Clinical, histopathological and phylogenetic analysis of Myxobolus lentisturalis (Myxozoa: Myxobolidae) infecting the musculature of farmed population of goldfish (Carassius auratus) in Iran: 2021-2022.

There is a claimed increase in the global prevalence and incidence of emerging diseases observed in many organisms. Myxozoa represents an essential group of metazoan parasites that hold both economic and ecological significance. In the current study, 1% of the fish population at two commercial goldfish (Carassius auratus) farms in Tehran and Ghom province, Iran, developed cavitating muscular lesions resembling humps in February 2021 and January 2022. Fish displaying pathological abnormalities were transported to the Ornamental Fish Clinic and subjected to clinical examination. Light microscopy was subsequently used to examine wet smears of skin and gills, as well as whitish exudate. In addition, tissue homogenates were collected for more precise identification and molecular confirmation. The study discovered that individuals from the goldfish farms were infected with the pathogenic myxozoan Myxobolus lentisuturalis, which caused significant damage to the epaxial muscles. The spores collected from the humps had a lack of uniformity and were primarily ellipsoidal in shape. Histopathological analysis also revealed parasites in various stages of development, such as plasmodia and spores, as well as inflammatory cell infiltration (macrophage, giant cell and lymphoplasmacytic infiltration) between skeletal muscle fibers. Phylogenetic analysis of M. lentisuturalis was performed by using MEGA 11 and the maximum likelihood method. M. lentisuturalis is a myxozoan parasite that has been sparsely recorded and lacks widespread recognition. The current study is the first clinical, histopathological, and molecular characterization of M. lentisuturalis isolated from the skeletal musculature of goldfish (C. auratus) in Iran.

The Tilapia Cyst Tissue Enclosing the Proliferating Myxobolus bejeranoi Parasite Exhibits Cornified Structure and Immune Barrier Function.

Myxozoa, a unique group of obligate endoparasites within the phylum Cnidaria, can cause emerging diseases in wild and cultured fish populations. Recently, the myxozoan Myxobolus bejeranoi has been identified as a prevalent pathogen infecting the gills of cultured hybrid tilapia, leading to systemic immune suppression and considerable mortality. Here, we employed a proteomic approach to examine the impact of M. bejeranoi infection on fish gills, focusing on the structure of the granulomata, or cyst, formed around the proliferating parasite to prevent its spread to surrounding tissue. Enrichment analysis showed increased immune response and oxidative stress in infected gill tissue, most markedly in the cyst's wall. The intense immune reaction included a consortium of endopeptidase inhibitors, potentially combating the myxozoan arsenal of secreted proteases. Analysis of the cyst's proteome and histology staining indicated that keratin intermediate filaments contribute to its structural rigidity. Moreover, we uncovered skin-specific proteins, including a grainyhead-like transcription factor and a teleost-specific S100 calcium-binding protein that may play a role in epithelial morphogenesis and cysts formation. These findings deepen our understanding of the proteomic elements that grant the cyst its distinctive nature at the critical interface between the fish host and myxozoan parasite.

First report of Myxobolus neurofontinalis (Bivalvulida: Myxobolidae) infecting anadromous Brook Trout from Prince Edward Island, Canada.

OBJECTIVE: During routine histological examination of tissues from mortality events of anadromous Brook Trout Salvelinus fontinalis from Prince Edward Island (PEI), Canada, myxospores consistent with Myxobolus were observed infecting the central nervous system. The objective of this study was to identify the species of Myxobolus infecting the nervous system of anadromous Brook Trout from PEI, Canada. METHODS: Myxospore morphology, small subunit (SSU) ribosomal DNA (rDNA) sequence data, and histology were used to identify myxospores isolated from infected Brook Trout. RESULT: Myxospore measurements from the PEI samples matched those reported in the description of Myxobolus neurofontinalis from North Carolina. A 1057-bp fragment of the SSU rDNA from myxospores collected from Brook Trout in PEI was identical to an isolate of M. neurofontinalis (MN191598) collected previously from the type locality, New River basin, North Carolina. Histological sections confirmed infections were intercellular in the central nervous system. Minimal host response was observed, with only sparse mononuclear inflammatory infiltrates present at the periphery of and within dispersed myxospores, suggesting that infections are not pathogenic to Brook Trout. CONCLUSION: Myxospores were identified as M. neurofontinalis, which was previously described from the central nervous system of Brook Trout from the New River basin, North Carolina, USA. This constitutes the first time M. neurofontinalis has been documented outside of the New River basin in North Carolina.

