WEBVTT 00:25:21.000 --> 00:25:51.000 Okay. 00:25:54.000 --> 00:26:24.000 Okay. 00:27:29.000 --> 00:27:38.000 Thank you, Don, for that introduction and thank you all for coming out to my presentation defense early on a Friday morning, especially those on Zoom that are on the West Coast. 00:27:38.000 --> 00:27:41.000 I really appreciate it. 00:27:41.000 --> 00:27:53.000 And so today I'll be presenting my work for my dissertation, which looked at morphological plasticity and trophic ecology in Gordonian octopus. 00:27:53.000 --> 00:28:06.000 We're organisms can exhibit expensive intra-specific variation. And this phenotypic variation can be pathological, physiological, or behavioral, and can be caused by genetic variation. 00:28:06.000 --> 00:28:18.000 A classic example of intro specific variation in coloration is in the peppered moths where during the industrial period black individuals were selected for as they could remain cryptic against darker backgrounds. 00:28:18.000 --> 00:28:40.000 More so than the classic white or lighter color. Another classic example is in coloration in male transatlantic guppies where the color pattern in nails has been extensively studied and is maintained due to natural selection, as well as through sexual selection. 00:28:40.000 --> 00:28:53.000 However, intro specific variation can also be due to the plasticity. And we define scientific plasticity as when one GAN type expresses a different phenotype in different environments. 00:28:53.000 --> 00:29:02.000 One classic example of scientific plasticity. Is the increase of spines and helmet size in Dafnia under increased predation. 00:29:02.000 --> 00:29:11.000 Another example that's interesting is in the tropical Mycalessian butterflies were under the dry season. 00:29:11.000 --> 00:29:18.000 They exhibit the wing patterns more similar to those on the left of these individuals where they're able to remain cryptic in the leaf litter. 00:29:18.000 --> 00:29:29.000 We're doing the what see then the exhibit wing patterns more similar to the right wings on the individuals where they are able to evade predation by developing eyespot. 00:29:29.000 --> 00:29:36.000 Many studies on corals have documented intro specific variation between populations in inshore shallow environments. 00:29:36.000 --> 00:29:48.000 And compared to those in offshore deeper environments. These environments differ in many different environmental factors including sunlight, water movement, and nutrient availability. 00:29:48.000 --> 00:29:55.000 And previous studies on symbiotic Nigerians or Nigerians that host ended symbiotic algae and their tissues like this one. 00:29:55.000 --> 00:30:09.000 I've found morphological and physiological differences. Green individuals in these different environments. And these studies have been done on toxic, such as the nevenes. 00:30:09.000 --> 00:30:17.000 But what are gorgonian octoporals? Despite being a toxo that exists in all seeds and ocean around the globe. 00:30:17.000 --> 00:30:27.000 This octopus are often frequently under-studied or overlooked. One common order within octoporalia are Gorgonian octopus. 00:30:27.000 --> 00:30:39.000 This diverse group exists from shallow to and are characterized by its central protonation access that gives them their structure, which is surrounded by calories. 00:30:39.000 --> 00:30:53.000 And squares are these calterious structures similar to spectral sponges and that they provide support and structure for the individual as well as that they're shape and composition is species specific. 00:30:53.000 --> 00:31:04.000 So octopus are prominent members in both tropical and temporary communities and they provide 3D habitat for both invertebrate. 00:31:04.000 --> 00:31:07.000 Those are an example. 00:31:07.000 --> 00:31:17.000 So for my dissertation, I looked at interest specific variation in both temperate It's antibiotic octopus as well as tropical symbiotic octopus. 00:31:17.000 --> 00:31:23.000 And today I'm going to tell you about some of my work from all 3 of my chapters. 00:31:23.000 --> 00:31:32.000 The 1st 2 parts of this talk will focus on work that I conducted in the Gulf of Mexico on an asymptotic species of octoporal, left the gorge of her gulada. 