Myxosporean Spores Research Articles

New data on wild grey mullet (mugil cephalus linnaeus, 1758) myxosporean (myxobolus episquamalis egusa et al., 1990) in the black sea

Flathead grey mullet, Mugil cephalus, with mass whitish cystlike plasmodia on their scales were collected at Kerchensky preglass of the Black Sea in 2015. The prevalence of infected fish varied from 15% in spring, reached 100% in summer, and declined to 2.5% in autumn. No fish mortality was detected. The spores were oval in frontal view, tapering to a blunt apex. Two unequal polar capsules were pyriform and extended over the anterior half of spore. Spores were 8.2±0.03 µm (7.9-8.4) long, 5.9±0.23 µm (5.2-7.3) wide, and 4.4±0.17 µm (4.0-4.7) thick. Two pyriform and unequal polar capsules were observed (4.0±0.07 µm (3.3-4.5) long, and 1.5±0.24 µm (1.1-1.8) wide). The investigation of nucleotide sequences of the 18S rDNA gene of the myxosporean spores from scales with universal primer А (5′-ACCTGGTTGATCCTGCCAGT-3′) and В (5′-TGATCCTTCTGCAGGTTCACCTAC-3′) showed 100% identity with episquamalis, and 99 % with M. bizerti, M. ichkeulensis, M. spinacurvata and Myxobolus sp. previously detected in mullets. The results obtained from the present study reveal that M. episquamalis, found on the scales of M. cephalus from Kerchensky preglass of the Black Sea, is new record for Russian waters.

Intraspecific morphometric variation in myxosporeans.

Morphometric data from spores of ten myxosporean species were statistically analysed to explore myxosporean intraspecific variation in measurements when obtained from a sample from: (1) the same plasmodium, (2) different plasmodia from the same host and (3) different host individuals and localities. In some cases, significant differences in spore dimensions were found between samples from the same plasmodium, highlighting the difficulty of obtaining representative measurements of myxosporean spore. In addition, significant differences in spore dimensions were found when plasmodia from the same site of infection were compared, suggesting that measurements of spores should come from several different plasmodia of the sampling to increase the reliability of the morphology data. Moreover, significant differences in spore dimensions were observed for most spore dimensions when data were compared between localities. In all cases, there was clear overlap in ranges of dimensions even when means differed significantly. The present statistical analysis shows that intraspecific morphometric variation of myxosporean species commonly occurs, highlighting the importance of reporting ranges of measurements for a species, not just the mean dimensions, and taking into account all evidence when assigning or describing myxosporean species.

청자갈치(Bothrocara hollandi)의 근육에 기생하는 점액포자충Myxobolus aeglefini (Myxozoa: Myxobolidae)

동해안에서 어획된 청자갈치 (Bothrocara hollandi)의 체측 근육에 유백색의 불투명한 시스트를 형성하고 있는 점액포자충이 발견되었다. 시스트를 마쇄하여 광학현미경으로 관찰해본 결과, 원형에 가까운 점액포자충의 성숙 포자가 관찰되었다. 성숙 포자의 평균 길이는 <TEX>$11.9(11.0{\sim}13.5){\mu}m$</TEX>, 평균 폭은 <TEX>$11.6(10.7{\sim}13.6){\mu}m$</TEX>, 평균 두께는 <TEX>$7.8(6.9{\sim}8.8){\mu}m$</TEX>이었다. 극낭의 평균 길이는 <TEX>$4.4(3.2{\sim}5.3){\mu}m$</TEX>이었으며, 평균 폭은 평균 <TEX>$3.3(2.4{\sim}4.2){\mu}m$</TEX>이었다. 감염숙주와 성숙포자의 형태학적 특징, 각 부위의 측정값으로부터 본 연구에서 발견된 점액포자충은 Myxobolus aeglefini Auerbach 1906으로 동정하였다. 또한 18S rDNA sequences를 이용한 계통분석 결과 M. albi와 M. groenlandicus와 같은 분지에 속하는 것이 확인되었으며 각각 97.7%와 96.9%의 상동성을 나타내었다. A specimen of porous-head eelpout Bothrocara hollandi (Zoarcidae: Perciformes) caught from the East Sea was found to harbour a myxosporean parasite. Numerous whitish pseudocysts were scattered throughout the body musculature of this individual specimen. Fresh myxosporean spores were found from the squashed pseudocysts under light microscopy. They were subspherical in frontal view with a length of <TEX>$11.9(11.0{\sim}13.5){\mu}m$</TEX>, width of <TEX>$11.6(10.7{\sim}13.6){\mu}m$</TEX>, and thickness of <TEX>$7.8(6.9{\sim}8.8){\mu}m$</TEX>. Two polar capsules were almost equally pyriform with a length of <TEX>$4.4(3.2{\sim}5.3){\mu}m$</TEX> and width of <TEX>$3.3(2.4{\sim}4.2){\mu}m$</TEX>. Morphometric and host ecology analysis revealed that this myxosporean parasite could be identified as Myxobolus aeglefini Auerbach 1906. Phylogenetic analysis based on 18S rDNA sequences also revealed that M. aeglefini was clustered with M. albi and M. groenlandicus in the same branch, sharing 97.7% and 96.9% sequence similarities with M. albi and M. groenlandicus, respectively.

