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Systematic studies on 70 MACC isolates previously identified as ‘Chlamydomonas’, a unicellular flagellate, were carried out based on partial 18S rRNA. The aim of this study was to determine the phylogenetic affiliations of Chlamydomonas strains in the MACC collection. The study found that most of the strains were not Chlamydomonas. Nine clusters of phylogenetically similar taxa were identified. The previous determinations were completed with their new phylogenetic affiliations (partly due to changes in green algae classification). Molecular data revealed that 3 of the 70 strains are from Arenicolinia, 14 are members of the phylogroup Stephanosphaerinia, 11 are Oogamochlamydinia, 1 is Chloromonadinia, 19 are Reinhardtinia, 2 are Polytominia, 9 are Scenedesmaceae, 5 are Moewusinia, and 6 are Chlorella. Clades were established by 18S rRNA similarity and p-distances. This study reveals the need to revise established culture collections whose isolates are solely identified with morphology.

Polybrominated diphenyl ethers (PBDEs), a class of brominated flame retardants, have been extensively applied and eventually leached into the surrounding environment. Marine microalgae are not only the dominant primary producers of marine ecosystem, but also food source for aquaculture. PBDEs have been found to remarkably inhibit growth, photosynthesis and metabolism of marine microalgae. However, whether they also affect swimming behavior of marine motile microalgae remains unknown. We chose BDE-47, BDE-99 and BDE-153 as model PBDEs, and the unicellular marine green flagellate, Platymonas subcordiformis, as test organism to figure out this issue. After two-hour exposure, motile cells proportion (MOT), swimming velocity (VCL, VAP and VSL), and swimming pattern (LIN and STR) of P. subcordiformis were measured via computer assisted cell movement tracking. Results suggest that the three PBDEs not only reduced motile cells proportion and swimming velocity, but also altered swimming pattern. BDE-47 was more toxic than BDE-99, followed by BDE-153, indicating their toxicity decreased as bromination degree increases. Swimming ability of P. subcordiformis was even completely arrested when BDE-47 and BDE-99 at 32 μg/L. The impairment of swimming ability by PBDEs might thereby hinder growth and survival of marine microalgae, and subsequently threaten marine ecosystem and aquaculture industry. More importantly, this study implies that marine microalgae swimming behavior test is more efficiency and sensitive than traditional marine microalgal bioassays, like growth and photosynthesis tests. We suggest that although future work is needed, swimming behavior analysis of P. subcordiformis with MOT, VCL and VAP as endpoints can be developed as a low-cost, convenient, fast, reliable and sensitive method for seawater quality assessment.

Contaminating organisms in mass cultivation present one of the major challenges that must be overcome for successful commercialization of algal biofuels. The present study identified a range of contaminating organisms in Chlorella cultures cultivated in outdoor raceway ponds at the Arizona Center for Algae Technology and Innovation (Mesa, AZ). Nineteen organisms or Operational Taxonomic Units (OTU) in the Chlorella culture were identified by a combination of microscopic observation and 18S rRNA denaturing gradient gel electrophoresis (DGGE). More detailed analyses identified these contaminating organisms as 2 fungi, 7 flagellates, 3 amoebae, 4 ciliates, 1 rotifer, and 2 large insects. Among them Poterioochromonas sp., a small flagellate, appeared to be one of the most harmful causing culture collapse. In order to control Poterioochromonas sp., various operational parameters and application strategies of ultrasonic treatment were investigated. During sixteen-day consecutive Chlorella cultivation in batch mode, the ultrasonication conditions of 6 L min−1 flow rate with the power of 495 W at 100% amplitude and a treatment frequency of once for 1 h every day, was proven to be the most effective in preventing Poterioochromonas outbreak in Chlorella culture with volume of 60 L. The above ultra-sonication method was also effective at destroying an unknown fungus, an amoeba (Acanthocystis sp.), and ciliates (a member of the family Orchitophryidae). Our findings can serve as a technical foundation for the application of ultrasonication to control some of the contaminating microorganisms in mass cultivation of microalgae including Chlorella.

Although the abyssal seafloor represents the most common benthic environment on Earth, eukaryotic microbial life at abyssal depths is still an uncharted territory. This is in striking contrast to their potential importance regarding the material flux and bacteria consumption in the deep sea. Flagellate genotypes determined from sedimentary DNA deep‐sea samples might originate from vital deep‐sea populations or from cysts of organisms sedimented down from surface waters. The latter one may have never been active under deep‐sea conditions. We wanted to analyze the principal ability of cultivable heterotrophic flagellates of different phylogenetic groups (choanoflagellates, ancyromonads, euglenids, kinetoplastids, bicosoecids, chrysomonads, and cercozoans) to survive exposure to high hydrostatic pressure (up to 670 bar). We summarized our own studies and the few available data from literature on pressure tolerances of flagellates isolated from different marine habitats. Our results demonstrated that many different flagellate species isolated from the surface waters and deep-sea sediments survived drastic changes in hydrostatic pressure. Barophilic behavior was also recorded for several species isolated from the deep sea indicating their possible genetic adaptation to high pressures. This is in accordance with records of heterotrophic flagellates present in environmental DNA surveys based on clone libraries established for deep‐sea environments.

The potential of using Chlorella for biomass production for biofuels and other products on a large scale has been explored for many years. However, Chlorella cells are vulnerable to predation by protozoa, resulting in productivity loss. Poterioochromonas malhamensis, a mixotrophic predator flagellate, is regarded as one of the most dangerous grazers in Chlorella cultures on a commercial scale, as it can cause deterioration or collapse of microalgal mass cultivation in just a few days. There is a need, therefore, for effective and safe methods for protecting microalgal crops from grazing organisms. In this study, a chemical compound, ammonium bicarbonate (NH4HCO3), was used to control P. malhamensis in cultures of Chlorella sorokiniana under both laboratory and pilot-scale conditions. The results showed that 400 to 800 mg L−1 NH4HCO3 could effectively control P. malhamensis, and the 24-h mortality of P. malhamensis was as high as 94% in indoor experiments and 90% in outdoor ponds. Compared with the control groups, the biomass of Chlorella increased by 77% in indoor cultures and 95% in outdoor ponds when treated with NH4HCO3. In addition, microscopic observation indicated that NH4HCO3 could also effectively control other contaminants in outdoor cultures, such as some ciliates, amoebae and cysts. Molecular sequencing further revealed that NH4HCO3 might be effective in controlling other flagellates. Experiments assessing the influence of pH on toxicity of NH4HCO3 to P. malhamensis revealed, for the first time, that it was the NH3 of NH4HCO3, not the NH4+, or the total NH3-N (NH3 and NH4+), that killed P. malhamensis. Owing to its high toxicity to P. malhamensis and other contaminants, its relative safety to C. sorokiniana, and its low commercial price and ecological acceptability, addition of NH4HCO3 is considered to be a highly effective, safe, and cost-efficient way to control P. malhamensis contamination in mass cultivation of Chlorella.