Comparative evaluation of three real-time polymerase chain reaction assays to detect Myxobolus cerebralis.

OBJECTIVE: We sought to evaluate accurate and reproducible detection of Myxobolus cerebralis (Mc), the causative agent of whirling disease, by using nested polymerase chain reaction (nPCR) and three previously established real-time quantitative PCR (qPCR) assays: K18S (Kelley 18S), C18S (Cavender 18S), and Hsp70 (heat shock protein 70). We used a "fit for purpose" approach combined with intra- and interlaboratory testing to identify a molecular testing method that would be equivalent to the currently accepted nPCR procedure for Mc. METHODS: Assay performance was compared using a combination of intra- and interlaboratory testing that used synthetic gBlocks along with naturally and experimentally infected fish tissue. North American isolates representing geographically distinct locations were also tested using all three assays. RESULT: The K18S and C18S assays exhibited high assay sensitivity, intra- and interlaboratory repeatability of sample replicates, and reproducible identification of all test samples across multiple laboratories. In contrast, the Hsp70 assay failed to detect several positive samples at low DNA concentrations during intra- and interlaboratory testing. The K18S assay was the only procedure that demonstrated perfect detection accuracy when testing geographically distinct Mc isolates. Results demonstrated the K18S assay is robust under variable test conditions, is more accurate than the C18S and Hsp70 assays, and provides detection capabilities equivalent to those of the currently accepted nPCR confirmation assay "gold standard" that is described in the American Fisheries Society-Fish Health Section (AFS-FHS) Blue Book. CONCLUSION: The "fit for purpose" approach and preliminary completion of the World Organization for Animal Health validation pathway demonstrate that the K18S assay provides an alternate method for Mc testing. This work provides the foundation for acceptance of the K18S assay into the AFS-FHS Blue Book as a standardized test procedure for Mc.

Myxobolus acanthogobii Hoshina, 1952 and Myxobolus selari n. sp. (Myxosporea: Myxobolidae) infecting brain of commercial fishes in Terengganu, Malaysia.

Myxosporean infection in marine water fishes has drawn less attention than in freshwater fishes, which resulted in a higher taxonomic variety in freshwater in Malaysia. This study aimed to address the gap by conducting a myxosporean survey on two commercially significant marine fish species, Nemipterus furcosus (Valenciennes) (Eupercaria incertae sedis: Nemipteridae) and Selar crumenophthalmus (Bloch) (Carangiformes: Carangidae), collected from the northeastern part of peninsular Malaysia. During the examination of the organs, two distinct Myxobolus Bütschli, 1882 species were discovered in the brain tissue of these fishes, despite the absence of any observable pathological signs. The two Myxobolus species were characterized through morphometry, morphology, and analysis of partial small subunit ribosomal RNA (18S rDNA) gene. As a result, Myxobolus acanthogobii Hoshina, 1952, which infects 2.3% of N. furcosus, is synonymous with a myxobolid species commonly found in Japanese waters, based on its morphological traits, tissue tropism, and molecular diagnostics. Furthermore, a novel species, Myxobolus selari n. sp., was described, infecting the brain of one (11%) individual S. crumenophthalmus. This unique species displayed distinctive features, placing it within a well-supported subclade primarily comprising brain-infecting myxobolids. Maximum likelihood analysis further revealed the close relationships among these brain-infecting myxobolids, underscoring the significance of tissue tropism and host taxonomy for myxobolids. This study represents the initial documentation of Myxobolus species within the southern South China Sea, shedding light on the potential diversity of marine myxosporean in this region. This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as urn:lsid:zoobank.org:pub:7C400E35-7CB8-4DEE-92B7-F75FF3926441.

