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The Paramecium caudatum stock GR L-1, isolated from a freshwater sample of a natural habitat in Greece, Livadia, was grown in a modified 0.25% Cerophyl infusion [ 62] inoculated with Pseudomonas fluorescens, as described previously [ 63].
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This result is not surprising because GS20 is the only freshwater sample, GS11 and GS12 are two of the three estuary samples, and GS33 is the only hypersaline sample.
One freshwater sample contains no metadata on sample depth collection.
The freshwater sample was collected in the anoxic zone at 90 m depth, where the highest methanogen diversity was available in the lacustrine environment.
The precision and detection limit of FCM for measuring total virus counts was determined with a serial dilution of freshwater samples.
The approach allowed the identification of 45 elements and their simultaneous quantification in a single run of (preconcentrated) natural freshwater samples.
Marine samples showed a higher abundance of Cyanobacteria, Alphaproteobacteria while freshwater samples were dominated by Betaproteobacteria (Figure 3).
The three marine libraries grouped together showing a high similarity with each other, whereas freshwater samples were dispersed in the plot and seem to be different among them.
Preconditioning of the conductivity of the samples may be needed when applying the method to the concentration of viruses from freshwater samples [ 86] and a variation of the method has also been reported for sewage samples [ 87].
The results obtained in this study suggest that when studying the abundance of viruses in freshwater, samples should be filtered with a 0.22 μm pore size membrane, fixed with glutaraldehyde (0.25% final concentration) for 15 min at 4°C, frozen in liquid nitrogen, and stored at −80°C.
These include examination of protein expression profiles from an estuary transect [7], infant fecal samples [8], freshwater samples following exposure to heavy metals [9] and contaminated soil and groundwater [10].
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