Science Stories

January 27, 2023

By Rosemary Hartman

Delta Smelt

Small fish with large eye, small pectoral fin, and adipose fin. — Picture of Delta Smelt, photo by Rene Reyes, US Bureau of Reclamation

Wakasagi

Small fish with short pectoral fin, adipose fin, and upright dorsal fin. Very similar to the Delta Smelt. — Picture of a Wakasagi, photo by Rene Reyes, US Bureau of Reclamation

You’ve probably heard of Delta Smelt (Hypomesus transpacificus), and you may have heard of their cousin, the Longfin Smelt (Spirinchus thaleichthys), but there is a third osmerid in the estuary. The Wakasagi (Hypomesus nipponensis), also known as Japanese Smelt, is in the same genus as Delta Smelt, and was once thought to be the same species. It is native to Japan, but was introduced to reservoirs in California by the California Department of Fish and Game in the 1950s, and now it is established throughout the watershed, including the Delta.

But what does this brother of the Delta Smelt do? Is there sibling rivalry? A group of IEP scientists was curious, so they decided to look at all of our existing data to see when, where, how big, and how many Wakasagi are in the Delta and how their environmental tolerances and diet compares to Delta Smelt. A paper about their analysis recently came out in the Journal San Francisco Estuary and Watershed Sciences.

In order to compare Delta Smelt and Wakasagi, they looked at all the data from thirty different fish datasets from San Francisco Bay, Suisun Marsh/Suisun Bay, the Delta, and the watershed (see map below). This resulted in a dataset with over 250,000 individual Wakasagi! They also looked at data from special studies of Wakasagi and Delta Smelt growth and diets in the Yolo Bypass.

Map of the San Francisco Estuary showing hundreds of sampling points in the San Francisco Bay and Delta with scattered points upstream. — Maps of the San Francisco Bay-Delta Estuary. (A) Four long-term CDFW monitoring surveys and region assignments used for the comparative Delta Smelt and Wakasagi analysis. (B) Sampling locations for a subset of additional surveys used to assess Wakasagi catch, as well as Yolo Bypass surveys used to assess life-history traits including growth, phenology, and diet. Map reproduced from Davis et al, 2022, with permission.

They found some similarities between delta smelt and Wakasagi – both fish really like hanging out in the Sacramento Deep Water Ship Channel and both like eating calanoid copepods (a particularly tasty variety of zooplankton). They spawn at about the same time, but Wakasagi are usually a little earlier (though this varies from year to year), and Wakasagi usually grow a little faster. They are similar enough that they sometimes interbreed and produce hybrid offspring.

Wakasagi aren’t the same as Delta Smelt though. Wakasagi aren’t actually very common in the Delta, instead finding their homes further upstream in reservoirs (they especially seem to like the Feather River, the screw trap there catches tens to hundreds of thousands of Wakasagi per year!). In the Delta they are mostly in the northern region, which might be just them washing in from upstream. Though they were mostly found in freshwater reaches of the Delta, Wakasagi can actually tolerate a wider range of salinity and temperatures than Delta Smelt, but they seem to prefer cooler temperatures.

So, are Wakasagi competing with Delta Smelt for limited food resources? Maybe a bit, but while they play a similar ecological role when they do overlap, they don’t overlap spatially very often, and both Delta Smelt and Wakasagi are rare in the Delta. However, they overlap enough that areas that are good for Wakasagi are probably good for Delta Smelt too. Delta Smelt are becoming more and more endangered, so we can use Wakasagi as indicators of good Delta Smelt conditions and as substitutes for smelt in some laboratory experiments.

**Major similarities and differences between Delta Smelt and Wakasagi**
Delta Smelt	Wakasagi	Comparison
Annual life span	Annual life span
Spawn later	Spawn earlier
Eat calanoid copepods	Eat calanoid copepods
Grow slower	Grow faster
Narrower tolerances	Wider tolerances
Endangered	More common
Native	Non native
Mostly semi-anadromous	Mostly freshwater
Small and silver	Small and silver
Loves the North Delta	Loves the North Delta
Smells like cucumber	Smells like fish

What is eDNA?