00:31:32.000 --> 00:31:45.000 I'll start by briefly discussing the color more distribution in this species as well as my findings from a study that I conducted on their reproduction as well as a genetic analysis between color and location. 00:31:45.000 --> 00:31:52.000 And then I'll move into talking about my findings, looking at phenotypic plasticity and genotype by environment interactions. 00:31:52.000 --> 00:32:00.000 Finally, I'll transition to some of my work on 2 symbiotic Caribbean octopus species in the genus Pseudoplexora. 00:32:00.000 --> 00:32:11.000 Alright, documented intra specific variation in fault morphology and investigated whether these interest specific differences are correlated with variation in tropic stress. 00:32:11.000 --> 00:32:19.000 So to start off, let me introduce you to the populations of what the Virgilata in the northeastern Gulf of Mexico. 00:32:19.000 --> 00:32:30.000 Okay, Gilaudet is an asymptotic octopus world species, meaning that it does not host Algol and dots and and instead derive its nutrients purely from heterootropic beta. 00:32:30.000 --> 00:32:40.000 It has a wide range spanning from the Chesapeake Bay on the east side of the US, down that coast into the Gulf of Mexico and down to Brazil. 00:32:40.000 --> 00:32:46.000 Alberta is commonly referred to as the colorful seaweed. I'm sure from these photos you can see why. 00:32:46.000 --> 00:32:51.000 In the Gulf of Mexico, there are 3 predominant color morphs that can commonly sound. 00:32:51.000 --> 00:32:56.000 And their magenta, yellow, and orange. 00:32:56.000 --> 00:33:07.000 In the northeastern Gulf of Mexico, Elver Gilada is patches to hard substrates such as limestone reefs and outproppings like this one shown here as well as artificial reefs. 00:33:07.000 --> 00:33:14.000 And the transact shown here, it's taken at limestone reefs offshore at Goliath Superhole. 00:33:14.000 --> 00:33:29.000 You can see that octoporals including, Elvis, which is here. Here, provide a lot of that 3D structure that exists off of the, in the tab. 00:33:29.000 --> 00:33:34.000 Cause that's our marine and vertebrates are actually some of the most colorful marine ordnance. 00:33:34.000 --> 00:33:42.000 It's a embryonic octopus especially are known from being for being more pigmented and polymorphic and coloration than symbiotic octoporals. 00:33:42.000 --> 00:33:51.000 And then this has been attributed to selection for an increase in segmentation to prevent UV damage in the app. 00:33:51.000 --> 00:34:01.000 Coloration and left of Gorgilata is produced in their square rights and is genetically determined hypotheticalically through one gene with at least 4 alleles. 00:34:01.000 --> 00:34:09.000 In several closely related left to gorgeous species that are similar in coloration. Collaration has been attributed to the compounds. 00:34:09.000 --> 00:34:19.000 CITY. Interestingly, it's a couple of them prior to being identified in October, had previously only been found in Paris. 00:34:19.000 --> 00:34:32.000 All the function in active world has not been studied. In parrots, this compound has been attributed to decreases in disease as well as arasian resistance to their products. 00:34:32.000 --> 00:34:40.000 Anecdotally, it has been noted that in Elver Gilotta, the color distribution of of the species is dependent upon the region that they're located. 00:34:40.000 --> 00:34:48.000 So for example, in the trust and as well as off of Pensacola, Florida, they tend to be predominantly in Genta. 00:34:48.000 --> 00:34:56.000 However, populations off the coast of Maryland and the orange those off of North Carolina tend to be yellow and orange. 00:34:56.000 --> 00:35:10.000 And so because of this variation, dependent upon region. I wanted to start by quantifying the color more distribution at inshore and offward sites in the northeastern Gulf of Mexico in this area. 00:35:10.000 --> 00:35:16.000 Using 3 35 meters squared belt transects similar to the one that I showed you this video. 00:35:16.000 --> 00:35:17.000 At 4 ensured sites shown here in the cooler colors and for offshore sites shown here in the warmer colors. 00:35:17.000 --> 00:35:28.000 I documented the color more distribution in this region. 