Chlorella is a strong candidate as a potential production organism for future biofuels and other algal-derived products, but contamination by grazing protozoa is a major constraint to its commercial production on a large scale. In this study, a mixotrophic flagellate chrysophyte that targeted Chlorella in mass culture was observed in samples from outdoor massive photobioreactor systems. Data from field studies demonstrated that once the predatory flagellate was observed in the algal culture, there was a dramatic reduction in Chlorella cell concentration. The grazer was identified as Poterioochromonas malhamensis, based on both morphology and molecular barcoding employing 18S rDNA gene sequences. Transmission electron microscopy revealed that P. malhamensis digested the ingested Chlorella sorokiniana cells within a food vacuole, with residual cell wall from the prey being observed in the final phase. It was noted that during the process of ingesting and digesting the C. sorokiniana cells, the chloroplast of P. malhamensis reduced in size, whereas the number and volume of mitochondria increased. After the prey had been completely digested, the chloroplast and mitochondria of P. malhamensis returned to their pre-feeding status. Feeding experiments in the laboratory demonstrated that cell concentrations of C. sorokiniana and P. malhamensis were negatively correlated. Furthermore, once P. malhamensis reached 5 × 105 cells mL− 1 and the ratio of C. sorokiniana to P. malhamensis was in the range 15:1 to 30:1, the cell concentration of the C. sorokiniana culture decreased dramatically. The optimal temperature for P. malhamensis grazing on C. sorokiniana was 25 °C; the grazing ability of P. malhamensis in light conditions was higher than that in dark conditions; and P. malhamensis preferred weakly acidic conditions. In addition, it was demonstrated that this flagellate could graze on other common microalgae of commercial value. On the basis of its capacity to ingest a wide spectrum of phytoplankton, and its wide environmental range and global distribution, this contaminant has the potential to be a common problem in microalgal mass cultures and warrants further study.

The microalga Poterioochromonas malhamensis was found to be capable of accumulating the storage β-1,3-glucan in soluble form under heterotrophic conditions. In this study, the highest biomass yield of 32.8 g L−1 was achieved by combining the utilization of ammonium chloride as the nitrogen source, simultaneous addition of vitamins B1 and B12 and maintenance of pH at 6.0. Sugar profiling and nuclear magnetic resonance analysis indicated that the P. malhamensis β-1,3-glucan was composed of glucose with the β-(1 → 3) main chain and the β-(1 → 6) side chain. Under the optimal cultivation conditions, the cellular β-1,3-glucan content was up to 55% of the cell dry weight. Moreover, the P. malhamensis β-1,3-glucan could significantly promote the fin regeneration and improve the in vivo antioxidative activity of zebrafish. This study underpins the feasibility of culturing P. malhamensis under heterotrophic conditions for producing the highly water-soluble bioactive β-1,3-glucans for food and pharmaceutical applications.

Symbiogenesis, literally ‘becoming by living together’, refers to the crucial role of symbiosis in major evolutionary innovations. The term usually is reserved for the major transition to eukaryotes and to photosynthesising eukaryotic algae and plants by endosymbiosis. However, in some eukaryote lineages endosymbionts have been lost secondarily, showing that symbiosis can trigger a major evolutionary innovation, even if symbionts were lost secondarily. This leads to the intriguing possibility that symbiosis has played a role in other major evolutionary innovations as well, even if not all extant representatives of such groups still have the symbiotic association. We evaluate this hypothesis for two innovations in termites (Termitoidae, also known informally as “Isoptera”): i) the role of flagellate gut protist symbionts in the transition to eusociality from cockroach-like ancestors, and ii) the role of non-gut associated symbionts in the transition to ‘higher’ termites, characterized by the absence of flagellate gut protists. In both cases we identify a crucial role for symbionts, even though in both cases, subsequently, symbionts were lost again in some lineages. We also briefly discuss additional possible examples of symbiogenesis. We conclude that symbiogenesis is more broadly applicable than just for the endosymbiotic origin of eukaryotes and photosynthetic eukaryotes, and may be a useful concept to acknowledge the important role of symbiosis for evolutionary innovation. However, we do not accept Lynn Margulis's view that symbiogenesis will lead to a paradigm shift from neoDarwinism, as the role of symbiosis in evolutionary change can be integrated with existing theory perfectly.

The rice rhizosphere is a hotspot for microorganisms, where heterotrophic protists are the major group of microeukaryotes and probably function as microbial predators. In this work, the microscale distribution of soil protists in a rice rhizosphere was studied in a microcosm experiment. Rice seedlings (Oryza sativa L. Nipponbare) were grown in “mini-rhizoboxes” made of thin glass plates and containing a nutrient solution and a diluted suspension of a rice-field soil in soft agar. This enabled nondestructive microscopic observations and the enumeration of heterotrophic protists in the rhizosphere. Different morphotypes of protists (ciliates, flagellates, and amoebae) densely populated the rhizosphere within 3 days after root development with an extended area at the root tips. There was a clear boundary between the rhizosphere and non-rhizosphere in terms of protistan density. Flagellates had the highest frequency and abundance in the colonization. Different protist morphotypes differed in their microscale distribution patterns. Flagellates including amoeboflagellates dominated near the roots, while amoebae inhabited sites adjacent to the flagellates, at a slight distance from the roots and with a clearly segregated habitat. Ciliates were exclusively observed at the tips of the rice roots. Denaturing gradient gel electrophoresis demonstrated that the microeukaryotic community in the rice rhizosphere was distinct from that in the non-rhizosphere. Protistan species inhabiting the rice rhizosphere were affiliated with Cercozoa (Nucleocercomonas sp. and Gymnophrys sp.), Amoebozoa (Echinamoeba sp.), and Ciliophora (Oxytricha sp.). Our results demonstrate that the rhizosphere effect of rice roots on heterotrophic protists differs at a microscale depending on the root segment. They also suggest a close relationship between Cercozoa flagellates and the roots.

The unicellular freshwater flagellate Euglena gracilis has a highly developed sensory system. The cells use different stimuli such as light and gravity to orient themselves in the surrounding medium to find areas for optimal growth. Due to the ability to produce oxygen and consume carbon dioxide, Euglena is a suitable candidate for life support systems. Participation in a long-term space experiment would allow for the analysis of changes and adaptations to the new environment, and this could bring new insights into the mechanism of perception of gravity and the associated signal transduction chain. For a molecular analysis of transcription patterns, an automated system is necessary, capable of performing all steps from taking a sample, processing it and generating data. One of the developmental steps is to find long-term stable reagents and materials and test them for stability at higher-than-recommended temperature conditions during extended storage time. We investigated the usability of magnetic beads in an Euglena specific lysis buffer after addition of the RNA stabilizer Dithiothreitol over 360 days and the lysis buffer with the stabilizer alone over 455 days at the expected storage temperature of 19 °C. We can claim that the stability is not impaired at all after an incubation period of over one year. This might be an interesting result for researchers who have to work under non-standard lab conditions, as in biological or medicinal fieldwork.

Chinese researchers have made substantial progresses in research on the harmful algal bloom (HAB) species Phaeocystis globosa since the first P. globosa bloom outbreak in the Chinese coastal waters in 1997. However, as many results, especially the earlier ones, were published in non-English literature, much of the research on P. globosa biology, ecology, and biogeochemistry made by Chinese researchers have been unknown to colleagues from other countries. We review current knowledge on taxonomy, morphology, genetics, physiology, survival strategies and mitigation of P. globosa gained by Chinese researchers from the past two decades. P. globosa is the only Phaeocystis species that causes blooms in the Chinese waters, although other Phaeocystis species including P. jahnii and P. cordata have been detected in Chinese coastal regions. P. globosa has a complex life history with at least two morphotypes including a haploid flagellate and a diploid colonial cell. Colonial P. globosa blooms typically occur in winter after a diatom bloom in coastal waters of the South China Sea. P. globosa in Chinese coastal waters usually has extremely large colonial sizes, up to 3 cm in diameter, an order of magnitude greater than that observed in European coastal waters. The development of giant colonies is associated with enhanced sinking rate, limited nutrient diffusion, as well as decreased stability of colonies. The Chinese P. globosa strains also showed strong genetic diversity and physiological plasticity, being able to grow and develop into colonies at higher temperature and irradiance relative to that in European waters. High genetic diversity of P. globosa was revealed by developing high-resolution and high-specificity molecular markers including Phaeocystis globosa chloroplast 1 (pgcp1). Due to the severe impact of P. globosa on ecology and economy, much effort has been made to mitigate P. globosa blooms including the application of modified clays. Overall, P. globosa in the Chinese waters demonstrate unique genetic, phenotypical and physiological features that differ from P. globosa in other ocean regions. Further studies are needed to explore how environmental factors trigger the occurrence of P. globosa blooms and ascertain the impact of P. globosa blooms on the environment.

