Free-living amoebae promote Candida auris survival and proliferation in water

Candida auris is an emerging species responsible for life-threatening infections. Its ability to be resistant to most systemic antifungal classes and its capacity to persist in a hospital environment have led to health concerns. Currently, data about environmental reservoirs are limited but remain essential in control of C. auris spread. The aim of our study was to explore the interactions between C. auris and two free-living amoeba (FLA) species, Vermamoeba vermiformis and Acanthamoeba castellanii, potentially found in the same water environment. Candida auris was incubated with FLA trophozoites or their culture supernatants. The number of FLA and yeasts was determined at different times and transmission electron microscopy (TEM) was performed. Supernatants of FLAs promoted yeast survival and proliferation. Internalization of viable C. auris within both FLA species was also evidenced by TEM. A water environmental reservoir of C. auris can therefore be considered through FLAs and contamination of the hospital water networks would consequently be possible.     

Siliceous protozoan plates in lake sediments

The silicified plates of certain testate amoebae have the necessary characteristics to be useful paleoindicators. These plates are usually abundant and well preserved in sedimentary deposits, where they can be studied using the same preparation techniques developed for other siliceous microfossils. Individual plates can usually be identified to the generic and, in some cases, to the species level. A preliminary survey of plates recorded in the surficial (top centimetre) sediments of 38 Adirondack Mountain (NY) lakes suggests that these testaceans may reflect catchment features (e.g. bog development) more closely than lakewater characteristics. The siliceous, species-specific scales of heliozoans are also shown to preserve in sedimentary profiles.     

Cultivation of the sapropelic ciliate Plagiopyla nasuta Stein and isolation of the endosymbiont Methanobacterium formicicum

The sapropelic ciliate Plagiopyla nasuta was isolated and cultured in monoculture. Optimal conditions for growth were: 15–20°C, pH about 7, and about 2% of oxygen in the headspace. Cultures of P. nasuta produced methane. Epifluorescence microscopy revealed the presence of methanogenic bacteria as endosymbionts. An endosymbiont of the ciliate was isolated and identified as Methanobacterium formicicum. In the ciliate cell these methanogens were found to be closely associated with microbody-like organelles. No mitochondria could be detected.     

Predation rates of flagellate and ciliated protozoa on bacterioplankton in a river

Abstract Predation rates of flagellate and ciliate protozoa on the bacterioplankton of Butrón River (Spain) were determined from FLB (fluorescently labelled bacteria) uptake rates. Bacterial and ciliate protozoa counts were higher when higher water temperature was recorded. Flagellate counts did not show this pattern, which suggested predation of flagellates by other organisms, or some other different nutritional mode besides phagotrophy. Average individual ciliate predation rates were up to 40-times higher than those of flagellates. These results were compared with similar data obtained from other authors in several aquatic systems. However, the population predation rates of flagellate protozoa were on average 6-times higher than that of ciliate protozoa, due to the low population numbers of the latter. Thus, flagellate protozoa can be considered as more important bacterial consumers than ciliates in this aquatic system.     

Export of species to islands from sources of differing maturity

Export of species from sources (epicenters) of differing ages and complexities was examined using laboratory microcosms. Polyurethane foam (PF) artificial substrates were colonized by protozoans for different time periods in a small pond. Substrates were returned to the laboratory and used as epicenters for protozoan colonization of barren PF islands in initially sterile microcosms. Islands were exposed to epicenters for either 24 h or continuously for 28 d. Islands from pairs of microcosms exposed to epicenters of identical ages were sampled on 1, 3, 7, 14, 21, 28 and 46 d after initial epicenter exposure. Colonization parameters were estimated by fitting numbers of colonizing species to the MacArthur-Wilson equilibrium model. Islands exposed continuously to epicenters were colonized by significantly more species than those exposed for only 24 h. Islands exposed to immature, species poor epicenters were colonized by a greater proportion of the source community than those exposed to more mature, species rich epicenters. All islands were depauperate compared to epicenters except those exposed to the most immature (1 d old) epicenter. Colonization continued at a reduced rate in spite of the absence of the epicenter. Results from communities with rapid species turnover and rapidly reproducing species suggest that the continuous presence of a species source is less important for colonization of a new habitat. Dispersal of potential colonists occurs rapidly in these communities. Less mature communities dominated by pioneer forms are more effective at producing colonists than more mature communities.     

