Scoliosis and kyphosis in blue-spotted and marbled flathead fish associated with a Myxobolus acanthogobii-like parasite.

Spinal deformities in finfish have the potential to impact aquaculture industries and wild populations by increasing morbidity, mortality, and reducing growth rates. Myxobolus acanthogobii has been implicated in causing scoliosis and lordosis in various aquatic species in Japan. We investigated 4 cases of spinal deformity in 2 flathead (Platycephalus) species that were submitted to the Elizabeth Macarthur Agricultural Institute (EMAI) in New South Wales (NSW), Australia, between 2015 and 2021. Flathead are commercially significant species that are popular among Australian consumers, and are also sought-after species targeted by recreational fishers. Gross deformities are concerning to the community and may impact the quality and quantity of specimens available for consumption. Three blue-spotted flathead (P. caeruleopunctatus) and one marbled flathead (P. marmoratus) were submitted, all with marked scoliosis and kyphosis; 1-2-mm cysts were present on the dorsum of the brain, most often over the optic lobe or cerebellum. Cytology and differential interference microscopy of cyst material revealed numerous oval spores, x̄ 14 ± SD 0.75 µm × x̄ 11.5 ± SD 0.70 µm, with 2 pyriform polar capsules, the morphology of which is consistent with a Myxobolus sp. PCR assay and 18S rDNA sequencing of the cyst material identified a Myxobolus sp. with 96% identity to M. acanthogobii. The identification of this Myxobolus sp. confirms the presence of parasites with the potential to cause spinal deformity in significant aquatic species in NSW waterways.

Myxozoan survey of thicklip grey mullet Chelon labrosus reinforces successful radiation of Myxobolus in mugiliform hosts.

A myxozoan survey was performed on specimens of thicklip grey mullet Chelon labrosus (Risso) captured from the Douro River estuary, northern Portugal. Eleven new species, all belonging to the genus Myxobolus Bütschli, 1882 (M. abdominalis n. sp., M. aestuarium n. sp., M. caudalis n. sp., M. chelonari n. sp., M. cucurbitiformis n. sp., M. douroensis n. sp., M. intestinicola n. sp., M. invictus n. sp., M. labicola n. sp., M. peritonaei n. sp., and M. pinnula n. sp.) are described based on microscopic and molecular data, confirming the known high radiation of these myxozoans in mullets. Additionally, Myxobolus pupkoi Gupta et al., 2022 is reported for the first time from C. labrosus, bringing forth a novel case of morphological plasticity between geographic isolates. We consider that molecular-based comparisons are imperative for the description of mugiliform-infecting Myxobolus, with distance estimation further matching two of the novel Myxobolus spp. with sphaeractinomyxon types previously reported from another Portuguese estuary. This finding supports sphaeractinomyxon as specific life cycle counterparts of Myxobolus that infect mullets. Phylogenetic analyses of 18S rDNA retrieved a monophyletic clade of mugiliform-infecting myxobolids comprising well-supported lineages of species parasitizing mullets from the genera Chelon, Mugil, Crenimugil, and Planiliza. The existence of more than one Chelon- and Planiliza-infecting lineage reveals that myxobolids parasitized members of these genera multiple times during their evolution. Lastly, the elevated number of unmatched sphaeractinomyxon sequences included in the Chelon-infecting lineages clearly shows that Myxobolus diversity hosted by this genus remains underrated.

