Microsporidia: Pervasive natural pathogens of Caenorhabditis elegans and related nematodes.

The nematode Caenorhabditis elegans is an invaluable host model for studying infections caused by various pathogens, including microsporidia. Microsporidia represent the first natural pathogens identified in C. elegans, revealing the previously unknown Nematocida genus of microsporidia. Following this discovery, the utilization of nematodes as a model host has rapidly expanded our understanding of microsporidia biology and has provided key insights into the cell and molecular mechanisms of antimicrosporidia defenses. Here, we first review the isolation history, morphological characteristics, life cycles, tissue tropism, genetics, and host immune responses for the four most well-characterized Nematocida species that infect C. elegans. We then highlight additional examples of microsporidia that infect related terrestrial and aquatic nematodes, including parasitic nematodes. To conclude, we assess exciting potential applications of the nematode-microsporidia system while addressing the technical advances necessary to facilitate future growth in this field.

SEM and LM insights: Spore morphology and its taxonomic significance in Sino Himalayan, Malesian, and European elements of Pteridaceae.

The Pteridaceae family, known for its taxonomic complexity, presents challenges in identification due to high variability among its species. This study investigates the spore morphology employing both SEM and LM techniques in 10 Pteridaceae taxa phytogeographicaly Sino-Himalayan, Malesian, and European elements in Pakistan. The taxa include Adiantum capillus-veneris, A. incisum, A. venustum, Aleuritopteris bicolor, Oeosporangium nitidulum, O. pteridioides, Onychium cryptogrammoides, O. vermae, Pteris cretica, and P. vittata. The objective is to assess their taxonomic relevance and develop a spore-based taxonomic key. Findings indicate differences in spore shape, sizes, exospore thickness, and in surface ornamentation highlighting the potential for taxonomic differentiation. Spores are trilete, and notable differences are observed in the dimension of spores in both distal and proximal sides. Equatorial dimensions vary between 35 and 50 µm, while the polar diameter ranges from 29 to 50 µm. SEM revealed different spore ornamentation types that show several useful characteristics establishing valuable taxonomic variations. The studied Adiantum taxa feature a perispore with tubercules and a micro-granulose surface. The spores of examined Oeosporangium and Aleuritopteris taxa shows cristate sculptures with variable ornamentations. Both species of Onychium have tuberculate-pleated tubercles with sinuous folds on both distal and proximal sides. The surface ornamentation among examined Pteris taxa show variability. PCA analysis indicated that spore quantitative data identified distinct groups, underscoring taxonomic significance. Nevertheless, there was variation observed in surface ornamentation and spore shape, indicating the potential for discrimination among taxa. RESEARCH HIGHLIGHTS: Spore morphology of 10 Pteridaceae taxa has been investigated through LM and SEM. Investigated species shows differences in spore shape, sizes, exospore thickness, and in surface ornamentation. Ornamentation on the perispore provides several valuable characteristics, establishing useful taxonomic distinctions. Spore morphological analysis is effective at the generic level, with minor distinctions discernible at the species level.

Wind and small mammals are complementary fungal dispersers.

Following a disturbance, dispersal shapes community composition as well as ecosystem structure and function. For fungi, dispersal is often wind or mammal facilitated, but it is unclear whether these pathways are complementary or redundant in the taxa they disperse and the ecosystem functions they provide. Here, we compare the diversity and morphology of fungi dispersed by wind and three rodent species in recently harvested forests using a combination of microscopy and Illumina sequencing. We demonstrate that fungal communities dispersed by wind and small mammals differ in richness and composition. Most wind-dispersed fungi are wood saprotrophs, litter saprotrophs, and plant pathogens, whereas fungi dispersed in mammal scat are primarily mycorrhizal, soil saprotrophs, and unspecified saprotrophs. We note substantial dispersal of truffles and agaricoid mushrooms by small mammals, and dispersal of agaricoid mushrooms, crusts, and polypores by wind. In addition, we find mammal-dispersed spores are larger than wind-dispersed spores. Our findings suggest that wind- and small-mammal-facilitated dispersal are complementary processes and highlight the role of small mammals in dispersing mycorrhizal fungi, particularly following disturbances such as timber harvest.

