Small Subunit rRNA Gene Sequences Research Articles

Study on genetic characteristics of Cryptosporidium isolates and first report of C. parvum IIdA24G2 subtype in dairy cattle in China.

Cryptosporidium is an important gastrointestinal parasite that can cause mild to severe diarrhea in various vertebrates, including humans and domestic animals. Infection is prevalent in dairy cattle, particularly calves, resulting in diarrhea and increased mortality with significant production losses. However, the prevalence and identity of Cryptosporidium spp. in cattle in Heilongjiang Province is still poorly known. Our study aimed to investigate the prevalence and species and subtype distribution of Cryptosporidium in cattle in the region. In addition, we evaluated the zoonotic potential of Cryptosporidium isolates and assessed possible transmission routes and health effects of this organism. We collected 909 fecal samples from five different farms in Heilongjiang Province between August and September 2022. The samples underwent Cryptosporidium detection by nested PCR and small subunit (SSU) rRNA gene sequence analysis. Four Cryptosporidium species were identified, including C. parvum, C. bovis, C. ryanae, and C. andersoni, with an overall prevalence of 4.4% (40/909). Based on sequence analysis of the 60 kDa glycoprotein gene of C. parvum and C. bovis, three subtypes of C. parvum were identified, namely two previously known subtypes (IIdA19G1 and IIdA20G1), and one novel subtype (IIdA24G2). Two distinct subtype families were identified in C. bovis (XXVId and XXVIe). The high diversity of Cryptosporidium in dairy cattle and the emergence of a novel subtype of C. parvum in Heilongjiang Province suggest that dairy cattle may serve as a significant source of zoonotic cryptosporidiosis infection in this region.

Molecular detection and assessment of risk factors for Theileria lestoquardi in sheep from Balochistan, Pakistan.

This study aimed to determine the molecular prevalence and associated risk factors of theileriosis in sheep from Balochistan, Pakistan. For this purpose, a total of 408 blood samples were collected from tick-infested sheep in three different zones of Balochistan (i.e., Quetta, Zhob, and Loralai). All the collected samples were analyzed using conventional microscopy techniques, polymerase chain reaction (PCR), 18S small subunit rRNA gene sequencing, and phylogenetic analysis. The results of the microscopy and PCR confirmed the highest prevalence of Theileria species in district Zhob (14.22% and 15.68%) followed by district Loralai (11.52% and 13.97%) and district Quetta (10.29% and 12.00%), respectively. In addition, the prevalence of T. lestoquardi was higher in female sheep (84.12%), followed by adult sheep (74.71%) and the Hernai breed of sheep (28.23%) in the studied area. Similarly, the prevalence of theileriosis was higher in the summer season (40.59%), followed by the spring, autumn, and winter seasons. However, numerous risk factors such as age, sex, area, season, and breeds of the sheep were not significantly correlated (P > 0.05) with the presence of T. lestoquardi, except tick abundance and feeding pattern of animals (P < 0.05). Furthermore, sequencing and phylogenetic analyses of the isolated T. lestoquardi displayed 99% sequence similarity with isolates from Germany, Egypt, Iraq, India, Iran, and Pakistan. Altogether these results showed that T. lestoquardi is the main species causing ovine theileriosis in Balochistan. As a result, large-scale studies are required to design practical control approaches to reduce the risk of theileriosis infection in Balochistan, Pakistan.

Morphology and molecular phylogeny of two new species of Psammophaga (Rhizaria, Foraminifera) from the west coast of India