Histopathological evaluation of the infection elicited by three species of Myxobolus (Cnidaria: Myxosporea), with identification of Myxobolusxiaoganensis n. sp. isolated from the silver carp Hypophthalmichthys molitrix.

Several parasites infecting the Asian carp species have been broadly spread along with the global fish trade. However, the diversity of specific parasite groups and their pattern of parasitism remain insufficiently elucidated even in their native regions. Here, we conducted a holistic identification and histological analysis of three Myxobolus species. The oblate myxospores of the first isolates were found infecting the spleen of silver carp Hypophthalmichthys molitrix, measuring 6.4 ± 0.4 (5.4-7.1) µm in length, 8.2 ± 0.4 (7.5-9.0) in width, and 5.9 ± 0.3 (5.2-6.2) in thickness. They were morphologically distinct from other congeners and regarded as a novel species, Myxobolus xiaoganensis n. sp. For the second isolates, we associated the formation of round plasmodia on the gill raker of silver carp with Myxobolus allotypica Chen, 1998. The third isolates, encapsulated in the intestinal serosa membrane of the grass carp Ctenopharyngodon idella, was proved to be conspecific to Myxobolus huasaensis Chen, 1998. While M. xiaoganensis n. sp. and M. huasaensis exhibited distinct origins, with genetic differences exceeding 4% from other congeners, M. allotypica displayed complete genetic congruence with an item available in Genbank. Histologically, myxospores of M. xiaoganensis n. sp. were scattered in the spleen, and the branchial and intestinal infections of M. allotypica and M. huasaensis were determined, respectively. Phylogenetic analysis demonstrated a non-monophyletic origin of both the Thelohanellus and Myxobolus genera, with a remarkable association of host affinity with myxosporean clustering.

Myxobolus nekrasovae n. sp. (Cnidaria, Myxozoa) is a new species parasitizing the gills of the gibel carp, Carassius auratus gibelio.

A new Myxobolus species, Myxobolus nekrasovi n. sp., was found in the gill arch of the gibel carp Carassius auratus gibelio during investigation of fish myxosporean fauna of ponds of Lake Baikal basin. The parasites were studied on the basis of spore morphology, as well as with histological and molecular methods. Mature spores of M. nekrasovi n. sp. are ellipsoidal in frontal view and lemon-shaped in lateral view, measuring 13.84 ± 0.4 (12.2-15) µm in length, 9.73 ± 0.2 (8.5-10.7) µm in width, 6,75 ± 0.1 (6.0-7.6) µm in thickness. Polar capsules are unequal and pyriform, measuring: length 6.31 ± 0.1 (5.4-7.4), width 3.49 ± 0.04 (3.12-4) µm and length 2.88 ± 0.1 (2.1-3.5), width 1.4 ± 0.03 (1-1.6) µm. Phylogenetic analysis with the SSU rDNA gene shows Myxobolus nekrasovae n. sp. as a sister species of the subclade formed by Thellohanellus sinensis, Myxobolus acutus, M. zhaltsanovae that infect gibel carp Carassius auratus gibelio.

The myxozoan parasite Myxobolus bejeranoi (Cnidaria: Myxozoa) infection dynamics and host specificity in hybrid tilapia aquaculture.