Title: Un inventaire des myxozoaires du mulet lippu Chelon labrosus confirme le rayonnement réussi de Myxobolus chez les hôtes mugiliformes. Abstract: Un inventaire des myxozoaires a été réalisé sur des spécimens de mulets lippus Chelon labrosus (Risso) capturés dans l'estuaire du fleuve Douro, au nord du Portugal. Onze nouvelles espèces, toutes appartenant au genre Myxobolus Bütschli, 1882 (M. abdominalis n. sp., M. aestuarium n. sp., M. caudalis n. sp., M. chelonari n. sp., M. cucurbitiformis n. sp., M. douroensis n. sp., M. intestinicola n. sp., M. invictus n. sp., M. labicola n. sp., M. peritonaei n. sp. et M. pinnula n. sp.) sont décrites sur la base de données microscopiques et moléculaires, confirmant le rayonnement connu de ces myxozoaires chez les mulets. De plus, Myxobolus pupkoi Gupta et al., 2022 est signalé pour la première fois chez C. labrosus, démontrant un nouveau cas de plasticité morphologique entre des isolats géographiques. Nous considérons que les comparaisons moléculaires sont impératives pour la description des Myxobolus infectant les mugiliformes, l'estimation de la distance correspondant en outre à deux des nouveaux Myxobolus spp. avec des types de sphaeractinomyxons précédemment signalés dans un autre estuaire portugais. Cette découverte soutient les sphaeractinomyxons en tant que contreparties spécifiques du cycle de vie de Myxobolus qui infectent les mulets. Les analyses phylogénétiques de l'ADNr 18S ont montré un clade monophylétique de Myxobolidae infectant les mugiliformes, comprenant des lignées robustes d'espèces parasitant les mulets des genres Chelon, Mugil, Crenimugil et Planiliza. L'existence de plusieurs lignées infectant Chelon et Planiliza révèle que les Myxobolidae ont parasité des membres de ces genres plusieurs fois au cours de leur évolution. Enfin, le nombre élevé de séquences de sphaeractinomyxons non appariées incluses dans les lignées infectant Chelon montre clairement que la diversité de Myxobolus hébergée par ce genre reste sous-estimée.

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.

Immunosuppression and apoptosis activation mediated by p53-Bcl2/Bax signaling pathway -The potential mechanism of goldfish (Carassius auratus Linnaeus) gill disease caused by Myxobolus ampullicapsulatus.

Myxobolus, a major harmful type of myxospora, is one of the main parasitic pathogens of freshwater fish. Once myxoboliosis occurs, treatment can be extremely difficult. Therefore, clear understandings of the etiology of myxoboliosis and its pathological mechanism are keys for prevention and control. Here, histology, transmission electron microscopy, transcriptome study, tunel assay, and immunohistochemistry were carried out, revealing the morphology, pathological effects as well as host response mechanism of goldfish gill to Myxobolus ampullicapsulatus. Histological studies showed that the mature spores of Myxobolus ampullicapsulatus were composed of three parts, the spore shell, sporoplasm and bottle shaped polar capsule containing double S-shaped polar filaments. Transcriptome analysis revealed that Myxobolus ampullicapsulatus -infected (Myx) goldfish gills were characterized by apoptosis activation mediated by "p53 signaling pathway" with significantly up-regulated apoptosis-related differential genes dominated by p53-Bcl2/Bax signaling pathway. In addition, tunel assay revealed severe gill apoptosis in the Myx group. Transcriptome analysis also revealed that Myx group showed changes in immune response and significantly down-regulated immune-related differential genes. Beyond that, immunohistochemistry showed that there was no significant increase in the number of gill lymphocyte after parasite infection. These results suggest that the pathological mechanism of Myxobolus ampullicapsulatus infection on gills of goldfish may be related to apoptosis and immunosuppression. Subsequent qRT-PCR showed that apoptosis-related genes (Caspase3,Bad, Bax) and anti-inflammatory gene IL-10 were significantly increased, while immune-related pro-inflammatory genes (IL-1ß, IL-8) were markedly down-regulated, further verifying the transcriptome results. Based on the above results, we concluded that p53-Bcl2/Bax related networks that dominant the expression of apoptosis genes were activated while immunity was suppressed in the gills of Myxobolus ampullicapsulatus infected goldfish. Our study is not only of benefit to enrich the taxonomy of Myxobolus but also clarifies its pathogenic mechanism, thus providing targets for prevention and control of myxoboliosis.

Two new species of Myxobolus Bütschli, 1882 (Cnidaria: Bivalvulida: Myxobolidae) infecting the gill of the black redhorse, Moxostoma duquesnei (Lesueur) (Cypriniformes: Catostomidae) in the Little Tennessee River Basin, North Carolina.