Whirling disease revisited: pathogenesis, parasite biology and disease intervention

Whirling disease (WD) is an ecologically and economically debilitating disease of rainbow trout Oncorhynchus mykiss caused by the actinosporean spores of the parasite Myxobolus cerebralis. M. cerebralis has a complex, 2-host life cycle alternating between salmonid fish and the oligochaete host Tubifex tubifex. The parasite alternates between 2 spore forms as transmission stages: an actinosporean triactinomyxon spore that is produced in the oligochaete host and a myxosporean spore that develops in the salmonid host. Waterborne triactinomyxon spores released from infected T. tubifex oligochaetes attach to the salmonid host by polar filament extrusion elicited by chemical (nucleoside) and mechanical (thigmotropy) stimuli-a process which is rapidly followed by active penetration of the sporoplasms into the fish epidermis. Upon penetration, sporoplasms multiply and migrate via peripheral nerves and the central nervous system to reach the cartilage where they form trophozoites which undergo further multiplication and subsequent sporogenesis. M. cerebralis myxospores are released into the aquatic environment when infected fish die and autolyse, or when they are consumed and excreted by predators. Myxospores released into the water are ingested by susceptible T. tubifex where they develop intercellularly in the intestine over a period of 3 mo through 4 developmental stages to give rise to mature actinospores. In this article, we review our current understanding of WD-the parasite and its alternate hosts, life cycle and development of the parasite in either host, disease distribution, susceptibility and resistance mechanisms in salmonid host and strategies involved in diagnosis, prevention and control of WD.

Proliferation and resistance difference of a liver-parasitized myxosporean in two different gynogenetic clones of gibel carp

Some myxosporeans have been demonstrated to be harmful to worldwide aquaculture. However, the proliferation information has remained unclear in the fish hosts. In this study, we utilized the mix-culturing equality to reveal significant difference in disease assistance between two different clones of gibel carp, in which clone D had been cultured for nearly 40 years, whereas clone A(+) was a newly created clone. According to morphological and genetic analysis of isolated spores, the diseasing pathogen was identified as Myxobolus wulii of the genus Myxobolus in Myxosporea. Subsequently, a polyclonal antibody specific to soluble proteins of the purified spores was generated. Using the antibody, we performed immunofluorescence observation of the liver lump sections sampled from the heavily diseased clone D individuals, and found that the liver lumps were completely composed of numerous honeycomb-like cysts, full of maturing and mature myxosporean spores, and almost all of liver tissues were destroyed. Comparative co-localization detection revealed a significantly inducing expression of apo-14 protein around the infected myxosporean sporoplasms and plasmodia, and the inducing level was much stronger in clone A(+) than in clone D. Furthermore, a primarily screening of 15 different major histocompatibility complex class Iα variants also excavated major variants that respectively belong to clones D and A(+). Therefore, these data provide significant information for differences in myxosporean proliferation and disease resistance in fish clone hosts with different genetic background. Further studies on myxosporean development and the mechanism for disease resistance will be very important for preventing and controlling the parasitic myxosporean disease.