Nile × blue tilapia hybrid (Oreochromis niloticus × O. aureus) has become an important food fish in intensive freshwater aquaculture. Recently, the parasite Myxobolus bejeranoi (Cnidaria: Myxozoa) was found to infect hybrid tilapia gills at high prevalence, causing immune suppression and high mortality. Here, we explored additional characteristics of M. bejeranoi­tilapia interaction, which enable efficient proliferation of this parasite inside its specific host. Highly sensitive quantitative polymerase chain reaction (qPCR) and in situ hybridization analyses of fry collected from fertilization ponds provided evidence to an early-life infection of fish by a myxozoan parasite, occurring less than 3 weeks post-fertilization. Because Myxobolus species are highly host-specific, we next compared infection rates in hybrid tilapia and in both its parental species following a 1-week exposure to infectious pond water. Analysis by qPCR and histological sections showed that while blue tilapia was as susceptible to M. bejeranoi as the hybrid, Nile tilapia appeared to be resistant. This is the first report of differential susceptibility of a hybrid fish vs its parental purebreds to a myxozoan parasite. These findings advance our understanding of the relationship between M. bejeranoi and tilapia fish and raise important questions regarding the mechanisms that allow the parasite to distinguish between very closely related species and to infect a specific organ at very early-life stages.

A new species of Myxobolus (Cnidaria: Myxosporea: Myxobolidae) from the gibel carp Carassius gibelio (Cypriniformes: Cyprinidae).

Myxobolus zhaltsanovae n. sp., is described from the gills of gibel carp Carassius gibelio found during a survey of myxozoans from the watershed of Lake Baikal, Russia, based on morphological and molecular characterizations. Plasmodia of M. zhaltsanovae n. sp. develop extravascularly and measure 500-1000 µm long, 25-100 µm wide. The myxospore is circular to oval, measuring 13.23 ± 0.09 (11.3-14.8) µm (mean ± SD, range) in length, 10.19 ± 0.07 (9.1-11.4) µm in width, and 6.49 ± 0.12 (5.4-7.2) µm in thickness. Polar capsules are unequal and subspherical; measurements of polar capsules are: length 5.62 ± 0.06 (4.7-6.7), width 3.44 ± 0.04 (2.4-4.4) µm and length 3.42 ± 0.05 (2.5-4.1), width 1.94 ± 0.04 (1.3-3.3) µm. Phylogenetic analysis with the 18S rDNA gene shows M. zhaltsanovae n. sp. as a sister species of the subclade formed by M. musseliusae, M. tsangwuensis, and M. basilamellaris, which infect common carp Cyprinus carpio.

Myxobolus lentisuturalis infection in a farmed population of goldfish Carassius auratus from the USA.

Myxobolus lentisuturalis is a myxozoan parasite of piscine muscle that has been described in goldfish Carassius auratus and Prussian carp Carassius gibelio. This report documents a naturally occurring infection of M. lentisuturalis in a population of farmed goldfish in the USA. Postmortem examination was performed on 4 affected goldfish. Gross findings included large cystic cavities along the dorsal midline filled with caseous exudate. Histopathology revealed myxozoan plasmodia and spores in the epaxial muscles with varying degrees of granulomatous and necrotizing myositis accompanied by lymphohistiocytic meningoencephalitis. Spore morphology and dimensions were consistent with M. lentisuturalis, as observed by light microscopy. PCR and sequence analysis of the small subunit ribosomal DNA of infected muscle samples from 2 goldfish confirmed the parasite to have 99-100% nucleotide identity to M. lentisuturalis sequences recovered from similar cases of this parasite infecting goldfish in China and Italy and Prussian carp in China. This is the first reported case of M. lentisuturalis in the USA and furthers the understanding of the pathogenicity of this under-described parasite.

The first detection of Myxobolus lentisuturalis Dyková, Fiala et Nie, 2002, a highly pathogenic muscle-infecting parasite of gibel carp (Carassius auratus gibelio Berg, 1932) in Hungary.