Benthic foraminifera are unicellular eukaryotic protists that construct an organic, agglutinated, or calcareous test wall. Although single-chambered (monothalamous) foraminifera are ubiquitous in marine habitats worldwide, they are poorly known compared to their multi-chambered relatives, notably from the tropical marine environments of India. In this study, we describe two new species of marine monothalamid genus Psammophaga Arnold, 1982, from the Rajapuri Creek, coastal Maharashtra, India (Arabian Sea). Psammophaga holzmannae sp. nov. is ovoid to spherical shaped, 103–246 µm in length, single aperture, translucent to orange color cytoplasm, outer surface is composed of agglutinated fine clay particles, and ingested mineral grains are concentrated near its aperture. Psammophaga sinhai sp. nov. is oblong, elliptical, or droplet-shaped, 279–448 µm in length, single aperture, yellow olive color cytoplasm, the exterior surface formed of agglutinated fine clay particles, and the ingested mineral grains are dispersed throughout the body. Phylogenetic analyses based on partial small subunit rRNA gene sequences position new species within the Clade E of monothalamids and are genetically distinct from other Psammophaga. Elemental (SEM–EDS) analysis of engulfed mineral grains revealed preferential selection and uptake of heavy opaque titaniferous minerals from the ambient environment in the cytoplasm.

Bradyrhizobium commune sp. nov., isolated from nodules of a wide range of native legumes across the Australian continent.

Bradyrhizobia are particularly abundant in Australia, where they nodulate native legumes growing in the acidic and seasonally dry soils that predominate in these environments. They are essential to Australian ecosystems by helping legumes to compensate for nutrient deficiencies and the low fertility of Australian soils. During a survey of Australian native rhizobial communities in 1994-1995, several Bradyrhizobium genospecies were identified, among which genospecies B appeared to be present in various edaphic and climatic conditions and associate with a large range of leguminous hosts across the whole continent. We took advantage of the recent sequencing of the genome of strain BDV5040T, representative of Bradyrhizobium genospecies B, to re-evaluate the taxonomic status of this lineage. We further characterized strain BDV5040T based on morpho-physiological traits and determined its phylogenetic relationships with the type strains of all currently described Bradyrhizobium species using both small subunit (SSU) rRNA gene and complete genome sequences. The digital DNA-DNA hybridization relatedness with any type strain was less than 35 % and both SSU rRNA gene and genome phylogenies confirmed the initial observation that this strain does not belong to any formerly described species within the genus Bradyrhizobium. All data thus support the description of the novel species Bradyrhizobium commune sp. nov. for which the type strain is BDV5040T (=CFBP 9110T=LMG 32898T), isolated from a nodule of Bossiaea ensata in Ben Boyd National Park in New South Wales, Australia.

A New Algivorous Heterolobosean Amoeba, Euplaesiobystra perlucida sp. nov. (Tetramitia, Discoba), Isolated from Pilot-Scale Cultures of Phaeodactylum tricornutum.

The diatom Phaeodactylum tricornutum is regarded as a prospective "cell factory" for the high-value products fucoxanthin and eicosapentaenoic acid (EPA). However, contamination with grazing protozoa is a significant barrier to its commercial cultivation. Here, we describe a new species of heterolobosean amoeba, Euplaesiobystra perlucida, which caused the loss of Phaeodactylum tricornutum in pilot-scale cultures. Morphological and molecular characteristics distinguish E. perlucida from the other species in the genus Euplaesiobystra. E. perlucida is 1.4 to 3.2 times larger than other Euplaesiobystra species in terms of average length/width and maximum length/width of the trophozoites. Unlike Euplaesiobystra salpumilio, E. perlucida has no cytostome; E. perlucida lacks a flagellate stage, whereas Euplaesiobystra hypersalinica and E. salpumilio both display a flagellate stage in their life cycle. The small-subunit rRNA gene sequence of E. perlucida shared only 88.02% homology with that of its closest relative, Euplaesiobystra dzianiensis, and had two distinctive regions. Its phylogenetic branch was clustered with one uncultured heterolobosean clone (bootstrap support/posterior probability = 100%/1.00). Results of feeding experiments demonstrated that E. perlucida could graze on various unicellular and filamentous eukaryotic microalgae (chlorophytes, chrysophytes, euglenids, and diatoms) and cyanobacteria. E. perlucida's ingestion rate declined exponentially with increasing size of unicellular prey, and E. perlucida attained the highest growth rates on P. tricornutum. On the basis of its strong ability to graze on microalgae, capacity to form large populations in a short period of time, and capacity to form resistant resting cysts, this contaminant has the potential to cause severe problems in large-scale microalgal culture and merits further attention. IMPORTANCE Heteroloboseans have garnered considerable interest because of their extraordinary ecological, morphological, and physiological diversity. Many heteroloboseans have adapted to various extensive habitats, including halophilic, acidophilic, thermophilic, psychrophilic, and anaerobic habitats. Most heteroloboseans are bacterivores, with a few algivorous species reported. In this study, a new species of algivorous heterolobosean amoeba, Euplaesiobystra perlucida, is described as a significant grazer that causes losses in outdoor industrial Phaeodactylum cultures. This study provides phenotypic, feeding, and genetic information on a previously unknown heterolobosean, emphasizes the impact of contaminating amoebae in commercial microalgal cultures, and will contribute to the management strategies for predicting this kind of contaminant in large-scale microalgal cultivation.

