit in northern Iberian Peninsula is a biodiversity hotspot for groundwater oligochaetes, due to (1) the presence of a high number of stygobiotic species (corresponding to 18% of the total stygobionts known in southern Europe); (2) the comparatively high number of oligochaete species collected (corresponding to 35% of the total epigean and hypogean oligochaete species in the region); and (3) the presence of 5 species endemic to the region. A list of the oligochaete taxa found in the karst unit is presented and a conservation ranking of the cavities in the karst is proposed based on the application of four biodiversity indices (Species richness, Rarity, Vulnerability and Complementarity) to the oligochaete taxa. Vulnerability was evaluated for the first time for groundwater oligochaete taxa and it provided a useful tool to assess the protection status of oligochaetes in karstic systems. Groundwater conservation management strategies could incorporate biodiversity data from the present study.     

Detection of myxozoan parasites in oligochaetes imported as food for ornamental fish

To determine the potential for dissemination of myxozoan parasites by transfer of their alternate oligochaete hosts, shipments of tubificid worms obtained from an overseas commercial aquarium supplier were screened for actinospore stages of myxozoan parasites. At least 7 different triactinomyxon types were identified. The morphological characteristics of the actinospores recovered from these tubificids shared characteristics with triactinomyxons characterized in other surveys, particularly from eastern Europe. Analysis of the screened samples by polymerase chain reaction and comparison of morphological data indicated that these actinospores did not correspond to the triactinospore of Myxobolus cerebralis. Although identification of these triactinomyxon types was beyond the scope of this study, these data suggest that the unregulated import-export and exchange of live organisms for ornamental fish food may result in accidental introduction or dissemination of myxozoan parasites.     

Molecular characterization of myxozoan parasites from Lake Sasajewun, Algonquin Park, Ontario, by riboprinting

The small subunit-rRNA genes of 18 myxozoans from Lake Sasajewun, Algonquin Park were amplified and digested with restriction endonucleases for riboprinting analysis. Identical riboprints were not found between the myxosporeans and the actinosporeans. The distinct riboprinting patterns observed among these myxozoans indicate considerable genetic diversity within this group. Identical riboprints were found between Myxobolus pendula and Myxobolus pellicides, and between triactinomyxon 'C' and Triactinomyxon ignotum. Parsimony analysis of the riboprints demonstrated that neither the myxosporeans nor the actinosporeans formed a monophyletic group. Some species of Myxobolus are more closely related to forms of triactinomyxon, echinactinomyxon or raabeia than to other Myxobolus species. These results are consistent with the two-host life cycle hypothesis of myxozoans that myxosporeans and actinosporeans are alternating stages of the same organisms.     

Prevalence and susceptibility of infection to Myxobolus cerebralis,and genetic differences among populations of Tubifex tubifex

The prevalence of infection and susceptibility of the aquatic oligochaete Tubifex tubifex to Myxobolus cerebralis, was examined in 2 studies on the upper Colorado River, Colorado, USA, where whirling disease occurs in wild trout populations. In the first study, the prevalence of infection ranged from 0.4 to 1.5%, as determined by counting the number of T. tubifex releasing triactinomyxons of M. cerebralis directly following their collection from the field. The susceptibility of those T. tubifex not releasing triactinomyxons was assessed by the number of these oligochaetes releasing triactinomyxons 3 mo following experimental exposures to spores of M. cerebralis. The prevalence of infection following experimental exposures of these T. tubifex ranged from 4.2 to 14.1%. In a second study, all T. tubifex collected at 2 different times directly from the 2 field sites in Colorado were exposed to spores of M. cerebralis. Individual oligochaetes representing those groups of T. tubifex releasing and those groups not releasing triactinomyxons at 3 mo were screened with molecular genetic markers. T. tubifex populations found at the 2 study sites consisted of 4 genetically distinct lineages that varied with respect to their susceptibility to experimental exposure to M. cerebralis. Lineages I and III contained the most oligochaetes susceptible to M. cerebralis and were the most prominent lineages at Windy Gap Reservoir, a site of high infectivity for wild rainbow trout on the upper Colorado River. In contrast, at the Breeze Bridge site which is below Windy Gap Reservoir and where M. cerebralis infections are less severe in wild trout, oligochaetes in lineages V and VI that are resistant to M. cerebralis were more prominent. These results suggest that certain habitats, such as Windy Gap Reservoir, are conducive to large and more homogenous populations of susceptible T. tubifex lineages that may serve as point sources of infection for M. cerebralis. Although not a direct objective of this study, there was no evidence of M. cerebralis infections among any oligochaetes other than those that would be classified as T. tubifex by standard morphological characteristics.     

