Myxosoma cerebralis (Myxozoa: Myxosporea) Etiologic Agent of Salmonid Whirling Disease Requires Tubificid Worm (Annelida: Oligochaeta) in its Life Cycle

Studies of the life cycle of Myxosoma cerebralis showed that development of infectivity did not occur endogenously but that the spore “aging” process required participation of an aquatic tubificid oligochaete. Data suggestive of such involvement were derived from trials in which spores were “aged” in an array of inert, sterilized, pasteurized, or natural aquatic substrates and from examination of aquatic soils from trout hatcheries in which whirling disease was epizootic. The role of the aquatic oligochaete was confirmed two ways. First, signs of whirling disease developed, and M. cerebralis spores were produced in young rainbow trout (Salmo gairdneri) that had been fed oligochaetes harvested from pond soil taken from two hatcheries where whirling disease was epizootic. Second, when containers of pasteurized soil were populated with four genera of oligochaetes–Aeolosoma, Dero, Stylaria, or Tubifex– from a biological supply house, or with tubificid worms from trout hatcheries free of whirling disease, and then seeded with M. cerebralis spores and “aged” for 4 months, whirling disease occurred only in trout held with Tubifex and with hatchery tubificids.     

Models of stream habitat characteristics associated with tubificid populations in an intermountain watershed

Habitat assessments were conducted in an intermountain watershed at three spatial extents to explore ways to predict the presence of tubificid oligochaetes likely to support the parasite Myxobolus cerebralis, cause of salmonid whirling disease. Stream reaches with six different reach slope characteristics were selected using GIS. The aquatic habitat in 60 reaches selected at random was measured and classified into distinct habitat units. Within the habitat units, areas of microhabitat with depositional fine sediments were chosen, measured, and core samples were removed to characterize the sediments and benthic oligochaetes. Two tubificids, Tubifex spp. and Limnodrilus hoffmeisteri, were abundant and co-occurred in silt-clay and fine sand sediments in these habitats. Models were posed and tested to predict the presence and relative abundance of tubificids using habitat characteristics from the three spatial extents: reach, habitat unit, and microhabitat. At the reach extent, tubificids were associated with low-reach slope and with slow water habitats. Within habitat units, tubificids were associated with higher percentages of fine sediments and higher stream width:depth ratios. In microhabitat cores, the presence of silt-clay sediments was positively associated with higher average stream width:depth ratios. Since ecological relationships are often scale dependent and stream systems have a natural hierarchy, predictive habitat models such as these that use measures from several scales may help researchers and managers more efficiently identify and quantify aquatic communities at highest risk of infection by the M. cerebralis parasite. Handling editor: J. Trexler     

Mysterious `crystals': found attached to the epidermal peritoneum of marine tubificid (Annelida,Clitellata) species

Marine tubificids are abundant and diverse in the carbonate sediments of Bermuda, as well as in many other tropical and subtropical locations. Recently, during microscopic observations of living specimens, crystal-like structures were observed attached to the coelomic peritoneum and in the coelomic cavity of some Bermuda species, including phallodrilines of the genera Aktedrilus and Pectinodrilus,and a rhyacodriline of the genus Heterodrilus. Similar structures were not seen in tubificid species of Thallasodrilides and other limnodriloidines, a second species of Heterodrilus, a tubificine of the genusTubificoides, a phallodriline of the genus Bathydrilus,nor in a number of marine enchytraeid genera and species found in Bermuda. The crystal-like structures have two needle arms, each about 5–10 m long and about 0.5 m in diameter, meeting at an obtuse angle. At the junction of the arms, there is a small membrane-bound `knob', about 1 m in diameter, which may be continuous with the coelomic peritoneum. The numbers of `crystals' per individual worm are estimated at 100–400 per body segment, or well over 2 × 10³ in an adult worm. `Crystals' are found: throughout the length of the worms, in all individuals of species in which `crystals' occur, and over the range of environmental conditions where these species are found in Bermuda. Simple digestions with hypochlorite, weak and dilute acids, and staining with nuclear and cytoplasmic stains indicate that the composition of the knob is organic and the arms inorganic. The fluorescent tracer Calcein (Sigma) was not incorporated into any structures during a 24-h bath incubation of living worms, and the `crystals' do not show birefringence when viewed between crossed polarizing filters. These last two results do not support an hypothesis that these are calcium carbonate `crystals'. Geographically, the crystal-like structures are widespread, and have also been observed in a species of immature (unidentified) marine tubificid from Rottnest Island, Western Australia.     

