Protist Biodiversity and Biogeography in Lakes From Four Brazilian River-Floodplain Systems.

The biodiversity and biogeography of protists inhabiting many ecosystems have been intensely studied using different sequencing approaches, but tropical ecosystems are relatively under-studied. Here, we sampled planktonic waters from 32 lakes associated with four different river-floodplains systems in Brazil, and sequenced the DNA using a metabarcoding approach with general eukaryotic primers. The lakes were dominated by the largely free-living Discoba (mostly the Euglenida), Ciliophora, and Ochrophyta. There was low community similarity between lakes even within the same river-floodplain. The protists inhabiting these floodplain systems comprise part of the large and relatively undiscovered diversity in the tropics.

Morphological and molecular characterisation of Notosolenus urceolatus Larsen and Patterson 1990, a member of an understudied deep-branching euglenid group (petalomonads).

Petalomonads are particularly important for understanding the early evolution of euglenids, but are arguably the least studied major group within this taxon. We have established a culture of the biflagellate petalomonad Notosolenus urceolatus, and conducted electron microscopy observations and molecular phylogenetic analysis. Notosolenus urceolatus has eight pellicular strips bordered by grooves and underlain by close-set microtubules. There are ventral and dorsal Golgi bodies. Mitochondria apparently contain fibrous inclusions, as in Petalomonas cantuscygni. A previously undocumented type of large, globular extrusome is present instead of the tubular extrusomes characteristic of Euglenozoa. The feeding apparatus lacks rods and vanes, and is partly supported by an "MTR". The flagella have complex transition zones that are extremely elongated but unswollen. Only the emergent portion of the anterior flagellum has an organised paraxonemal rod, and also has very fine mastigonemes. The basal bodies are offset and lack connecting fibres. 18S rRNA gene phylogenies show that N. urceolatus is closely related to Petalomonas sphagnophila and P. cantuscygni, not Notosolenus ostium, confirming that current generic assignments based on the number of emergent flagella are phylogenetically unreliable, and making it difficult to infer whether features shared by N. urceolatus and P. cantuscygni (for example) are general for petalomonads.

Combining molecular data with classical morphology for uncultured phagotrophic euglenids (Excavata): a single-cell approach.

Phagotrophic euglenids are one of the most diverse and important forms of heterotrophic flagellates in sediment systems, and are key to understanding the evolution of photosynthetic euglenids and 'primary osmotrophs', yet relatively little is known about their biodiversity and phylogenetic relationships. A wealth of light microscopy-based information is available, but little progress has been made in associating this with molecular sequence data. We established a protocol to obtain light microscopy data and molecular data from single euglenid cells isolated from environmental samples. Individual cells from freshwater and marine benthic samples were isolated and rinsed by micropipetting, documented using high-resolution photomicroscopy, then subjected to single-cell nested PCR using taxon-specific primers in combination with universal eukaryotic primers, generating > 75% or full-length SSU rDNA sequences. As a proof-of-principle eight individuals were characterised and subjected to phylogenetic analyses. Many of these cells were identified as Anisonema or Dinema, and grouped with existing sequences assigned to these taxa, and with a 'Peranema sp.' sequence that we could now clearly demonstrate was misidentified or misannotated. Another cell is Heteronema c.f. exaratum, the first 'skidding heteronemid' for which sequence data are available. This is not closely related to Heteronema scaphurum, and intriguingly, branches as the sister group to primary osmotrophs. A cell similar to Ploeotia vitrea (the type of this genus), shows no particular phylogenetic affinity to Ploeotia costata, the best studied Ploeotia species. Our experimental protocol provides a useful starting point for future analyses on euglenid biodiversity (including environmental sequence surveys), and their evolution and systematics.

Ultrastructure and molecular phylogenetic position of Heteronema scaphurum: a eukaryovorous euglenid with a cytoproct.

Euglenids comprise a distinct clade of flagellates with diverse modes of nutrition, including phagotrophy, osmotrophy and phototrophy. Much of the previous research on euglenids has focused on phototrophic species because of their ecological abundance and significance as indicators for the health of aquatic ecosystems. Although largely understudied, phagotrophic species probably represent the majority of euglenid diversity. Phagotrophic euglenids tend to be either bacterivorous or eukaryovorous and use an elaborate feeding apparatus for capturing prey cells. We characterized the ultrastructure and molecular phylogenetic position of Heteronema scaphurum, a eukaryovorous euglenid collected in freshwater. This species was equipped with a distinct cytoproct through which waste products were eliminated in the form of faecal pellets; a cytoproct has not been reported in any other member of the Euglenida. Heteronema scaphurum also had a novel predatory mode of feeding. The euglenid ensnared and corralled several green algal prey cells (i.e. Chlamydomonas) with hook-like flagella covered in mucous before engulfing the bundle of prey cells whole. Molecular phylogenetic analyses inferred from small subunit rDNA sequences placed this species with other eukaryovorous euglenids, which was consistent with ultrastructural features associated with the feeding apparatus, flagellar apparatus, extrusomes, and pellicle.