Trichomes are prevalent on the surfaces of many organs in Rosa and have an important impact on the edibility of the fruit. Here, the diversity, distribution, anatomical structure, and genetic regulation of trichomes in Rosa roxburghii Tratt. (RR) and R. sterilis S. D. Shi (RS) were explored. RS and RR are important commercial crops in China due to their nutritional and medicinal value; however, their consumption and utilization are limited, in part, by the abundance of trichomes on the fruit. Two main forms of trichomes were present in both germplasms, namely glandular and non-glandular trichomes. Non-glandular flagellate and acicular trichomes were observed on the sepals, fruit, major leaf veins, and pedicels of both germplasms, whereas non-glandular branched trichomes were present only on RS. Flagellate trichome exfoliation and acicular trichome lignification occurred gradually on the developing fruit. Capitate glandular and bowl-shaped glandular trichomes were abundant in RS, but were only observed on the major leaf veins of RR. Furthermore, some capitate glandular trichomes on the pedicels were found to develop into prickles in RS. Transmission electron microscopy indicated that vacuolation and in the extraplasmic space occurred in the glandular cells of the capitate trichomes at the late secretory stage. The prickles in RR and RS mainly consisted of lignin, suberin, cellulose, and hemicellulose, though the ratios of these constituents varied between species. The expression levels of several well-known core trichome regulatory genes varied depending on the trichome type. The transcript abundance of GL1, GL2, and TTG1 was significantly higher in organs covered with acicular trichomes in both RR and RS, while the highest mRNA level of TRY was observed in the glabrous organs, suggesting a negative effect of this gene on trichome formation. Notably, the negative regulatory factor CPC was only highly expressed in the leaf mesophyll of RR, where no glandular trichomes were present, and the GL3, PDF2-like, and CPC transcription factors co-regulated glandular trichome formation in RS. The expression of these genes peaked in the Fb3-stage buds of RR and B3-stage buds of RS, indicating different key phases for the regulation of trichome initiation. These data provide new insights into the genetic control of trichome formation in two Rosa species.

Poterioochromonas malhamensis is a flagellate grazer that readily contaminates Chlorella cultures and can have a devastating impact on them, especially in large-scale culture. Here, we developed a quantitative real-time polymerase chain reaction (qPCR) method to detect and quantify P. malhamensis from high-density Chlorella cultures. A set of primers specific to the mitochondrial cytochrome oxidase subunit I gene (COI) of P. malhamensis was screened and characterized comprehensively for specificity, sensitivity and applicability to the field samples. The results showed that the primers were sufficiently specific to distinguish P. malhamensis from a wide variety of micro-eukaryotes which often coexist with P. malhamensis in microalgal cultures, and could detect a very low concentration of P. malhamensis from high concentrations of Chlorella sorokiniana (106, 107, and 108 C. sorokiniana cells mL−1). Moreover, the qPCR method was applied successfully to the monitoring of P. malhamensis population dynamics in Chlorella cultures in 100 L open ponds. Finally, the qPCR method was applied to the detection of this flagellate in the wider environment of the culture facility. It showed that the flagellate was widely distributed in the environment, and we speculate that Poterioochromonas in Chlorella cultures could originate from the air, the water, and the seed cultures. The qPCR method developed therefore proved to have the specificity and efficiency to be useful for early detection of P. malhamensis in industrial-scale Chlorella cultivation.

Recent progress in understanding the early evolution of eukaryotes was tied to morphological identification of flagellates and heliozoans in the natural samples, isolation of their cultures and genomic and ultrastructural investigations. These protists are the smallest and least studied microbial eukaryotes but play an important role in functioning of microbial food webs. Using light and electron microscopy, we have studied the diversity of heterotrophic flagellates and centrohelid heliozoans from marine waters of Curacao (The Netherlands Antilles), and provide micrographs and morphological descriptions of observed species. Among 86 flagellates and 3 centrohelids encountered in this survey, five heterotrophic flagellates and one Centrohelid heliozoan were not identified even to the genus. Some flagellate protists have a unique morphology, and may represent undescribed lineages of eukaryotes of high taxonomic rank. The vast majority (89%) of identified flagellates are characterized by wide geographical distribution and had been reported previously from all hemispheres and various climatic regions. More than half of the species were previously observed not only from marine, but also from freshwater habitats. The parameters of the species accumulation curve indicate that our species list obtained for the Curacao study sites is far from complete, and each new sample should yield new species.

High hydrostatic pressure (HHP) is a promising method for the inactivation of cells and enzymes in many applications (e.g. food industry, pharmaceutical industry, chemistry). The effects of various high-hydrostatic pressures (20 MPa, 50 MPa, 55 MPa, 60 MPa, 75 MPa, 100 MPa, 150 MPa, 200 MPa, 400 MPa) on the unicellular flagellate Euglena gracilis were determined: (a) life/dead staining with ethidium bromide (EtBr), (b) loss of flagellum, (c) movement behavior, (d) recovery after 7 d, (e) photosystem II quantum yield (Y(PSII)). Pressure was applied for 300 s at room temperature (pressure increment and decrement: 10 MPa s −1 ). The EC 50 value of vitality (EtBr-positive cells) directly after pressure application was 109 MPa. Cell vitality was not impaired below 100 MPa. No recovery of cells after 7 d-cultivation in fresh medium was observed after pressure treatment above 100 MPa. Flagellum-based free swimming of cells was already impaired at pressures above 50 MPa, where some cells started metabolic movement behavior. From 75 MPa to 100 MPa all cells moved metabolically, while at higher-pressure all cells became immotile. In the pressure range between 75 MPa and 200 MPa, cells lost their flagellum. Interestingly, cells pressurized with 400 MPa retained their flagella. Directly after HHP-treatment EC 50 of Y(PSII)-inhibition was about 101 MPa, after 6 h about 102 MPa. The use of fluorescence-based methods is considerably fast, easy and reliable. Because of the different easily recordable parameters, we believe that Euglena gracilis is a promising test organism to determine HHP-effects on eukaryotes. The effects on photosynthesis indicate impacts on intracellular membrane and protein complexes.

Among the freshwater cyclopoids, small and medium-sized species can be propagated in the laboratory with a flagellate Chilomonas as the food. By contrast, large species need to be given, in addition to the flagellate, a larger food organism. For this, although a metazoan food is preferable, a ciliate Stentor may in part serve as a substitute. Methods for infection experiment involving copepods are given, and photographs of some parasite larvae in the copepod host are shown. Usage of carbonated water is helpful to immobilize cyclopoids and thus recommended for infection experiments.

Bleomycin induced flagellate dermatitis is an uncommon and unique adverse effect. With the declining use of bleomycin, this complication is becoming increasingly infrequent in day-to-day clinical practice. We herein describe a case of a 13 year old male patient with left thalamic mixed germ cell tumour treated by multimodality approach, who developed flagellate erythema after two cycles of combination chemotherapy with bleomycin, etoposide and cisplatin (BEP). This brief report highlights the importance of awareness and timely identification and management of this dermatological toxicity in patients undergoing bleomycin based combination chemotherapy.