00:35:28.000 --> 00:35:40.000 Here we found that in short sites tended to have greater color more variation than offward sites where insurance sites are comprised mostly of purple and yellow morphs as well as the occasional orange marks. 00:35:40.000 --> 00:35:51.000 Whereas offshore sites are almost entirely. And very rarely yellow or important. And so given this, this difference in distribution between the color marks. 00:35:51.000 --> 00:36:03.000 I wanted to look at mechanisms that may be maintaining this difference in distribution, including barriers to reproduction as well as genetic 00:36:03.000 --> 00:36:13.000 So reproductive isolation of broadcast spawning. She is that live in can occur either through differences in timing for their spawning. 00:36:13.000 --> 00:36:20.000 As well as through Canadian incompatibility. The 1st to test for reproductive isolation between the color morphs of Alberta. 00:36:20.000 --> 00:36:32.000 I conducted a spawning study in the summer of 2022. Overgalata is gonacaristic and spawns daily at sunrise from April through September. 00:36:32.000 --> 00:36:39.000 And so I collected both magenta and yellow color works as they're most abundant at the end for sites. 00:36:39.000 --> 00:36:45.000 And I collected them from a singular in short site, this purple site here, Wilson Beach. 00:36:45.000 --> 00:36:51.000 And so just like as anecdotally these colonies are living in very close proximity. 00:36:51.000 --> 00:36:56.000 Send on the meeting directly next to each other. 00:36:56.000 --> 00:37:04.000 So 1st I determined to call and need sex using light microscopy. We're, they were, they were. 00:37:04.000 --> 00:37:11.000 Delineated by the presence of either eggs or sperm, like eggs shown in this yellow. Individual and sperm. 00:37:11.000 --> 00:37:17.000 Do you want to individual? And then each day at sunrise I paired females and males together. 00:37:17.000 --> 00:37:31.000 And allow them to spawn. All these that spawn. I documented the time at which female eggs began spawning each day because they were visible in the water column. 00:37:31.000 --> 00:37:40.000 And then I waited a few hours until fertilized egg began to cleave and I scored fertilization success. 00:37:40.000 --> 00:37:52.000 After a few hours, eggs became swimming, And eventually some of them even successfully deter, develop into follow-up after. 00:37:52.000 --> 00:38:07.000 So 1st I calculated adjusted female. Spawning time for each day. Where the 1st female that spawned that day was assigned time 0 and each female that spawned subsequently was assigned number of minutes from time 0 that day. 00:38:07.000 --> 00:38:21.000 And I found that both magenta and yellow females did not differ from one another. Indicating that there is no evidence for barriers to cross fertilization due to differences. 00:38:21.000 --> 00:38:32.000 Additionally, I found no evidence for incompatibility as eggs were equally as successfully fertilized by sperm of the same work as well as they were from sperm of different parts. 00:38:32.000 --> 00:38:39.000 And therefore we found no detectable barriers to hospitalization between, or magenta and yellow color morphs. 00:38:39.000 --> 00:38:43.000 That occur in the same population. 00:38:43.000 --> 00:38:51.000 Additionally, we wanted to look for evidence of genetic divergence between both color morphs as well as in short offshore sites. 00:38:51.000 --> 00:39:08.000 And so we sequence 2 mitochondrial, I mute S and N 2. And 2 nuclear loci ITS 2 and We found variation present at US and EF one A, but not for ND 2 or ITS. 00:39:08.000 --> 00:39:18.000 And so we wanted to look for whether or not these this variation in mute s. Was due to differences in color more or location. 00:39:18.000 --> 00:39:32.000 And so 1st the phased EF one A sequences. The variable sites of the EFNA sequences and then concatenated them to the variable sites of S to create a have a tight map. 00:39:32.000 --> 00:39:41.000 And so this haplotype map, the nodes are colored by the proportion of individuals. That are different colors and in their different locations. 00:39:41.000 --> 00:39:50.000 And these colored curves connect. Couple of types that come from the same individual, but are different types. 