Morphologic and Genetic Evidence for Mixed Infection with Two Myxobolus Species (Myxozoa: Myxobolidae) in Gray Mullets, Mugil cephalus, from Korean Waters

The present study was performed to trace the decisive evidence for mixed infection of 2 Myxobolus species, M. episquamalis and Myxobolus sp., in the gray mullet, Mugil cephalus, from Korean waters. Mullets with whitish cyst-like plasmodia on their scales were collected near a sewage plant in Yeosu, southern part of Korea, in 2009. The cysts were mainly located on scales and also found in the intestine. The spores from scales were oval in a frontal view, tapering anteriorly to a blunt apex, and measured 7.2 µm (5.8-8.0) in length and 5.3 µm (4.7-6.1) in width. Two polar capsules were pyriform and extended over the anterior half of the spore, measuring 3.5 µm (2.3-4.8) in length and 2.0 µm (1.5-2.2) in width. In contrast, the spores from the intestine were ellipsoidal, 10.4 µm (9.0-11.9) in length and 8.4 µm (7.3-10.1) in width. The polar capsules were pyriform but did not extend over the anterior half of the spore, 3.7 µm (2.5-4.5) in length and 2.2 µm (1.8-2.9) in width. The nucleotide sequences of the 18S rDNA gene of the 2 myxosporean spores from scales and intestine showed 88.1% identity to each other and 100% identity with M. episquamalis and 94.5% identity with M. spinacurvatura from mullet, respectively. By the above findings, it is first confirmed that mullets from the Korean water are infected with 2 myxosporean species, M. episquamalis and Myxobolus sp.

Infection of wild mullet (Mugil cephalus) with Myxobolus episquamalis in Korea

Wild mullet (Mugil cephalus) with white cysts on their scales were obtained from Yeosu on the south coast of Korea in 2009. Cyst-like plasmodia consisted of a large number of mature myxosporean spores and numerous sporogonic stages. Spores were oval-shaped in their front view, tapering anteriorly to a blunt apex, and lenticular in their lateral view. They measured 7.0μm (6.2-7.6) in length, 5.2μm (4.0-6.2) in width, and 4.9μm (3.8-6.0) in thickness. Polar capsules contained a polar filament with five to six turns and measured 3.5μm (2.5-4.5) in length and 2.0μm (1.6-2.3) in width. Nucleotide sequences of the 18S rRNA gene of the myxosporean parasites in our study showed 99.8% identity with Myxobolus episquamalis Egusa, Maeno and Sorimachi, 1990 from mullet in Tunisia. These results suggest that the Myxobolus sp. found on the scales of wild mullet is M. episquamalis. In the histopathological examination, spores were observed not only in the plasmodia on the scales, but also in the intestine, pancreas, heart, kidney, stomach, gill, skin, spleen, and liver, suggesting the possibility of the coinfection by different Myxobolus species.

A new host record of Sphaerospora epinepheli (Myxosporea: Bivalvulida) occurring on orange-spotted grouper Epinephelus coioides from Thailand: Epidemiology, histopathology and phylogenetic position

In 1991, the first record of Sphaerospora epinepheli was described as a kidney parasite of wild and cultured malabar grouper, Epinephelus malabaricus, along coastlines of Thailand, the Gulf of Thailand and the Andaman Sea. However, the present study detected high infection of this parasite in kidney renal tubes of orange spotted grouper, Epinephelus coioides, collected from Andaman Sea. The highest infection rate of 36.82% was observed during the rainy season in 2009 in Phang-Nga Bay, in the north of Andaman Sea, which is an important grouper production site in Thailand. The biological and histopathological data of the parasite in this new host record are presented. Species classification is described based on morphological data of mature spore and molecular analysis of myxosporean 18S rDNA phylogeny including that of S. epinepheli which infected E. malabaricus. The genetic position of this parasite found in two host species was also studied. The phylogenetic tree analysis of small-subunit rDNA sequences of S. epinepheli from both infected hosts was constructed using two algorithms, maximum likelihood (ML) and Bayesian inference (BI). They were placed in the clustered basal sphaerosporid clade that contain four long SSU rDNA sphaerosporid species including Sphaerospora truttae, Sphaerospora elegans, Sphaerospora ranae, Sphaerospora fugu and Bipteria formosa with strong bootstrap supports. Histopathologically, renal intratubular myxosporean spores were associated with tubulonephosis, tubular necrosis, chronic interstitial nephritis and mimic membranoproliferative glomerulonephritis. This myxosporean parasite appears to be a significant pathogen on the basis of pathological changes in the renal tubules and is highly distributed in orange-spotted grouper.

