Palynological analysis of the genus Ctenitis (C. Chr.) C. Chr. (Dryopteridaceae) in the Southern Cone of America.

The spore morphology and wall ultrastructure of 12 species of Ctenitis from Southern Cone of America were studied using light microscope, scanning and transmission electron microscope. The study was carried out with herbarium material from Argentine and Brazilian institutions. Equatorial diameters, polar diameters and laesura length were measured. The spores are monolete with echinate or folded ornamentation. In the echinate type, the spines are conical, with broad base and attenuate apex. In the rugate type, the folds are inflated, linear, sinuous, subglobose or handle-shape. The perispore surface is scabrate, rugulate, microverrucose or psilate. Stratification and ultrastructure in the species analyzed are very similar. The exospore is smooth and two-layered in section. Simple and branched channels are observed mainly in the outer exospore. The perispore is composed of two layers, the inner one forms the ornamentation and the outer covers all the outer and inner surfaces. Immature spores were found in all samples of C. fenestralis. The characteristics of the studied spores like macro-ornamentation, color and fold length provide relevant information to differentiate some species or groups of species within the genus.

Identification of a new species, Unicapsula aequilobata n. sp., and Unicapsula seriolae (Myxozoa: Myxosporea: Multivalvulida) in carangid fish from the South China Sea.

An examination of 18 fishes caught in the South China Sea detected two Unicapsula spp. in the myofibers of the trunk muscles of carangid fishes: Unicapsula aequilobata n. sp. in the Japanese scad, Decapterus maruadsi, and Unicapsula seriolae in the yellowstripe scad, Selaroides leptolepis. They formed thin filamentous pseudocysts of 0.9-2.0 (mean 1.4) mm by 0.03-0.06 (0.04) mm (n = 5) and 0.9-3.4 (2.1) mm by 0.02-0.05 (0.04) mm (n = 12), respectively. Myxospores of U. aequilobata n. sp. are composed of three equal shell valves and measured 6.7-8.5 (7.3) µm in length and 7.1-8.8 (7.6) µm in width, and contained a prominent polar capsule (PC) 3.2-3.8 (3.6) µm in diameter (n = 18) and two rudimentary PCs. A nucleotide sequence (5127 bp) of the ribosomal RNA gene (rDNA) array was obtained for the genetic characterization of this new species. Based on morphological and phylogenetic criteria, we erect U. aequilobata n. sp. as the sixteenth species in the genus Unicapsula. Nucleotide sequences of the 18S and 28S rDNA obtained from U. seriolae from the yellowstripe scad were almost identical (99.6-100% or 99.0-99.6%, respectively) to those from fish found in the seawaters around Australia and Japan. Consequently, this is a new host and geographical distribution records for U. seriolae. In addition, we illustrated the predicted secondary structure of the available 5.8S rDNA sequences of multivalvulid species, including those obtained from U. aequilobata n. sp., to assess the significance of interspecific nucleotide variations in this short rDNA unit.

Morphological and phylogenetic characterisation of Unicauda tavaresii n. sp. (Myxosporea: Myxobolidae): a parasite of the circumorbital tissue of the eye of two characiform fishes from the Amazon region of Brazil.

Myxozoans of the family Myxobolidae are common parasites in fish. The diversity and ecology of the species of the genus Unicauda are poorly known, which hampers the understanding of the distribution and prevalence of this group of parasites. In the present study, cysts containing parasites whose morphology was consistent with the genus Unicauda were found in the circumorbital region of the ocular conjunctiva of the freshwater fish Moenkhausia grandisquamis Müller & Troschel, 1845 (Characiformes: Characidae) and Triportheus angulatus Spix & Agassiz, 1829 (Characiformes: Triportheidae). The spores have an oval body and long caudal appendage, with a mean total length of 65.2 ± 5.9 µm and width of 5.2 ± 0.7 µm, with two oval and symmetrical polar capsules of 4.9 ± 0.5 µm in length and 1.4 ± 0.2 µm in width, containing polar filaments with five or six coils. An integrated comparative analysis of the morphological characteristics of this parasite and partial sequences of the SSU rDNA gene supported the identification of a new species of histozoic parasite of the genus Unicauda found in fish from the Tocantins River basin, in the eastern Brazilian Amazon region. The new species was denominated by Unicauda tavaresii n. sp.