00:39:50.000 --> 00:40:01.000 Each individual break in the line is a singular And so what we see is no detectable. Differentiation between color or location. 00:40:01.000 --> 00:40:12.000 As they're intermixed. And so therefore, genetic differences in color or differences in distribution of color are unlikely due to genetic divergence in the system. 00:40:12.000 --> 00:40:22.000 However, for future experiments, we opted just to use the magenta morphs. Just due to their difference in distribution. 00:40:22.000 --> 00:40:31.000 They're next to the speeches, I looked for phenotypic plasticity and genotype by environment interactions at inshore and offshore sites. 00:40:31.000 --> 00:40:40.000 Scientific plasticity is especially important for sessile organisms as they're unable to move in response to environmental change. 00:40:40.000 --> 00:40:50.000 Costicity can therefore buffer individuals from seasonal environmental fluctuation as well as help by time in the event of long term environmental change. 00:40:50.000 --> 00:41:01.000 Well, plasticity is often thought of as having positive influences on individuals. Under unpredictable environmental variation or environmental variation with unreliable fuse. 00:41:01.000 --> 00:41:05.000 Paste City can also be. 00:41:05.000 --> 00:41:16.000 Additionally, plastic responses can be variable and this is due to both variation in the environment. As well as variation in the response to the environment by different genotypes. 00:41:16.000 --> 00:41:26.000 Known as genotype by environment interactions. And so, you know, type by environment interactions or do by E interactions can therefore provide a genetic basis. 00:41:26.000 --> 00:41:36.000 For plasticity as a trait by which selection can act upon. To the 00:41:36.000 --> 00:41:42.000 It's a big foster city in symbiotic. Sexile Nigerians is expensive. 00:41:42.000 --> 00:41:59.000 It's embryonic octave worlds. Colonies at insured sites tend to have a higher density of polyps as well as a higher density of branches or as well as smaller square rights as a reminder that square it's are similar to spatial open sponges. 00:41:59.000 --> 00:42:10.000 And typically these plastic trends are attributed to changes in light of availability, which can influence the photosynthetic ability of the symbiotic algae. 00:42:10.000 --> 00:42:24.000 As well as differences in water movement. Which can influence the hydrodynamics of heterotrophic feeding their respiration as well as the potential for breakage and displacement of the colonies. 00:42:24.000 --> 00:42:43.000 Pages in both light and water movement. Often co occur, making them difficult to parts apart. In the field and these plastic responses in multiple species of octave world may also be driven not only by the needs of the host but also by the needs of their 00:42:43.000 --> 00:42:50.000 Well, previous studies on plastic or on plasticity in octic worlds have focused on species with endants. 00:42:50.000 --> 00:42:59.000 The majority of octopural species around the globe are actually asymptotic. So by examining plasticity in an asymptotic species of octic world. 00:42:59.000 --> 00:43:17.000 We're not only able to increase our understanding of philosophy and octic worlds. In general, but we're also able to eliminate the demands of light availability, allowing us to parse apart morphological responses that may be due to light availability. 00:43:17.000 --> 00:43:26.000 Sorry, chose to study plasticity in Elver, ilata in the Gulf of Mexico as the Gulf of Mexico is known for being a highly variable body of water. 00:43:26.000 --> 00:43:32.000 And I go to leave this might look like someday being able to see your buddy and the work that you're working on. 00:43:32.000 --> 00:43:39.000 And some days you're just hoping that you've descended on the correct site and you're holding your buddy's hands so that they don't lose you. 00:43:39.000 --> 00:43:47.000 But in all seriousness, inshore and offered sites in the northeastern Gulf of Mexico can vary from one another quite a bit. 00:43:47.000 --> 00:43:59.000 One main difference between these 2 locations. Our differences in variable water flow. And so at in short sites, they tend to have greater turbulence and water movement that's driven by both wind and tides. 