Description of a new nematode species, Chromadorinacommunis sp. nov. (Nematoda, Chromadoridae), from Changdao Island, China and phylogenetic analysis of Chromadorida based on small subunit rRNA gene sequences.

Chromadorinacommunissp. nov. is described from Changdao Island at the confluence of the Yellow and the Bohai seas. The new species is characterized by its medium-sized body; finely striated cuticle with homogeneous punctations; absence of ocelli; buccal cavity with three equal-sized, solid teeth; four cephalic setae; oval amphidial fovea which is positioned between cephalic setae; curved spicules with tapered distal ends; simple, boat-shaped gubernaculums; five or six cup-shaped precloacal supplements; and conical tail with a very short spinneret. A phylogenetic analysis of small subunit rRNA gene sequences using maximum-likelihood and Bayesin inference confirmed the taxonomic position of Chromadorinacommunissp. nov. within Chromadorinae. Tree topology in Chromadorida shows six morphological families clustered into a monophyletic clade and verifies the taxonomic position of the family Neotonchidae based on morphological and molecular analysis.

Molecular phylogeny of Spirotrichonymphea (Parabasalia) with emphasis on Spironympha, Spirotrichonympha, and three new genera Pseudospironympha, Nanospironympha, and Brugerollina.

Spirotrichonymphea, one of the six classes of phylum Parabasalia, are characterized by bearing many flagella in spiral rows, and they occur exclusively in the guts of termites. Phylogenetic relationships among the 13 described genera are not well understood due to complex morphological evolution and a paucity of molecular data. One such understudied genus is Spironympha. It has been variously considered a valid genus, a subgenus of Spirotrichonympha, or an "immature" life cycle stage of Spirotrichonympha. To clarify this, we sequenced the small subunit rRNA gene sequences of Spironympha and Spirotrichonympha cells isolated from the hindguts of Reticulitermes species and Hodotermopsis sjostedti and confirmed the molecular identity of H. sjostedti symbionts using fluorescence in situ hybridization. Spironympha as currently circumscribed is polyphyletic, with both H. sjostedti symbiont species branching separately from the "true" Spironympha from Reticulitermes. Similarly, the Spirotrichonympha symbiont of H. sjostedti branches separately from the "true" Spirotrichonympha found in Reticulitermes. Our data support Spironympha from Reticulitermes as a valid genus most closely related to Spirotrichonympha, though its monophyly and interspecific relationships are not resolved in our molecular phylogenetic analysis. We propose three new genera to accommodate the H. sjostedti symbionts and two new species of Spirotrichonympha from Reticulitermes.

Detection and identification of fungi in the lower airway of children with and without cystic fibrosis.