Sublittoral and profundal Oligochaeta fauna of the Lake Constance (Bodensee-Obersee)

Fourteen tubificid species and one lumbriculid make up the oligochaete population of the northern half of Lake Constance, a meso- to mesoeutrophic lake. Tubifex tubifex makes up 72% of the total abundance, the clean water species Stylodrilus heringianus only 0.1%. No correlation exists between total worm abundance and organic carbon content of the sediment, only 4% of the organic carbon being used by the worms. A highly significant correlation between particulate organic matter loading and its organic carbon content from 10 tributaries and total oligochaete abundance is demonstrated. The ecological index I_PA is defined and used to assess the biologically relevant trophic status of the lake bed. This integrates information on the percent representation of three groups of worm species classified on atrophic basis and total abundance of the worms. Despite control of the increase in phosphorus loadings progressive eutrophication of sediments was observed after an 8 year interval.     

A preliminary study of oligochaetes in Poyang Lake,the largest freshwater lake of China,and its vicinity,with description of a new species of Limnodrilus

The oligochaete fauna of the largest freshwater lake of China, Poyang Lake, has never been investigated before. On the basis of a preliminary survey in the lake and its vicinity in 1997–1999, 25 species belonging to 20 genera and 5 families are recorded. Among them, one genus, Cernosvitoviella (Enchytraeidae), and two species, Bratislavia unidentata (Naididae), Potamothrix bedoti (Tubificidae), are recorded from China for the first time. Another tubificid, Limnodrilus paramblysetus sp. nov., is new to science. It is similar to L. amblysetus Brinkhurst et al., 1990 in chaetal shape but differs in having short hooded penis sheaths.     

Present knowledge on the life cycle,taxonomy, pathology,and therapy of some Myxosporea spp. important for freshwater fish

In this review some important myxosporean species of fish are depicted and described, with special reference to Myxobolus cerebralis (causing whirling disease of salmonids), Sphaerospora renicola (causing Swimbladder inflammation of common carp), Hoferellus carassii (causing kidney enlargement disease of goldfish and proliferative kidney disease [PKD] of salmonids). Detailed information on the life cycles of M. cerebralis, M. cotti, and M. pavlovskii is presented proving that these myxosporeans develop in an intermediate host, the oligochaete worm, Tubifex tubifex to actinosporeans which represent the only infective stage for susceptible fish. These findings imply that the present taxonomy for the phylum Myxozoa has to be revised. Trends for treatment of myxosporean associated diseases are summarized with special emphasis on the antibiotic Fumagillin DCH.     

The effect of tubificids and chironomids on particle redistribution of lake sediment

The effects of chironomid larvae,Chironomus plumosus, and tubificid worms,Limnodrilus spp., on particle redistribution in lake sediment were investigated experimentally using pots containing sediments obtained from Lake Suwa, Japan. The chironomids and tubificids increased the water content of surface sediment. The chironomid larvae had no effect on particle size distribution, while tubificids continuously accumulated small particles on the surface sediment through their selective feeding activity. Particles larger than 0.125 mm were buried at a sediment depth of 6 cm. In Lake Suwa, long diatom frustules, large plant debris and blue-green algal flocs were found to accumulate in the deeper layer of the lake sediment inhabited by tubificids at high density.     