Summary The kinetoplastids are a widespread and important group of single-celled eukaryotes, many of which are devastating parasites of animals, including humans [1–3]. We have discovered a new insect trypanosomatid in the gut of Culex pipiens mosquitoes. Glyceraldehyde-3-phosphate dehydrogenase- and SSU rRNA-based phylogenetic analyses show this parasite to constitute a distinct branch between the free-living Bodo saltans and the obligatory parasitic clades represented by the genus Trypanosoma and other trypanosomatids. From draft genome sequence data, we identified 114 protein genes shared among the new flagellate, 15 trypanosomatid species, B. saltans , and the heterolobosean Naegleria gruberi , as well as 129 protein genes shared with the basal kinetoplastid Perkinsela sp. Individual protein phylogenies together with analyses of concatenated alignments show that the new species, here named Paratrypanosoma confusum n. gen., n. sp., branches with very high support at the base of the family Trypanosomatidae. P. confusum thus represents a long-sought-after missing link between the ancestral free-living bodonids and the derived parasitic trypanosomatids. Further analysis of the P. confusum genome should provide insight into the emergence of parasitism in the medically important trypanosomatids.

Soil protists are commonly suggested being solely bacterivorous, serving together with bacterivorous nematodes as the main controllers of the bacterial energy channel in soil food webs. In contrast, the fungal energy channel is assumed to be controlled by arthropods and mycophagous nematodes. This perspective accepted by most soil biologists is, however, challenged by functional studies conducted by taxonomists that revealed a range of mycophagous protists. In order to increase the knowledge on the functional importance of mycophagous protists we isolated and initiated cultures of protist taxa and tested eight for facultative feeding on diverse fungi in microcosm experiments. Two different flagellate species of the genus Cercomonas, the testate amoeba Cryptodifflugia operculata and four genera of naked amoebae (Acanthamoeba sp., Leptomyxa sp., two Mayorella spp. and Thecamoeba spp.) fed and grew on yeasts with four taxa (Cercomonas sp., Leptomyxa sp., Mayorella sp., and Thecamoeba sp.) also thriving on spores of the plant pathogenic hyphal-forming fungus Fusarium culmorum. To identify the potential importance of mycophagous protists in the environment we applied a data-mining approach targeting small subunit (SSU) rRNA data obtained in metatranscriptomes of five fundamentally different terrestrial samples. We focused our analyses on the distribution and relative abundances of two well-studied mycophagous protist groups, vampyrellid amoebae and grossglockneriid ciliates. Both groups were detected in all of the highly contrasting terrestrial samples, comprising up to 3% of all protist SSU rRNA transcripts. SSU transcripts of these two groups, in contrast to all remaining protist SSU transcripts, showed strong correlations with the relative abundance of fungal sequences indicating close direct trophic interactions. Taken together, this study provides evidence that mycophagy among soil protists is common and might be of substantial but hitherto overlooked ecological importance in terrestrial ecosystems. Future studies should aim at evaluating taxon-specific (facultative) mycophagy, decipher changes caused in the fungal community and quantitatively evaluate the functional importance of this trophic position in soil ecosystems.

Heterotrophic flagellates (HF) play an essential function in the microbial food loops as it is an agent of biochemical cycling of limiting nutrients and being a useful bio-indicator of environmental changes. However, in spite of its profound role in aquatic ecosystem studies regarding the evaluation of the spatial and seasonal patterns of the flagellate communities are lacking in Bangladeshi water, and therefore, study on community pattern of HF in a tropical river like Padma River could be a best example for other tropical rivers of the world. The present investigation was performed with the primary objectives to evaluate flagellate community structure, their spatial and seasonal distribution patterns, and the role of hydro-biological and physicochemical variables in determining their variability. The study was conducted at selected four study sites in Padma river, Bangladesh for a period of two distinct season (dry and wet season) in 2018. Standard procedures were followed in collecting, analyzing and interpreting of sampled data. The study revealed a total of 18 genera with different degrees of specificity to hydro-biological and physicochemical variables. It further establishes significant seasonal patterns, rather than spatial differences in the distribution of the flagellate communities. We found that the higher precipitation intensity during monsoon was the critical determinant in hydrological, biological, and physicochemical fluctuations which caused a significant decline in the value of the above variables. Likewise, total phytoplankton, zooplankton, and total heterotrophic bacterial abundance was also declined five-folds during the wet season. Similarly, total abundance of HF showed a significant decline in the community during the wet season. Genus wise relative abundance (RA%) of Euglena (23.81 %) was the highest during the dry season. In conclusion, this research and data analysis techniques propose unique insights on the flagellate links to environmental and hydro-biological regime in a tropical river with global implications.

Highlights: • New predatory protist (Tunicaraptor) with unique morphology is related to animals • Tunicaraptor calls into question many of the well-accepted relationships in Holozoa • Tunicaraptor possesses a unique combination of “animal-specific” gene products • Eukaryovorous flagellates may represent a major share of unicellular animal relatives The origin of animals is one of the most intensely studied evolutionary events, and our understanding of this transition was greatly advanced by analyses of unicellular relatives of animals, which have shown many “animal-specific” genes actually arose in protistan ancestors long before the emergence of animals [1–3]. These genes have complex distributions, and the protists have diverse lifestyles, so understanding their evolutionary significance requires both a robust phylogeny of animal relatives and a detailed understanding of their biology [4, 5]. But discoveries of new animal-related lineages are rare and historically biased to bacteriovores and parasites. Here, we characterize the morphology and transcriptome content of a new animal-related lineage, predatory flagellate Tunicaraptor unikontum. Tunicaraptor is an extremely small (3–5 μm) and morphologically simple cell superficially resembling some fungal zoospores, but it survives by preying on other eukaryotes, possibly using a dedicated but transient “mouth,” which is unique for unicellular opisthokonts. The Tunicaraptor transcriptome encodes a full complement of flagellar genes and the flagella-associated calcium channel, which is only common to predatory animal relatives and missing in microbial parasites and grazers. Tunicaraptor also encodes several major classes of animal cell adhesion molecules, as well as transcription factors and homologs of proteins involved in neurodevelopment that have not been found in other animal-related lineages. Phylogenomics, including Tunicaraptor, challenges the existing framework used to reconstruct the evolution of animal-specific genes and emphasizes that the diversity of animal-related lineages may be better understood only once the smaller, more inconspicuous animal-related lineages are better studied.

Phytoplankton community' dynamics were studied in Salgados coastal lagoon in order to evaluate the effects of excessive organic loads and also physical stress caused by the irregular opening of the lagoon. Salgados is a hypereutrophic intermittently open coastal lagoon, which received freshwater inputs from small rivers and from a wastewater treatment plant. Cyanophyceae dominated the phytoplankton communities most of the time; Bacillariophyceae became the main taxonomic group in winter when the lagoon was closed; Chlorophyceae was the major class in early summer; pico-nano flagellate algae accounted for a high percentage of total phytoplankton during spring. Potentially harmful taxa were observed during most of the sampling periods, forming blooms and accounting for a considerable percentage of total phytoplankton abundance. A strong differentiation among dry and wet seasons could be noticed. The dry season was dominated by Microsystis aeruginosa , Rhodomonas sp., pico-nano flagellate algae, Cyclotella spp. and Planktothrix sp., while the wet season, although still with the presence of Microsystis aeruginosa , was dominated by Dolichospermum spiroides . The best environmental variables explaining stations patterns and based on phytoplankton taxa were days of isolation, pH, and salinity. Temperature, cumulative rain and total phosphorus were also related with species and stations patterns. The high nutrient load in Salgados lagoon promoted the development and persistence of harmful algae blooms. Proper management of coastal lagoons involves not only the control of direct discharges of nutrients, but also of other factors, including water level and communication with the sea.