Two new species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) are described from the gill of the black redhorse (Moxostoma duquesnei [Leueur][Cypriniformes: Catostomidae]) from the Little Tennessee River Basin, North Carolina, United States. Myxobolus branchiofilum n. sp. infects lumen of the lamellar arterioles and Myxobolus branchiopecten n. sp. infects the bone and cartilage at the tip of the gill rakers. They differ from all congeners by a combination of myxospore dimensions and the presence or absence of an iodinophilic vacuole in the sporoplasm, mucous envelope, intercapsular process, and sutural markings. A phylogenetic analysis of the small subunit ribosomal DNA recovered M. branchiopectin sister to Myxobolus sp. (AF378343) in a clade composed of 6 species of Myxobolus, which infect predominately cypriniform intermediate hosts. Myxobolus branchiofilum was recovered sister to Myxobolus ictiobus Rosser, Griffin, Quiniou, Alberson, Woodyard, Mischker, Greenway, Wise & Pote, 2016 in a clade composed of 8 species of Myxobolus, which predominately infect catostomid intermediate hosts. Histological sections of infected gill revealed intra-lamellar plasmodia of M. branchiofilum in the lumen of the lamellar arterioles and foci of M. branchiopecten developing in the bone and cartilage of the gill raker tip. These are the first myxozoans reported from the black redhorse. Given that these two new species are morphologically congeneric but recovered in distantly related clades, we discuss the persistent issue of myxobolid genera paraphyly/polyphyly.

Streptococcus, Centrocestus formosanus and Myxobolus tilapiae concurrent infections in farmed Nile tilapia (Oreochromis niloticus).

Stress triggered concurrent microbial/parasitic infections are prevalent in earthen pond based farmed Nile tilapia Oreochromis niloticus. In the current study, a total of thirty five O. niloticus were collected from a commercial fish farm with a history of severe mortalities at Port Said, Egypt. Nile tilapia samples were subjected to bacteriological, parasitological and pathological examinations. Twenty one Enterococcus fecalis and 15 Streptococcus agalactiae isolates were presumptively identified utilizing the semi-automated API 20 Strept test kit. The identities of the retrieved bacteria were confirmed by the sequencing of 16 S rRNA gene. Moribund O. niloticus were found to be heavily infected by one or both of Centrocestus formosanus encysted metacercariae (EMC) and/or Myxobolus tilapiae spores presenting a unique form of synergistic and/or symbiotic relationship. The identities of both parasites were confirmed through morphological and molecular characterization. Variable circulatory, degenerative, necrotic and proliferative changes were also noticed in hematopoietic organs. Interestingly, multiple myxobolus spores and EMC were noticed in some histological sections. It was obvious that the current concurrent bacterial and parasitic infections are triggered by the deleterious effects of some stressing environmental conditions. The unfavorable climatic conditions (high temperature and high relative humidity) recorded at the surge of mortalities are probable predisposing stress factors.

A NEW SPECIES OF MYXOBOLUS (CNIDARIA: MYXOSPOREA: MYXOBOLIDAE) FROM THE BLUE SUCKER, CYCLEPTUS ELONGATUS (LESUEUR) (CYPRINIFORMES: CATOSTOMIDAE: CYCLEPTINAE), FROM ARKANSAS.

During 9-10 February 2018 and 21-22 February 2020, 7 adult Blue Suckers, Cycleptus elongatus, were collected by hoop nets from the Red River, Little River County (n = 3), and the Black River, Lawrence County (n = 4), Arkansas, and their gills, gallbladders, fins, integument, other major organs, and musculature were examined for myxozoans. All 7 (100%) were infected with an unknown species of gill-infecting Myxobolus sp. Twenty formalin-fixed plasmodia (cysts) of Myxobolus cloutmani n. sp. were elliptoidal, 407 µm long × 270 µm wide. Formalin-fixed myxospores were orbicular to broadly elliptoidal, 8.7 µm long × 7.8 µm wide. Two polar capsules were pyriform and subequal in size, extending over halfway in the myxospore. The larger polar capsule was 5.5 µm long × 3.1 µm wide, while the shorter was 5.1 × 2.9 µm. A coiled polar filament possessed 5 or 6 coils. The myxospore was 3.7 µm thick in sutural view, with a distinct sutural ridge. Qualitative and quantitative morphological data were from formalin-fixed as well as ethanol-preserved spores, while molecular data consisted of a 2,010 base pair sequence of the partial 18S ribosomal RNA gene and a 2,502 base pair sequence of the partial 28S ribosomal RNA gene. Phylogenetic analysis grouped M. cloutmani n. sp. with the other catostomid-infecting myxobolids. This is the first myxozoan reported from C. elongatus.