Myxobolus lentisuturalis is a myxosporean parasite infecting the musculature both of goldfish (Carassius auratus auratus) and gibel carp (Carassius auratus gibelio). The species was originally described in China from gibel carp that is a common fish for sport fishing in Hungary meanwhile is one of the most popular farmed fish in China due to its high demand. Eighteen gibel carp with distortions were collected from a barrage pond in southern Hungary. All fish had large humps in the dorsolateral region due to infection of the muscle between the head and the dorsal fin. The swollen degenerated tissue was filled with myxozoan spores, which were collected for morphological and molecular studies. By size and morphology, the spores were consistent with morphological description of M. lentisuturalis. Histopathological examination showed that the formation of plasmodia containing myxospores leads to severe destruction of muscle tissue. The 18S ribosomal DNA and 28S ribosomal DNA data of the samples presented matched with previous sequences of M. lentisuturalis in GenBank. Phylogenetic analyses confirmed that our sequences belong to a monophyletic group with them supported by a high bootstrap. This study highlights the occurrence of a highly pathogenic myxozoan, M. lentisuturalis in Hungary as a new geographical location.

A myxozoan genome reveals mosaic evolution in a parasitic cnidarian.

BACKGROUND: Parasite evolution has been conceptualized as a process of genetic loss and simplification. Contrary to this model, there is evidence of expansion and conservation of gene families related to essential functions of parasitism in some parasite genomes, reminiscent of widespread mosaic evolution-where subregions of a genome have different rates of evolutionary change. We found evidence of mosaic genome evolution in the cnidarian Myxobolus honghuensis, a myxozoan parasite of fish, with extremely simple morphology. RESULTS: We compared M. honghuensis with other myxozoans and free-living cnidarians, and determined that it has a relatively larger myxozoan genome (206 Mb), which is less reduced and less compact due to gene retention, large introns, transposon insertion, but not polyploidy. Relative to other metazoans, the M. honghuensis genome is depleted of neural genes and has only the simplest animal immune components. Conversely, it has relatively more genes involved in stress resistance, tissue invasion, energy metabolism, and cellular processes compared to other myxozoans and free-living cnidarians. We postulate that the expansion of these gene families is the result of evolutionary adaptations to endoparasitism. M. honghuensis retains genes found in free-living Cnidaria, including a reduced nervous system, myogenic components, ANTP class Homeobox genes, and components of the Wnt and Hedgehog pathways. CONCLUSIONS: Our analyses suggest that the M. honghuensis genome evolved as a mosaic of conservative, divergent, depleted, and enhanced genes and pathways. These findings illustrate that myxozoans are not as genetically simple as previously regarded, and the evolution of some myxozoans is driven by both genomic streamlining and expansion.

The Immune Response to the Myxozoan Parasite Myxobolus cerebralis in Salmonids: A Review on Whirling Disease.

Salmonids are affected by the economically significant whirling disease (WD) caused by the myxozoan parasite Myxobolus cerebralis. In the past, it was endemic to Eurasia, but it has now spread to different regions of North America, Europe, New Zealand, and South Africa. Among salmonids, rainbow trout is considered the most highly susceptible host. Upon entering to the host's body, the parasite invades the spine and cranium, resulting in whirling behaviour, a blackened tail, and destruction of cartilage. The disease is characterized by the infiltration of numerous inflammatory cells, primarily lymphocytes and macrophages, with the onset of fibrous tissue infiltration. Several efforts have been undertaken to investigate the role of various immune modulatory molecules and immune regulatory genes using advanced molecular methods including flow cytometry and transcriptional techniques. Investigation of the molecular and cellular responses, the role of STAT3 in Th17 cell differentiation, and the inhibitory actions of suppressors of cytokine signaling (SOCS) on interferons and interleukins, as well as the role of natural resistance-associated macrophage proteins (Nramp) in WD have significantly contributed to our understanding of the immune regulation mechanism in salmonids against M. cerebralis. This review thoroughly highlights previous research and discusses potential future directions for understanding the molecular immune response of salmonids and the possible development of prophylactic approaches against WD.