In this study, we investigated the effect of land use intensity, soil parameters and vegetation on protistan communities in grassland soils. We performed qualitative (T-RFLP) and quantitative (qPCR) analyses using primers specifically targeting the 18S rRNA gene for all Eukarya and for two common flagellate groups, i.e. the Chrysophyceae and the Kinetoplastea. Both approaches were applied to extracted soil DNA and RNA, in order to distinguish between the potentially active protists (i.e. RNA pool) and the total protistan communities, including potentially inactive and encysted cells (i.e. DNA pool). Several environmental determinants such as site, soil parameters and vegetation had an impact on the T-RFLP community profiles and the abundance of the quantified 18S rRNA genes. Correlating factors often differed between quantitative (qPCR) and qualitative (T-RFLP) approaches. For instance the Chrysophyceae/Eukarya 18S rDNA ratio as determined by qPCR correlated with the C/N ratio, whereas the community composition based on T-RLFP analysis was not affected indicating that both methods taken together provide a more complete picture of the parameters driving protist diversity. Moreover, distinct T-RFs were obtained, which could serve as potential indicators for either active organisms or environmental conditions like water content. While site was the main determinant across all investigated exploratories, land use seemed to be of minor importance for structuring protist communities. The impact of other parameters differed between the target groups, e.g. Kinetoplastea reacted on changes to water content on all sites, whereas Chrysophyceae were only affected in the Schorfheide. Finally, in most cases different responses were observed on RNA- and DNA-level, respectively. Vegetation for instance influenced the two flagellate groups only at the DNA-level across all sites. Future studies should thus include different protistan groups and also distinguish between active and inactive cells, in order to reveal causal shifts in community composition and abundance in agriculturally used systems.

Chagas disease, the American trypanosomiasis, is an important neglected tropical illness caused by the flagellate protozoan Trypanosoma cruzi (Kinetoplastida, Trypanosomatidae) and transmitted by insects of the subfamily Triatominae (Hemiptera: Reduviidae). Here we provide an overview on the current knowledge about Triatominae ecology, its association with human, T. cruzi infection and the immediate consequences of habitat fragmentation. We also discuss the geographic distribution of the species and the importance of predicting their distributions to control programs.

In 2010, a massive bloom of the raphidophycean flagellate Chattonella occurred in the Ariake Sea and Tachibana Bay. Bloom dynamics and hydrographical conditions were examined by field survey. The development and decline of the bloom occurred three times in Tachibana Bay. First and third bloom developments synchronized with precipitation, and the second bloom developed in synchronization with a salinity decrease which occurred in relation to an increase of river discharge from the Chikugo River which takes several days to flow from the Ariake Sea. These results imply that the bloom was transported with the low salinity water from the Ariake Sea to Tachibana Bay. During blooms along the northern coast of Shimabara Peninsula, the predominant phytoplankton species changed from Chattonella to Skeletonema. Low salinity water intrusion induced an interregional difference of the Chattonella and Skeletonema bloom spatially-differentiated by the salinity in the Ariake Sea and Tachibana Bay.

A novel free-living heterotrophic stramenopile, Platysulcus tardus gen. nov., sp. nov. was isolated from sedimented detritus on a seaweed collected near the Ngeruktabel Island, Palau. P. tardus is a gliding flagellate with tubular mastigonemes on the anterior short flagellum and a wide, shallow ventral furrow. Although the flagellar apparatus of P. tardus is typical of stramenopiles, it shows novel ultrastructural combinations that are not applied to any groups of heterotrophic stramenopiles. Phylogenetic analysis using SSU rRNA genes revealed that P. tardus formed a clade with stramenopiles with high support. However, P. tardus did not form a subclade with any species or environmental sequences within the stramenopiles, and no close relative was suggested by the phylogenetic analysis. Therefore, we concluded that P. tardus should be treated as a new genus and species of stramenopiles and have proposed a new family, Platysulcidae fam. nov., for this phylogenetically distinct organism.

Red tides – discolorations of the sea surface due to dense plankton blooms – occur regularly in coastal and offshore waters along much of the world's coastline. Red tides often cause large-scale mortalities of fish and shellfish and significant losses to the aquaculture and tourist industries of many countries. Therefore, understanding and predicting the mechanisms controlling the outbreak, persistence, spread, and decline of red tides are important concerns to scientists, officials, industry, and the public. With increasing knowledge of red-tide species and red-tide events, new mechanisms have been discovered. Based on the nutrition and behaviors of red-tide organisms and biological interactions among them, red-tide outbreaks can be categorized into a hierarchy of four generation mechanisms (GM1–GM4). In the simplest, GM1, all phototrophic red-tide species were treated as exclusively autotrophic organisms without the ability to swim. However, this GM cannot explain red-tide outbreaks in oligotrophic surface waters offshore. Vertical migration (considered in GM2) and mixotrophy (GM3) enable red-tide flagellates to acquire growth factors from nutrient-rich deep waters or co-occurring prey, respectively. In natural environments, all red tides occur by those species outgrowing co-occurring organisms; red-tide species dominate communities by eliminating other species or reducing their abundances. Thus, GM4 contains the direct biological interactions (i.e., inhibition by physical contact or chemical effects) and indirect biological interactions (i.e., acquiring resources faster than others) that can affect the dominance of red-tide species under given conditions. Correctly choosing one of these four GMs for red tides dominated by one causative species is important because the accuracy of predictions may be outweighed by the costs and time required to acquire the relevant information. In this study, mechanisms describing the outbreak, persistence, and decline of red tides were reviewed, the advantages and limitations of each mechanism were evaluated, and insights about the evolution of the mechanisms were developed.

A countrywide surveillance of sand flies was performed to obtain information on their geographical distribution and natural infection by Leishmania protozoa in Ecuador. A total of 18,119 sand flies were collected by human landing collections during 32 years from 1982 to 2014, and 29 species were recognized. The most prevalent 10 species were Lutzomyia gomezi, Lu. robusta, Lu. hartmanni, Lu. shannoni, Lu. trapidoi, Lu. panamensis, Lu. maranonensis, Lu. ayacuchensis, Lu. tortura and Lu. yuilli yuilli, and their topographical and vertical distributions were identified. Among all the sand flies, only 197 (1.09%) flies of four Lutzomyia species, Lu. gomezi, Lu. trapidoi, Lu. tortura and Lu. ayacuchensis, were positive for Leishmania. Endotrypanum, a flagellate parasite not pathogenic to humans, were detected in five Lutzomyia species, Lu. robusta, Lu. hartmanni, Lu. trapidoi, Lu. panamensis and Lu. yuilli yuilli, suggesting wide vector-ranges of Endotrypanum species. These data on the genus Lutzomyia and their natural infections with Leishmania and Endotrypanum will be useful for transmission studies and surveillance of leishmaniasis.

Animals and fungi independently evolved from the protozoan phylum Choanozoa, these three groups constituting a major branch of the eukaryotic evolutionary tree known as opisthokonts. Opisthokonts and the protozoan phylum Amoebozoa (amoebae plus slime moulds) were previously argued to have evolved independently from the little-studied, largely flagellate, protozoan phylum, Sulcozoa. Sulcozoa are a likely evolutionary link between opisthokonts and the more primitive excavate flagellates that have ventral feeding grooves and the most primitive known mitochondria. To extend earlier sparse evidence for the ancestral (paraphyletic) nature of Sulcozoa, we sequenced transcriptomes from six gliding flagellates (two apusomonads; three planomonads; Mantamonas). Phylogenetic analyses of 173–192 genes and 73–122 eukaryote-wide taxa show Sulcozoa as deeply paraphyletic, confirming that opisthokonts and Amoebozoa independently evolved from sulcozoans by losing their ancestral ventral groove and dorsal pellicle: Apusozoa (apusomonads plus anaerobic breviate amoebae) are robustly sisters to opisthokonts and probably paraphyletic, breviates diverging before apusomonads; Varisulca (planomonads, Mantamonas, and non-gliding flagellate Collodictyon) are sisters to opisthokonts plus Apusozoa and Amoebozoa, and possibly holophyletic; Glissodiscea (planomonads, Mantamonas) may be holophyletic, but Mantamonas sometimes groups with Collodictyon instead. Taxon and gene sampling slightly affects tree topology; for the closest branches in Sulcozoa and opisthokonts, proportionally reducing missing data eliminates conflicts between homogeneous-model maximum-likelihood trees and evolutionarily more realistic site-heterogeneous trees. Sulcozoa, opisthokonts, and Amoebozoa constitute an often-pseudopodial ‘podiate’ clade, one of only three eukaryotic ’supergroups’. Our trees indicate that evolution of sulcozoan dorsal pellicle, ventral pseudopodia, and ciliary gliding (probably simultaneously) generated podiate eukaryotes from Malawimonas-like excavate flagellates.