00:43:59.000 --> 00:44:05.000 Resulting in multi-directional water flow. The more I know what you might experience while swimming at the beach. 00:44:05.000 --> 00:44:13.000 Comparatively offshore sites tend to have weaker bottom currents that tend to be unidirectional and are driven by tides. 00:44:13.000 --> 00:44:23.000 Well, initially, you might think that variation of water flow might primarily affect the colonies mechanically by or dislodgement. 00:44:23.000 --> 00:44:30.000 However, variation in water movement can also result in physiological 00:44:30.000 --> 00:44:38.000 Changes in water movement can influence various aspects of octoporal lights, including feeding, respiration, and metabolism. 00:44:38.000 --> 00:44:46.000 Under low flow. Branches like those shown in A. When they have a food particle that is approaching them. 00:44:46.000 --> 00:44:58.000 The food particles likely to be captured by either that front polyp or a polyp on the side. However, under moderate flow or high flow, that food particle is likely to bounce off of that front polyp and even side polyps. 00:44:58.000 --> 00:45:07.000 And may as likely to be captured if it's captured by either the bulb on the back side of the branch or subsequent call-ups downstream of that call. 00:45:07.000 --> 00:45:19.000 And this is one hypothesis for why under increased water, flow, we get, ranching and pallop density as it improves heterotrophic feeding efficiency. 00:45:19.000 --> 00:45:24.000 Additionally, flow speed can also influence the boundary layer. 00:45:24.000 --> 00:45:32.000 This layer exists around the coral colony and causes a decrease in the diffusion of both gases and metabolites. 00:45:32.000 --> 00:45:50.000 And so the flaws changes in metabolism as well as respiration. And so under high flow, that boundary layer is actually shrink it shrinks and therefore they can have an increase in metabolism and respiration which might contribute to increases in growth. 00:45:50.000 --> 00:45:58.000 So while many environmental factors that ensured sites and offshore sites, can influence choral physiology and phenotype. 00:45:58.000 --> 00:46:20.000 Water Movement, direction, and velocity is one that can have a multitude of effects. Since it's difficult to parse apart light and water movement in the field in symbiotic octoporal species, I aim to investigate how flow independent of the demands of light availability can influence colony more quality. 00:46:20.000 --> 00:46:34.000 Given findings from previous studies on intra-specific variation in octopurals, we expected that the morphology of Elver Gilotta would likely differ between inshore and offshore populations, but we wanted to start off by documenting that. 00:46:34.000 --> 00:46:52.000 Before testing whether or not it was due if it was present the genetic differences or specific plasticity. Again, because of the differences in distribution and the color morphs in this region, I only compared the morphology between magenta colonies that in shore and offward sites. 00:46:52.000 --> 00:47:04.000 So at each site, again, you think those transects that I showed you before. I surveyed magenta colonies, measuring the height, counting the number of branches to get a metric, but pushiness. 00:47:04.000 --> 00:47:12.000 And taking tissue samples to measure the average branch. The polypensity and the square length. 00:47:12.000 --> 00:47:20.000 How's your reminder, in previous studies on symbiotic octic worlds, what we tend to find is that those inshore sites we see an increase in bushiness. 00:47:20.000 --> 00:47:26.000 All of density and a decrease in square rate size. 00:47:26.000 --> 00:47:35.000 Using a, I compared the bushiness between inshore sites shown here in block and offshore sites shown here in block and offshore sites shown here in, right? 00:47:35.000 --> 00:47:45.000 And I found a significant difference in bushiness between those 2 locations, where colonies at ensured sites tended to be bushier than colonies at offshore sites. 00:47:45.000 --> 00:47:50.000 And this difference occurred even in the absence. 00:47:50.000 --> 00:47:58.000 However, comparing all the other morphological traits that we looked at, we saw no difference in morphology between any other trait. 