Ecological Significance of Nitrogen Cycling by Tubificid Communities in Shallow Eutrophic Lakes of the Danube Delta

Importance of tubificid populations on nitrogen cycle in two categories of shallow eutrophic lakes in the Danube Delta was quantitatively assessed for the 1992-1993 period. The structure of the primary producers in the studied lakes was used to discriminate between the two categories:(i) lakes dominated by macrophytes (A₁) and (ii) lakes dominated by phytoplankton (A₂). In both categories tubificid worms represented important fraction of the entire benthic community (35 and 32%, respectively, as number of individuals). They influence the sediment-water exchange of nutrients. The main processes involved are excretion of nutrients and their continuous release from sediments by molecular diffusion or through channels created by bioturbation. Inorganic nitrogen released from bottom sediments may regulate nitrogen load in the water body and thus, phytoplankton production. In 1992-1993, nitrogen stocks in tubificid biomass accounted for 5.3% in A₁ lakes and 15.6% in A₂ lakes of the amount stocked in phytoplankton, and only for 1.2 and 2.9% respectively, of the nitrogen load in water body. Nitrogen excretion rates ranged between 60.52 and 153.74 mg N m^–2 year^–1, and release rates from sediments between 378.26 and 960.87 mg N m^–2 year^–1, the lowest values being recorded for A₂ category. Differences are related to tubificid biomass, structure and abundance of primary producers and to nutrient load in different ecosystems. Ratios between release rate of inorganic nitrogen by tubificid worms and sedimentation rate of organic nitrogen in the two categories of lakes were 8.3 and 6.4% respectively. Contribution of nitrogen released daily from sediments to the dissolved inorganic nitrogen load in the water column was less than 0.5%. However, in A₁ and A₂ lakes, the released nitrogen had a potential to sustain 24.74 and 8.01%, respectively, of the annual phytoplankton production. These values suggest the significance of tubificids in keeping the eutrophication process at a high level, especially during the periods when nitrogen is the main limiting factor for phytoplankton production.     

Disappearance of deep profundal zoobenthos in Lake Ikeda, southern Kyushu, Japan, with relation to recent environmental changes in the lake

A benthological survey in a deep caldera, Lake Ikeda, southern Kyushu, Japan, in 1998 revealed that no zoobenthos were found in the deep profundal, although two tubificid oligochaetes, Tubifex tubifex and Limnodrilus hoffmeisteri, and a chironomid, Procladius sp., were distributed in the upper profundal zone. This is the first record of oligochaete composition in the lake. Lake Ikeda had been typically oligotrophic until the 1940s, and zoobenthic assemblages were recorded throughout the profundal bottom in the 1920s and 1970s. Recent disappearance of the deep profundal zoobenthos could be caused by the stagnation of anoxic waters in the hypolimnion, in connection with eutrophication triggered by nutrient loading, as well as change in the thermal circulation system presumably caused by global warming.     

The effects of the oligochaete Branchiura sowerbyi Beddard (Tubificidae) on the biological and chemical characteristics of overlying water and soil in a submerged ricefield soil system

Submerged ricefield soils, outdoors, and with and without the tubificid Branchiura sowerbyi Beddard, were used to study the effects of tubificids on some biotic and abiotic variables. Measurements of chemical and biological features showed that the tubificids accelerated the diffusion of dissolved substances, increased the release of soil bacteria and enhanced the production of algae and a duckweed (Lemna paucicostata) in the overlying water. In the presence of tubificids, increased numbers of Cladocera (Moina sp., Simocephalus sp.) and ostracods in the overlying water, and higher number of sulfate-reducing bacteria in the soil were observed. The tubificids further enhanced the microbial activity in the upper soil layer, which in turn produced large amounts of organic matter.     

The effect of organic pollution on the abundance and distribution of aquatic oligochaetes in an urban water basin, Taiwan

Aquatic oligochaetes are abundant in polluted areas and are, therefore, commonly used as bioindicators to study organic pollution in rivers and streams. In order to develop a species-level oligochaete biotic index to reflect the River Pollution Index (RPI) in the Taichung Water Basin in Taiwan, we conducted a systematic sampling scheme to collect aquatic oligochaetes from the sediment samples of watercourses in the Taichung Water Basin, Taiwan. We evaluated the relationships between aquatic oligochaetes and the sewage pollution using statistical methods. The distribution of aquatic oligochaetes in relation to environmental variables, such as water quality and sediment characteristics of the regional urban contaminated streams was expressed by Canonical Correspondence Analysis (CCA). We identified 17 species of aquatic oligochaetes (Annelida, Clitellata) including 3 species of Tubificidae, 13 species of Naididae, and 1 species of Enchytraeidae from the watercourses of an urban region in the Taichung Water Basin in Taiwan, during the summer and winter of 2005 and 2006. A positive correlation was found between the total abundance of aquatic oligochaetes and the RPI (r = 0.58, P < 0.05). However, only population density of the most abundant tubificid, Limnodrilus hoffmeisteri, increased with increasing RPI values and a significantly negative correlation was found between the population density of the naidid, Nais communis, and RPI values. The results of CCA indicated that certain naidids, such as Aulophorus furcatus and Allonais gwaliorensis also tolerated extremely polluted environments in upper stream or stony habitats, implying that tubificids should not be the sole representation of simple biotic indices but should also include pollution-tolerant naidids. We found that the community structure of aquatic oligochaetes was influenced by short-term variations in microhabitat rather than according to seasonal factors in our study region. The results proved that aquatic oligochaetes were sensitive enough to provide a supplement for the regional urban pollution assessment applications for biotic indicators at the species-level.     