Taxonomic importance of spore morphology of selected taxa of Asplenium (Aspleniaceae) from Pakistan.

Taxonomic importance of spore morphology of seven taxa of genus Asplenium (Aspleniaceae) from Pakistan is illustrated with SEM. Spore morphology of the taxa were monolete with bilateral symmetry, distal and proximal view were ellipsoidal, spheroidal, equatorial view were hemicircular; perispore surface were alate, costate, costate-alate, with reticulate and tuberculate ornamentation. Taxonomic importance of spore morphology of seven taxa of Asplenium (Aspleniaceae, polypodiales) from Pakistan. Basionym, Synonyms with updated nomenclature. Spores are monolete, distal and proximal view ellipsoidal, spheroidal, equatorial view hemicircular, perispore with alate, costate, reticulate, and tuberculate surface. Habitat ecology with specimens examined from different areas of Pakistan and their worldwide general distribution.

Inactivation of fungal spores from clinical environment by silver bio-nanoparticles; optimization, artificial neural network model and mechanism.

The present study aimed to assess the efficiency of silver bio-nanoparticles (Ag-NPs) in inactivating of the Aspergillus fumigatus, A. parasiticus and A. flavus var. columnaris and A. aculeatus spores. The AgNPs were synthesized in secondary metabolic products of Penicillium pedernalens 604 EAN. The inactivation process was optimized by response surface methodology (RSM) as a function of Ag NPs volume (1-10 µL/mL); time (10-120 min); pH (5-8); initial fungal concentrations (log10) (3-6). The artificial neural network (ANN) model was used to understand the behavior of spores for the factors affecting inactivation process. The best conditions to achieved SAL 10-6 of the fungal spores were recorded with 3.46 µl/mL of AgNPs, after 120 min at pH 5 and with 6 log of initial fungal spore concentrations, at which 5.99 vs. 6.09 (SAL 10-6) log reduction was recorded in actual and predicted results respectively with coefficient of 87.00%. The ANN revealed that the timehas major contribution in the inactivation process compare to Ag NPs volume. The fungal spores were totally inactivated (SAL 10-6, 6 log reduction with 99.9999%) after 110 min of the inactivation process, 10 min more was required to insure the irreversible inactivation of the fungal spores. The absence of protease and cellulase enzymes production confirm the total inactivation of the fungal spores. FESEM analysis revealed that the AgNPs which penetrated the fungal spores leading to damage and deform the fungal spore morphology. The AFM analysis confirmed the total spore surface damage. The bands in the range of the Raman spectroscopy from 1300 to 1600 cm-1 in the inactivated spores indicate the presence of CH3, CH2 and the deformation of lipids released outside the spore cytoplasm. These finding indicate that the AgNPs has high potential as a green alternative inactivation process for the airborne fungal spores.

Spores morphology of the family Pteridaceae from Shandong Province, China.

In this article, we explored systematically the spore morphology of Pteridaceae by observation of the species distributed in Shandong Province using scanning electron microscopy (SEM). The results showed that the spore morphology of all the species in the family is tetrahedral and trilete. The characters of spore ornamentation are intraspecies stable, but significantly different among species and genera. Spore morphology is significant in exploring the phylogenetic relationships of Pteridaceae as well as in generic and specific delimitations.

Taxonomy and spore morphology of selected taxa of Cheilanthoideae and Pteridoideae (Pteridaceae) from Pakistan.