The Molecular Mechanisms Employed by the Parasite Myxobolus bejeranoi (Cnidaria: Myxozoa) from Invasion through Sporulation for Successful Proliferation in Its Fish Host.

Myxozoa is a unique group of obligate endoparasites in the phylum Cnidaria that can cause emerging diseases in wild and cultured fish populations. Recently, we identified a new myxozoan species, Myxobolus bejeranoi, which infects the gills of cultured tilapia while suppressing host immunity. To uncover the molecular mechanisms underlying this successful parasitic strategy, we conducted transcriptomics analysis of M. bejeranoi throughout the infection. Our results show that histones, which are essential for accelerated cell division, are highly expressed even one day after invasion. As the infection progressed, conserved parasitic genes that are known to modulate the host immune reaction in different parasitic taxa were upregulated. These genes included energy-related glycolytic enzymes, as well as calreticulin, proteases, and miRNA biogenesis proteins. Interestingly, myxozoan calreticulin formed a distinct phylogenetic clade apart from other cnidarians, suggesting a possible function in parasite pathogenesis. Sporogenesis was in its final stages 20 days post-exposure, as spore-specific markers were highly expressed. Lastly, we provide the first catalog of transcription factors in a Myxozoa species, which is minimized compared to free-living cnidarians and is dominated by homeodomain types. Overall, these molecular insights into myxozoan infection support the concept that parasitic strategies are a result of convergent evolution.

Myxobolus xiushanensis n. sp. (Myxozoa: Myxobolidae) infecting the gill arches of Yellowhead Catfish Tachysurus fulvidraco from China.

OBJECTIVE: Myxosporidiosis of bagrid fishes has been a focus of aquaculture research in recent years. The purpose of this study is to characterize a novel myxobolid, named Myxobolus xiushanensis n. sp., infecting Yellowhead Catfish Tachysurus fulvidraco in China. METHODS: We used molecular biology, morphology, phylogeny, and histopathology in the present study. RESULT: Mature myxospores were circular to ellipsoidal in valve view, measuring 12.2 ± 0.4 µm (mean ± SD; range = 11.2-13.2 µm) in length and 10.6 ± 0.4 µm (9.5-11.1 µm) in width. Two oval polar capsules were equal in width (3.4 ± 0.2 µm; 3.0-3.8 µm) but slightly unequal in length: 5.6 ± 0.3 µm (5.3-6.1 µm) and 4.7 ± 0.2 µm (4.4-5.5 µm). The polar capsule was packed with five to seven spirals of polar tubules. Histopathological investigation demonstrated that the plasmodium under the cuticular layer of the gill arch only induced a local inflammatory response and did not cause serious damage to the gill arch's internal structure. The two small subunit (SSU) ribosomal DNA sequences of M. xiushanensis n. sp. showed 100% similarity and uniqueness, and the highest similarity with other myxosporean sequences in GenBank was 90.27% (query coverage = 94%). The secondary structures of the SSU ribosomal RNA revealed that the present species was distinctly different from related species in regions V4 and V7. Phylogenetic analysis showed that M. xiushanensis n. sp. clustered independently within a branch. CONCLUSION: These results enrich our understanding of the biodiversity of myxobolids infecting bagrid fishes and provide fundamental data for the diagnosis of myxosporidiosis.

Synopsis of myxosporean species (Cnidaria: Myxozoa) parasitizing fishes from Vietnam.

This paper provides an updated checklist of species-level identified myxosporeans from marine and freshwater fishes in Vietnam. The list includes 51 nominal species (38 marine and 13 freshwater) belonging to 9 genera: Myxobolus Bütschli, 1882 (26 species); Kudoa Meglitsch, 1947 (6 species); Henneguya Thélohan, 1892 (6 species); Thelohanellus Kudo, 1933 (5 species); Unicapsula Davis, 1924 (2 species); Ceratomyxa Thélohan, 1892 (2 species), Zschokkella Auerbach, 1909 (2 species); Auerbachia Meglitsch, 1960 (1 species), and Meglitschia Kovaleva, 1988 (1 species). For each parasite species, information on myxospore morphology, line drawings, fish hosts, infection sites, and collection locality in Vietnam are reported. Where available, we also provide GenBank accession numbers for nucleotide sequence data. In addition, taxonomic status of several species was discussed and Myxobolus eszterbaueri nom. nov. is proposed as a junior homonym for Myxobolus hakyi Baska, Voronin, Eszterbauer, Müller, Marton & Molnár 2009, which is preoccupied.