Structure and population dynamics of myxobolus infections in wild and cultured Oreochromis niloticus Linnaeus, 1758 in the Noun division (West-Cameroon)

Myxosporidian parasites of Oreochromis niloticus Linnaeus, 1758 from the Noun River at Kouoptamo and the Foumban fish ponds in west Cameroon, were investigated from May 2008 to June 2009. Out of 537 Tilapia (267 cultivated and 270 wild) examined, 64.8% (n=173) specimens from the fish farming and 61.1% (n=165) from the Noun River harbored Myxosporean parasites. A total of ten parasite species were found. Myxobolus kainjiae, Myxobolus sarigi were scarce in both study sites; Myxobolus Tilapiae, Myxobolus equatorialis scarce in Foumban and Kouoptamo, respectively. M. agolus, M. brachysporus, M. camerounensis, M. equatorialis, Myxobolus Heterosporus, Myxobolus israelensiswere secondary in the two sites. M. Tilapiae, M. equatorialis appeared secondary in the Noun River and the fish ponds respectively. Myxosporean spores were most encountered in the kidney (61.3 and 49.0%, respectively in cultured and wild fish) and the spleen (50.5% in Foumban and 47.5% in Kouoptamo) but no host sex preference was found. In the Foumban fish farm site, high significant infection rate was observed for M. tilapiae, M. camerounensis and M. israelensis during the rainy season, while in the Noun River, no significant seasonal effect was found. Older hosts were significantly most infected at the fish ponds while youngs Tilapia were most commonly infected in the River. Key words: Myxosporean, Oreochromis niloticus, prevalence, fish-farm, Noun River, Cameroon.

Flavobacterium columnare / Myxobolus tilapiae Concurrent Infection in the Earthen Pond Reared Nile Tilapia (Oreochromis niloticus) during the Early Summer

Flavobacterium columnare (F. columnare), the dermotropic Gram negative yellow pigmented bacteria was isolated from different sites of skin ulcerations in the Nile tilapia (Oreochromis niloticus) and Nile catfish (Clarias gariepinus) collected from an earthen pond located at an aquaculture station in Sharkiya Province, Lower Egypt during an acute episode of mass kills during the early summer of 2009. An acute infection with F. columnare was behind the emergent event of mass mortalities among both populations. Many of the Nile tilapias exhibited typical signs of hole - in- the head like lesions from which F. columnare together with the myxosporean spore, Myxobolus tilapiae (M. tilapiae) were retrieved. Most of the cohabitating infected Nile catfishes exhibited severe form of saddle back like ulcer. The identities of the retrieved isolates were confirmed using morphological, biochemical and molecular tools. The research lead us to conclude that the two diverse etiological agents (F. columnare and M. tilapiae) under the triggering effect of the abrupt change in the water quality measures (abrupt rise in the water temperature, ammonia, pH, sharp decrease in dissolved oxygen) have synergized together to induce the above mentioned pathology with the consequent reemergence of fish mass mortalities.

