Journal of Eukaryotic Microbiology最新文献

Thecate amoebae play important roles in terrestrial and aquatic ecosystems. This study introduces a novel thecofilosean amoeba from Arctic and Antarctic sea sediments. Phylogenetic analysis based on the 18S rDNA sequence places it in the family Chlamydophryidae (order Tectofilosida, class Thecofilosea). However, the novel organism exhibits a significant genetic divergence and distinct morphology from its closest relatives, prompting us to erect the novel genus Katarium with its type species Katarium polorum. K. polorum is a consumer of diatoms and prokaryotes, indicating an important role in nutrient cycling in the polar marine food webs.

As chloroplast-stealing or “kleptoplastidic” lineages become more reliant on stolen machinery, they also tend to become more specialized on the prey from which they acquire this machinery. For example, the ciliate Mesodinium rubrum obtains > 95% of its carbon from photosynthesis, and specializes on plastids from the Teleaulax clade of cryptophytes. However, M. rubrum is sometimes observed in nature containing plastids from other cryptophyte species. Here, we report on substantial ingestion of the blue-green cryptophyte Hemiselmis pacifica by M. rubrum, leading to organelle retention and transient increases in M. rubrum's growth rate. However, microscopy data suggest that H. pacifica organelles do not experience the same rearrangement and integration as Teleaulax amphioxeia's. We measured M. rubrum's functional response, quantified the magnitude and duration of growth benefits, and estimated kleptoplastid photosynthetic rates. Our results suggest that a lack of discrimination between H. pacifica and the preferred prey T. amphioxeia (perhaps due to similarities in cryptophyte size and swimming behavior) may result in H. pacifica ingestion Thus, while blue-green cryptophytes may represent a negligible prey source in natural environments, they may help M. rubrum survive when Teleaulax are unavailable. Furthermore, these results represent a useful tool for manipulating M. rubrum's cell biology and photophysiology.

Amyloodinium ocellatum is a protozoan parasite that causes amyloodiniosis in marine and brackish water fish, threatening global aquaculture. The present study investigates the morphology and ultrastructure of the free-living stages of A. ocellatum (tomont and dinospore) using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Dinospores measured 13.03–19.66 μm in length, 12.32–18.71 μm in width, and were laterally flattened. Dinospores had a transverse flagellum for propulsion and a longitudinal flagellum for direction control. The cyst wall had three distinct layers and included cellulose. The outer wall was coated with numerous bacteria. The orange-red speckled eyespot was observed all tomont developmental stages and in the dinospore of A. ocellatum. Tomonts proliferation required successive nuclear division, the formation of new cyst walls, and cytoplasmic segregation. The cytoplasm comprises mainly the matrix, organelles, and inclusions. The matrix was grainy and evenly distributed. In addition to organelles, including mitochondria with tubular cristae, Golgi apparatus, and endoplasmic reticulum, the cytoplasm had starch grains and lipid droplets as inclusions. The A. ocellatum cells lacked chloroplasts. This study provides the first ultrastructural view of the cytoplasmic structure of the free-living stages of A. ocellatum.

Human babesiosis is a malaria-like, tick-borne infectious disease with a global distribution. Babesiosis is caused by intraerythrocytic, apicomplexan parasites of the genus Babesia. In the United States, human babesiosis is caused by Babesia microti and Babesia duncani. Current treatment for babesiosis includes either the combination of atovaquone and azithromycin or the combination of clindamycin and quinine. However, the side effects of these agents and the resistance posed by these parasites call for alternative approaches for treating human babesiosis. Proteases play several roles in the context of parasitic lifestyle and regulate basic biological processes including cell death, cell progression, and cell migration. Using the SYBR Green-1 assay, we screened a protease inhibitor library that consisted of 160 compounds against B. duncani in vitro and identified 13 preliminary hits. Dose response assays of hit compounds against B. duncani and B. microti under in vitro conditions identified five effective inhibitors against parasite growth. Of these compounds, we chose ixazomib, a proteasome inhibitor as a potential drug for animal studies based on its lower IC₅₀ and a higher therapeutic index in comparison with other compounds. Our results suggest that Babesia proteasome may be an important drug target and that developing this class of drugs may be important to combat human babesiosis.

Protists show diverse lifestyles and fulfill important ecological roles as primary producers, predators, symbionts, and parasites. The degradation of dead microbial biomass, instead, is mainly attributed to bacteria and fungi, while necrophagy by protists remains poorly recognized. Here, we assessed the food range specificity and feeding behavior of the algivorous flagellate Orciraptor agilis (Viridiraptoridae, Cercozoa) with a large-scale feeding experiment. We demonstrate that this species is a broad-range necrophage, which feeds on a variety of eukaryotic and prokaryotic algae, but fails to grow on the tested fungi. Furthermore, our microscopic observations reveal an unexpected flexibility of O. agilis in handling food items of different structures and biochemistry, demonstrating that sophisticated feeding strategies in protists do not necessarily indicate narrow food ranges.

