What we're going to be talking about today are ways in which we can quantify. So plankton composition and abundance within me and estuaries. You take a moment to think about Nain and iconic Maine in particular. Fisheries are very closely tied to mains, economy and cultural identity. At a time where we're seeing fisheries management having more and more questions to deal with climate change and changing waters. So when we start thinking about zooplankton, let's zoom out a little bit. Zooplankton play a pivotal role in aquatic ecosystems and global biogeochemical cycles. They function as prey for economically important fish. There grazers of primary production and their drivers of carbon and nitrogen cycles, their population and community dynamics, including their growth, mortality distribution, and diversity, while structure and shape the ecosystem, these dynamics are incredibly susceptible to changing environmental conditions. At a time when climate change is profoundly impacting marine ecosystems. Research suggests that the Gulf of Maine is warming faster than 99% of other marine systems would further intensifies the need for better understanding of zooplankton community structure and coastal me. So as we said, zooplankton perform critical ecosystem functions by transferring energy between trophic levels. And they provide key forage for research for species. So what does that mean? Research suggests that the amount of primary productivity that is channeled upwards to mezzo zooplankton is actually more closely related with fishery yields than primary production. Long-term changes in zooplankton biomass and species composition had been closely linked with large shifts in biomass and recruitment success. Upper trophic level species and species of economic value here and name zooplankton have been proven to be an effective ecosystem indicator, fisheries productivity, and it's been suggested that changes in zooplankton productivity may also be one of the most important pathways for climate to impact fishery sources in the Atlantic. Much of Mainz economy and cultural identity is integrally linked to marine resources. And resource suggests that the Gulf of Maine is exceptionally vulnerable to overfishing and climate change. So what are we going to do? Environmental DNA technology can reveal how marine food webs are responding to anthropogenic, such as the one face, the main metabarcoding could divide an in-situ snapshot of species composition and diversity and is poised to change the use of traditional trawl surveys to inform stock assessments. However, despite the critical need for more data to inform management decisions, resource managers are still rely on traditional labor intensive surveys and research cruises. The development and calibration of species detection and quantification through noninvasive techniques could significantly alter marine species surveillance at a time when we're facing greater environmental and climate uncertainty. So to better understand how these coastal systems And these coastal communities of zooplankton are shaped and how they're changing. We'd like to use ED and a TA in across, integrate across the main eating a framework to examine zooplankton community structures across multiple estuaries in Maine while developing and calibrating an 80 and a survey for future use. A method for future use. We would like to provide a seasonal scale characterization of zooplankton that will likely provide data relevant deficiency EECS, since most species have spawning and juvenile migration windows, which are narrowly defined in both space and time. For example, if we look at the recent declines in the winter spawning Codd and the northeast continental shelf been closely associated with a reduced the abundance of the CocoaPod soup pseudo colonise in spring, while the decrease of the copepods entropys in autumn has affected the spring spawning caught. So clearly, a coast Wide seasonable assessment of the temporal and spatial variability in zooplankton community composition is required to further our mechanistic understanding of the observed shifts in both fish assemble is and their distribution here in the Gulf of Maine. This could very easily help to inform Ecosystem Assessment initiatives. And we'd like to see this survey have an impact on fisheries management. And there might actually be a commercial application when selecting agriculture. So who are we going to partner with? Us m primarily because this is going to be funded as a graduate proposal here at USF. I'm working in the Lashley rash rational lap. We'd like to partner with you, Maine, on the new DNA grant. So taking a step back prior to its completion in January, 20-20 mains F Score C-net grant, which was a track one grant, had collected an identified zooplankton and five coastal estuaries over three years. Providing baseline community structure was actually involved in the identification and analysis of the zooplankton. So we'd like to add an additional years worth of data onto what we have there. And recently, last August, the main environmental DNA, or main DNA was awarded its a new five-year F Score grant. And it's can we focused on understanding coastal ecosystems and monitoring environmental conditions through the use of DNA technology. So we're hoping to kind of slot into what they're doing to use our technique to help inform other surveys. And we're gonna do most of our metabarcoding and molecular work, either hear us m or sending it out to Bigelow depending on what's available at the time to charge the qPCR machines. And we're hoping to work with cache estuary for some data analysis. So how are we going to do this? Well, we're going to start by collecting zooplankton from five estuaries in Maine, which I mentioned earlier. These are going to be to the bioregions identified in previous unit grants, soko, new meadows, the mayor, Scott, the baggies, and Matthias. And then we're going to match the existing C-net zooplankton library to what we find today. So if anyone wants to know, zooplankton are collected using a traditional thrall method, where we use are really tiny 200 micron net. We bring him back to the lab. I'm usually identify them. So we're going to do that. But we're also going to collect water samples. So we can filter these water samples and run them for DNA analysis. And we're going to extract them and either send that, as I said before, you do it here or at Bigelow. And then we'll do some sequence alignment and look for unknown Africans and start putting together what kind of community structure we have. The data's going to be used to expand that Sina zooplankton survey that we had previously. And we're hoping that as a final step, we can incorporate this collective information with geo referenced sample locations, with hydro morphological elements, and hopefully some phytoplankton and environmental data to kinda see really what's going on in terms of who's the driver is and what's happening within HHS humidity assemblages. So it's a pretty basic plan in terms of how we're going to implement this. We're going to collect samples, we're going to visually identify and run through the metabarcoding. And then we'll move into the analysis and comparison face. We've actually done this over the past year in the lab where we've gone through step 123, the only thing that will be adding at this point in time is adding metabarcoding. So we understand how this works and how the project flop. Who are the people going to be? Dr. Rachel lives. Erasure is going to be the PI project oversight and everywhere. I don't really anticipate her being super hands-on, but my name is Kevin again said earlier, I'll be the project lead. I'll