Environmental DNA (abbreviated eDNA) describes the genetic material that an organism sheds or excretes into its environment (e.g., skin cells, hair, mucus, blood, gametes, waste products, pollen, leaves, fungal spores). Once released, eDNA can be collected and extracted from environmental samples such as soil, sediment, water, snow or air. Once extracted, eDNA can be analyzed by several genetic methods. Depending on the method used, researchers can choose to target a single species (e.g., invasive or endangered), a particular community (e.g., fishes), or even multiple communities (e.g., all animals). For studies targeting multiple species, a common approach is to use a reference database to link the genetic sequence obtained in the eDNA sample to a particular species or taxonomic group. By scanning a reference sequence database for a match, scientists can identify which organism(s) the DNA in their sample came from. This is akin to scanning the barcode on any item in your local grocery store. When the item is scanned, information in the database links the barcode with the price of the item, etc. If the item isn’t listed, they won’t know how much to charge you.

Diagram showing pictures of animals with DNA coming off of them into the environment and aquatic organisms shedding DNA into a stream.
Figure 1. eDNA is the genetic material of living things shed into the environment. We can collect from different substrates including soil, water, and even air.

How is it used?

In recent years, eDNA has been used in a wide array of applications.

Some closely related species can be difficult to differentiate in the field, and accurate identification often requires collecting tissue from the organism, followed by several days of processing in a molecular biology lab. A new technique called SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) is a sensitive, rapid method that can provide species identification (such as the difference between endangered Delta Smelt and the visually similar, non-native Wakasagi) without invasive sampling and can be done in the field in an hour or less. Pairing eDNA or other non-invasive DNA sampling with SHERLOCK allows managers to make rapid and accurate decisions, a critical necessity for protecting listed species.

To understand the migratory dynamics of salmonids in a creek just south of the SFE, researchers from the Monterey Bay Aquarium Research Institute (MBARI) deployed an autonomous robotic sampler (Video) to collect 750 eDNA samples over the course of one year. The creek provides habitat for endangered Coho Salmon and Steelhead Trout but is at risk from non-native species including Striped Bass. This sampler will not only filter eDNA samples but perform the molecular reactions and transmit the results, ultimately giving scientists the ability to monitor species of concern in near real time.

California encompasses many unique ecosystems considered to be biodiversity “hotspots,” or areas with high biodiversity. To protect these areas, California recently developed a “30 x 30” initiative to conserve 30% of California’s lands and coastal waters by 2030. Cataloging the biodiversity of an entire ecosystem, especially for the many diverse hotspots of California, is a tall order. To aid this effort, scientists are enlisting the help of community members and students to participate in “Bioblitz” events. Teams collect eDNA samples from diverse areas throughout the state, including the SFE. This is part of the CALeDNA program run by the University of California Conservation Genomics Consortium. By involving the community in monitoring efforts, scientists not only increase their ability to collect precious data, but the community gets to learn about these innovative new technologies and gain a deeper appreciation of their local ecosystems.

The previous three examples provide a snapshot of the different types of applications for using eDNA to monitor the diverse ecosystems of the SFE and beyond. Outside of the ecological applications of eDNA, there are additional important uses. For example, to aid efforts to mitigate and monitor COVID-19 dynamics, human health researchers have begun incorporating eDNA techniques to track the virus in wastewater. Far from being a comprehensive list of eDNA applications, these studies are just the tip of the iceberg.

How eDNA can help management?

Environmental DNA sampling can be an effective management tool in the SFE. Due to its sensitivity, eDNA sampling methods can be used to help find rare and hard to find species that are difficult to detect with traditional sampling methods, like trawls or seines. Further, it can be used to detect invasive species that may be present in low numbers or to track an invasion front. Anywhere that you can collect water, you can collect eDNA. This approach can be used to sample hard to access areas or where traditional surveys cannot be utilized. An additional benefit of eDNA sampling is that it provides a way to obtain information without the need to visually observe or handle organisms. Collection and handling can have negative effects, particularly for sensitive species. Using genetic identification through eDNA sampling can also help when species are challenging to identify visually. Different scales of analysis allow managers to ask different questions or carry out different management tasks, such as: habitat use of a rare species over space and time; initial site evaluation for a species of interest to see if further, more intensive sampling is needed; and to evaluate habitat restoration on a community wide scale. The utility and ease of sampling for eDNA make it a good compliment to traditional sampling methods and can even be more efficient in terms of time, labor, and expense. For more information about eDNA sampling in estuaries and how it can help with management needs, with a focus on those of the SFE, please see Nagarajan et al. (2022).