Myxobolus freitasi n. sp. (Myxozoa: Bivalvulida), a parasite of the brain of the electric knifefish in the Brazilian Amazon region / Myxobolus freitasi n. sp. (Myxozoa: Bivalvulida), parasita do cérebro do peixe-faca elétrico na Amazônia brasileira

Abstract A total of 30 specimens of the Amazonian electric knifefish, Brachyhypopomus beebei Schultz, 1944 (Gymnotiformes: Hypopomidae), were collected from the Peixe-Boi River in the state of Pará, Brazil (1°06'59" S; 47°18'26" W). Fragments of the brain tissue were extracted for analysis via optical microscopy, and 18 specimens (60%) presented microparasites of the genus Myxobolus, with unequal capsules. The spores were 18.6 µm (17.7-19.8 µm) long and 8.6 µm (8.4-9.0 µm) wide; the largest polar capsule was 13.0 µm (12.4-13.4 µm) long and 5.6 µm (5.3-6.0 µm) wide, and the smallest capsule was 5.0 µm (4.5-5.3 µm) long and 2.5 µm (2.3-2.6 µm) wide. Infected brain fragments were extracted for histological processing and staining with hematoxylin-eosin and Ziehl-Neelsen. Some fragments were conserved in ethanol for molecular genetics analysis. A partial sequence of the 18S DNA gene was obtained from the spores, which did not correspond to any other sequences deposited in GenBank, although it did form a clade with other Myxobolus parasites of the nervous system. The morphological data, together with molecular phylogeny, supported the designation of a new species Myxobolus freitasi n. sp.

Resumo Um total de 30 espécimes do peixe-faca elétrico da Amazônia, Brachyhypopomus beebei Schultz, 1944 (Gymnotiformes: Hypopomidae), foram coletados no rio Peixe-Mani, no estado do Pará, Brasil (1 ° 06'59 "S; 47 ° 18 ' 26 "W). Fragmentos de tecido cerebral foram extraídos para análise em microscopia óptica, sendo que 18 espécimes (60%) apresentavam microparasitos do gênero Myxobolus, com cápsulas desiguais. Os esporos apresentavam 18,6 µm (17,7-19,8 µm) de comprimento e 8,6 µm (8,4-9,0 µm) de largura; a maior cápsula polar tinha 13,0 µm (12,4-13,4 µm) de comprimento e 5,6 µm (5,3-6,0 µm) de largura, e a menor cápsula tinha 5,0 µm (4,5-5,3 µm) de comprimento e 2,5 µm (2,3-2,6 µm) de largura. Fragmentos cerebrais infectados foram extraídos para processamento histológico e coloração com hematoxilina-eosina e Ziehl-Neelsen. Alguns fragmentos foram conservados em etanol para análise genética molecular. Dos esporos, foi obtida uma sequência parcial do gene 18S do DNA, que não correspondeu a nenhuma outra sequência depositada no GenBank, embora tenha formado um clado com outros parasitas do gênero Myxobolus do sistema nervoso. Os dados morfológicos, juntamente com a filogenia molecular, apoiaram a designação de uma nova espécie Myxobolus freitasi n. sp.

Morphological and molecular characterization of Myxobolus dibombensis sp. n. (Myxozoa: Myxobolidae), a parasite of the African carp Labeobarbus batesii (Teleostei: Cyprinidae) from Dibombe River, Cameroon.