00:47:58.000 --> 00:48:03.000 Offward sites including all density and square. 00:48:03.000 --> 00:48:12.000 Therefore, while studies on symbiotic species have found differences in pushiness, pulp, morphology, and square it size, between inshore and offered sites. 00:48:12.000 --> 00:48:19.000 The only found differently than bushy. It's possible then that differences in pushing it may be driven more. 00:48:19.000 --> 00:48:26.000 You too, water movement, then light availability. Given the differences in pushing us between inter and offshore sites though. 00:48:26.000 --> 00:48:32.000 I wanted to investigate whether this difference would be the specific process. 00:48:32.000 --> 00:48:41.000 So to investigate this, I conducted a reciprocal transplant study where I collected 8 colonies from 3 different sites. 00:48:41.000 --> 00:48:47.000 I brought them into a lab. I fragmented them into 9 branch tips, roughly 14 cm in size. 00:48:47.000 --> 00:48:56.000 And I transplanted 3 replicates of each colony. Back out to all 3 original sites for a fully reciprocal design. 00:48:56.000 --> 00:49:02.000 And so that's total 96 fragments going to each site and a total of 288 across the whole study. 00:49:02.000 --> 00:49:08.000 And so these call, these fragments were attached. The PDC frames which were anchored to the Ventos. 00:49:08.000 --> 00:49:12.000 And you can see in this photo, the 00:49:12.000 --> 00:49:21.000 So in total they were out for a total of 70 to 71 days between July and September in 2022. 00:49:21.000 --> 00:49:28.000 The 3 sites that we use to this experiment varied in their distance from shore, where site one, the purple, site. 00:49:28.000 --> 00:49:35.000 Was the closest to short? It was around 2 meters in depth and less than a kilometer short. 00:49:35.000 --> 00:49:41.000 And since it's so close to shore in these freshwater inputs, this site likely has the most environmental variation. 00:49:41.000 --> 00:49:47.000 Site 2 is Turkey point show east. Which was around 3 meters deep and 2 kilometers from short. 00:49:47.000 --> 00:49:54.000 And site 3, which was Turtle Towers. Was around 14 metres deep and 20 kilometers from shore. 00:49:54.000 --> 00:50:01.000 So that's our offshore site that likely experiences less environmental variation than the 00:50:01.000 --> 00:50:11.000 How's the brief reminder? Under no plasticity, we expect that an individual when under different environments will continue to express the same Okay. 00:50:11.000 --> 00:50:20.000 However, plasticity is occurring. We expect that that individual will respond and its phenotype and change phenotypes with changes in the environment. 00:50:20.000 --> 00:50:28.000 And so for this experiment, I looked to search the treat relative growth and branching as we saw the differences between branching it into an awkward. 00:50:28.000 --> 00:50:34.000 Site. Okay. Relative growth, I included both positive growth as well as no growth. As well as a reduction in growth or shrinkage. 00:50:34.000 --> 00:50:45.000 The species have been shown to shrink, especially in the winter time, and recede its tissue. 00:50:45.000 --> 00:50:50.000 So let me start by orienting you to this figure as I'm about to show you several of them like this. 00:50:50.000 --> 00:50:55.000 So along the x-axis, we have the destination site that the colony was transplanted to. 00:50:55.000 --> 00:51:04.000 With the distance from shore of that site as a reminder. Along the y-axis, we will have the trait that we measured, in this case the proportion of individuals that branched. 00:51:04.000 --> 00:51:14.000 And the color of the point within each destination site. Will be the site that that colony is collected for, selected from, or its origin site. 00:51:14.000 --> 00:51:19.000 And so what we found is a significant effect of transplant destination on the proportion of colonies that branched, but not origin nor an interaction of origin and destination. 00:51:19.000 --> 00:51:37.000 So this is an example here actually of the same genotype that was transplanted to an site one versus offshore site. 00:51:37.000 --> 00:51:45.000 In terms of relative growth, we again found a significant effect of destination, but neither origin or the interaction of origin and destination. 