Airway infection and inflammation lead to the progression of obstructive lung disease in persons with cystic fibrosis (PWCF). However, cystic fibrosis (CF) fungal communities, known drivers of CF pathophysiology, remain poorly understood due to the shortcomings of traditional fungal culture. Our objective was to apply a novel small subunit rRNA gene (SSU-rRNA) sequencing approach to characterize the lower airway mycobiome in children with and without CF. Bronchoalveolar lavage fluid (BALF) samples and relevant clinical data were collected from pediatric PWCF and disease control (DC) subjects. Total fungal load (TFL) was measured using quantitative PCR, and SSU-rRNA sequencing was used for mycobiome characterization. Results were compared across groups, and Morisita-Horn clustering was performed. 161 (84%) of the BALF samples collected had sufficient load for SSU-rRNA sequencing, with amplification being more common in PWCF. BALF from PWCF had increased TFL and increased neutrophilic inflammation compared to DC subjects. PWCF exhibited increased abundance of Aspergillus and Candida, while Malassezia, Cladosporium, and Pleosporales were prevalent in both groups. CF and DC samples showed no clear differences in clustering when compared to each other or to negative controls. SSU-rRNA sequencing was used to profile the mycobiome in pediatric PWCF and DC subjects. Notable differences were observed between the groups, including the abundance of Aspergillus and Candida. Fungal DNA detected in the airway could represent a combination of pathogenic fungi and environmental exposure (e.g., dust) to fungus indicative of a common background signature. Next steps will require comparisons to airway bacterial communities.

Morphological and genetic analysis of Ceratomyxa saurida Zhao et al. 2015 and Ceratomyxa mai sp. nov. (Myxozoa: Ceratomyxidae) from the East China Sea.

We describe Ceratomyxa saurida Zhao et al. 2015 and Ceratomyxa mai sp. nov. (Myxozoa: Ceratomyxidae) from the East China Sea. C. saurida was found in the gallbladders of 3/13 specimens of its type host, Saurida elongata Temminck and Schlegel 1846 (Aulopiformes). Myxospore characters were consistent with the original description to which we have added small subunit (SSU) rRNA gene data. C. mai sp. nov. was found in gallbladders of 3/13 specimens of S. elongata and 5/13 specimens of Neobythites sivicola Jordan and Snyder 1901 (Ophidiiformes). Mature myxospores of C. mai sp. nov. were crescentic in sutural view, with a deeply concave posterior angle 142.2±8.2° (125.8‒158.2°) and an arched anterior side. Shell valves were smooth and equal, 20.9±1.9 (17.3‒24.7) µm thick and 9.2±0.5 (8.1‒9.9) µm long, and joined at a straight, thin sutural plane passing between two nematocysts (polar capsules). The nematocysts were equal-sized, pyriform, 2.6±0.2 (2.4‒2.9) µm long and 2.7±0.2 (2.4‒3.3) µm wide, with their tapered ends pointed toward each other, located in the anterior third of the spore. Sequences of the SSU rRNA gene and internal transcribed spacer 1 showed that the isolates of C. mai sp. nov. obtained from S. elongata and N. sivicola were identical. The SSU rRNA gene sequence of C. mai sp. nov. was distinct from all known myxosporeans and clustered with C. saurida, and then with Ceratomyxa filamentosi Kalatzis, Kokkari and Katharios 2013, both of which also infect Aulopiformes fishes.

Kinopus chlorellivorus gen. nov., sp. nov. (Vampyrellida, Rhizaria), a New Algivorous Protist Predator Isolated from Large-Scale Outdoor Cultures of Chlorella sorokiniana.