Abollifer is a little-known genus of marine heterotrophic flagellates with no ultrastructural and molecular information, and its taxonomic position remains uncertain. In this study, we report a new species of Abollifer, Abollifer globosa sp. nov., isolated from a seawater sample collected at Tokyo Bay. To reveal the taxonomic position and morphological characteristics of A. globosa, we performed light and electron microscopic observations and a phylogenetic analysis using small subunit ribosomal DNA sequences. A. globosa cells were 29.5 μm in length and 22.4 μm in width, oval or ovoid in shape with an apical projection. Two unequal flagella emerged from a deep subapical flagellar pit. The rim of the flagellar pit except for the ventral side was swollen. Electron microscopic observations showed that A. globosa possessed mitochondria with tubular cristae, a Golgi apparatus, microbodies, extrusomes, and many endosymbiotic bacteria. Basal bodies were arranged in parallel. The flagellar apparatus of A. globosa showed affinity with common gliding cercozoan flagellates. Our phylogenetic tree showed that A. globosa branched as the sister position of order Marimonadida (Imbricatea, Cercozoa). On the basis of the morphological and molecular phylogenetic analysis, we conclude that A. globosa is a new member of the order Marimonadida.

The biochemical composition and abundance variation of zooplankton (copepods and Artemia salina) were determined in four ponds of increasing salinity (A5, A16, C41 and M2) in the Sfax solar saltern (Tunisia). The zooplankton community was dominated by copepods in the ponds A5, A16 and C41. The pond M2 was marked by the presence of only Artemia salina. Our results showed the dominance of total saturated fatty acids (SFA), which made up 57%–95% of total fatty acids (TFA). SFA 16:0 and 18:0 dominate in all ponds. A. salina showed the highest amounts of the total monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA), this indicates that this species could be employed in hatcheries and used as food source for some aquarium species. Fatty acids of herbivory, proportion of all diatom markers to all flagellate markers (D/F), were negatively correlated with the total zooplankton (r = −0.998, p

Phenol, a monosubstituted aromatic hydrocarbon with various commercial uses, is a major organic constituent in industrial wastewaters. The ecotoxic action of phenol for aquatic environment is well known. In this study, rapid phenol toxicity tests (1 h) were developed based on chlorophyll a (Chl a) fluorescence and the movement parameters of the freshwater flagellate, Euglena agilis Carter. Phenol significantly reduced the maximum quantum yield (Fv/Fm) of photosystem II (PS II) and the maximum photosynthetic electron transport rate (rETRmax) with median effective concentration (EC50) values of 8.94 and 4.67 mM, respectively. Phenol reduced the motility and triggered change in the swimming velocity of the test organism. Among the parameters tested, velocity was the most sensitive biomarker with an EC50 of 3.17 mM. The EC50 values for Fv/Fm, motility, and velocity appear to overlap the permitted levels of phenol. In conclusion, the photosynthesis and movement of E. agilis can be fast and sensitive risk assessment parameters for the evaluation of phenol toxicity in municipal and industrial effluents.

We report the light-microscopic morphology and ultrastructure of a novel free-living, heterotrophic protist, Creneis carolina gen. et sp. nov. isolated from marine anoxic sediments. C. carolina is a heterotrophic, obligately anaerobic amoeboid flagellate, and superficially resembles Mastigamoeba (Amoebozoa: Archamoebae) or Breviata (Breviatea) by possessing a single anterior flagellum closely associated with the nucleus, and because it appears to be an anaerobe. However, its life cycle contains multiflagellate cells with an unusual morphology. The structure of the mastigont of C. carolina is unique and not readily comparable with any eukaryotic group. Unexpectedly, phylogenetic analyses of SSU rDNA and of a concatenate of α- and β-tubulin genes with SSU rDNA convincingly showed that C. carolina is a member of Heterolobosea and belongs to the taxon Tetramitia.

Diverse wild animal species can be reservoirs of zoonotic flagellate parasites, which can cause pathologic Chagas disease. The present study aimed to detect the natural occurrence of flagellate parasites through direct microscopic examination of the parasites in blood samples and through PCR of whole blood and blood culture (haemoculture) samples from 38 captive and 65 free-living wild animals in the Centre for Conservation of Wild Fauna (CCWF), an area endemic for leishmaniasis. For this study, PCR was accomplished using primers for the ribosomal region (ITS-1) of the flagellate parasites. The amplified fragments were cloned and sequenced to identify DNA of the Trypanosomatid parasite species, observed in blood cultures from 3.9% (04/103) of the animals. Through these techniques, Trypanosoma cruzi was identified in haemoculture samples of the following three free-living species: common agouti (Dasyprocta aguti), white-eared opossum (Didelphis albiventris), and nine-banded armadillo (Dasypus novemcinctus). Furthermore, Trypanosoma minasense was identified in whole blood samples from 01 (0.9%) captive animal (black howler monkey-Alouatta caraya). These results demonstrated the first report of T. cruzi isolation in wild species from the CCWF using blood culture, which can be applied in addition to molecular tools for epidemiological studies and to identify trypanosomatids in wild animals.

The biodiversity of oxymonadid flagellates in termite hindguts is not fully explored. Many species have been differentiated only by morphological features, and small-celled species have been overlooked or ignored. Our analysis of the dry wood termite Neotermes jouteli by light and electron microscopy revealed the presence of two distinct morphotypes of oxymonads. The larger one matched the morphology of Oxymonas jouteli, the only oxymonad species described from this termite. Although it generally lacks the typical anterior rostellum of the genus Oxymonas, its SSU rRNA gene sequence clusters among other members of this genus, including novel phylotypes that we obtained from Incisitermes tabogae. The second morphotype was a tiny oxymonad that showed the typical traits of the genus Opisthomitus, including a pointed anterior prolongation (lappet). However, the four equal flagella were much longer than those of Opisthomitus avicularis from Kalotermes flavicollis, the only species of the genus and so far described only by light microscopy. We provide a detailed description of Opisthomitus longiflagellatus sp. nov. and demonstrate that despite ultrastructural similarities to members of the Polymastigidae, its SSU rRNA gene sequences form a separate family-level lineage with a slight affinity to the Pyrsonymphidae.

Primary amoebic meningoencephalitis is a rare fatal meningitis caused by free living amoeba Naegleria fowleri, found in freshwater ponds and lakes. It infects children and young adults with exposure due to swimming or diving. We report a case of N. fowleri meningitis in a 6-year-old boy who presented with signs and symptoms of acute bacterial meningitis. No history of travelling or swimming was present. However, the boy frequently played with water stored from a "kuhl" (diversion channels of water). Wet mount of cerebrospinal fluid (CSF) revealed amoeboid and actively motile flagellate forms of trophozoites. CSF culture done on 1.5% non-nutrient agar plates with a lawn culture of Escherichia coli kept at 37°C for 15 days did not reveal any growth. The test of flagellation on passing CSF in distilled water was however positive in 3 h. Water of the "kuhl" from the stored tank also showed actively motile trophozoites similar to the forms obtained from the CSF. Based on our reports, the boy was immediately treated with amphotericin B, rifampicin and fluconazole for 21 days. Repeat CSF examination after 14 days did not reveal any trophozoites in wet mount and patient was discharged after 3 weeks of successful treatment.