An approach to analyzing spatial patterns of protozoan communities for assessing water quality in the Hangzhou section of Jing-Hang Grand Canal in China.

INTRODUCTION: In order to evaluate water quality of a canal system, the spatial pattern of protozoan communities in response to physicochemical variables was studied in the Hangzhou section of the Grand Canal, northern China during a 1-year cycle (February 2008-January 2009). MATERIALS AND METHODS: Protozoan samples were monthly collected at six sampling stations with a spatial gradient of environmental status. Physicochemical parameters, e.g., water temperature, dissolved oxygen, chemical oxygen demand (COD), biological oxygen demand, total nitrogen (TN), and total phosphorus (TP), were measured synchronously for comparison with biotic parameters. RESULTS: The protozoan community structures represented significant differences among the six sampling stations. The spatial patterns of protozoan communities were significantly correlated with the changes of chemical variables, especially COD, either alone or in combination with TP and/or TN. Of 88 protozoan taxa recorded over the study period, ten species (e.g., Carchesium polypinum, Colpidium campylum, Prorodon teres, Vorticella putrina, Zoothamnium arbuscula, Euglena spp., and Phacus spp.) were significantly related to COD, either alone or in combination with TP and/or TN. CONCLUSION: These findings suggest that protozoa can be used as a robust bioindicator of water quality in freshwater river systems.

Phytobenthos and phytoplankton community changes upon exposure to a sunflower oil spill in a South African protected freshwater wetland.

The occurrence of a sunflower oil spill in 2007 in the Con Joubert Bird Sanctuary freshwater wetland, South Africa, inhibited the growth of sensitive phytoplankton species and promoted that of tolerant species. The algal divisions Chlorophyta and Euglenophyta were well represented in the sunflower oil contaminated water, especially the species Euglena sociabilis, Phacus pleuronectes and Chlamydomonas africana. Young and mature resting zygotes of Chlamydomonas africana were recorded in high abundance at all the sunflower oil contaminated sampling sites. The phytobenthos diversity and abundance were significantly suppressed and negatively associated with low Dissolved Oxygen concentrations and the negative redox potential of the bottom sediment. At the intracellular level, phytoplankton chlorophyll a and b concentrations as physiological variables were more sensitive indicators of the adverse effects of sunflower oil than the 72 h Selenastrum capricornutum algal bioassay conducted.

Performance evaluation of a simple wastewater treatment system comprised by UASB reactor, shallow polishing ponds and coarse rock filter.

The work investigates a small full-scale wastewater treatment system comprised by the following units in series: UASB reactor, three polishing ponds and one coarse rock filter. The overall performance of the system is analyzed based on three years of monitoring using physical-chemical and biological parameters. Good organic matter, suspended solids and ammonia removal is achieved, together with excellent coliform removal (5.70 log units). Mean effluent concentrations of the main parameters are: BOD: 39 mg/L; COD: 109 mg/L; SS = 41 mg/L; ammonia: 10 mg/L; E. coli: 540 MPN/100 mL, indicating compliance with many regulations for effluent discharge and reuse. Main algal classes found in the ponds and final effluent were chlorophyta and euglenophyta. The system is completely unmechanized and has a relatively small total hydraulic retention time (less than 13 days), compared with most natural treatment processes. No sludge removal from the ponds and filter has been necessary so far.

[Algal flora of Lake Amapá in Rio Branco, acre state, I: euglenophyceae]. / Ficoflórula do Lago Amapá em Rio Branco- Acre, I: Euglenophyceae.

The Euglenophyceae flora of Lake Amapá I, Rio Branco, Acre State, Brazil, constitutes a contribution to the phycological inventory of the State of Acre. It is based on the study of 15 samples collected with plankton net and by passing an open flask in areas with dense plankton concentrations. Samples were prepared and preserved with Transeau solution. Twenty five taxons were identified. Genera Euglena and Trachelomonas were the most frequently represented.