The large-scale culture of low-cost algal biomass can be significantly affected by microbial grazing on the algae. To minimize the impact, it is necessary to manage the predators. In this study, we describe a new genus and species of vampyrellid amoeba, Kinopus chlorellivorus, which caused the loss of Chlorella sorokiniana in large-scale cultures. We assigned it to the family Leptophryidae (Vampyrellida) based on morphology and small-subunit (SSU) rRNA gene sequence comparisons. Using transmission electron microscopy, we found spherical lucent inclusions, which have not been reported for any leptophryids or other vampyrellids. The gene sequence of SSU rRNA did not match any recognized genera or species and contained four characteristic regions. K. chlorellivorus preys on algae by engulfment. Laboratory feeding experiments confirmed that its grazing rate was as high as 131 Chlorella cells day-1 individual-1. Results of prey-range experiments demonstrated that it could consume other chlorophyte microalgae (e.g., Scenedesmus, Coelastrella, and Haematococcus) but with a strong feeding ability on Chlorella spp., with ingestion rates ranging from 2.67 to 3.15 prey predator-1 h-1 and growth rates of the amoeba ranging from 0.039 to 0.045 h-1. On the basis of its high grazing ability on Chlorella, capacity to form large populations in a short period of time, and capacity to form resistant resting stages, this contaminant has the potential to cause serious problems in large-scale Chlorella culture and should be of concern to operators of algal production facilities. IMPORTANCE The vampyrellids (Vampyrellida, Rhizaria) are a major group of predatory amoebae that have attracted significant attention because of their diversity of feeding strategies. The crucial roles they play in important processes such as suppressing soil disease and controlling aquatic algae, and as microbial contaminants in outdoor large-scale algal cultures, have also received increasing attention. In this study, a new genus and species of algivorous vampyrellid amoeba, Kinopus chlorellivorus, is described as a significant grazer responsible for losses in outdoor industrial Chlorella cultures. We found that the amoeba's detrimental effects on Chlorella cultures may be related to its specific feeding characteristics. This study provides phenotypic and genetic information on a previously unknown vampyrellid, emphasizes the impact of contaminating vampyrellids in commercial microalgal cultures, and will contribute to the development of management strategies for predicting this kind of contaminant in large-scale microalgal cultivation.

Taxonomy and molecular phylogeny of two new Blepharisma species (Ciliophora, Heterotrichea) from northeastern China

The morphology and ciliary pattern of two new heterotrich ciliates, Blepharisma orientale n. sp. and B. sinicum n. sp., collected from Sanjiang Plain Wetland, northeastern China, were investigated using living observation and protargol staining. Blepharisma orientale n. sp. is characterized by the following combination of characteristics: limnetic; body in vivo 280–380 × 120–190 μm, slender to elongate, cortical granules dark pink, ventral margin somewhat sigmoid; buccal field about 55–60% of body length; 30–39 somatic kineties; 70–76 adoral membranelles, each comprising two long rows of basal bodies; paroral membrane consists of dikinetids with the posterior third obliquely arranged, both basal bodies in each dikinetid are ciliated. Blepharisma sinicum n. sp. is recognized by the combination of the following characters: limnetic; body 190–260 × 100–170 μm in size in vivo, irregularly sigmoid and slightly bilaterally flattened, dorsal margin curved; cortical granules pale pink in color; buccal field about 60–75% of body length; 25–29 somatic kineties; 53–82 adoral membranelles; paroral membrane consisting entirely of dikinetids. Phylogenetic analyses based on small subunit rRNA (SSU rRNA) gene sequence data recovered the two new species as sister taxa, both the sister group of B. penardi.

Characterization of a Novel Hypotrich Ciliate From Heavy Metal-Contaminated Industrial Outlet in Onsan, Ulsan, South Korea.

Very few studies exist on the description of protozoan ciliates from industrially contaminated sites. In this study, we report a description of a novel hypotrich ciliate isolated from water samples collected from an industrially contaminated outlet in Onsan, Ulsan, South Korea. The oxytrichid ciliate, Histriculus tolerans n. sp., was investigated using live observation and protargol impregnation. The morphology, morphogenesis, and molecular phylogeny inferred from small-subunit (SSU) rRNA gene sequences were studied. The new species is mainly characterized by a cell size of about 70 × 40 μm in vivo, two elongate ellipsoidal macronuclear nodules and one or two micronuclei, adoral zone of about 51% of body length with 32 membranelles on average, about 34 cirri in the right and 24 cirri in the left marginal row, 18 frontoventral transverse cirri, six dorsal kineties including two dorsomarginal rows, and dorsal kinety 1 with 26 bristles. Morphogenesis is similar to that of the type species, i.e., Histriculus histrio, except that oral primordium does not contribute to anlage II of the proter. Phylogenetic analyses, based on small-subunit rRNA gene sequences, consistently place the new species within the family Oxytrichidae, clustering with H. histrio.