Myxobolus dibombensis sp. n. (Cnidaria: Myxosporea: Bivalvulida) is described from the fins of the African carp, Labeobarbus batesii, based on morphological and molecular data. Prevalence of infection was 51.9% (67/129). Ovoid to spherical cyst-like plasmodia were found in the intrasegmental region and among the fin rays. No pathological changes were found in the fish host tissue surrounding the cyst-like plasmodia. Mature myxospores were ovoid in frontal view and lenticular in lateral view, with slightly truncated anterior and rounded posterior ends. Myxospores measured 16.8 (15.8-18.0) µm long and 11.4 (10.0-13.0) µm wide. There was a triangular intercapsular appendix measuring 3.8 (2.6-4.5) µm long. Polar capsules were ovoid and slightly unequal in size, occupying approximately one-third of the myxospore length. The larger polar capsule measured 7 (6-8) µm long and 3.6 (3-4) µm wide, while the smaller one measured 5.8 (4.8-7.0) µm long and 3 (2-4) µm wide. The larger polar capsule contained nine to 11 filament coils, whereas the smaller one contained seven to nine coils. SSU rDNA gene sequence of M. dibombensis sp. n. did not match any sequences available in the GenBank. The similarity with available Myxobolus spp. sequences ranged from 65 to 81%. The novel species clustered with M. algonquinensis, which infects the cyprinid Luxilus cornutus from Canada.

Supplemental diagnosis and phylogeny of Myxobolus absonus (Cnidaria, Myxozoa) from the eye of the freshwater fish Pimelodus maculatus (Siluriformes, Pimelodidae).

Myxobolus absonusCellere et al., 2002 was originally described as having free cysts in the opercular cavity of the freshwater fish Pimelodus maculatus in Brazil. The present study provides a supplemental description of this parasite from the eye of its type host, with basis on morphological, ultrastructural, and molecular data of the SSU rRNA gene. The parasite formed spherical whitish cysts, which wall presented numerous microvilli that attached to the collagen layers of the corneal stroma. Myxospores were oval in valvular and sutural view, measuring 13.2 ± 0.4 µm in length, 8.5 ± 0.4 µm in width, and 6.6 ± 0.3 µm in thickness. Two asymmetric pyriform polar capsules were located at the anterior pole: the larger 6.2 ± 0.4 µm long and 3.6 ± 0.3 µm wide, containing a polar filament coiled in 6 turns; and the smaller 3.5 ± 0.3 µm long and 1.9 ± 0.1 µm wide, containing a polar filament coiled in 4 turns. At the posterior pole, the sporoplasm displayed two nuclei and numerous spherical sporoplasmosomes. Phylogenetic analysis using maximum likelihood, Bayesian inference and maximum parsimony revealed M. absonus clustering within a well-supported clade with poorly-resolved internal nodes, amongst the SSU rRNA sequences of other myxobolids that infect siluriform and characiform fish hosts, as well as the perciform-infecting species Myxobolus acanthogobii, Triangula percae and Cardimyxobolus japonensis. This clade appeared separated from the other clades comprising most of the species that infect siluriform and characiform hosts, showing that more than one myxobolid lineage evolved while parasitizing these taxonomic groups of fish.

First molecular evidence of Thelohanellus wallagoi Sarkar, 1985 (Myxozoa) from economically important food fish, freshwater shark Wallago attu (Siluridae) in India.

The freshwater shark Wallago attu (Bl. and Schn.) is a frequent silurid in the River Ganga and one of the most commercially exploit fish in India. In a survey on its infection with myxosporeans, spore type belongs to Thelohanellus species was found in the gills, kidney and intestine respectively. Through morphological and molecular investigations, we identified the spore as Thelohanellus wallagoi Sarkar, 1985. They were pyriform in valvular view and slim in sutural view, and had one pyriform polar capsule with four to five turns. The spores measured 8.0 × 4.0 × 2.3 µm. T. wallagoi developed in small cysts in the gill lamellae, whereas cyst and scattered spores of T. wallagoi were also found in the kidney and intestine respectively. The 18S rDNA sequence of T. wallagoi isolates recovered from gills, kidney and intestine were found similar to each other and differed from any other Thelohanellus species available in GenBank and validated its status after 32 years of original description. Phylogenetic analysis signified that T. wallagoi was placed sister to Myxobolus species in the clade that indicated the polyphyletic nature of the genus Thelohanellus.