A new Myxobolus (Cnidaria: Myxosporea: Myxobolidae) from the gills of the southern striped shiner, Luxilus chrysocephalus isolepis (Cypriniformes: Leuciscidae), from southwestern Arkansas, USA.

The southern striped shiner, Luxilus chrysocephalus isolepis (Hubbs & Brown) is a relatively large minnow belonging to the true minnow family Leuciscidae Bonaparte. Between May 2020 and January 2022, 55 L. c. isolepis were collected from watersheds in Montgomery (n = 6), Polk (n = 17) and Sevier (n = 32) counties, Arkansas, USA, and their gills, gallbladders, urinary bladders, fins, integument, other major organs, and musculature were examined for myxozoans. Gills of 11 (34%) individual southern striped shiners from Sevier County were infected with a new myxozoan, Myxobolus carlhubbsi n. sp. A qualitative and quantitative morphological description was based on formalin-fixed preserved myxospores, and molecular data consisted of a 1,970 base pair sequence of the partial small subunit rRNA gene from ethanol-preserved specimens. Histologically, plasmodia filled and expanded interlamellar troughs. Hyperplastic epithelial and goblet cells filled interlamellar troughs adjacent to plasmodia, but inflammatory response was limited to scattered lymphocytes. Phylogenetic analysis revealed that M. carlhubbsi n. sp. is a member of a clade of species with pyriform myxospores parasitizing North American Pogonichthyinae Girard and North American and Eurasian Leuciscinae Bonaparte. This is the first report of a myxozoan from L. c. isolepis. This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as urn:lsid:zoobank.org:pub:D10D71C2-2C75-4A1C-80ED-B98FF36CB509.

Myxobolus dumerilii sp. n. (Myxozoa: Myxobolidae) infecting the brain of Chinese longsnout catfish Tachysurus dumerili (Bleeker) in China.

During an investigation of Myxobolus diversity in the Chinese longsnout catfish Tachysurus dumerili (Bleeker), a new species infecting the intracranial epidermis of the host was discovered. Upon opening the cranial cavity, several round whitish plasmodia measuring 0.55-0.80 mm in diameter were observed. Fresh spores (n= 50) were pyriform in the frontal view and fusiform in the sutural view, with a length of 15.4±0.6 (13.9-16.5) µm, width of 9.1±0.4 (8.3-9.8) µm, and thickness of 7.0±0.4 (6.3-7.9) µm. The spores had smooth shell surfaces and transparent membrane sheaths in the posterior. No folds, intercapsular appendix, and caudal appendages were observed. Two equal polar capsules were pyriform and measured 7.5±0.5 (6.7-8.7) µm in length and 3.2±0.3 (2.5-3.6) µm in width. The polar filaments were coiled with five to six turns and perpendicular to the polar capsule length. A BLAST search indicated M. dumerilii sp. n. was closely related to five Myxobolus species (with sequences similarities ranging from 90.54% to 96.52%) found in different organs of yellow catfish Tachysurus fulvidraco (Richardson), rather than the T. dumerili-infecting species M. branchiola Dong and Zhao, 2014 (with 90.5% sequence similarity). Phylogenetic analysis showed that M. dumerilii sp. n. didn't form sister clade with brain-infecting Myxobolus spp, but clustered with M. jianlinensis Gao et Zhao, 2020 and M. voremkhai Akhmerov, 1960 within the Siluriformes-clade with highly supported values, indicating that the host specificity may play a stronger signal than site infections during the evolution of Myxobolus species. Based on the morphological, ecological, and molecular differences observed between the newly discovered species and other available Myxobolus species, M. dumerilii sp. n., is proposed and described in this study.