History of myxozoan character evolution on the basis of rDNA and EF-2 data

BackgroundPhylogenetic relationships among myxosporeans based on ribosomal DNA data disagree with traditional taxonomic classification: a number of myxosporeans with very similar spore morphology are assigned to the same genera even though they are phylogenetically distantly related. The credibility of rDNA as a suitable marker for Myxozoa is uncertain and needs to be proved. Furthermore, we need to know the history of myxospore evolution to understand the great diversity of modern species.ResultsPhylogenetic analysis of elongation factor 2 supports the ribosomal DNA-based reconstruction of myxozoan evolution. We propose that SSU rDNA is a reliable marker for inferring myxozoan relationships, even though SSU rDNA analysis markedly disagrees with the current taxonomy. The analyses of character evolution of 15 morphological and 5 bionomical characters show the evolution of individual characters and uncover the main evolutionary changes in the myxosporean spore morphology and bionomy. Most bionomical and several morphological characters were found to be congruent with the phylogeny. The summary of character analyses leads to the simulation of myxozoan ancestral morphotypes and their evolution to the current species. As such, the ancestor of all myxozoans appears to have infected the renal tubules of freshwater fish, was sphaerosporid in shape, and had a spore with polar capsules that discharged slightly sideways. After the separation of Malacosporea, the spore of the common myxosporean ancestor then changed to the typical sphaerosporid morphotype. This species inhabited the marine environment as a parasite of the gall bladder of marine fish and ultimately separated into the three main myxosporean lineages evident today. Two of these lineages re-entered the freshwater environment, one as a myxosporean with Chloromyxum and another with a primitive sphaerosporid morphotype. The common ancestor of all marine myxosporeans had a ceratomyxid shape of spore.ConclusionsWe support rDNA based myxozoan phylogeny by the analysis of a protein coding gene and demonstrate the reliability of rDNA as a marker explaining myxozoan relationships. Our tracing the history of myxozoan character evolution discloses ancestral morphotypes and shows their development over the course of evolution. We point out several myxozoan characters that are to a certain extent congruent with the phylogeny and determined that the discrepancy between phylogeny and current taxonomy based on spore morphology is due to an extreme myxospore plasticity occurring during myxozoan evolution.

Morphological and molecular redescription of the myxozoan Unicapsula pflugfelderiSchubert, Sprague &amp; Reinboth 1975 from two teleost hosts in the Mediterranean. A review of the genus Unicapsula Davis 1924

Elongate plasmodia with myxosporean spores belonging to the genus Unicapsula, Davis, 1924 were found in the skeletal muscle of the striped seabream, Lithognathus mormyrus (L.), a candidate for the mediterranean aquaculture. The only species of Unicapsula described from the Mediterranean is Unicapsula pflugfelderi Schubert et al. 1975, which occurs in the picarel, Spicara smaris (L.). For morphological and molecular comparison of U. pflugfelderi from S. smaris with Unicapsula sp. from L. mormyrus measurements of plasmodia and spores, ultrastructural details and 18S and 28S rDNA sequences were analysed. Whereas plasmodia were 2-3 times larger in S. smaris than in L. mormyrus (length 2.47-0.81 mm; width 0.22-0.09 mm; P = 0.000), spore morphology showed minor differences and both 18S and 28S rDNA sequences were 100% identical identifying the myxozoan as U. pflugfelderi. Scanning electron microscopy of the spores revealed a different shell valve distribution than the one used for the diagnosis of the genus Unicapsula. This resulted in a review of the genus Unicapsula dividing it into two morphological groups of different spore valve arrangement. TEM revealed the presence of a yet undescribed crystalline structure in the sporoplasm of the spores.

Structure and Dynamics of Myxosporean Parasites Component Communities in Two Freshwater Cichlids in the Chari River (Republic of Chad)

Myxosporean parasites of two freshwater Tilapia species from the Chari River, Chad Republic, Oreochromis niloticus and Sarotherodon galilaeus, were investigated from November 2001 to October 2002. A total of 360 specimens per Cichlid species were examined. Eleven parasite species were found in both cases with different prevalences. Myxobolus agolus, M. brachysporus, M. clarii, M. cichlidarum, M. heterosporus, M. tilapiae and M. camerounensis in O. niloticus) appeared common while M. equatorialis, M. nyongana (and M. camerounensis in S. galilaeus) were secondary; lastly M. israelensis (and M. kainjiae in O. niloticus) were rare. The gills, fins, eyes and teguments were preferential locations of cysts building pathogens while the kidneys then the gall bladder were most commonly infected by myxosporean spores. In the Chari ecosystem, no significant host sex and size effects were found for the parasite cystic load. A clear seasonal occurrence was observed for most of these pathogens. In the view of pathogenic control, this study raised the necessity in a farm fish station to identify the most important myxosporean species and the period of their potential demographic explosion.