Taxonomy and spore morphology of 12 taxa of Cheilanthoideae and Pteridoideae (Pteridaceae, Polypodiales) from Pakistan is illustrated with scanning electron microscopy images based upon the specimens collected from various localities. A total of six genera belong to 12 taxa viz. Actiniopteris radiata, Aleuritopteris albomarginata, A. ancepes, Notholaena himalaica, Oeosporangium nitidulum, O. pteridioides subsp. acrosticum, Onychium cryptogrammoides subsp. cryptogrammoides, O. vermae, Pteris cretica subsp. cretica, P. cretica subsp. laeta, P. vittata subsp. emodi, and P. vittata subsp. vittata were reported. Spore morphology of the taxa was trilete, triangular in proximal and distal view, ellipsoidal and hemicircular in equatorial view, polar proximal and distal surface with cristate, granulose, reticulate, perforate and tuberculate ornamentation.

Taxonomy and spore morphology of genus Adiantum (Vittarioideae; Pteridaceae) from Pakistan.

Taxonomy and spore morphology of genus Adiantum (Vittarioideae; Pteridaceae) from Pakistan is illustrated with scanning electron microscopy images based upon the specimens examined in herbaria of Pakistan, United Kingdom, and United States. A total of five species viz. Adiantum capillus-veneris, A. incisum subsp. incisum, A. pedatum subsp. pedatum, A. tibeticum, and A. venustum were reported with the habitat ecology of the species from Pakistan. Spore morphology of the taxa was trilete, triangular in proximal and distal view, ellipsoidal and hemicircular in equatorial view, polar proximal and distal surface was with tuberculate and granulose ornamentation.

Taxonomic revision of the Asplenium wrightii complex (Aspleniaceae) with reinstatement of A. alatulum and A. subcrenatum.

The Asplenium wrightii complex is morphologically variable and difficult in species delimitation. Owing to lack of comprehensive sampling in phylogenetic studies, the taxonomy of this complex remains unresolved. Based on extensive field observations, specimen examination and our recent molecular data, the present study aims to clarify the identities of three species of Asplenium in this complex from Asia. Our study revealed that A. alatulum and A. subcrenatum, previously treated as synonyms of A. wrightii, should be reinstated. A taxonomic revision of the three species, including their type information, detailed descriptions, voucher specimens, distribution, ecology, as well as taxonomic notes, is carried out.

Spore morphology and leaf epidermal anatomy as a taxonomic source in the identification of Asplenium species from Malakand division Pakistan.

Several studies have demonstrated the usefulness of leaf epidermal, and spore morphological characters in the taxonomy of genus Asplenium. However, works on the Pakistani species of Asplenium are not existent. With the objective to verify the efficacy of leaf epidermal and spore morphological traits, the leaf epidermis and spore morphology in nine Asplenium species occurring in Malakand division was studied. The spores were studied under light microscope (LM) and scanning electron microscope (SEM), whereas for leaf epidermal anatomy, the LM was used. The spores are monolete, ellipsoidal in shape, the equatorial and polar diameter ranges between 28.3 and 50.2 × 27.6 and 45.8 µm. The exospore thickness varied from 0.5 to 3.8 µm. The perispore is 0.8-3.5 µm thick, ornamented, and morphologically variable. The spores characters specifically the perispore ornamentation are useful in distinguishing species within the genus. The irregular spore shape and unusual development of perispore wall surface characterize aborted spores in the species of Asplenium × alternifolium. The most informative quantitative characters appeared to be length and width of epidermal cells, and length and width of stomata were useful to distinguish species. The most significant qualitative characters to distinguish species were the anticlinal wall pattern. Our study has shown that considerable variations exist in the leaf epidermis of Asplenium species, at least some of which has taxonomic significance. We confirmed the prevalent taxonomic value of stomatal and epidermal cells traits. The leaf epidermal anatomy and spore morphological features showed to be a good source of information for taxonomy of the genus Asplenium.

Phylogenetic relationships of three Kudoa spp. with morphologically similar myxospores (K. iwatai, K. lutjanus, and K. bora), with the redescription of K. uncinata and K. petala and description of a new species (K. fujitai n. sp.) in fishes in the South China Sea.