A New Species of ThelohanellusKudo, 1933 (Myxozoa: Bivalvulida) Infecting Skeletal Muscle of Blacktail Shiner, Cyprinella venusta Girard, 1856 (Cypriniformes: Cyprinidae) in the Chattahoochee River Basin, Georgia.

Thelohanellus magnacysta n. sp. (Bivalvulida: Myxobolidae) infects the skeletal muscle of blacktail shiner, Cyprinella venusta Girard, 1856 (Cypriniformes: Cyprinidae) in Bull Creek, Chattahoochee River Basin, eastern Georgia. Although numerous members of ThelohanellusKudo, 1933 have overlapping myxospore dimensions with the new species, it differs from all nominal congeners by polar filament coil number and polar capsule width as well as by lacking a mucous envelope, iodinophilic vacuole, and sutural markings. With the use of novel primers for Myxozoa, a phylogenetic analysis of the small subunit ribosomal DNA (SSU rDNA) suggests that the new species shares a recent common ancestor with a clade of cyprinid-infecting species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) and Thelohanellus. Consistent with other published research concerning the systematics of Thelohanellus, this result suggested that Thelohanellus and Myxobolus are polyphyletic and need revision. Histological sections of infected blacktail shiners confirmed that myxospores were only found within a plasmodium and only infected skeletal muscle and that plasmodia were encapsulated by a granuloma comprising varying degrees of acute granulomatous inflammation. The new species is the fourth of Thelohanellus reported from North America and the first reported from Cyprinella, as well as the first myxozoan described from the blacktail shiner.

Morphological and Molecular Characterisation of Myxidium kudoi Meglitsch, 1937 from the Blue Catfish Ictalurus furcatus, Valenciennes in Oklahoma, USA.

BACKGROUND: Myxidium kudoi Meglitsch, 1937 has been described from the type host, blue catfish Ictalurus furcatus, with no additional host records or molecular data available for this species. PURPOSE: To provide molecular data and a novel host locality for this species and carry out phylogenetic analyses to infer the evolutionary relationship of the species to other members of the family Myxidiidae for which DNA sequence data is available. METHODS: These data were collected using myxospores from the gallbladder of a blue catfish, Ictalurus furcatus collected from Lake Texoma, Oklahoma, USA. Myxospores were morphologically consistent with the only other account of this species and not any other Myxidium species described from siluriform fishes. RESULTS: Myxospores were oblong with rounded ends and were 10.8-12.6 (11.6 ± 0.5) µm in length and 4.7-6.6 (5.7 ± 0.5) µm in width. Polar capsules were subspherical and 2.7-3.9 (3.4 ± 0.3) µm in length and 2.4-3.5 (3.1 ± 0.3) µm in diameter, with each capsule containing a polar filament with 3-4 coils. Molecular data consisted of a 2918-bp sequence of the partial 18S, complete ITS1, 5.8S, ITS2, and partial 28S ribosomal rRNA regions as well as a 2455-bp sequence of partial 28S ribosomal RNA. The partial 18S and 28S data was used in a concatenated Bayesian phylogenetic analysis to further infer the evolutionary relationships of the Myxidiidae. Additionally, the partial 18S data was used in a separate phylogenetic analysis. CONCLUSIONS: The present work reports novel morphological and molecular data for Myxidium kudoi as well as a novel locality of occurrence for this species. In concatenated phylogenetic analysis using 18S and 28S data and other molecular data from Myxozoa, M. kudoi grouped with other freshwater Myxidiidae. In the single-locus, 18S analysis, M. kudoi grouped with Myxidium rhodei from Rutilus rutilus and Myxidium amazonense from Corydoras melini, the only other Myxidium species of catfish for which molecular data are available.

Two Novel Myxozoans from Pirate Perch Aphredoderus sayanus (Gilliams, 1824) in the Upper Mississippi River, Including the First North American Species of Hennegoides Lom, Tonguthai, & Dyková, 1991.