Mixed-function oxidase activity in enriched populations of ciliated cells freshly isolated from rabbit tracheas

A method is described for the preparation of enriched populations of ciliated cells from rabbit tracheas. Following protease digestion of tracheal lumen tissue, cells were subjected to centrifugal elutriation. This produced two cell fractions of interest: an 8 µm diameter fraction believed to be composed largely of basal cells, and a 15 µm diameter fraction containing a mixture of ciliated cells and Clara cells. Further treatment of the 15 µm cells with a dextran/polyethylene glycol/phosphate buffer system resulted in separation of a highly enriched ciliated cell fraction (84.3 ± 2.7% ciliated cells with 6.5 ± 1.5% Clara cells) from a fraction containing both ciliated cells (42.0 ± 2.1%) and Clara cells (27.0 ± 3.5%). The yield of cells in the enriched ciliated cell fraction was 0.68 ± 0.09 × 10⁶ cells/ trachea. Analysis of mixed-function oxidase activity in tracheal cells showed 7-ethoxycoumarin deethylase and coumarin hydroxylase activities to be present in the 8 µm cells as well as in ciliated cells and Clara cells. Enzyme activities measured in the ciliated cells (152 ± 66 pmol/ min/ mg protein or 51.2 ± 20.5 pmol/ min/ 10⁶ cells for 7-ethoxycoumarin deethylase and 31.7 ± 15.4 pmol/ min/ mg protein or 10.5 ± 4.8 pmol/ min/ 10⁶ cells for coumarin hydroxylase) were not attributable to contamination with Clara cells.Abbreviations CD cell digest - DNase deoxyribonuclease I - E-1 first elutriator fraction - E-2 second elutriator fraction - E-3 third elutriator fraction - 7-Ec 7-ethoxycoumarin - FCS fetal calf serum - HEPES N-2-hydroxy-ethylpiperazine-N-2-ethanesulfonic acid - HpBS HEPES-buffered salt solution - NADH reduced nicotinamide adenine dinucleotide - NADPH reduced nicotinamide adenine dinucleotide phosphate - NBT nitro blue tetrazolium - PEG Carbowax polyethylene glycol 6000     

Estimation of the size of freshwater ciliate populations by a sub-sampling technique

A technique is described for the estimation of the size of ciliate populations based on replicated counts. Sample drops were taken by automatic pipettes of different volume from activated sludge and from a small oligotrophic stream. Specific aims were: 1) the estimation of the number of species; 2) the estimation of the number of individual in each species; 3) the selection of a suitable sub-sample size and number of replicates. For each volume the cumulative increase in species taken in successive sub-samples was determined. For each species the minimum permissible sub-sample volume and the number of replicates required for an allowable error of 10% and 25% were determined. Lastly, the relationship between the mean number of individuals counted for each species and the relative coefficient of variation was visualized.     

Protein phylogeny of translation elongation factor EF-1α suggests microsporidians are extremely ancient eukaryotes

Partial regions of the mRNA encoding a major part of translation elongation factor 1 (EF-1) from a mitochondrion-lacking protozoan,Glugea plecoglossi, that belongs to microsporidians, were amplified by polymerase chain reaction (PCR) and their primary structures were analyzed. The deduced amino acid sequence was highly divergent from typical EF-1's of eukaryotes, although it clearly showed a eukaryotic feature when aligned with homologs of the three primary kingdoms. Maximum likelihood (ML) analyses on the basis of six different stochastic models of amino acid substitutions and a maximum parsimony (MP) analysis consistently suggest that among eukaryotic species being analyzed,G. plecoglossi is likely to represent the earliest offshoot of eukaryotes. Microsporidians might be the extremely ancient eukaryotes which have diverged before an occurrence of mitochondrial symbiosis. Sequence availability: The nucleotide sequence data reported here appear in the GSDB, DDBJ, EMBL, and NCBI databases with the accession number D32139     

A comparative study of ciliated protozoa communities in activated-sludge plants

Abstract: Ciliated protozoa present in ten activated sludge plants at Madrid (Spain) were identified. The abundance of key groups of ciliates was determined at each plant; attached ciliates made up the most abundant and representative group. Multivariate statistical analysis was employed to study relationships between ciliates and both the physico-chemical and operational parameters of the plants. Partial correlation analysis revealed: (1) The indicator value of attached ciliates in assessing management and performance of the activated sludge process, (2) the relationship of swimming ciliates with short-aged sludges and lower quality effluents and (3) the direct association between swimming-crawling ciliates and bad settlement conditions of the sludge. Factor analysis showed the associations of the most frequent species of ciliates with the operational parameters of the plants, suggesting the indicator value of some of the species: Vorticella striata was related with poor quality of effluent; Aspidisca cicada with stable plant conditions, and Litonotus lamella with a deficiently settling sludge.     

Microbial-feeding nematodes and protozoa in soil: Their effectson microbial activity and nitrogen mineralization in decomposition hotspots and the rhizosphere

Food web studies from a range of ecosystems have demonstrated that the fauna contributes about 30% of total net nitrogen mineralization. This results mainly from the activities of microbial-feeding microfauna (nematodes and protozoa). Microbial and microfaunal activity is concentrated at spatially discrete and heterogeneously distributed organic substrates, including the rhizosphere. The dynamics of microfauna and their effect on nutrient cycling and microbial processes at these sites is reviewed. The potential manipulation of microfauna, either as an experimental tool to further understand soil microbial ecology or as a practical means of managing nutrient flows in agroecosystems, is discussed.