Identification of Myxobolus distalisensis n. sp. (Cnidaria: Myxozoa) infecting yellow catfish Tachysurus fulvidraco (Richardson), with a supplement description of M. voremkhai (Akhmerov, 1960) Landsberg and Lom, 1991.

With growing scale of intensive fish cultivation, the risk of parasite infection in commercial fish is increased. Precisely identifying and characterizing the parasites that infect the farmed fish is critical to understanding the dynamics of their communities. Here, two species of Myxobolus were identified in farmed yellow catfish Tachysurus fulvidraco (Richardson) in China. Myxobolus distalisensis n. sp. developed plasmodia in the gill filaments, with oval to elliptical myxospores measuring 11.3 ± 0.6 (10.4-12.6) × 8.1 ± 0.3 (7.5-8.6) × 5.5 ± 0.2 (5.2-5.8) µm. Two pyriform polar capsules of equal size were measured 5.3 ± 0.4 (4.5-6.3) × 2.7 ± 0.1 (2.3-3) µm. Myxobolus voremkhai (Akhmerov, 1960) Landsberg and Lom, 1991 developed plasmodia in the gill arch and had a myxospore morphology similar to the conspecific isolates described in previous studies. The consensus sequences of M. distalisensis was remarkably distinct from those deposited in the GenBank, with exception of whereas M. voremkhai showing 99.84% identity. The genetic data on both isolates differed considerably from each other, revealing only 86.96% molecular identity. Histologically, M. distalisensis resided in the filament cartilage, and the aggressive proliferation of the sporogenic stages led to lytic cartilage corrosion. In contrast, plasmodia of M. voremkhai grossly observed at the base of the gill filament were embedded by the connective tissue in the gills arch. Phylogenetically, both isolates were separately placed in different subclades, indicating difference in their evolutionary history. Besides, the taxon under the family Myxobolidae was demonstrated non-monophyletic origins, and parasite radiation largely followed their host affinity.

Synopsis of the aurantiactinomyxon collective group (Cnidaria, Myxozoa), with a discussion on the validity of morphotype definition and demise of guyenotia.

Aurantiactinomyxon is one of the most diverse myxozoan collective groups, comprising types that mostly infect freshwater and marine oligochaetes belonging to the family Naididae Ehrenberg, 1828, but also Lumbriculidae Claus, 1872. In this study, a comprehensive revision of all known aurantiactinomyxon types is performed and highlights the fallibility of using the form and length of the valvular processes as main criterion for differentiating among style-less actinospore morphotypes. The demise of the guyenotia collective group is proposed based on the ambiguous features of several types that allow conformity with both the aurantiactinomyxon and guyenotia definitions. Nonetheless, the information presently available clearly shows that a general shift is needed in our approach to actinospore grouping, which should probably be based on actinospore functionality relative to environment and host ecology, rather than on morphology. Life cycle studies based on experimental transmission and molecular inferences of the 18S rDNA have linked aurantiactinomyxon (including former guyenotia) to myxozoans belonging to a diverse array of genera, including Chloromyxum, Henneguya, Hoferellus, Myxobolus, Paramyxidium, Thelohanellus and Zschokkella. This undoubtedly shows a high capacity of the aurantiactinomyxon morphotype to promote infection in intrinsically distinct vertebrate hosts and environmental habitats, consequently increasing interest in its study for attaining a better understanding of myxozoan-host interactions. The identification of novel and known types, however, is impeded by the lack of concise information allowing a comprehensive analysis of biological, morphological, and molecular criteria. In this sense, the compilation of data presented in this study will ultimately help researchers seeking to perform reliable identifications.