The pirate perch Aphredoderus sayanus is a relatively small fish species found in rivers throughout much of the eastern United States. Due to their cryptic nature, relatively little is known regarding their parasite fauna. A survey of pirate perch from the upper Mississippi River revealed 2 novel myxozoans. Hennegoides flockae n. sp. was observed in heavily infected gills where the lamellae featured irregular expansion by bulbous myxozoan polysporic plasmodia, typically affecting the middle to distal half of the filaments. When severe, infection of sequential filaments was such that the filaments were fused, forming what appeared as multicystic/lobular parasitic aggregates subdivided by fine epithelial cords. The total myxospore length of Hennegoides flockae was 35.4-46.4 (41.3 ± 3.3) and the spore body, asymmetrically ovoid in valvular view, was 15.4-18.7 (17.0 ± 0.7) × 7.1-8.7 (7.9 ± 0.4). Henneguya marcquenskiae n. sp. was observed in the liver with plasmodia present randomly and infrequently in the hepatocellular parenchyma. The total myxospore length for Henneguya marcquenskiae was 39.5-55.9 (48.4 ± 4.2), with the spore body being lanceolate, 13.9-16.5 (15.4 ± 0.7) × 7.1-9.0 (8.3 ± 0.5). Phylogenetic analysis of the SSU rRNA gene placed both Hennegoides flockae and Henneguya marcquenskiae as sisters to each other in a clade containing other Myxozoans known to infect the gills of esocids, percids, and centrarchids. These parasites represent the first reports of Henneguya and Hennegoides from pirate perch, with the latter being the first report of this genus outside of the Asian continent.

Morphology of the megaspore Lagenoisporites magnus (Chi and Hills 1976) Candilier et al. (1982), from the Carboniferous (lower Mississippian: mid-upper Tournaisian) of Bolivia.

The morphology and structure of megaspores assigned to Lagenoisporites magnus from the Toregua Formation, Retama Group, mid-upper Tournaisian of Bolivia were studied. The analysis was performed with light, fluorescence and scanning electron microscopy. Megaspores were laterally compressed and presented a spherical body with a proximal gula, of the hologula type. Gula had verrucae ornamentation and the spore body presented complex processes consisting of a bulbous base and an internally partitioned projection with sharp apex. In addition to this main ornamentation, perforations were present throughout the spore surface. Megaspores showed well marked curvaturae perfectae due to the abrupt transition existing between the gula ornamentation and the spore body processes. These megaspores were assigned to heterosporous arborescent lycopsids of the Lepidocarpaceae family, as in section view, exospore structure presented a three-dimensional network of fused elements. Likewise, due to a similarity found between sporoderm and Isoetes L. structure, it is evident that megaspores structure has remained intact inside the heterosporous lycopsids. Therefore; the L. magnus structure not only would confirm its affinity with the Lycophyta fossils but also with the living ones.

Studies on the gametophytic development, mating behaviour and reproductive biology of Macrothelypteris torresiana (Gaudich.) Ching (Thelypteridaceae).

Macrothelypteris torresiana is a terrestrial fern belongs to the family Thelypteridaceae. It is very imperative to know the germination and growth pattern of this fern to make strategy for ex- situ conservation. In the present study germination of spore, developmental pattern of prothallium with particular emphasis on emergence of antheridia as well as archegonia, their fertilization and development of sporophytes were studied. The spores were spheroidal or reniform with broadly winged fimbriate perispore having average size of 48 ± 3.6 × 39 ± 4.2 µm. The spores started germinating just after 6 days of plating and percentage of germination was 94%. The germination pattern of spore was Vittaria type and development of gametophyte was Aspidium type. After 27 days of plating, a very distinct cordate-shaped adult gametophyte with deep apical notch and unicellular papillate hairs throughout the gametophyte was developed. The rhizoids appeared away from the apical notch of the gametophyte. The gametophyte first developed archegonia on 32nd day, whereas antheridia were developed on 42nd day. In isolate culture the gametophytes did not develop any sporophyte whereas in composite culture juvenile sporophytes were emerged on 77th day. This suggests the plant has ability to reproduce only through inter-gametophytic selfing and crossing, hence decreasing the plant's capacity for colonization in a particular locality.

