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Long-term survival of pathogenic microorganisms in streams enables long-distance disease transmission. In order to manage water-borne diseases more effectively we need to better predict how microbes behave in freshwater systems, particularly how they are transported downstream in rivers. Microbes continuously immobilize and resuspend during downstream transport owing to a variety of processes including gravitational settling, attachment to in-stream structures such as submerged macrophytes, and hyporheic exchange and filtration within underlying sediments. We developed a stochastic model to describe these microbial transport and retention processes in rivers that also accounts for microbial inactivation. We used the model to assess the transport, retention, and inactivation of Escherichia coli in a small stream and the underlying streambed sediments as measured from multitracer injection experiments. The results demonstrate that the combination of laboratory experiments on sediment cores, stream reach-scale tracer experiments, and multiscale stochastic modeling improves assessment of microbial transport in streams. This study (1) demonstrates new observations of microbial dynamics in streams with improved data quality than prior studies, (2) advances a stochastic modeling framework to include microbial inactivation processes that we observed to be important in these streams, and (3) synthesizes new and existing data to evaluate seasonal dynamics.

Free-living protozoa are thought to be of fundamental importance in aquatic ecosystems, but there is limited understanding of their diversity and ecological role, particularly in surface-associated communities such as biofilms. Existing eukaryote-specific PCR primers were used to survey 18S rRNA gene sequence diversity in stream biofilms but poorly revealed protozoan diversity, demonstrating a need for protozoan-targeted primers. Group-specific PCR primers targeting 18S rRNA genes of the protozoan phylum Ciliophora were therefore designed and tested using DNA extracted from cultured protozoan isolates. The two most reliable primer combinations were applied to stream biofilm DNA, followed by cloning and sequencing analysis. Of 44 clones derived from primer set 384F/1147R, 86% were of probable ciliate origin, as were 25% of 44 clones detected by primer set 121F/1147R. A further 29% of 121F/1147R-detected clones matched sequences from the closely related phylum Apicomplexa. The highly ciliate-specific primer set 384F/1147R was subsequently used in PCRs on biofilm DNA from four streams exhibiting different levels of human impact, revealing differences in ciliate sequence diversity in samples from each site. Of a total of 240 clones, 73% were of probable ciliate origin; 54 different putative ciliate sequences were detected from throughout seven taxonomic ciliate classes. Sequences from Oligohymenophorea were most commonly detected in all samples, followed by either Spirotrichea or Phyllopharyngea. Restriction fragment length polymorphism profile-based analysis of clones suggested a potentially higher level of diversity than did sequencing. Nevertheless, newly designed PCR primers 384F/ 1147R were considered to provide an effective molecular basis for characterization of ciliate diversity in stream biofilms.

ABSTRACT. Recent freshwater policy reforms in New Zealand promote increased community involvement in freshwater decision making and management. Involving community members in scientific monitoring increases both their knowledge and their ability to discuss this knowledge with professionals, potentially increasing their influence in decision-making processes. However, these interactions rarely occur because, in particular, of perceptions that volunteer-collected data are unreliable. We assessed the agreement between volunteer (community group) and local government (regional council) data at nine stream sites across New Zealand. Over 18 months, community groups and regional council staff monitored, in parallel, a common set of water quality variables, physical habitat, periphyton and benthic macroinvertebrates that are routinely used by regional councils for statutory state of environment reporting. Community groups achieved close agreement (correlations ≥ 0.89, bias < 1%) with regional councils for temperature, electrical conductivity, visual water clarity, and Escherichia coli. For dissolved oxygen, nitrate, and pH, correlations were weaker (0.2, 0.53, and 0.4, respectively). Volunteer assessments of physical habitat were as consistent over time as those of councils. For visual assessments of thick periphyton growths (% streambed cover), volunteers achieved a correlation of 0.93 and bias of 0.1% relative to councils. And for a macroinvertebrate biotic index that indicates water and habitat quality, correlation was 0.88, bias was < 5%, and the average difference was 12% of the index score. Volunteers showed increased awareness of local freshwaters, understanding of stream ecosystems, and attentiveness to local and national freshwater issues. Most volunteers had shared their knowledge and interest with others in their community. Most groups had developed relationships with their regional council, and some volunteers became more interested in engaging in freshwater decision making. Given adequate professional support, community-based water monitoring can provide data reliable enough to augment professionally collected data, and increase the opportunities, confidence, and skills of community members to engage in freshwater decision making.