Myxobolus infection in the cornea of the roach (Rutilus rutilus) in Lake Balaton.

Infection of the cornea in fishes by Myxobolus plasmodia is a common but still little known site preference of myxosporeans. A sporadic but striking infection in the cornea of the roach (Rutilus rutilus) was observed in Lake Balaton, Hungary. Relatively small, round plasmodia 250 to 500 µm in diameter developed in the dense connective tissue of the cornea. Morphological and molecular biological examination of spores collected from cysts in the cornea demonstrated that this infection is caused by Myxobolus fundamentalis, a species hitherto reported only from the cartilaginous gill arch of the roach. The 18S rDNA sequences of spores from the cornea showed 99.9% identity to the sequences of spores from the gill arch, and they also shared 99.9% identity with the sequences of triactinomyxon actinospores obtained from the oligochaete Isochaetides michaelseni.

Morphological and molecular characterization of a novel myxosporean parasite Myxobolus bejeranoi n. sp. (Cnidaria: Myxosporea) from hybrid tilapia in Israel.

Myxosporean infections can cause severe damage to commercially grown tilapia. Here, we report a novel myxosporean that was found in gills of Oreochromis aureus male × Oreochromis niloticus female, which is an important aquaculture tilapia hybrid in Israel. Three-month-old fish were found to have cysts located in gill muscle tissue, which were filled with both immature and mature spores. Affected fish displayed higher mortality rate. Spore dimensions (10.8 ± 0.7 µm length × 6.8 ± 0.6 µm width) and molecular characterization using 18S ribosomal DNA revealed that the unknown parasite belongs in the Myxobolus clade. Based on the infection site, spore morphology and molecular characterization, we describe this parasite as Myxobolus bejeranoi n. sp. (MF401455). Phylogenetic analysis showed that the new species is most closely related to two Myxobolus spp. from O. niloticus in Egypt and Ghana.

Myxobolus marajoensis sp. n. (Myxosporea: Myxobolidae), parasite of the freshwater catfish Rhamdia quelen from the Brazilian Amazon region / Myxobolus marajoensis sp. n. (Myxosporea: Myxobolidae), parasita do bagre de água doce Rhamdia quelen da região da Amazônia brasileira

Abstract This study provides morphological and molecular data of a new parasite species found in the muscle layer of the intestinal tract of the South American silver catfish, Rhamdia quelen from Marajó Island region (Pará State, Brazil), an important fishery resource with recognized potential for fish farming. The morphology of these parasites was reanalyzed and phylogenetic analyses were run on their 18S rDNA gene sequences. The spores were morphologically distinct from those of other Myxobolus species described previously. The obtained partial sequence of the 18S rDNA gene sequences of the new species were compared to those of 24 other Myxobolus and Henneguya species available in GenBank. The results of morphological and molecular analyses indicated clearly the existence of a new species, Myxobolus marajoensis sp. n.

Resumo Este estudo fornece dados morfológicos e moleculares de um novo parasita encontrado na parede intestinal do jandiá, Rhamdia quelen coletado na região da ilha do Marajó (Estado do Pará, Brasil), um importante recurso pesqueiro com potencial para aquicultura. Foram realizadas comparações morfológicas deste parasita e análises filogenéticas da região do gene 18S rDNA sequenciada. Os esporos foram morfologicamente distintos das espécies de outros Myxobolus descritos anteriormente. A sequência parcial obtida do gene 18S rDNA da nova espécie foi comparada com outras 24 espécies de Myxobolus e Henneguya retiradas do GenBank. Os resultados de análises morfológica e molecular indicaram claramente a existência de uma nova espécie, Myxobolus marajoensis sp. n.

A novel myxosporean parasite Myxobolus klamathellus n. sp. (Cnidaria: Myxosporea) from native blue chub (Gila coerulea) in Klamath Lake, Oregon.