Myxidium mackiei (Myxosporea) in Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii: parasite-host interaction and ultrastructure

Myxosporeans are common parasites of fish, and uncommon parasites of amphibians, reptiles and invertebrates, that can cause significant morbidity and mortality. The common genus Myxidium infects the excretory system of turtles, yet knowledge of its pathogenicity in these hosts is limited. We offer new knowledge of morphological and ultrastructural aspects of host-parasite interactions in Myxidium infections from our recent diagnostic investigations on captive freshwater turtles listed in CITES (Appendix II). We investigated the cause of death of 2 adult Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii from a zoo collection. After post-mortem examination, tissues were processed for histopathology, and special stains were used to demonstrate morphology of myxosporean spores. Additional kidney tissue, immersion-fixed in formalin, was processed for transmission electron microscopy. Both turtles were infected with a myxosporidian, Myxidium mackiei, in the kidney, which occluded 5 to 10% of the renal proximal convoluted tubules. The polysporic plasmodia contained pairs of developing and mature spores. Each mature, spindle-shaped spore had 2 asymmetric valves (1 overlapping, 1 overlapped), with 10 to 13 and 10 to 14 longitudinal ridges per valve, and 2 polar capsules each containing a polar filament with 4 to 5 turns. A pair of spores, each surrounded by a membrane-bound electron-lucent matrix, lay in an enclosing cell within the plasmodium. Regions of the parasite-host interface consisted of undulations of the parasite surface, with intense pinocytotic activity beneath, intermingled with the hosts' microvilli, and endocytotic channels at the apex of renal epithelial cells. The microvilli of the renal epithelial cells of infected tubules were frequently sheared or compressed, or occasionally missing; we did not detect other pathology induced by the parasite. Our report of M. mackiei in L. punctata is a new host record. Both individuals also had disseminated pale yellow nodules (bacterial granulomas) present in lung, heart, kidney, and skeletal muscle, and both were infected with coccidia (tentatively identified as Eimeria sp.) in multiple organs. The cause of death for one turtle was septicemia, but remained unknown for the other individual.

A vital staining technique with fluorescein diacetate (FDA) and propidium iodide (PI) for the determination of viability of myxosporean and actinosporean spores

A vital staining technique with fluorescein diacetate (FDA) and propidium iodide (PI) was used for determination of viability of myxosporean stage spores of Myxobolus artus and actinosporean stage spores of M. cultus. Viable spores stained green with FDA and non‐viable spores stained red with PI were clearly distinguishable in M. artus myxosporean spores released from live infected fish, while the spores collected from pseudocysts were not well stained with FDA. Immediately after release from the oligochaete, Branchiura sowerbyi, actinosporean spores of M. cultus, stained bright green in the sporoplasms and red in the spore processes. In vitro survivability tests revealed that the longevity of M. artus spores was 5 months at 25°C and 15 months at 18°C. Drying and ultraviolet irradiation (36 000 mW s cm–2 for M. artus myxosporean spores, 600 mW s cm–2 for M. cultus actinosporean spores) were most effective as sporicidal treatments.

Proliferative kidney disease and Sphaerospora oncorhynchi in wild-caught salmonids from the Puntledge River system, Vancouver Island, British Columbia

Proliferative kidney disease (PKD), caused by the PKX myxosporean, was observed in kokanee salmon (non-anadromous sockeye salmon) (Oncorhynchus nerka) and chinook salmon (O. tshawytscha) collected from the Puntledge River, Vancouver Island, British Columbia in July 1993. This is the first report of a natural infection of PKX in either captive or wild sockeye salmon. All 14 underyearling kokanee salmon and the one underyearling chinook salmon exhibited numerous PKX organisms and associated chronic inflammation in the renal interstitium. Thirty-eight percent of sexually mature kokanee salmon collected in the autumn of 1993 from Comox Lake (which drains into the Puntledge River) were infected with Sphaerospora oncorhynchi, Kent, Whitaker and Margolis, 1993 in the renal tubules, while immature cutthroat trout (O. clarki) and coho salmon (O. kisutch) from the same collection did not exhibit myxosporean spores in the kidney. The kidneys of threespine stickleback (Gasterosteus aculeatus) and prickly sculpin (Cottus asper) collected from the Puntledge River during the summer were all infected with a Myxobilatus sp., but no organisms suggestive of PKX or Sphaerospora were observed. This study further supports the hypothesis that PKX is a developmental stage of S. oncorhynchi, which may sporulate only in sexually mature salmonids.