Arbuscular Mycorrhizal Communities in the Roots of Sago Palm in Mineral and Shallow Peat Soils

Communities of arbuscular mycorrhizal fungi (AMF) in plant roots improve host plant growth. In this study, AMF communities in the roots of the sago palm (Metroxylon sagu Rottb.) were investigated in mineral soil (MS) and shallow peat soil (SPS) in Sarawak, Malaysia. MS exhibited lower moisture content (MS, 38.1; SPS, 79.8%), higher pH (H2O) (MS, 4.6; SPS, 4.1), higher soil bulk density (MS, 1.03; SPS, 0.20 g cm−3), and higher nitrogen content (MS, 16.9; SPS, 2.7 kg m−3) than SPS at the same soil depth, while the phosphorus (P) content (Bray II) (MS, 1.6; SPS, 1.9 g P2O5 m−3) was similar. The AMF colonization rate was significantly lower in SPS (39.2 ± 12.5%) than in MS (73.2 ± 4.6%). The higher number of AMF operational taxonomic units (OTUs) was detected by amplicon sequencing of the partial small-subunit rRNA gene (MS, 78; SPS, 50). A neighbor-joining tree of obtained OTUs revealed that they belonged to Acaulosporaceae, Ambisporaceae, Claroideoglomeraceae, Gigasporaceae, and Glomeraceae. The lower abundance and diversity of AMF in SPS are possibly caused by abiotic factors, including soil physicochemical properties. Glomus and Acaulospora species detected in SPS might have strong tolerance against acidity and high soil moisture content.

Detection of planktonic coenzyme factor 430 in a freshwater lake: small-scale analysis for probing archaeal methanogenesis

We present the first data describing molecular evidence of planktonic coenzyme factor 430 (hereafter referred to as F430), a key signature of methanogenic archaea, obtained from a water column sample in a freshwater lake in Japan. We report concentrations of native F430 ranging from 6.8–35 × 102 femto mol g-wet−1 and 8.5–86 × 102 femto mol g-wet−1 in the water column and core-top sediment, respectively. Among these profiles, the epimer F430 of the deactivated chemical formula was very low overall in both planktonic and benthic environments. Small subunit (SSU) rRNA gene sequencing revealed that the planktonic microbial community was clearly different from that of the benthic sediment. Planktonic cyanobacteria (e.g., Microcystis) were predominant in the water column, whereas a wide variety of bacteria and archaea, including methanogens (e.g., Methanobacterium, Methanoregula, Methanothrix, Methanosarcina, and Methanocella), were observed in the core-top sediment. These results suggest that the planktonic cyanobacterial community may be rapidly degraded at the water–sediment interface driven by activated benthic microbial heterotrophic processes.Graphic abstract

A New Protocruzia Species (Ciliophora: Protocruziida) Isolated From the Mariana Trench Area.

A new species of Protocruzia, isolated from the deep-sea Pacific Ocean (>3,000-m depth) in the vicinity of the Mariana Trench, is described based on morphological and molecular data. The systematic status of the ciliate genus Protocruzia has long been highly ambiguous, and Protocruzia species have been assigned to an independent class until recently. In the present study, we described Protocruzia marianaensis sp. n. as a small (25–32 × 14–17 μm in vivo) drop-shaped ciliate, with longitudinal furrows along the ciliary rows on the right side, six adoral membranelles, eight somatic kineties, and one macronucleus comprising 7–11 nuclear globules. Phylogenetic analyses inferred from small subunit rRNA gene sequences revealed that seven Protocruzia species in the phylogenetic tree formed a fully supported clade representing an independent class. Protocruzia marianaensis sp. n. was established to be most closely related to Protocruzia adhaerens, with a sequence similarity of 96.64%, and was found to be able to survive at both atmospheric pressure and hydrostatic pressure of 320 bar, thereby indicating effective barotolerance.