Blue chub, Gila coerulea Girard, 1856 is a freshwater cyprinid fish native to inland drainages of western North America. It has not previously been recorded as a host of any myxosporean parasite (Cnidaria: Myxosporea), despite myxosporeans being cosmopolitan in freshwater and marine fishes worldwide and sympatric with this host. Herein, we describe a novel myxosporean from subcutaneous cysts in native blue chub from Klamath Lake, Oregon. Myxospores were consistent with genus Myxobolus, being obovoid but compressed in thickness, length 14.3 ± 0.4 (13-15) µm, width 9.7 ± 0.4 (9-10) µm, thickness 7.7 ± 0.3 (7-8) µm; two polar capsules ovoid slightly dissimilar in size, length 6.4 ± 0.4 (6-7) µm, width 3.8 ± 0.3 (3-4) µm, with four (3-5) turns of the polar filament (tubule); capsule openings apical, one in each valve cell. The small subunit ribosomal DNA sequence was up to 97 % similar to Myxobolus spp. from other cyprinids from North America and Europe. Given the novel host, unique myxospore morphometrics, and DNA sequence, we describe this as Myxobolus klamathellus n. sp.

Myxozoa + Polypodium: A Common Route to Endoparasitism.

Recent evidence places the problematic Polypodium, a parasite of fish eggs, firmly as sister taxon to Myxozoa within the Cnidaria. This resolution suggests a single route to endoparasitism in Cnidaria, with larval stages of a common ancestor exploiting fish as first hosts. It also enables new interpretations and insights regarding evolutionary transitions associated with endoparasitism.

Genomic insights into the evolutionary origin of Myxozoa within Cnidaria.

The Myxozoa comprise over 2,000 species of microscopic obligate parasites that use both invertebrate and vertebrate hosts as part of their life cycle. Although the evolutionary origin of myxozoans has been elusive, a close relationship with cnidarians, a group that includes corals, sea anemones, jellyfish, and hydroids, is supported by some phylogenetic studies and the observation that the distinctive myxozoan structure, the polar capsule, is remarkably similar to the stinging structures (nematocysts) in cnidarians. To gain insight into the extreme evolutionary transition from a free-living cnidarian to a microscopic endoparasite, we analyzed genomic and transcriptomic assemblies from two distantly related myxozoan species, Kudoa iwatai and Myxobolus cerebralis, and compared these to the transcriptome and genome of the less reduced cnidarian parasite, Polypodium hydriforme. A phylogenomic analysis, using for the first time to our knowledge, a taxonomic sampling that represents the breadth of myxozoan diversity, including four newly generated myxozoan assemblies, confirms that myxozoans are cnidarians and are a sister taxon to P. hydriforme. Estimations of genome size reveal that myxozoans have one of the smallest reported animal genomes. Gene enrichment analyses show depletion of expressed genes in categories related to development, cell differentiation, and cell-cell communication. In addition, a search for candidate genes indicates that myxozoans lack key elements of signaling pathways and transcriptional factors important for multicellular development. Our results suggest that the degeneration of the myxozoan body plan from a free-living cnidarian to a microscopic parasitic cnidarian was accompanied by extreme reduction in genome size and gene content.

Supplementary studies and the first molecular data on Myxobolus scardinii Reuss, 1906 (Myxozoa: Myxosporea) infecting the gill filaments of rudd, Scardinius erythrophthalmus (L.).

