Zeta diversity patterns in metabarcoded lotic algal assemblages as a tool for bioassessment.

Assessments of the ecological health of algal assemblages in streams typically focus on measures of their local diversity and classify individuals by morphotaxonomy. Such assemblages are often connected through various ecological processes, such as dispersal, and may be more accurately assessed as components of regional-, rather than local-scale assemblages. With recent declines in the costs of sequencing and computation, it has also become increasingly feasible to use metabarcoding to more accurately classify algal species and perform regional-scale bioassessments. Recently, zeta diversity has been explored as a novel method of constructing regional bioassessments for groups of streams. Here, we model the use of zeta diversity to investigate whether stream health can be determined by the landscape diversity of algal assemblages. We also compare the use of DNA metabarcoding and morphotaxonomy classifications in these zeta diversity-based bioassessments of regional stream health. From 96 stream samples in California, we used various orders of zeta diversity to construct models of biotic integrity for multiple assemblages of diatoms, as well as hybrid assemblages of diatoms in combination with soft-bodied algae, using taxonomy data generated with both DNA sequencing as well as traditional morphotaxonomic approaches. We compared our ability to evaluate the ecological health of streams with the performance of multiple algal indices of biological condition. Our zeta diversity-based models of regional biotic integrity were more strongly correlated with existing indices for algal assemblages classified using metabarcoding compared to morphotaxonomy. Metabarcoding for diatoms and hybrid algal assemblages involved rbcL and 18S V9 primers, respectively. Importantly, we also found that these algal assemblages, independent of the classification method, are more likely to be assembled under a process of niche differentiation rather than stochastically. Taken together, these results suggest the potential for zeta diversity patterns of algal assemblages classified using metabarcoding to inform stream bioassessments.

Implementing an Operational Framework to Develop a Streamflow Duration Assessment Method: A Case Study from the Arid West United States.

Streamflow duration information underpins many management decisions. However, hydrologic data are rarely available where needed. Rapid streamflow duration assessment methods (SDAMs) classify reaches based on indicators that are measured in a single brief visit. We evaluated a proposed framework for developing SDAMs to develop an SDAM for the Arid West United States that can classify reaches as perennial, intermittent, or ephemeral. We identified 41 candidate biological, geomorphological, and hydrological indicators of streamflow duration in a literature review, evaluated them for a number of desirable criteria (e.g., defensibility and consistency), and measured 21 of them at 89 reaches with known flow durations. We selected metrics for the SDAM based on their ability to discriminate among flow duration classes in analyses of variance, as well as their importance in a random forest model to predict streamflow duration. This approach resulted in a "beta" SDAM that uses five biological indicators. It could discriminate between ephemeral and non-ephemeral reaches with 81% accuracy, but only 56% accuracy when distinguishing 3 classes. A final method will be developed following expanded data collection. This Arid West study demonstrates the effectiveness of our approach and paves the way for more efficient development of scientifically informed SDAMs.

Classifying Streamflow Duration: The Scientific Basis and an Operational Framework for Method Development.

Streamflow duration is used to differentiate reaches into discrete classes (e.g., perennial, intermittent, and ephemeral) for water resource management. Because the depiction of the extent and flow duration of streams via existing maps, remote sensing, and gauging is constrained, field-based tools are needed for use by practitioners and to validate hydrography and modeling advances. Streamflow Duration Assessment Methods (SDAMs) are rapid, reach-scale indices or models that use physical and biological indicators to predict flow duration class. We review the scientific basis for indicators and present conceptual and operational frameworks for SDAM development. Indicators can be responses to or controls of flow duration. Aquatic and terrestrial responses can be integrated into SDAMs, reflecting concurrent increases and decreases along the flow duration gradient. The conceptual framework for data-driven SDAM development shows interrelationships among the key components: study reaches, hydrologic data, and indicators. We present a generalized operational framework for SDAM development that integrates the data-driven components through five process steps: preparation, data collection, data analysis, evaluation, and implementation. We highlight priorities for the advancement of SDAMs, including expansion of gauging of nonperennial reaches, use of citizen science data, adjusting for stressor gradients, and statistical and monitoring advances to improve indicator effectiveness.

The importance of open science for biological assessment of aquatic environments.