Distribution,Abundance and Life Cycles of Oligochaeta from the Marine Hygropsammal with Special Reference to the Phallodrilinae (Tubificidae)

Tubificid and enchytraeid oligochaetes are common members of the interstitial fauna of sandy beaches. Their abundance dynamics, life cycles and distribution patterns were investigated at two beaches on the western Baltic Sea and at a North Sea beach (Isle of Sylt). Populations of 115,000 ind./m² were found at protected Baltic beaches. Reproduction of the tubificids, Phallodrilus monospermathecus and Spiridion insigne takes place within a well defined breeding period once a year. The market horizontal and vertical distribution patterns of various tubificid and enchytraeid species are elucidated by preference reactions to several physiographic parameters, such as oxygen and water content of the sediment, and salinity.     

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.     

Coexistence of Bacterial Sulfide Oxidizers, Sulfate Reducers, and Spirochetes in a Gutless Worm (Oligochaeta) from the Peru Margin

Olavius crassitunicatus is a small symbiont-bearing worm that occurs at high abundance in oxygen-deficient sediments in the East Pacific Ocean. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization, we examined the diversity and phylogeny of bacterial symbionts in two geographically distant O. crassitunicatus populations (separated by 385 km) on the Peru margin (water depth, ~300 m). Five distinct bacterial phylotypes co-occurred in all specimens from both sites: two members of the γ-Proteobacteria (Gamma 1 and 2 symbionts), two members of the δ-Proteobacteria (Delta 1 and 2 symbionts), and one spirochete. A sixth phylotype belonging to the δ-Proteobacteria (Delta 3 symbiont) was found in only one of the two host populations. Three of the O. crassitunicatus bacterial phylotypes are closely related to symbionts of other gutless oligochaete species; the Gamma 1 phylotype is closely related to sulfide-oxidizing symbionts of Olavius algarvensis, Olavius loisae, and Inanidrilus leukodermatus, the Delta 1 phylotype is closely related to sulfate-reducing symbionts of O. algarvensis, and the spirochete is closely related to spirochetal symbionts of O. loisae. In contrast, the Gamma 2 phylotype and the Delta 2 and 3 phylotypes belong to novel lineages that are not related to other bacterial symbionts. Such a phylogenetically diverse yet highly specific and stable association in which multiple bacterial phylotypes coexist within a single host has not been described previously for marine invertebrates.     

Propagation of the myxozoan parasite Myxobolus cerebralis by different geographic and genetic populations of Tubifex tubifex: An Oregon perspective

Tubifex tubifex are obligate invertebrate hosts in the life cycle of Myxobolus cerebralis, the myxozoan parasite that causes whirling disease in salmonid fishes. This exotic parasite is established to varying degrees across Oregon’s Columbia River system (Pacific Northwest, USA) and characteristics of local T. tubifex populations likely play a role in the pattern of disease occurrence. To better understand these patterns, we collected T. tubifex from three Oregon river basins (Willamette, Deschutes, and Grande Ronde), determined their genotype (mitochondrial 16S rDNA lineage and RAPD genotype) and exposed 10 different populations to M. cerebralis in the laboratory. Four mt lineages were identified: I, III, V and VI. Lineage III was found in all river basins but dominated both central and eastern sites. The RAPD assay further divided these lineages into geographic sub-populations; no RAPD genotype was common to all basins. There was a significant difference in prevalence of infection and level of parasite production among the populations we exposed to M. cerebralis that was attributed to genotypic composition. Only lineage III worms released actinospores and only populations dominated by this lineage amplified the parasite. These populations had the lowest survival, however, the lineage dominant before exposure remained dominant despite the high prevalence of infection. The distribution and infection dynamics of susceptible T. tubifex throughout Oregon may contribute to the differences in M. cerebralis occurrence; our studies further support the influence of oligochaete genotypes on the manifestation of whirling disease in salmonid populations.     