What researchers are currently doing with eDNA

Environmental DNA sampling is currently being instituted by different agencies and institutions to answer management questions. For example, the Washington Department of Fish and Wildlife has implemented eDNA detection for several projects to evaluate streams after wildfires for Rocky Mountain Tailed Frog, to identify whether redds belong to Coho Salmon or Steelhead, and early detection of invasive mussels and snails in water bodies. Here in the SFE, Ann Holmes, a graduate student in the Genomic Variation Lab (GVL) at UC Davis, studied eDNA detection patterns of Delta Smelt in cages during the first cage deployment experiments at Rio Vista and the Deep Water Ship Channel (manuscript in prep). The California Department of Water Resources (DWR) has several active studies currently underway focused on endangered Longfin Smelt and other listed species. The Longfin Smelt team at the California Department of Water Resources (DWR), in collaboration with Cramer Fish Sciences, has also been investigating whether eDNA sampling could be used to detect larval Longfin Smelt entrained at the Barker Slough Pumping Plant, filtering water that was collected from vegetation. DWR and Cramer Fish Sciences are also planning on utilizing eDNA sampling to monitor Chinook Salmon during droughts. A collaborative group lead by the GVL at UC Davis is working to build a database of reference sequences for fish and invertebrates in the SFE to be used for eDNA based studies. They are also comparing eDNA metabarcoding, or identifying a large group of taxa, with long term fish and invertebrate catch data around the SFE to evaluate the success of eDNA sampling as survey method in these monitoring locations.

Four water bottles with tubing connected to a pump and filtration system that is sitting on a benchtop. The setup is designed to filter the water for eDNA.
Figure 2. eDNA filtration set up in the lab. Replicate water grab samples are filtered using a peristaltic pump. Sites with lower turbidity take only a few minutes to filter. Filters can be stored in ethanol, frozen, or dried before analysis.

Challenges of eDNA in estuaries

If eDNA detection can be a helpful tool in our management toolbox, what is hindering us from applying it further in the SFE? There are a few challenges when it comes to utilizing eDNA captured in an estuarine system. Tidally influenced systems make sampling eDNA more difficult; although unidirectional flows found in streams or rivers mean eDNA is transported in the same direction, the sloshing of the tides can make it hard to know how eDNA is being moved around. This means that sampling location (shore vs. boat) and sample spacing will likely vary by species and habitat. A better understanding of hydrodynamics in estuaries would help us to understand how best to capture and interpret eDNA results.

Additionally, the higher turbidities that are found in our estuary can cause filters that capture eDNA to quickly clog, decreasing the amount of water that is filtered and thus decreasing the probability of capturing the eDNA of our species of interest. Larger pore sized filters can be used to compensate for less volume being filtered at higher turbidities, but there is a risk of allowing smaller eDNA particles to slip through the larger pores. Turbidity also can inhibit reactions in the lab, leading to incorrectly assuming eDNA was not found in the samples (false negatives). An additional clean up step to remove inhibitors may be helpful.

Used filter paper sitting on a table top that has a smiley face drawn onto the filter paper.
Figure 3. eDNA is happy to help with your monitoring questions!

Considerations for eDNA applications

The field of eDNA is still relatively new and is most powerful when used to answer questions for which it is well-suited. There are many factors that can impact the distribution and detection of eDNA in the environment such as life stage, temperature, flow rate (in aquatic environments), and degradation by microbes. Scientists are still working to understand how these factors play into the ecology of eDNA, and how to optimize their methods to account for them. Another important consideration for any study involving eDNA is the availability of reference databases. The reference databases for various organisms are constantly expanding and improving, but many species are still missing. While eDNA has been used to determine species presence or absence for some time, there is still uncertainty about its ability to estimate biomass or abundance. As the field of eDNA research continues to expand, many of these limitations will likely be overcome, which will improve the utility of the tool.

Future plans

The California Department of Water Resources (DWR) has initiated a new Genetic Monitoring (GeM) lab that will conduct genetic monitoring and molecular ecological studies using eDNA for water management decision-making within the SFE. As part of the State Water Project and Interagency Ecological Program (IEP), GeM research will prioritize the needs identified within the Incidental Take Permit, Biological Opinions, and water rights decisions for the State Water Project. The new lab will use innovative technology and collaborative partnerships to advance management decision-making critical to the State's water supply operation and planning.

Female scientist bending down next to a running stream collecting a water sample.
Figure 4. Sampling for eDNA is non-invasive and can allow managers to monitor hard to access locations.

Science Stories: Adventures in Bay-Delta Data

Further reading