Taxonomic importance of spore morphology in Thelypteridaceae from Northern Pakistan.

Spore morphology of Thelypteridaceae species growing in Malakand Division, Northern Pakistan, was studied using both light microscopy and scanning electron microscopy. The taxa are Christella dentata and Glaphyropteridopsis erubescens in the subfamily Thelypteridoideae, and Phegopteris connectilis, Pseudophegopteris pyrrhorhachis, and Pseudophegopteris levingei in the subfamily Phegopteridoideae. The studied species exhibit differences in spore size, exospore thickness, color, and ornamentation. Spores of the studied species are monolete and medium-sized, and shape is ellipsoidal in both polar and equatorial views. The average measurement of the polar diameter ranges from 27 µm to 31 µm, whereas in the equatorial direction it varied from 20 µm to 40 µm. The exospore thickness ranges from 1.2 µm to 2.4 µm. Reticulate, laevigate with microgranules, cristate, and coarsely echinate surface ornamentation are observed among the species. Multivariate analysis including unweighted pair group method with arithmetic mean and principal component analysis was used for the grouping and discrimination of species and genera.

Systematic approach to the correct identification of Asplenium dalhousiae (Aspleniaceae) with their medicinal uses.

In present study, multiple microscope techniques were used for the systematics identification of the species Asplenium dalhousiae. The plant was collected from different phytogeographical and its natural habitat of Pakistan, where it shows higher diversity. Morphology, foliar epidermal anatomy, and spore morphological characters of the species were studied in detailed using multiple microscopic techniques through light microscopy (LM) and scanning electron microscopy (SEM). LM and SEM were used for the systematics identification of the species. Traditionally, the species is used in the ailment of many diseases, so the spore morphology, anatomical features, and morphological characters are relevant to describe the species taxonomy. The importance of multiple methods of taxonomic study (e.g., documentation and morphological characteristics) for characterizing herbs are important step in systematic certification to maintain the efficacy of herbal medicines. The aim of the present study is to examine the morphological, anatomical, and spore morphology of the species A. dalhousiae in more detailed for the correct taxonomic identification and their medicinal validation from Pakistan.

Contributions of crust proteins to spore surface properties in Bacillus subtilis.

Surface properties, such as adhesion and hydrophobicity, constrain dispersal of bacterial spores in the environment. In Bacillus subtilis, these properties are influenced by the outermost layer of the spore, the crust. Previous work has shown that two clusters, cotVWXYZ and cgeAB, encode the protein components of the crust. Here, we characterize the respective roles of these genes in surface properties using Bacterial Adherence to Hydrocarbons assays, negative staining of polysaccharides by India ink and Transmission Electron Microscopy. We showed that inactivation of crust genes caused increases in spore relative hydrophobicity, disrupted the spore polysaccharide layer, and impaired crust structure and attachment to the rest of the coat. We also found that cotO, previously identified for its role in outer coat formation, is necessary for proper encasement of the spore by the crust. In parallel, we conducted fluorescence microscopy experiments to determine the full network of genetic dependencies for subcellular localization of crust proteins. We determined that CotZ is required for the localization of most crust proteins, while CgeA is at the bottom of the genetic interaction hierarchy.

Studies on the complementary relationship of surface ornamentations between megaspores and microspores of Selaginella P. Beauv. (Selaginellaceae).

In the present study, the surface ornamentations of both megaspores and microspores of 20 species of Selaginella were studied using scanning electron microscopy. The results of the present study showed that the megaspore surface ornamentations are complementary to the corresponding microspore surface ornamentations. In detail, reticulate surface ornamentation of megaspores is complementary to granulate and baculate ornamentations of microspores; verrucate ornamentation of megaspores is complementary to scabrate ornamentation of microspores; granulate ornamentation of megaspores is complementary to verrucate-rugulate or verrucate ornamentations of microspores; verrucate-rugulate ornamentation of megaspores is complementary to spinulate, verrucate, or verrucate-rugulate ornamentations of microspores; tuberculate ornamentation of megaspores is complementary to lamellate, tuberculate or verrucate ornamentations of microspores. Complementary ornamentation between megaspores and microspores allows microspores to adhere to megaspores. Our observations support the "synaptospory" hypothesis, that is, spore surface ornamentation plays a part in keeping or bringing spores together during their dispersal in the pteridophytes.