Settling Rates and Density of Spores of Henneguya doori (Myxosporea) in Water

Using Henneguya doori from Perca flavescens (yellow perch), the current study provides a method for examining settling rates, attitudes, and densities of myxosporean spores. Fresh and fixed spores settled similarly at 8.34 x 10(-6) m/sec and 8.53 x 10(-6) m/sec, respectively. The spores settled in the anterior end first, did not rotate, and had a specific gravity of 1.062. Comparison of settling rates proved more effective for determining spore density than direct chemical estimation. The slow rate of settling permits effective dispersal in open water.

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

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

Salmonid whirling disease: myxosporean and actinosporean stages cross‐react in direct fluorescent antibody test

Abstract. Serologic relatedness of the two life stages of the salmonid whirling disease parasite Myxosoma cerebralis Hofer, 1903 — myxosporean spores from fish cartilage and actinosporean triactinomyxon spores from aquatic tubificids — were investigated. When the direct fluorescent antibody technique was used, anti‐triactinomyxon and anti‐M. cerebralis rabbit sera conjugated with fluorescein isothiocyanate cross‐reacted with the respective heterologous life stage. Both stages showed similar locations of specific fluorescence with conjugates of either homologous or heterologous serum. Thus, serology supports the relatedness of the myxosporean M. cerebralis and the actinosporean triactinomyxon stages.

“Unidentified” Mobile Protozoans from the Blood of Carp and Some Unsolved Problems of Myxosporean Life Cycles1

ABSTRACTHeavy infections with enigmatic mobile organisms have recently been found in the blood of carp (Cyprinus carpio) in Central Europe. The organisms measure up to 15 μm, are variable in shape, and exhibit an unceasing twitching or dancing movement. Their developmental cycle starts with a primary cell enclosing a secondary cell. The former grows while the latter produces inside itself by a series of binary fissions and internal cleavages up to eight secondary cells, each of which encloses an inner (tertiary) cell of its own. In addition, up to four tiny cells with compact nuclei (“residual bodies”) also result from divisions of the secondary cells. Primary cells containing the products of the division of secondary cells finally disintegrate, releasing the secondary cells, which in their turn become new primary cells and repeat the cycle all over again. The structure and behavior of these organisms were so incompatible with existing ideas on myxosporean development that their myxosporean affinity was at first unrecognized. The final proof of their identity–appearance of myxosporean spores in sterile, experimentally infected hosts–is still to be presented. The interpretation of the myxosporean features of their life cycle (i.e., [1] the pericyte nature of the primary cell, [2] proliferation by disintegration of the pseudoplasmodial primary cell, [3] no rigidly fixed pattern in vegetative development), their ultrastructure (i.e., [1] characteristic bundles of microtubules and numerous free ribosomes in secondary cells, [2] lack of centrioles, [3] membranes enclosing the secondary cells within the primary cells), and facts on their epizootiology (i.e., [1] no success at transmission via leeches, [2] the occurrence of these organisms along with Sphaerospora renicola Dykova and Lom) suggest that they are stages of S. renicola from the kidney of carp. Similar mobile organisms were found in the blood of fry of two other fishes (Gobio gobi and Tinca tinca) which are also hosts for a Sphaerospora that infects the kidney. This suggests that these organisms represent an early phase in the developmental cycle in the genus Sphaerospora. The existence of cells enveloped one within the other (secondary and tertiary cells) in the developmental cycle, a characteristic myxosporean feature itself, is an intriguing parallel to similarly enclosed cells in sporogenesis of Paramyxea (Ascetospora).