Open science principles that seek to improve science can effectively bridge the gap between researchers and environmental managers. However, widespread adoption has yet to gain traction for the development and application of bioassessment products. At the core of this philosophy is the concept that research should be reproducible and transparent, in addition to having long-term value through effective data preservation and sharing. In this article, we review core open science concepts that have recently been adopted in the ecological sciences and emphasize how adoption can benefit the field of bioassessment for both prescriptive condition assessments and proactive applications that inform environmental management. An example from the state of California demonstrates effective adoption of open science principles through data stewardship, reproducible research, and engagement of stakeholders with multimedia applications. We also discuss technical, sociocultural, and institutional challenges for adopting open science, including practical approaches for overcoming these hurdles in bioassessment applications.

Using co-occurrence network topology in assessing ecological stress in benthic macroinvertebrate communities.

Ecological monitoring of streams has often focused on assessing the biotic integrity of individual benthic macroinvertebrate (BMI) communities through local measures of diversity, such as taxonomic or functional richness. However, as individual BMI communities are frequently linked by a variety of ecological processes at a regional scale, there is a need to assess biotic integrity of groups of communities at the scale of watersheds. Using 4,619 sampled communities of streambed BMIs, we investigate this question using co-occurrence networks generated from groups of communities selected within California watersheds under different levels of stress due to upstream land use. Building on a number of arguments in theoretical ecology and network theory, we propose a framework for the assessment of the biotic integrity of watershed-scale groupings of BMI communities using measures of their co-occurrence network topology. We found significant correlations between stress, as described by a mean measure of upstream land use within a watershed, and topological measures of co-occurrence networks such as network size (r = -.81, p < 10-4), connectance (r = .31, p < 10-4), mean co-occurrence strength (r = .25, p < 10-4), degree heterogeneity (r = -.10, p < 10-4), and modularity (r = .11, p < 10-4). Using these five topological measures, we constructed a linear model of biotic integrity, here a composite of taxonomic and functional diversity known as the California Stream Condition Index, of groups of BMI communities within a watershed. This model can account for 66% of among-watershed variation in the mean biotic integrity of communities. These observations imply a role for co-occurrence networks in assessing the current status of biotic integrity for BMI communities, as well as their potential use in assessing other ecological communities.

Using alpha, beta, and zeta diversity in describing the health of stream-based benthic macroinvertebrate communities.

Ecological monitoring of streams has frequently focused on measures describing the taxonomic, and sometimes functional, α diversity of benthic macroinvertebrates (BMIs) within a single sampled community. However, as many ecological processes effectively link BMI stream communities there is a need to describe groups of communities using measures of regional diversity. Here we demonstrate a role for incorporating both a traditional pairwise measure of community turnover, ß diversity, in assessing community health as well as ζ diversity, a more generalized framework for describing similarity between multiple communities. Using 4,395 samples of BMI stream communities in California, we constructed a model using measures of α, ß, and ζ diversity, which accounted for 71.7% of among-watershed variation in the mean health of communities, as described by the California Streams Condition Index (CSCI). We also investigated the use of ζ diversity in assessing models of stochastic vs. niche assembly across communities of BMIs within watersheds, with the niche assembly model found to be the likelier of the two.

Selecting Comparator Sites for Ecological Causal Assessment Based on Expected Biological Similarity.

Sites in poor ecological condition often require causal assessment to determine appropriate follow-up actions. Site-specific causal assessments can be time consuming. To streamline the process, we describe a quantitative method that expedites a key component of causal assessment: identifying a group of ecologically similar (comparator) sites that are used to compare and contrast biological condition and stressor exposure at the site of interest. A good set of comparator sites should: 1. Be capable of supporting similar biota to the impaired site in the absence of disturbance; 2. Comprise a gradient of biotic condition; and 3. Contain enough sites to assess variability. We used expected biological similarity to select good sets of comparator sites from a large pool of potential sites. Expected biological similarity was measured as Bray-Curtis dissimilarity values (BC) calculated from the expected benthic macroinvertebrate taxa lists produced by a predictive biotic index of stream health. Sets of comparator sites were created for 15 demonstration sites across Southern California in poor condition. We examined the stressor and biological data collected at the 15 sites and their comparators to assess the likelihood that four example stressors - total nitrogen, ammonia, specific conductivity, and bifenthrin - contribute to the poor biotic conditions that were observed. We were able to select more than 100 comparator sites for all but 1 of the 15 demonstration sites at a BC <0.1. These sets of comparator sites were then used to evaluate the four example stressors using two commonly used causal assessment types of evidence. Elevated conductivity was the most frequently supported likely cause among the demonstration sites, though total nitrogen and bifenthrin were also indicated at some sites. Though our specific approach was tailored for application in California's stream bioassessment framework, the concepts could be adapted for any bioassessment program with a large amount of sample data and an associated predictive index of biotic condition. Furthermore, this approach lays the groundwork for developing a novel approach to causal assessment that begins with a rapid, screening-level evaluation of stressors common in a region using these data-rich groups of comparator sites, which then informs follow-up management actions.