Spores potentially dispersed to longer distances are more tolerant to ultraviolet radiation: A case study in the moss genus Orthotrichum.

PREMISE OF THE STUDY: Ultraviolet (UV) radiation influences the viability of algal spores and seed-plant pollen depending on the species, the dose, and the wavelength. In bryophytes, one of the dominant groups of plants in many habitats, UV radiation could determine their spore dispersal strategy, and such data are critical for reconstructing the ancestral state in plants and for determining the distribution range and persistence of bryophyte species. METHODS: Spores of four bryophyte species of the moss genus Orthotrichum that were either hygrochastic or xerochastic (spores dispersed under wet or dry conditions, respectively) were exposed to realistic doses of UV radiation under laboratory conditions. Spore viability was evaluated through germination experiments and, for the first time in bryophytes, ultrastructural observations. Given that the UV-B doses used were relatively higher than the UV-A doses, the UV effect was probably due more to UV-B than UV-A wavelengths. KEY RESULTS: All four species reduced their spore germination capacity in a UV dose-dependent manner, concomitantly increasing spore ultrastructural damage (cytoplasmic and plastid alterations). Most spores eventually died when exposed to the highest UV dose. Interestingly, spores of hygrochastic species were much more UV-sensitive than those of xerochastic species. CONCLUSIONS: UV tolerance determines moss spore viability, as indicated by germination capacity and ultrastructural damage, and differs between spores of species with different dispersal strategies. Specifically, the higher UV tolerance of xerochastic spores may enable them to be dispersed to longer distances than hygrochastic spores, thus extending more efficiently the distribution range of the corresponding species.

Biliary Tract-Infecting Myxosporeans from Estuarine and Reef Stonefish (Scorpaeniformes: Synanceiidae) Off Eastern Australia, with Descriptions of Sphaeromyxa horrida n. sp. and Myxidium lapipiscis n. sp. (Myxosporea: Bivalvulida).

Two new species of myxosporeans are described from the gallbladders of estuarine stonefish, Synanceia horrida, and reef stonefish, Synanceia verrucosa, from localities off Cairns, in tropical north Queensland and in Moreton Bay in southern Queensland, Australia. Sphaeromyxa horrida n. sp. can be distinguished from congeners in the morphologically distinct "balbianii" species group within Sphaeromyxa on the basis of morphometric differences in length and width of mature spores, length and width of polar capsules, and unique small-subunit (SSU) ribosomal (rDNA) sequence composition relative to other taxa. Replicate SSU rDNA sequences generated from Sph. horrida n. sp. collected from Sy. horrida and Sy. verrucosa in tropical north Queensland and from Sy. horrida in Moreton Bay were identical, suggesting that this species is widely distributed along the east coast of Australia. Myxidium lapipiscis n. sp. can be distinguished from the majority of described Myxidium species on the basis of its relatively small mature spore size (6.1-7.9 µm long × 3.1-3.9 µm wide), and its unique SSU rDNA sequence. Specimens putatively identified as M. lapipiscis n. sp. were found in Sy. horrida from both tropical north Queensland and Moreton Bay, suggesting that this taxon is also widely distributed along the east coast of Australia. However, no molecular data were available for the specimens from tropical north Queensland for comparative genetic analyses. Bayesian inference and maximum likelihood analysis of the SSU rDNA sequences for these 2 new species revealed that Sph. horrida n. sp. formed a strongly supported clade with Sphaeromyxa zaharoni Diamant, Whipps, and Kent, 2004, which was described from the scorpaeniform, Pterois miles, from the Red Sea. This is the first report of myxozoans infecting stonefish (Synanceiidae).

Palynological analysis of Dennstaedtiaceae taxa from the Paranaense Phytogeografic Province that produce monolete spores and its systematic implications (I): Blotiella lindeniana , Histiopteris incisa and Paesia glandulosa.