Trypanosomes are parasitic flagellate protozoans that are important disease causing agents in humans and animals including economically significant livestock. Many blood-sucking insects are known to be vectors of trypanosomes but there is no data for the haematophagous black fly species in Asia. In this study, a molecular approach based on the small subunit ribosomal RNA (SSU rRNA) gene was used to detect trypanosomes in black flies from Thailand. A total of 470 wild-caught adult black flies representing nine morphological species were examined. Ten (2%) specimens of two ornithophilic black fly species, Simulium asakoae complex (n = 4) and S. chumpornense (n = 6), were positive for trypanosomes. The SSU rRNA sequences revealed that all trypanosome DNA found in black flies from Thailand is closely related to Trypanosoma avium with >99% sequence similarity. This is also supported by a phylogenetic analysis in which all trypanosomes from Thai black flies were resolved in the clade of T. avium. This is the first report for trypanosomes in Asian black flies and it is suspected that these insects are potential vectors of avian Trypanosoma.

Contamination of large-scale microalgal cultures by grazers can cause huge losses in biomass productivity. Here we propose the use of a quaternary ammonium salt cetyltrimethylammonium bromide (CTAB) to eradicate three types of commonly occurring grazers in microalgal cultures: the rotifer Brachionus, the ciliate Sterkiella and the flagellate Paraphysomonas. Low, premicellar doses (≤3 μM) of CTAB rapidly eradicated (within 1–2 d) all three tested grazers from microalgal cultures without significant losses (p

The protozoan flagellate Poterioochromonas malhamensis is one of the main contaminants in Chlorella culture; however, few effective and affordable methods have been developed to control it. Unexpectedly, one strain of an unknown green alga was observed to be capable of contaminating P. malhamensis culture and defending itself against the protozoan. Based on cell morphology and molecular information, the green microalga was identified as Chlorella sorokiniana CMBB-146. Feeding experiments showed that the addition of the predator P. malhamensis not only had no negative effect on the growth of C. sorokiniana CMBB-146, but also stimulated an increase in the biomass of C. sorokiniana CMBB-146. The grazing resistance of C. sorokiniana CMBB-146 against P. malhamensis was also verified on a pilot scale with 100-L raceway ponds. Further experiments revealed that P. malhamensis showed a strong ability in ingesting C. sorokiniana CMBB-146 cells, but the ingested cells were hard to digest. Through comparison with other species of Chlorella without grazing resistance, we found that the differences on cell size, cell morphology, and biochemical composition had no relation with the ability of C. sorokiniana CMBB-146 to resist grazing by P. malhamensis. Finally, it is speculated that variation in cell wall composition, especially the content of galactosamine, empowers C. sorokiniana CMBB-146 to resist P. malhamensis. Therefore, our study confirms that C. sorokiniana CMBB-146 is a strong candidate as a potential production organism for outdoor massive cultivation, and as a potential microalgal strain for studying how microalgae resist grazing by protozoa.

Lentinula edodes, or shiitake mushroom (SM), is typically grown in Eastern Asia and used in traditional Asian medicine and cuisine. Recently, SM became popular in Western culture and is now the second most commonly consumed mushroom in the world.1, 2 Although rare, adverse reactions to SM have been previously reported. In mushroom farm workers, contact dermatitis, contact urticaria, rhinitis, and hypersensitivity pneumonitis have been described. In the general population, oral intake of raw SM has been associated with a typical flagellate eruption, known as shiitake dermatitis (SD).3 Shiitake dermatitis, also termed shiitake toxicoderma or flagellate mushroom dermatitis, classically manifests 12 hours to 5 days after the ingestion of SM.1, 4 Since its first description in 1977, SD has been described in approximately 100 patients, predominantly Japanese.3, 5 Recently, a few cases were reported in Europe and in the United States.1 The mechanism underlying SD has not been fully elucidated. Although the eruption is usually considered nonallergic,6 5 cases of SD resulted in positive delayed skin prick testing, suggesting that delayed-type systemic hypersensitivity may be important in its pathogenesis.2, 3, 4, 7, 8, 9 We report the first case of SD in Canada and, to our knowledge, the first case of pustular SD. In our patient, positive delayed skin prick testing to SM was documented.

Cladocerans are important food animals in aquaculture, key grazers in freshwater ecosystems, and model animals for ecotoxicological investigations. Their epibiont community, extensively studied in Daphnia , includes filamentous bacteria, fungi, algae, peritrich ciliates, and rotifers; although epibionts are usually benign, heavy infections can be detrimental. During our laboratory culture of female Moina macrocopa Straus, we observed a novel flagellate infection associated with mortality. At day 10, all M. macrocopa were alive in uninfected cultures, whereas in untreated infected cultures, the survival was significantly lower: only 26% of cladocerans were alive. In infected cultures treated with humic substances (as 25 mg L − 1 dissolved organic carbon), mortalities were comparable to those in the untreated infected cultures; in contrast, in the infected cultures treated with 4 g L − 1 sea salt, mortalities were arrested, and 76% of the M. macrocopa were alive at day 10. Moribund cladocerans were transparent, had empty digestive tracts, and greatly reduced motor activity. Free-swimming flagellates moved forward with a wobbling motion, rotating around their long axis; they also attached to cladoceran tissue, the Petri dish, and the glass slide, by the tip of their posterior flagellum. Flagellates preserved for scanning electron microscopy were 6.9 ± 0.7 μm long and 2.1 ± 0.3 μm wide, with a short anterior flagellum (6.8 ± 1.1 μm) and long posterior flagellum (14.1 ± 1.5 μm). Multi-functionality of a flagellum, for locomotion and adhesion, is relatively rare, and previously reported from genera within the Kinetoplastea, suggesting that the flagellate on M. macrocopa may belong to this group. To combat flagellate mass occurrence in Moina cultures, we recommend a treatment with 4 g L − 1 sea salt.

Cyanobacterial toxins can cause damage in aquatic ecosystems worldwide, as well as the poisoning of livestock, plants and humans when ingested in large amounts. Although many studies investigated grazing of harmful cyanobacteria by metazoan plankton, grazing of cyanobacteria by hetertophic flagellates is largely unexplored. This laboratory study investigated grazing of toxic Microcystis aeruginosa by the heterotrophic flagellate Diphylleia rotans isolated from a Saudi hypertrophic lake. D. rotans was able to feed on M. aeruginosa with estimated ingestion (10.2 to 16.5 prey flagellate (-1)d(-1)) and specific growth rates (0.71 to 0.99d(-1)) differed with the increase in the initial density of the flagellate. Grazing increased microcystin production within Microcystis cells during first two days of incubation, and afterwards induced the release of these toxins into the medium. The concentrations of releasing microcystin were strongly reduced in grazing-treated cultures compared to controls, indicating the degradation of microcystins by D. rotans growing under axenic conditions. Taken these results, D. rotans can play an important role in the reduction of Microcystis biomass and microcystin toxins, and thus could be used as a safe bioagent for the biocontrol of harmful algal blooms in aquatic environments.