Correspondence of biological condition models of California streams at statewide and regional scales.

We used boosted regression trees (BRT) to model stream biological condition as measured by benthic macroinvertebrate taxonomic completeness, the ratio of observed to expected (O/E) taxa. Models were developed with and without exclusion of rare taxa at a site. BRT models are robust, requiring few assumptions compared with traditional modeling techniques such as multiple linear regression. The BRT models were constructed to provide baseline support to stressor delineation by identifying natural physiographic and human land use gradients affecting stream biological condition statewide and for eight ecological regions within the state, as part of the development of numerical biological objectives for California's wadeable streams. Regions were defined on the basis of ecological, hydrologic, and jurisdictional factors and roughly corresponded with ecoregions. Physiographic and land use variables were derived from geographic information system coverages. The model for the entire state (n = 1,386) identified a composite measure of anthropogenic disturbance (the sum of urban, agricultural, and unmanaged roadside vegetation land cover) within the local watershed as the most important variable, explaining 56% of the variance in O/E values. Models for individual regions explained between 51 and 84% of the variance in O/E values. Measures of human disturbance were important in the three coastal regions. In the South Coast and Coastal Chaparral, local watershed measures of urbanization were the most important variables related to biological condition, while in the North Coast the composite measure of human disturbance at the watershed scale was most important. In the two mountain regions, natural gradients were most important, including slope, precipitation, and temperature. The remaining three regions had relatively small sample sizes (n ≤ 75 sites) and had models that gave mixed results. Understanding the spatial scale at which land use and land cover affect taxonomic completeness is imperative for sound management. Our results suggest that invertebrate taxonomic completeness is affected by human disturbance at the statewide and regional levels, with some differences among regions in the importance of natural gradients and types of human disturbance. The construction and application of models similar to the ones presented here could be useful in the planning and prioritization of actions for protection and conservation of biodiversity in California streams.

Identifying reference conditions and quantifying biological variability within benthic macroinvertebrate communities in perennial and non-perennial northern California streams.

Identification of minimally disturbed reference sites is a critical step in developing precise and informative ecological indicators. We tested procedures to select reference sites, and quantified natural variation (inter-site and -annual variability) among reference conditions using a macroinvertebrate data set collected from 429 mediterranean-climate stream reaches in the San Francisco Bay Area, California (USA). We determined that a landscape GIS-based stressor screen followed by a local field-based stressor screen effectively identified least-disturbed reference sites that, based on NMS ordination results, supported different biological communities than sites identified with only landscape (GIS) or local (field) stressors. An examination of least-disturbed reference sites indicated that inter-site variability was strongly associated with stream hydrology (i.e., perennial vs. non-perennial flow) and annual precipitation, which highlights the need to control for such variation when developing biological indicators through natural gradient modeling or using unique biological indicators for both non-perennial and perennial streams. Metrics were more variable among non-perennial streams, indicating that additional modeling may be needed to develop precise biological indicators for non-perennial streams. Among 192 sites sampled two to six times over the 8-year study period, the biological community showed moderate inter-annual variability, with the 100 point index of biotic integrity scores varying from 0 to 51 points (mean = 11.5). Variance components analysis indicated that inter-annual variability explained only a fraction (5-18 %) of the total variation when compared against site-level variation; thus efforts to understand causes of natural variation between sites will produce more precise and accurate biological indicators.