The genera of Dennstaedtiaceae have sporophytes with very different morphological characteristics between each other, and this feature has made difficult the systematic circumscription of the family. This reason makes necessary the study of new characters that allow a better understanding of the relations within the group. The spore morphology and wall structure of Blotiella lindeniana, Histiopteris incisa and Paesia glandulosa from the Paranaense Phytogeographic Province were studied using light microscope, and scanning and transmission electron microscope. The exospore has two layers and, according to the species, the exospore surface bears pila, echinae, verrucae, bacula and tubercles. The perispore has two or three layers and its surface is psilate, baculate or rugulate. The variability found in the sculpture of the spores and their stratification and ultrastructure of perispore reflects the morphological differences observed in the sporophyte of the species studied. Additionally, while the stratification and ultrastructure of the exospore is shared by the Dennstaedtiaceae species, their ornamentation could be a character to distinguish species into the clade "hypolepidoide". The finding of spores with similar characteristics in phylogenetically unrelated families allows us to suggest that palynological features do not have an evolutionary value to establish relationships between groups above the genus level.

The light microscopy and ultrastructural characteristics of Myxobolus naffari (Myxosporea, Myxobolidae) infecting the Nile carp Labeo niloticus (Cyprinidae) and its histological impact

During a survey of myxosporean parasites infecting freshwater fishes from the River Nile at Giza Governorates, Egypt between March and September 2016, nine out of 30 specimens of the Nile carp Labeo niloticus (Cyprinidae) were found to be naturally infected with Myxobolus naffari (Myxobolidae). Small macroscopic plasmodia appeared embedded in the host gill tissue accompanied with fusion of the gill epithelia, and atrophy was observed at the site of infection. The host reaction was manifested by the encapsulation of the plasmodia with a thick layer of connective tissue. The plasmodia appeared as white, elongated rods between gill filaments with an intensity ranging from three to eight cysts/fish. The average dimensions of plasmodia were 1.2­2.0 (1.8 ± 0.2) mm long × 0.4­0.7 (0.6 ± 0.2) mm wide. The spores were oval, reaching 9.56­11.2 (10.2 ± 0.2) µm long and 6.5­7.7 (7.0 ± 0.4) µm wide with two equal-sized polar capsules regularly arranged at the anterior pole of each spore. They were 4.51­5.5 (5.1 ± 0.4) µm in length and 1.5­2.0 (1.7 ± 0.2) µm in width. Histological, semi-thin sections were taken through parasite plasmodia and transmission electron microscopic examination of ultrathin sections was performed to describe the developmental stages of the recorded parasite within the host fish.

Palynological analysis of Dennstaedtiaceae taxa from the Paranaense Phytogeografic Province that produce monolete spores and its systematic implications (I): Blotiella lindeniana, Histiopteris incisa and Paesia glandulosa

ABSTRACT The genera of Dennstaedtiaceae have sporophytes with very different morphological characteristics between each other, and this feature has made difficult the systematic circumscription of the family. This reason makes necessary the study of new characters that allow a better understanding of the relations within the group. The spore morphology and wall structure of Blotiella lindeniana, Histiopteris incisa and Paesia glandulosa from the Paranaense Phytogeographic Province were studied using light microscope, and scanning and transmission electron microscope. The exospore has two layers and, according to the species, the exospore surface bears pila, echinae, verrucae, bacula and tubercles. The perispore has two or three layers and its surface is psilate, baculate or rugulate. The variability found in the sculpture of the spores and their stratification and ultrastructure of perispore reflects the morphological differences observed in the sporophyte of the species studied. Additionally, while the stratification and ultrastructure of the exospore is shared by the Dennstaedtiaceae species, their ornamentation could be a character to distinguish species into the clade "hypolepidoide". The finding of spores with similar characteristics in phylogenetically unrelated families allows us to suggest that palynological features do not have an evolutionary value to establish relationships between groups above the genus level.

Myxobolus linzhiensis n. sp. (Myxozoa: Myxobolidae) from the gill filament of Schizothorax oconnori Lloyd (Cyprinidae: Schizothoracinae) in Tibet, China: morphological and molecular characterization.