A new potentially toxic dinoflagellate Fukuyoa koreansis sp. nov. (Gonyaulacales, Dinophyceae) from Korean coastal waters: Morphology, phylogeny, and effects of temperature and salinity on growth

To clarify an unspecified toxic Gambierdiscus-like species isolated from seawaters off Jeju Island, Korea, its morphology and molecular phylogeny based on the small subunit (SSU) and partial large subunit (LSU) rRNA gene sequences were examined. Cells were narrow in ventral view and broad in lateral view with a smooth surface. The round thecal pores were evenly distributed, with an average diameter of 0.41 µm. Cell depth, width and height were 51.7 ± 4.5 μm, 43.0 ± 4.2 μm and 55.0 ± 4.7 μm, respectively, and depth-to-width (D/W) and height-to-width (H/W) ratios were 1.1 ± 0.2 μm and 1.3 ± 0.02 μm, respectively. The nucleus was located in the hypotheca. Scanning electron microscope observations revealed that the cells displayed a plate formula of Po, 4′, 6′′, 6c, 6s, 5′′′ and 2′′′, and transmission electron microscope observation demonstrated that the cells contained crystal-like particles. Morphological features indicated that the unspecified Korean isolate belonged to the genus Fukuyoa, and based on the H/W and D/W ratios, the apical pore H/W ratio and thecal pore size, it could be differentiated from other Fukuyoa species. The phylogenetic analyses based on the SSU and LSU rRNA sequences revealed that the Korean isolate was nested within the genus Fukuyoa with high support, and it grouped with F. cf. yasumotoi isolated from Japan. Based on the morpho-molecular data, a new species, Fukuyoa koreansis sp. nov. is proposed. The maximum growth rate (0.254 d−1) of F. koreansis was observed at 25°C and a salinity of 25. The required levels of temperature and salinity for growth distinguished Fukuyoa koreansis from Gambierdiscus species.

Taxonomy and Phylogeny of Three Species of Dysteria (Ciliophora, Phyllopharyngea) Including the Description of Dysteria ozakii nom. nov.

In the last two decades, cyrtophorian ciliates have been revealed to demonstrate a high species diversity. But this group remains difficult to study, mainly because of their low abundance and relatively few taxonomically informative morphological characters. As a contribution to the taxonomy of cyrtophorians, here we investigate three Dysteria species based on their live morphology, ciliary pattern, and molecular phylogeny. Dysteria ozakii nom. nov. can be recognized by its elongate body shape and four right kineties, including three frontoventral kineties. A neotype has to be fixed for the species as no type materials were deposited. The other two species, D. brasiliensis Faria et al., 1922 and D. compressa (Gourret & Roeser, 1886) Kahl, 1931, are redescribed and supplementary information for each is supplied. Phylogenetic analyses based on small-subunit (SSU) rRNA gene sequences support the validity of the species. In addition, four species of bacterial epibionts were observed on the surface of the three Dysteria spp. The identities of these bacterial species are discussed based on the newly obtained 16S rRNA gene sequences.

Biologically facilitated precipitation of metals in low-Fe waters at the sulphidic Mount Chalmers mine, Queensland, Australia