Members of the myxozoan genus Kudoa (Myxosporea: Multivalvulida: Kudoidae) are characterized as having four or more shell valves in a myxospore, with a corresponding number of polar capsules. Certain Kudoa spp. are critical pathogens in fish, causing postmortem myoliquefaction, unmarketable fish musculature due to unsightly macroscopic cysts, and reduced aquaculture production due to the outbreaks of neurological symptoms or cardiac diseases. Molecular genetic techniques have enabled the differentiation of Kudoa spp. with morphologically similar myxospores. In the present study, we employed integrated taxonomic approaches on five Kudoa spp. forming cysts between the trunk muscle myofibers (K. bora from Osteomugil perusii and K. lutjanus from Acanthopagrus latus), or cysts in the gallbladder wall (K. petala from Sillago sihama), and pseudocysts in the trunk muscle myofibers (K. uncinata from Nuchequula nuchalis and K. fujitai n. sp. from O. perusii). These four host fishes, which originated in the South China Sea, were purchased in the wet markets in Zhanjiang City, Guangdong Province, China, between August 2016 and April 2018. We have redescribed the four Kudoa spp. (K. bora, K. lutjanus, K. petala, and K. uncinata) on which little data are available after their original descriptions. Particularly, genetic characterization of K. bora and K. lutjanus, which are known to have myxospores morphologically similar to those of K. iwatai, was performed based on the nuclear ribosomal RNA gene and partial mitochondrial DNA genes such as cytochrome c oxidase subunit I and small and large ribosomal genes, demonstrating the validity and independence of these three kudoid species. We also provide description of a new species-K. fujitai n. sp.-in the present study. Application of integrated taxonomic approaches to known species characterized solely based on morphological criteria, as well as unknown species (e.g., K. fujitai n. sp. in the present study), contributes to better understanding of the biodiversity of Kudoa and multivalvulid myxosporeans.

Placement of Triblidiaceae in Rhytismatales and comments on unique ascospore morphologies in Leotiomycetes (Fungi, Ascomycota).

Triblidiaceae is a family of uncommonly encountered, non-lichenized discomycetes. A recent classification circumscribed the family to include Triblidium (4 spp. and 1 subsp.), Huangshania (2 spp.) and Pseudographis (2 spp. and 1 var.). The apothecia of these fungi are persistent and drought-tolerant; they possess stromatic, highly melanized covering layers that open and close with fluctuations of humidity. Triblidialean fungi occur primarily on the bark of Quercus, Pinaceae and Ericaceae, presumably as saprobes. Though the type species of Huangshania is from China, these fungi are mostly known from collections originating from Western Hemisphere temperate and boreal forests. The higher-rank classification of triblidialean fungi has been in flux due in part to an overemphasis on ascospore morphology. Muriform ascospores are observed in species of Triblidium and in Pseudographiselatina. An intense, dark blue/purple ascospore wall reaction in iodine-based reagents is observed in species of Pseudographis. These morphologies have led, in part, to these genera being shuffled among unrelated taxa in Hysteriaceae (Dothideomycetes, Hysteriales) and Graphidaceae (Lecanoromycetes, Ostropales). Triblidiaceae has been placed within the monofamilial order Triblidiales (affinity Lecanoromycetes). Here, we demonstrate with a three-gene phylogenetic approach that triblidialean fungi are related to taxa in Rhytismatales (Leotiomycetes). We synonymize Triblidiales under Rhytismatales and emend Triblidiaceae to include Triblidium and Huangshania, with Pseudographis placed within Rhytismataceae. A history of Triblidiaceae is provided along with a description of the emended family. We discuss how the inclusion of triblidialean fungi in Rhytismatales brings some rarely observed or even unique ascospore morphologies to the order and to Leotiomycetes.

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.