Differential propagation of the metazoan parasite Myxobolus cerebralis by Limnodrilus hoffmeisteri, Ilyodrilus templetoni, and genetically distinct strains of Tubifex tubifex

Whirling disease, caused by the parasite Myxobolus cerebralis, has infected rainbow trout (Oncorhynchus mykiss) and other salmonid fish in the western United States, often with devastating results to native populations but without a discernible spatial pattern. The parasite develops in a complex 2-host system in which the aquatic oligochaete Tubifex tubifex is an obligate host. Because substantial differences in whirling disease severity in different areas of North America did not seem explainable by environmental factors or features of the parasite or its fish host, we sought to determine whether ecological or genetic variation within oligochaete host populations may be responsible. We found large differences in compatibility between the parasite and various laboratory strains of T. tubifex that were established from geographic regions with different whirling disease histories. Moreover, 2 closely related species of tubificids, Limnodrilus hoffmeisteri and Ilyodrilus templetoni, which occur naturally in mixed species assemblages with T. tubifex, were incompatible with M. cerebralis. Virulence of the parasite was directly correlated with the numbers of triactinomyxon spores that developed within each strain of T. tubifex. Thus, the level of virulence was directly related to the compatibility between the host strain and the parasite. Genetic analyses revealed relationships that were in agreement with the level of parasite production. Differences in compatibilities between oligochaetes and M. cerebralis may contribute to the spatial variance in the severity of the disease among salmonid populations.     

Description of raabeia,synactinomyxon and neoactinomyxum developing stages of myxosporeans (Myxozoa) infecting Isochaetides michaelseni Lastočkin (Tubificidae) in Lake Balaton and Kis-Balaton Water Reservoir,Hungary

Molecular and morphometric investigations were conducted on the actinosporean morphotypes of myxosporeans surveyed in oligochaetes of Lake Balaton and Kis-Balaton Water reservoir. Oligochaetes belonging to the species Isochaetides michaelseni Lasto?kin and Branchiura sowerbyi Beddard as well as to the genera Nais Dujardin, Dero Müller and Aeolosoma Ehrenberg were studied during an 18-month period. Actinosporeans were obtained exclusively from I. michaelseni (7,818 specimens) with very low prevalence (0.01–0.06%). Four new actinosporean morphotypes of the collective groups raabeia (2 types), synactinomyxon (1 type) and neoactinomyxum (1 type) were found and described, including the first synactinomyxon collective group from Hungarian biotopes and a new raabeia morphotype. Except for Synactinomyxon type 1, the 18S rDNA analysis revealed that the spores did not match any myxospore entity found in the GenBank.     

Distribution and ecology of Middle Atlantic Bight Oligochaeta

Distribution and ecology of continental shelf oligochaetes are poorly understood. From 1975 through 1977 baseline ecological studies were conducted in the Middle Atlantic Bight prior to oil exploration activities. A total of 38 species in three oligochaete families were collected. Of this total one family and 29 of the species were undescribed at the time of collection. The most abundant and widely distributed species were Limnodriloides monothecus, L. medioporus, Bathydrilus longus, Phallodrilus biprostatus, and Tubificoides diazi. No single species dominated the oligochaete fauna. All the oligochaete species responded to changes in physical parameters over the shelf similarly to the total macrofauna. Seasonal trends within the oligochaetes were not found. Species that preferred coarser sand sediments were all small interstitial forms. The finer sand and eurytopic species were all burrowers. Zoogeographically the Middle Atlantic Bight oligochaetes were predominantly temperate (16 species) with some southern species (8). Seven species were broadly distributed from the Caribbean to Massachusetts or around the world. Seven species were only known from the area studied.Contribution No. 1397 of the Virginia Institute of Marine Science.     

Factors influencing the distribution of Myxobolus cerebralis,the causative agent of whirling disease,in the Cache la Poudre River,Colorado

Oligochaetes, triactinomyxons (TAMs), and age-0 trout were sampled in the upper Cache la Poudre River, Colorado, to determine the distribution of Myxobolus cerebralis during 1997 and 1998. Densities of the intermediate host, the oligochaete Tubifex tubifex, were 3.5 orders of magnitude higher in the M. cerebralis-infected Poudre Rearing Unit (PRU) trout rearing ponds than at any of the river sampling reaches. Oligochaetes, including T. tubifex, were rare in the river (1 oligochaete m(-2)), except in a few stream side alcoves and eddies (50 oligochaete m(-2)). Species composition of oligochaetes in the river reaches was more diverse than in the PRU. Tubifex tubifex constituted 50% or less of the oligochaete community in the river and 98% in the PRU. Infection rates of T tubifex were 1% in the area above the PRU, 2% in the PRU, and 6% below the PRU. An increased M. cerebralis intensity of infection in age-0 trout below the PRU could not be attributed entirely to the high numbers of TAMs in its effluent (3.7 TAMs l(-1)). Low densities of TAMs ranging from 0 to 0.2 TAMs l(-1) were found in the river reaches, yet nearly all of the age-0 trout were infected soon after emergence. This suggests that very few TAMs, as measured by filtration, need be present in the water column to bring about infection in the majority of trout present. This also indicates that the parasite can persist and potentially cause reduced juvenile trout recruitment in cold, oligotrophic, sediment poor, high-gradient streams.