Within the present study, we offer new information on the morphology, histopathology, and 18S ribosomal DNA (rDNA) sequence of Myxobolus scardinii to supplement and revise the original description of Reuss (Bull Acad Imp Sci St-Pétersbourg, V Serie, XXV, No. 3: 199-205, 1906). M. scardinii was found infecting the gills of 6 of 125 specimens (4.8%) of Scardinius erythrophthalmus from northern Turkey. Large mature plasmodia, whitish, spherical, or ellipsoidal, measuring 0.9-1.0 mm in diameter, macroscopically occurred in the gills. Histopathological examination indicated that development of the cyst-like plasmodia was intrafilamental-vascular type. Mature spores are oval or shortly ellipsoidal. The spores are 12.1 ± 0.2 (11.5-12.6) µm long, 8.9 ± 0.3 (8.4-9.2) µm wide, and 6.1 ± 0.3 (5.9-6.2) µm thick. The two polar capsules are equal in size, 4.8 ± 0.2 (4.7-5.1) µm long and 2.8 ± 0.2 (2.7-3.1) µm wide, and are long pyriform in shape. A well-developed intercapsular appendix was found in the fresh spores. In this study, a specific gill myxosporean of S. erythrophthalmus, M. scardinii the identity of which was confirmed by careful morphological and histopathological examination, was sequenced with its 18S rDNA sequence for the first time and submitted to Genbank database (accession number KJ562362). Phylogenetic analysis of the 18S rDNA gene revealed that M. scardinii had the closest similarity to M. muelleri and M. bramae. This is the first molecular data for the validity of M. scardinii in S. erythrophthalmus. This new valid genetic data (KJ562362) has been used to establish phylogenetic relationships with other similar gills Myxobolus species from Eurasia.

Myxobolus anatolicus sp. nov. (Myxozoa) infecting the gill of Anatolian khramulya Capoeta tinca (Cyprinidae) in Turkey.

This work is part of an ongoing investigation into the characteristics of myxozoan parasites of freshwater fish in Turkey and was carried out using morphology, histopathology and molecular analysis. A new species of the genus Myxobolus (M. anatolicus sp. nov.) was found infecting the gills of 3 of 34 specimens (8.8%) of Anatolian khramulya Capoeta tinca from the Samsun Province, Northern Turkey. Both morphology and 18S rDNA sequence data revealed that M. anatolicus sp. nov. was distinct from other Myxobolus species found in the gills of cyprinid fishes. The small, white and round-shaped plasmodia, measuring 0.2 to 1.4 mm in diameter, were observed macroscopically in the gills. Histological analysis revealed that the cyst-like plasmodia have an intralamellar-vascular type development. Mature spores of M. anatolicus sp. nov. were oval in both frontal and sutural views, and tapered at the anterior poles. The spores were 10.1 ± 0.41 (9.4 to 10.7) µm long, 6.9 ± 0.28 (6.6 to 7.2) µm wide, and 4.5 ± 0.36 (4.4 to 4.6) µm thick. The 2 polar capsules were pyriform, equal in size, 4.6 ± 0.45 (4.4 to 4.8) µm long and 2.1 ± 0.12 (2 to 2.3) µm wide. Polar filaments within the polar capsules were coiled with 5 or 6 turns. Phylogenetic analysis placed M. anatolicus sp. nov. in a clade of gill-infecting myxobolids. This is the first record of a Myxobolus species infecting Anatolian khramulya Capoeta tinca, and the first record of this species from Eurasia.

Phylogeny, ultrastructure and histopathology of Myxobolus lomi sp. nov., a parasite of Prochilodus lineatus (Valenciennes, 1836) (Characiformes: Prochilodontidae) from the Peixes River, São Paulo State, Brazil.

This paper presents the morphological, histological, molecular and ultrastructural data on Myxobolus lomi sp. nov., a parasite of the gill filaments of Prochilodus lineatus from the Peixes River (48º06'38″W; 22º 49'53.1″S), São Paulo State, Brazil. From 20 P. lineatus specimens examined, 90.0% (n=18) were infected. The plasmodia were white and round, measuring 250 to 300µm in diameter and the development occurred in the base of the gill filament. The spores showed symmetrical and smooth valves, with the polar filament having 8 to 11 coils. A thorough comparison with all the Myxobolus species described so far is provided. A partial sequencing of the 18S rDNA gene revealed approximately 1600-bp. The Myxobolus species parasite of P. lineatus did not match any of the Myxozoa available in GenBank. In the phylogenetic analysis, M. lomi sp. nov. is clustered with ten other species and only four of these parasites were from gills. Histological analysis of P. lineatus gills infected by M. lomi sp. nov. revealed numerous well-delimited cysts at the base of the primary lamella, between connective tissue and bone, next to the gill arteries. However no pronounced inflammatory response was found at the infection site.