be involved in everything from sample collection to data analysis. We're going to partner with big loafer metabarcoding and hopefully get some there phytoplankton data from Dr. potent. And we'll be partnering with marine darling marine center for launch vessels, for research vessels, for they have a really good understanding of hydro morphological data. And Dr. Danny and Grady up there, and hopefully we get to use some of their lab so we don't have to transport everything to directly back to us. I'm after collection. So how are we going to evaluate this? There is a project management plan that includes benchmarks at all of our different stages. Their collection, identification and analysis. As we were developing a method that is going to be used to calibrate against traditional visual identification surveys. We anticipate that these benchmarks are going to be moving a little bit and will be refined based upon what the results are. But at the end of the day, our goal is to have a metabarcoding survey that is validated and calibrated and we can use it to compare against future results and past results. We're hoping to move this forward of the publication and a thesis. And really at the end of the day, what our driver of successes, can this be used by other, you know, other institutions, other individuals? We're looking to provide better management and better information about coastal ecosystems. So with that, I think I'm just about ten minutes. They got nine seconds left, so I'm going to stop here and it opened it up for questions like, this is Rob, your graphics are terrific. I, I want to see references in the slides. Has been going up some things that you're proposing. Your method is it I'll and and it needs to be standard in the literature, for example, your water quality sampling procedure, things like that. So you should address that and cite the source in cars like are using EPA methodologies, stuff like that. You're dissemination of your results, you've got publication and then thesis. Normally it's the other way around. So because I've seen people's theses and dissertations held up or something like that. So you should have your publication phase should be its own category, dissemination. So dissemination is not It's, it can be a part of a new evaluation phase, but it's much more that it's a scientific obligation, share this information. So I would, I would say it's worthy of its own strategy that learn to. I've met on some of those other ones, like you plan to present this at Ecological Society of America and various things like that main water conference, I would say that overall the structure is sound, the intellectual argument is sound. But you need that peer review, that literature review, to show why you're doing AS your benefits of it are going to be economic, research, environmental. And if you're looking at any federal agency, this kinda funded, they're going to want to see you hit all three of those components. Thanks. Great. Thank you. I really appreciate that. And cabinets. Richard, I have the same question that I had, I think for Leah, which is the budget seems really modest. Janae, given what you're hoping to accomplish, and I just want to make sure you've thought through really where all the potential cost streams might be in other places where you might need funding because I wouldn't want to see you get halfway through and a really modest budget. Realize that you have run out of money and have to raise more, et cetera, et cetera. So I just I don't know if you feel like you've really kick the tires and the budget really is sub $5 thousand. Or if there are other ways in which you might want to round that budget out a little bit more. Because it seems like you're doing a lot of research for not much money. Yeah. I mean, this was this was under the under the sea grant proposal for the old score track one. And there was a there was a cap in terms of how much value this grant could be, four. So what I did is I went through Main Street and procurement and got all the quotes for, you know, our primers and everything else. And then try to figure out like what's the most strategic way to do mileage. But I think that as we start going forward, you know, there's a lot of variables like extra for Berlin or like this needs to be sent off to Bigelow. And we need extra like FedEx cost that needs to be built in there. And that's just something I appreciate you bringing up. Say, I know that for moving us forward into something real that has to be fleshed out a little more. Thank you. Thank you. And Kevin, That was a really good explanation in response to Richard's question. And so I mean, even if you are under resourced that you went to that level of research to get at those figures, I think is merits some brief discussion. Your budget narrative. So I mean, I had my only question is, and again, I thought you did a really good job with the literature. And you know what, one of the things I just was kind of wondering about in your methods, you said that in the narrative 0s there was you have three years of baseline data, you get that correct and I get that right. Which lets now go ahead. Go ahead. I'm sorry. First at which was a previous main AP score grant. There was an election of zooplankton collected my colleague, Molly and myself over the past year, I've been doing the visual identification data of those samples. Ok. And so over from year from for those three baseline years, Have you seen any kind of dramatic drop off in zooplankton in those years are what we're seeing a lot of interesting trends within the estuary to estuary comparison. You know, I'm fortunate there hasn't been more done in terms of looking at it like large-scale coastal estuaries over time. Yeah, I was really hoping that prior to all this happening that we could really add another year this year just to keep that time series going. And then I was also looking forward to possibly looking at, Hey, what's going to happen, but we don't have as many humans running up and down with the virus got a river. And does that affect anything? But unfortunately, I was told recently that like, everything is shut down until at least July for collection. So I think we're going to skip this year. Okay, alright, well again, I enjoyed it and very good and good luck to you on this one. Thank you very much. Yeah, this is Rob. We're seeing environmental DNA is it's the sprouting, it's bursting out all across the academic landscape and everyone is excited about it. I think I would be interested in seeing your project and the context one of this overall use of ED. And a two, in the face of climate change, because we're looking at some of the, some major changes and coastal ecology as a result of that. And also as we begin to develop coastal Maine for aquaculture and other projects where, where different things are going to go into the environmental background. So you might be interested in thinking about your project in the context of that. And I think everyone should take Richard's comments to heart. And many of us are so eager to get these grants that like I said before, I actually get disappointed when I get one because I started myself on what I have to do for my match. But you can include matches if you all have dissemination sections that'll make you make that a budget item. And so that you can include PE meant to go to a conference and things like that and sharing these results because that's your number one. That's what makes this academic is finding out information and sharing that information. Because information that you find out that doesn't get shared doesn't have academic relevance. Thank you. Ever. I really appreciate the feedback.