Apusomonads are of evolutionary interest because they are close relatives to the supergroup Opisthokonta, which contains both animals and fungi. There are no detailed morphological studies of 'amastigomonad' type apusomonads, such as Thecamonas trahens, despite this species having a sequenced genome. We use serial-section transmission electron microscopy and 3D reconstruction to examine the cell architecture and complete microtubular cytoskeleton of Thecamonas. Thecamonas has two flagella and an anteriorly projecting 'tusk'. The anterior basal body associates with one microtubular root, which travels leftward, and a non-root 'ribbon' of six microtubules that travels down the right side of the cell. The posterior basal body associates with three roots: an eight-membered right root, a doublet left root, and an intermediate singlet root. These rearrange into two bands, both on the left side of the cell. One comprises the left and singlet roots plus one right root microtubule. The other comprises the remaining right root microtubules. A splitting right root and supernumerary singlet root are also present in breviates, ancyromonads, and 'typical excavates', suggesting that these characters are ancestral for much of eukaryote diversity. If so, opisthokonts, and most or all living eukaryotes, probably arose from cells with complex microtubular cytoskeletons.

Breviata anathema is an anaerobic amoeboid flagellate that does not branch within any established ‘supergroup’. Molecular phylogenies suggest affinities to Amoebozoa, Opisthokonta, or apusomonads. Here we describe its flagellar apparatus ultrastructure. Breviata has two basal bodies. The flagellated anterior basal body (AB) is associated with a fan of ∼18 microtubules and a short singlet microtubular root. Three microtubular roots associate with the posterior basal body. One, the right root (RR), is initially a triplet that splits into two parts. The other two are singlets: the left root (LR), and the middle root (MR), which arises on the posterior side of the basal body. The MR, LR and smaller part of RR support the left ventral side of the cell, while the larger part of RR runs down the right. Outer dynein arms were not observed on the flagellar axoneme. The mitochondrion-like organelle sometimes contains some tubular cristae. The posterior flagellar apparatus resembles that of several eukaryotic lineages, particularly apusomonads, ancyromonads, excavates, and myxogastrid amoebozoans. This comparison suggests that the complex flagellar apparatus of myxogastrids is actually plesiomorphic within Amoebozoa. The widely distributed splitting right root and posterior singlet (MR in Breviata) may be plesiomorphies in many eukaryotic lineages, and thus could be features of the last eukaryotic common ancestor.

A tetratrichomonad flagellate found in the diarrhoeic faeces of a 5 years-old male giant anteater (Myrmecophaga tridactyla) was characterised by morphological and genetic analysis. This protozoan presents four anterior flagella of unequal length and a recurrent flagellum attached to the undulating membrane without a free end portion, and a broad axostyle projection. Numerous vacuoles of different sizes containing bacteria and digestion products were found. The complete sequence of the DNA coding for the 16S rRNA-ITS1-5.8S rRNA-ITS2 region was also obtained in order to compare this isolate with other tetratrichomonad species. The sequence obtained was identical to others previously obtained by other researchers from bovines and turtles (Geochelone sp.). It is not easily explainable how the same organism could be found in such different hosts and locations; however these results indicate that some tetratrichomonad species could have a wide host range and could survive in a wide range of environmental conditions.

The vast majority of the marine environment consists of dark, cold, high-pressure environments into which microbes from the surface ocean are continuously transported by advection or by attachment to sinking particles. Little is known about how the low temperature and high pressure of the deep ocean affect the abundance and activity of surface organisms, in particular microbial eukaryotes. Cultures of two flagellate species, Cafeteria roenbergensis and Neobodo designis , both isolated from surface waters where they are considered cosmopolitan, were incubated in titanium chambers for one to two weeks under typical deep-sea temperature (2 °C) and pressure (50 MPa, representing 5000 m water depth). Samples were taken daily with minimal loss of pressure in the culture vessel, and flagellates were subsequently enumerated, along with prokaryotic prey when possible. The abundance of protists declined in all treatments, with a significantly greater rate of mortality under combined cold temperature and high pressure conditions than in the cold temperature-only conditions. However, 1.6 (SD = 3.8)% of C. roenbergensis and 10.0 (6.1)% of N. designis cells survived on average in the high pressure treatments, indicating that some fraction of sinking protists can survive transport to the deep ocean. In addition, after a period of acclimation, positive growth rates were measured in some cases, suggesting that surface-adapted flagellates cannot only survive under deep-sea conditions but are able to reproduce and potentially provide seed populations to cold, high-pressure environments.

A harmful bloom due to the raphidophycean flagellate, Chattonella antiqua, was found in the Yatsushiro Sea, western Kyushu, Japan, from the end of July to the beginning of August 2009. The bloom resulted in enormous economic damage to cultured finfish production in aquaculture farms concentrated in the southwestern area. To investigate the factors controlling the spatio-temporal distribution of the bloom, data analysis and numerical simulations were conducted using field monitoring data and a three-dimensional hydrodynamic model coupled to a Lagrangian particle-tracking model. Results of the monitoring data analysis showed that the initial development of the C. antiqua bloom occurred in Kusuura Bay and the northeastern area near the mouth of the Kuma River, and subsequently the bloom expanded rapidly to the whole area. The simulation results indicated that the source region of the widespread bloom was not Kusuura Bay but the northeastern area. The southwestward evolution of the bloom was primarily controlled by the passive transport due to the surface residual current driven by fresh water discharge from the Kuma River and northeasterly winds. On the favorable conditions of river discharge and wind, the massive bloom of C. antiqua that formed in the northeastern area was quickly transported southwestward within a few days.

Since 1998, the genus Pseudochattonella has formed massive blooms off the coasts of Norway, Sweden and Denmark, causing comprehensive kills of cultured and wild fish. Isolates from a bloom of Pseudochattonella sp. in 2001 were found to be a new heterokont flagellate Pseudochattonella farcimen (Dictyochophyceae). Since then recurrent blooms of Pseudochattonella sp. have appeared both in the early and late spring in Scandinavian waters. P. farcimen is the only species of Pseudochattonella documented to bloom in the cold waters and low light conditions characterizing winter and early spring. This study aims at determining why P. farcimen is able to bloom under these environmental regimes. The effect of temperature and irradiance on the growth of P. farcimen cells, isolated from a bloom in 2009, was examined. Furthermore a possible allelopathic effect of P. farcimen on the growth of coexisting algae (i.e. Heterocapsa triquetra, Chaetoceros decipiens and Chaetoceros diadema) was examined. Growth of P. farcimen was observed at irradiances as low as 2 μE m−2 s−1. Growth rates increased with temperature from 1 to 6 °C, while a further increase in growth was not seen when the temperature was elevated from 6 to 10 °C. At the highest tested temperature (10 °C), dense cultures of P. farcimen were observed to develop large multinucleate cells. Highest growth rates were observed at the pH range 7.7–8.4, while growth rates decreased above pH 8.4, until cultures stopped growing at pH 8.9–9.0. As pH increased and growth declined, the average shape of P. farcimen cells changed from elongated to round. This shape could serve as an indicator on the growth condition of the alga. P. farcimen did not affect the growth of any of the tested phytoplankton in the present experiment but the growth rate of P. farcimen was in the same range as the diatoms. Thus, this study has shown that P. farcimen is adapted to successfully compete and grow in a low light and temperature regime that it is exposed to in the late winter and early spring, making it able to form blooms under these conditions.

Polyprenyl-diphosphate synthase is a key enzyme in the biosynthesis of ubiquinone, a molecule considered essential for a typical eukaryotic cell. Its orthologue in the American stercorarian flagellate Trypanosoma cruzi, solanesyl diphosphate synthase, has been previously localized into the glycosomes. We wondered whether this unique cellular localization is shared by other trypanosome species. Using digitonin permeabilization, immunofluorescence and in situ tagging techniques, we show that in Trypanosoma brucei, the African salivarian flagellate, the enzyme localizes to the mitochondrion.