Biofilms enriched in copper and zinc have formed at the aqueous-air interface and in subaqueous environments at the abandoned Mount Chalmers volcanic-hosted massive sulphide mine in Queensland, Australia. Remedial activity on site has limited acid mine drainage and associated iron-rich discharges, and surface waters leave the site at pH ~ 7. Unlike in many other oxygenated metal-sulphide environments, (bio)precipitates at this site conspicuously lack iron and manganese. Hence, this site provides an opportunity to observe the interactions of microorganisms with other metals, unobscured by the usually dominant Fe and Mn processes. Small subunit rRNA gene sequencing analyses suggest that biofilms are dominated by neutrophilic species of moss, diatoms, fungi and rotifera, as well as minor cyanobacteria and photosynthesising microorganisms adapted to high metal loads (e.g., up to ~ 8 mg/L Zn and ~ 2 mg/L Cu) in surface waters. Microbial communities formed metal-bearing biofilms on a time-scale of months. Investigations of these biofilms using scanning electron microscopy with electron dispersive x-ray spectroscopy and synchrotron-based X-ray fluorescence microscopy indicate that biofilms are intimately associated with metals, especially Cu and Zn, with Cu concentrations of up to ~ 20 wt%. Microorganisms on site help to catalyse dissolution and precipitation processes, although abiotic precipitation and anthropogenic intervention (raised pH) are the main geochemical drivers of remediation. Our observations suggest that microorganisms catalysed metal accumulation processes by (1) providing a substrate to which metals can attenuate during wetting and drying cycles and (2) micro-fossilisation induced by microorganism-metal interactions. This site provides a unique view of the impacts of rehabilitation processes on the microbial community, and vice versa. However, small numbers of bacteria that can contribute to acid mine drainage remain in the biome, ready to proliferate should remediation activities wane. Processes identified at this site are also relevant for potentially economic, enhanced metal extraction from legacy mine sites.

Single-cell genomics unveils a canonical origin of the diverse mitochondrial genomes of euglenozoans

BackgroundThe supergroup Euglenozoa unites heterotrophic flagellates from three major clades, kinetoplastids, diplonemids, and euglenids, each of which exhibits extremely divergent mitochondrial characteristics. Mitochondrial genomes (mtDNAs) of euglenids comprise multiple linear chromosomes carrying single genes, whereas mitochondrial chromosomes are circular non-catenated in diplonemids, but circular and catenated in kinetoplastids. In diplonemids and kinetoplastids, mitochondrial mRNAs require extensive and diverse editing and/or trans-splicing to produce mature transcripts. All known euglenozoan mtDNAs exhibit extremely short mitochondrial small (rns) and large (rnl) subunit rRNA genes, and absence of tRNA genes. How these features evolved from an ancestral bacteria-like circular mitochondrial genome remains unanswered.ResultsWe sequenced and assembled 20 euglenozoan single-cell amplified genomes (SAGs). In our phylogenetic and phylogenomic analyses, three SAGs were placed within kinetoplastids, 14 within diplonemids, one (EU2) within euglenids, and two SAGs with nearly identical small subunit rRNA gene (18S) sequences (EU17/18) branched as either a basal lineage of euglenids, or as a sister to all euglenozoans. Near-complete mitochondrial genomes were identified in EU2 and EU17/18. Surprisingly, both EU2 and EU17/18 mitochondrial contigs contained multiple genes and one tRNA gene. Furthermore, EU17/18 mtDNA possessed several features unique among euglenozoans including full-length rns and rnl genes, six mitoribosomal genes, and nad11, all likely on a single chromosome.ConclusionsOur data strongly suggest that EU17/18 is an early-branching euglenozoan with numerous ancestral mitochondrial features. Collectively these data contribute to untangling the early evolution of euglenozoan mitochondria.

Morphology and molecular phylogeny of a new brackish pleurostomatid ciliate, Loxophyllum paludosum sp. n. (Ciliophora, Litostomatea, Haptoria), from a mangrove wetland in China

A new ciliate species of the genus Loxophyllum Dujardin, 1841, Loxophyllum paludosum sp. n., is described from a mangrove wetland near Daya Bay in Guangdong Province, southern China, based on morphological and molecular analyses. The new species can be distinguished from its congeners by a combination of the following characters: (1) 12–14 right kineties and 4–6 left kineties; (2) two macronuclear nodules and one micronucleus; (3) a single contractile vacuole located terminally; (4) extrusomes bar-shaped, evenly spaced along entire ventral margin, and clustered to form 5–7 warts along dorsal margin; and (5) presence of three ridges on the left side of cell. The new species is divergent from its congeners from 0.4% to 6.7% (5–104 nucleotide sites) based on the small subunit (SSU) rRNA gene sequence data. The validity of the new species is also supported by molecular phylogenetic trees inferred from SSU rRNA gene sequences.