Evaluating ethanol-based sample preservation to facilitate use of DNA barcoding in routine freshwater biomonitoring programs using benthic macroinvertebrates.

Molecular methods, such as DNA barcoding, have the potential to enhance biomonitoring programs worldwide. Altering routinely used sample preservation methods to protect DNA from degradation may pose a potential impediment to application of DNA barcoding and metagenomics for biomonitoring using benthic macroinvertebrates. Using higher volumes or concentrations of ethanol, requirements for shorter holding times, or the need to include additional filtering may increase cost and logistical constraints to existing biomonitoring programs. To address this issue we evaluated the efficacy of various ethanol-based sample preservation methods at maintaining DNA integrity. We evaluated a series of methods that were minimally modified from typical field protocols in order to identify an approach that can be readily incorporated into existing monitoring programs. Benthic macroinvertebrates were collected from a minimally disturbed stream in southern California, USA and subjected to one of six preservation treatments. Ten individuals from five taxa were selected from each treatment and processed to produce DNA barcodes from the mitochondrial gene cytochrome c oxidase I (COI). On average, we obtained successful COI sequences (i.e. either full or partial barcodes) for between 93-99% of all specimens across all six treatments. As long as samples were initially preserved in 95% ethanol, successful sequencing of COI barcodes was not affected by a low dilution ratio of 2∶1, transfer to 70% ethanol, presence of abundant organic matter, or holding times of up to six months. Barcoding success varied by taxa, with Leptohyphidae (Ephemeroptera) producing the lowest barcode success rate, most likely due to poor PCR primer efficiency. Differential barcoding success rates have the potential to introduce spurious results. However, routine preservation methods can largely be used without adverse effects on DNA integrity.

Bioassessment tools in novel habitats: an evaluation of indices and sampling methods in low-gradient streams in California.

Biomonitoring programs are often required to assess streams for which assessment tools have not been developed. For example, low-gradient streams (slopeor=450) to calculate the SCIBI in 28 of 45 samples and often collected fewer than 100 individuals, suggesting it is inappropriate for low-gradient streams in California; failures for the other methods were less common (TRC, 16 samples; MCM, 11 samples). Within-site precision, measured as the minimum detectable difference (MDD) was poor but similar across methods for the SCIBI (ranging from 19 to 22). However, RWB had the lowest MDD for O/E scores (0.20 versus 0.24 and 0.28 for MCM and TRC, respectively). Mantel correlations showed that assemblages were more similar within sites among methods than within methods among sites, suggesting that the sampling methods were collecting similar assemblages of organisms. Statistically significant disagreements among methods were not detected, although O/E scores were higher for RWB samples than TRC. Index scores suggested impairment at all sites in the study. Although index scores did not respond strongly to several measurements of disturbance in the watershed, percent agriculture showed a significant, negative relationship with O/E scores.

Long-term variability in bioassessments: a twenty-year study from two northern California streams.

Long-term variability of bioassessments has not been well evaluated. We analyzed a 20-year data set (1984-2003) from four sites in two northern California streams to examine the variability of bioassessment indices (two multivariate RIVPACS-type O/E scores and one multimetric index of biotic integrity, IBI), as well as eight metrics. All sites were sampled in spring; one site was also sampled in summer. Variability among years was high for most metrics (coefficients of variation, CVs ranging from 16% to 246% in spring) but lower for indices (CVs of 22-26% for the IBI and 21-32% for O/E scores in spring), which resulted in inconsistent assessments of biological condition. Variance components analysis showed that the time component explained variability in all metrics and indices, ranging from 5% to 35% of total variance explained. The site component was large (i.e., >40%) for some metrics (e.g., EPT richness), but nearly absent from others (e.g., Diptera richness). Seasonal analysis at one site showed that variability among seasons was small for some metrics or indices (e.g., Coleoptera richness), but large for others (e.g., EPT richness, O/E scores). Climatic variables did not show consistent trends across all metrics, although several were related to the El Niño Southern Oscillation Index at some sites. Bioassessments should incorporate temporal variability during index calibration or include climatic variability as predictive variables to improve accuracy and precision. In addition, these approaches may help managers anticipate alterations in reference streams caused by global climate change and high climatic variability.