Parvicapsula curvatura n. sp. in cultured olive flounder Paralichthys olivaceus and phylogenetic characteristics of the genus Parvicapsula.

Parvicapsula curvatura n. sp. (Myxozoa; Bivalvulida) was found in the urinary bladder of olive flounder Paralichthys olivaceus cultured in a fish farm on Jeju Island, ROK. When laterally viewed, the parasite has asymmetrical curved spores that measure 9.6-11.6 µm in length. Furthermore, it has 2 subspherical polar capsules at the apex. Based on the phenotypical traits, it is most similar to P. limandae but differs in the shape of polar capsule, locality, and host specificity (family level). BLAST analysis indicated that P. curvatura was closest to P. unicornis and P. petuniae via 18S and 28S rDNA sequences, respectively. The 18S rDNA from P. curvatura was used in molecular phylogenetic analyses of Parvicapsula spp. to examine the congruence of phylogeny with spore morphology, locality, and host specificity. The results demonstrated that the spore morphotype was correlated with the phylogeny of the genus Parvicapsula, and the parasites have speciated into an oblong and semicircular spore type.

Morphological and molecular diversity of arbuscular mycorrhizal fungi in revegetated iron-mining site has the same magnitude of adjacent pristine ecosystems.

Arbuscular mycorrhizal fungi (AMF) are important during revegetation of mining sites, but few studies compared AMF community in revegetated sites with pristine adjacent ecosystems. The aim of this study was to assess AMF species richness in a revegetated iron-mining site and adjacent ecosystems and to relate AMF occurrence to soil chemical parameters. Soil samples were collected in dry and rainy seasons in a revegetated iron-mining site (RA) and compared with pristine ecosystems of forest (FL), canga (NG), and Cerrado (CE). AMF species were identified by spore morphology from field and trap cultures and by LSU rDNA sequencing using Illumina. A total of 62 AMF species were recovered, pertaining to 18 genera and nine families of Glomeromycota. The largest number of species and families were detected in RA, and Acaulospora mellea and Glomus sp1 were the most frequent species. Species belonging to Glomeraceae and Acaulosporaceae accounted for 42%-48% of total species richness. Total number of spores and mycorrhizal inoculum potential tended to be higher in the dry than in the rainy season, except in RA. Sequences of uncultured Glomerales were dominant in all sites and seasons and five species were detected exclusively by DNA-based identification. Redundancy analysis evidenced soil pH, organic matter, aluminum, and iron as main factors influencing AMF presence. In conclusion, revegetation of the iron-mining site seems to be effective in maintaining a diverse AMF community and different approaches are complementary to reveal AMF species, despite the larger number of species being identified by traditional identification of field spores.

Phylogenetic study of the genus Kudoa (Myxozoa: Multivalvulida) with a description of Kudoa rayformis sp. nov. from the trunk muscle of Pacific sierra Scomberomorus sierra.

Kudoa rayformis n. sp. (Myxozoa; Multivalvulida) was observed in the trunk muscle of Pacific sierra Scomberomorus sierra caught off the coast of Tonosi, Panama. The species formed pseudocysts in myofibers and infection was subclinical. The myxospores possessed four polar capsules and spore valves, one of which had a distinct filamentous extension. This unique morphological characteristic of the myxospore validated this as a new species of Kudoa. Genetically, K. rayformis n. sp. is closest to K. inornata, with 98% and 91% similarity in 18S and 28S rDNA, respectively, but its spore shape was clearly distinct. The 18S rDNA and concatenated sequences from K. rayformis were used in molecular phylogenetic analyses of kudoids to examine the congruence of phylogeny with infection site tropism, spore morphology and cyst/pseudocyst formation. The results demonstrated that the phenotypic traits were correlated with the phylogeny of Kudoidae, and that the biological features of K. rayformis originated from the ancient Kudoidae as exhibited by the non-specific infection site tropism and the ability to infect muscle and form pseudocysts.