The Influence of Environmental Factors and Microhabitat Availability on the Distribution of an Aquatic Oligochaete Assemblage in a Mediterranean River Basin

Some aquatic oligochaete species, mainly certain naidids and tubificids, are often associated with polluted environments, with high levels of organic matter and oxygen deficit. However, in the stations sampled in the Guadaira river basin, which can be arranged on an axis of organic pollution, the main aquatic oligochaete species (Nais elinguis and Tubifex tubifex) inhabiting these environments do not show a distribution pattern significantly related to the water physico-chemical characteristics measured. Nor, have we registered any association between these species and any kind of microhabitat, defined here: water current, presence of different types of vegetation, substrate coarseness and presence of anoxic sediments. Paranais litoralis is the exception, being related to areas without water current, located at higher altitudes, with high levels of dissolved organic matter and an anoxic substrate made up of fine particles. These results can be explained by the highly fluctuating nature of Mediterranean ecosystems and by the marked generalist character and ubiquity of the oligochaete assemblage species, able to colonize any kind of environment.     

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.     

The bottom Oligochaeta communities in sand pits of different trophic status in Upper Silesia (Southern Poland)

Oligochaeta, occurring in the littoral zones of two sand pits, were sampled in order to determine the relationship between the oligochaete species and the waters’ trophic level. Differences between sand pits attributable to community structure and to an overall higher abundance of oligochaetes, especially tubificids, were observed in Dzierżno Małe. Multivariate analyses (correspondence analysis [CA] and canonical correspondence analysis [CCA]) reveal correlations between the distribution patterns of oligochaete species and the concentration of nitrates, phosphates and dissolved oxygen. Some species, e.g. Dero digitata, Limnodrilus hoffmeisteri, Potamothrix hammoniensis, Nais simplex, Limnodrilus claparedeanus, Limnodrilus udekemianus and Tubifex tubifex, were more abundant in eutrophic sites. The results of the study showed that the richness of Oligochaeta species is not a good method for estimating the trophic levels of the studied sand pits. Indicators based on the share of oligo-, meso- and eutrophic species in Oligochaeta communities are better for evaluating the water quality in the studied reservoirs.     

The chemical environment of tubificid and lumbriculid worms according to the pollution level of the sediment

In Lake Geneva (Switzerland), 14 tubificid and 2 lumbriculid worm species were collected in a total of 170 samples of sediment. The sediment was analysed through ten chemical variables: organic carbon, total phosphorus, Cd, Zn, Sn, Pb, Hg, Cu, Cr, Mn. The chemical environment characterizing the presence of every worm species was defined by the mean value of each of these ten variables in all samples where the species was found. The multivariate comparison of the chemical environment typical of every species enabled six groups of species to be distinguished, each characterized by the high value of one chemical variable: (1) Peloscolex ferox, Potamothrix hammoniensis, Limnodrilus claparedeanus, and Cd. (2) Psammoryctides barbatus, and Zn. (3) Limnodrilus hofmeisteri, L. udekemianus, L. profundicola, and total P. (4) Potamothrix heuscheri, Aulodriluspluriseta, A. limnobius, Tubifex tubifex, Ilyodrilus templetoni, Stylodrilus heringianus, and organic C. (5) Potamothrixvejdovskyi, and, Hg. (6) Peloscolex velutinus, Stylodrilus lemani, and Mn. Pollution level of the sediment decreased from group 1 to 6, so each of these groups may be used to define a different level of pollution. The pooled coefficient of variation of the ten chemical variables used to define the chemical environment of each species diminished from group 1 to 6: the most opportunistic species colonized the most polluted areas.