Bacteria are key players in biogeochemical cycles and control water quality in freshwater ecosystems. Nevertheless, little is known about the identity and ecology of riverine bacteria, especially during ice‐covered periods that are often mistakenly perceived as periods with negligible biological activities. Here, we analyzed in detail the effects of environmental and climatic conditions on freshwater bacterial community structure and diversity over a 2‐yr sampling campaign, targeting a seasonally ice‐covered river of the Quebec City (Canada) area, the Saint‐Charles River. Quantitative polymerase chain reaction and 16S ribosomal RNA gene high‐throughput sequencing demonstrated a strong seasonal cycle of the bacterial community composition with rapid successions of bacterial lineages, reflecting the harsh climatic condition of the region. During the summer, the bacterial community was dominated by typical freshwater microorganisms such as Limnohabitans, Sporichthyaceae hgcI/acI clade, and Pseudarcicella. In contrast, the results suggest that during the cold season, the low water temperatures, combined with other prevailing conditions such as reduced light availability and minimal particulate inputs from the catchment, created various environmental niches for potential methanotrophic Gammaproteobacteria, such as Crenothrix and Methylobacter, other Betaproteobacteria, such as Candidatus Methylopumilus, Candidatus Nitrotoga, and Rhodoferax, as well as Verrucomicrobia and Parcubacteria. The presence of these taxa in the winter suggests active carbon, iron, and nitrogen cycling under ice, whereas summer lineages are dormant or in a phase of reduced activity. These results increase our understanding of bacterial dynamic and potential metabolic processes occurring in seasonally ice‐covered inland waters, providing evidence that winter can be an especially important period for freshwater ecological processes.

We describe an inexpensive, reliable, and easily executed improvement for the extraction of DNA from SterivexTM filter units, that involves the separation of the SterivexTM filter from its casing. Our study demonstrates that our modification of the original extraction protocol significantly increased DNA yields, with an average increase of 4.1‐fold more DNA than with the standard approach. A comparison of the diversity after Illumina MiSeq sequencing of bacterial communities extracted with both the standard approach and the proposed one indicated that our modified protocol has no or little impact on the results. This protocol provides a relatively straight forward means to achieve higher yields of DNA from the extraction of SterivexTM cartridges without altering the community composition and will likely be of interest to a wide range of scientists that use techniques based on the recovery of DNA from filters.

Protists are key stone components of aquatic ecosystems, sustaining primary productivity and aquatic food webs. However, their diversity, ecology and structuring factors shaping their temporal distribution remain strongly misunderstood in freshwaters. Using high-throughput sequencing on water samples collected over 16 different months (including two summer and two winter periods), combined with geochemical measurements and climate monitoring, we comprehensively determined the pico- and nanoeukaryotic community composition and dynamics in a Canadian river undergoing prolonged ice-cover winters. Our analysis revealed a large protist diversity in this fluctuating ecosystem and clear seasonal patterns demonstrating a direct and/or indirect selective role of abiotic factors, such as water temperature or nitrogen concentrations, in structuring the eukaryotic microbial community. Nonetheless, our results also revealed that primary productivity, predatory as well as parasitism lifestyles, inferred from fine phylogenetic placements, remained potentially present over the annual cycle, despite the large seasonal fluctuations and the remodeling of the community composition under ice. In addition, potential interplays with the bacterial community composition were identified supporting a possible contribution of the bacterial community to the temporal dynamics of the protist community structure. Our results illustrate the complexity of the eukaryotic microbial community and provide a substantive and useful dataset to better understand the global freshwater ecosystem functioning.

Among the major issues linked with producing safe water for consumption is the presence of the parasitic protozoa Cryptosporidium spp. and Giardia spp. Since they are both responsible for gastrointestinal illnesses that can be waterborne, their monitoring is crucial, especially in water sources feeding treatment plants. Although their discovery was made in the early 1900s and even before, it was only in 1999 that the U.S. Environmental Protection Agency (EPA) published a standardized protocol for the detection of these parasites, modified and named today the U.S. EPA 1623.1 Method. It involves the flow-through filtration of a large volume of the water of interest, the elution of the biological material retained on the filter, the purification of the (oo)cysts, and the detection by immunofluorescence of the target parasites. Since the 1990s, several molecular-biology-based techniques were also developed to detect Cryptosporidium and Giardia cells from environmental or clinical samples. The application of U.S. EPA 1623.1 as well as numerous biomolecular methods are reviewed in this article, and their advantages and disadvantages are discussed guiding the readers, such as graduate students, researchers, drinking water managers, epidemiologists, and public health specialists, through the ever-expanding number of techniques available in the literature for the detection of Cryptosporidium spp. and Giardia spp. in water.

Objective This study was designed to find a method to enhance the recovery of Cryptosporidium spp. and Giardia spp. parasites from water samples for research purposes compared to the results that can be achieved with USEPA Method 1623.1. Four different approaches were used to test water samples that were artificially spiked with parasites. The approaches were: (i) Method 1623.1 itself, (ii) elution of Method 1623.1 combined with microfiltration, (iii) an elution technique based on grinding the filter membrane in a blender before the eluent was concentrated by immunomagnetic separation, and (iv) the blender elution followed by microfiltration. Fluorescence microscopy was used to determine which approach led to the highest parasite recovery rates. Results Method 1623.1 gave the best results for Giardia, while all four approaches were statistically equivalent for Cryptosporidium. We evaluated the costs and laboratory time requirements for each protocol to give readers a complete comparison of the methods tested. Elution of Method 1623.1 combined with microfiltration resulted in lower costs and less laboratory work time without compromising the recovery of the parasites.