In this report, a new myxosporean species, Myxobolus linzhiensis n. sp. (Cnidaria: Myxozoa: Bivalvulida), was described from the endemic cyprinid Schizothorax oconnori inhabiting the Yarlung Tsangpo River in the Tibetan plateau, China. The whitish ellipsoidal plasmodia of M. linzhiensis n. sp. grew under the epithelium of gill filaments, 1.7-mm long and 0.4-mm wide, and were found in the 3 of 8 examined S. oconnori (37.5%). Mature spores appeared suborbicular in frontal view, fusiform shaped in lateral view with tapering anterior, measuring 11.4 ± 0.7 (10.1-13.0) µm in length, 10.6 ± 0.4 (10.0-11.3) µm in width, and 6.6 ± 0.3 (6.2-7.0) µm in thickness. Two equal polar capsules were pyriform with an apophysis at its top end, measuring 5.7 ± 0.5 (4.8-6.7) µm in length, 3.8 ± 0.2 (3.2-4.2) µm in width, and polar filaments coiled 4 turns. Spore valves were symmetrical, and sutural ridge was straight and thin, running near the middle of the valves. Although Myxobolus linzhiensis n. sp. showed similar morphological characteristics with Myxobolus chushi Dar Kaur & Chishti 2017 from Schizothorax niger, Myxobolus kienweiensis Ma, 1976 from Schizothorax davidi, the new myxosporean was distinguished with them in spore shape, polar filament, and host-tissue tropism. Homology search by BLAST in GenBank indicated that the obtained 18S rDNA sequence of M. linzhiensis n. sp. (KY965935) did not match any available myxozoan sequence, most similar to Henneguya zikaweiensis, and showing less than 93% sequence similarity. Phylogenetical analyses demonstrated that M. linzhiensis n. sp. was firmly clustered in the clade consisting of myxosporeans Thelohanellus, Myxobolus, Henneguya from Asian cyprinids. This study is the first report on myxosporean parasitizing in endemic fish in the Tibetan Plateau, China.

Myxobolus sp. and Henneguya sp. (Cnidaria: Myxobolidae) natural co-infection in the kidney of Piaractus mesopotamicus (Characiformes: Serrasalmidae).

This study evaluated the myxozoan infection and histopathology of the kidney of freshwater fish Piaractus mesopotamicus from intensive fish farming in Brazil. A total of 55 fish were examined for myxozoan infection. Infected organs were processed by usual histology and stained with hematoxylin-eosin (H&E) and Ziehl-Neelsen (ZN). From the total of 55 fish analyzed, 47 (85.45%) presented myxospores, being 9.09% (5/55) only with Myxobolus sp., 5.45% (3/55) only with Henneguya sp., and 70.91% (39/55) presenting both parasites. The presence of myxospores was associated with histological alterations in both stromal and renal parenchyma. Myxospores were found mostly in the peritubular interstitial tissue and in low intensity in the glomerulus which caused nuclear hypertrophy and loss of Bowman space. An increase in the glomerular tuft and a reduction in the lumen of the collector tubules were also observed, besides the high number of melanomacrophage cells in the glomerulus. This study reports for the first time detection of myxozoan mixed infection in one organ of pacu and discuss the possible transportation of myxospores in the circulating blood.

In vitro effect of two commercial anti-coccidial drugs against myxospores of Kudoa septempunctata genotype ST3 (Myxozoa, Multivalvulida).

Kudoa septempunctata (Myxozoa: Multivalvulida) myxospores infect the trunk muscles of olive flounder (Paralichthys olivaceus). In this study, two popular commercially formulated anti-coccidial drugs (amprolium hydrochloride and toltrazuril) were serially diluted and incubated with purified mature Kudoa septempunctata myxospores. The viability of K. septempunctata spores was determined after a 2-day incubation followed by Hoechst 33342 and propidium iodide staining, and scanning electron microscopy. Amprolium hydrochloride significantly decreased spore viability (18% of control) at a concentration of 920 µg/mL, whereas toltrazuril showed almost no effect (83% of control). Viability of the control (untreated spores) was 90%. In vivo studies are required to confirm the efficacy of amprolium hydrochloride in fish infected with K. septempunctata myxospores on their growth and immune system performance.