The planktonic dinoflagellate genus Centrodinium has been understudied, with the type species C. elongatum remaining undocumented since the original description. Here, we report C. elongatum isolated from Mazatlán, Mexican Pacific. In the chains, the posterior daughter cell with an incomplete apical horn shows the morphology of C. elongatum, while the anterior daughter cell with complete epitheca corresponds to C. pulchrum. For the first time, a species of Centrodinium sensu stricto (highly laterally flattened species with horns) was cultured. An unarmored life stage, known as Murrayella ovalis, derived from the spheroplast after ecdysis. In the rDNA molecular phylogenies, C. elongatum (=C. pulchrum) nested as basal to morphologically similar species (C. eminens and C. intermedium) and as a sister group of a former Murrayella species, C. punctatum. C. elongatum differs from C. eminens and C. intermedium in the chain formation, second apical (2′) plate not being divided, horns with coarse poroid ornamentation, and missing prominent distal spinules. The taxonomy of Centrodinium sensu stricto is revised, with a discussion in the identities of C. complanatum, C. eminens, and C. maximum. The name C. deflexum is restored as a senior synonym of C. intermedium and C. ovale.

Pirsoniales is a stramenopile order composed of marine parasitoids of diatoms with unique life cycle. Until recently, a single genus, Pirsonia, uniting six species, was known. The recent identification of new free-living eukaryotrophic Pirsoniales Pirsonia chemainus, Feodosia pseudopoda, and Koktebelia satura changed our understanding of this group as exclusively parasitic. However, their cell ultrastructure and feeding preferences were not fully studied due to the death of the cultures. In this study, we re-isolated some of these Pirsoniales and established six new strains exhibiting predatory behavior, including a first freshwater representative. This allowed us to describe five new genera and species, as well as to emend the diagnosis of the order Pirsoniales. The 18S rRNA gene phylogenetic analysis revealed the position of new strains within Pirsoniales and their relationships with parasitoid relatives and environmental sequence lineages. Feeding experiments on novel Pirsoniales strains using diverse algal prey showed that they were not able to form trophosomes and auxosomes. The ability of cell aggregation in Pirsoniales was observed for the first time. One of the studied strains contained intracellular gammaproteobacteria distantly related to Coxiella. Ultrastructural analyses revealed a more complex cytoskeleton structure in Pirsoniales than previously thought and supported the monophyly of Bigyromonadea and Pseudofungi.

The Diphyllatea (CRuMs) are heterotrophic protists currently divided into three distinct clades (Diphy I–III). Diphy I are biflagellates in the genus Diphylleia, whereas Diphy II and III represent cryptic clades comprising Collodictyon-type quadriflagellates that were recently distinguished based on rRNA gene phylogenies. Here, we isolated Diphyllatea from freshwater crater lakes on two South Pacific islands and generated high-quality transcriptomes from species representing each clade, including the first transcriptomic data from Diphy III. Phylogenomic analyses support the separation of Diphy II and III, while transcriptome completeness highlights the utility of these data for future studies. Lastly, we discuss the biogeography and ecology of Diphyllatea on these remote islands.

Glugea sp. found infecting the liver of the teleost fish Carangoides bajad from the Red Sea, Egypt, is described based on light microscopy and ultrastructural characteristics combined with phylogenetic analyses. This microsporidium forms whitish xenomas up to ~4 mm in size. Xenomas display numerous parasitophorous vacuoles totally filled by mature spores, no other life cycle stages were observed. Mature spores ellipsoidal and measuring 6.3 × 4.0 μm in size. The polaroplast appears composed of two distinct regions: an electron-dense vesicular region and a densely packed lamellar region. The polar tubule forms approximately 24–27 coils arranged in three layers encircling the posterior vacuole. The small subunit (SSU) rRNA gene and its ITS region were sequenced and showed the highest similarity of 99.4% to other Glugea spp. Bayesian inference and maximum likelihood analyses place the novel isolate within the Glugea clade, more specifically within a subclade that predominantly grouped species described from fish inhabiting the Arabian Gulf or Red Sea. The results validate the parasite's classification in the Glugea genus. Nevertheless, until more detailed ultrastructural and molecular data are obtained, the identification of the current Glugea species is hampered by the absence of some developmental stages and the high degree of genetic similarity.