00:51:45.000 --> 00:51:54.000 This time there was more variation. Within each within the origins within a destination. And we again see a similar trend. 00:51:54.000 --> 00:51:59.000 Where those that were transplanted to inshore site one tended to grow as well as branch. 00:51:59.000 --> 00:52:12.000 More than when they were transplanted sites 2 and 3. And as a note, this dashed gray line here is indicative of no change in size, so anything below it actually exhibited shrinkage. 00:52:12.000 --> 00:52:20.000 And so again, in general, we're seeing those patterns of increased growth and branching. Specifically at that in short fight, which is likely the most variable and exhibiting the modes. 00:52:20.000 --> 00:52:25.000 Turbulent water. 00:52:25.000 --> 00:52:39.000 Next I wanted to examine whether there was genotype by environment interactions present. So again, as a reminder, with plasticity but no genotype by environment interactions, we expect that genotypes will respond to the environment but the same way as one another. 00:52:39.000 --> 00:52:48.000 However, when G by E is present, we expect that genotypes will respond differentially from other or interact. 00:52:48.000 --> 00:52:56.000 And so because we didn't find, we found very few individuals that actually branched when they were transplanted to site 2 and 3. 00:52:56.000 --> 00:53:02.000 We only looked for Dubai in relative growth. 00:53:02.000 --> 00:53:12.000 Go on this figure, each line is the average relative growth of a singular type. And the color of that line is the origin that the genotype is coming from. 00:53:12.000 --> 00:53:24.000 And so we see that in general, you know, types when transplanted site one. Tend to have the highest relative growth and that decreases with increasing distance from short. 00:53:24.000 --> 00:53:36.000 However, we do see a significant genotype by environment interaction here, those crossing. And so this tells us that there is a genetic basis on which selection could potentially act. 00:53:36.000 --> 00:53:44.000 On relative growth, leading potentially leading to the evolution of Relative growth plasticity in the system. 00:53:44.000 --> 00:53:53.000 But then I wanted to look at whether or not there was variation in GYE interactions that occur between origins or between genotypes from different origin. 00:53:53.000 --> 00:54:09.000 And so to do this, I calculated the individuals from the model that I used. To analyze G by interaction, to, That's so I calculated the residuals and took their absolute value to be able to compare the variation in Dbody interaction. 00:54:09.000 --> 00:54:15.000 Among origins at different destinations. 00:54:15.000 --> 00:54:21.000 So to briefly reorient you to this figure because this one is the opposite layout of the ones I've just shown you. 00:54:21.000 --> 00:54:28.000 This time along the x-axis. We have origin or the site that the colony came from with its distance from shore. 00:54:28.000 --> 00:54:36.000 And the colored points now are the destinations they're going to. And again, that y-axis is the absolute value of the residual or that variance term. 00:54:36.000 --> 00:54:49.000 The reason we flip this figure is because we wanted to directly compare how Oh, our genotypes from one origin performed differentially or had different variations in 00:54:49.000 --> 00:54:50.000 And so in general, we found that origin and destination both significantly influenced the variation in D. 00:54:50.000 --> 00:54:58.000 Ii interactions. 00:54:58.000 --> 00:55:14.000 More specifically though, using a 2 key code talk test, we see that unit types from site one. Tended to have the highest variance in G by interactions when they were transplanted to their home site and when they were transplanted to the 2 away sites. 00:55:14.000 --> 00:55:25.000 Typically we expect that genotype by environment interactions in theory. Should increase when they're once genotypes are transplanted to and away site. 00:55:25.000 --> 00:55:32.000 As you, it's pretend they have the potential to release. Kinetic variation are hidden in action forms. 00:55:32.000 --> 00:55:36.000 However, we found the inverse of this for genotypes from site one. 00:55:36.000 --> 00:55:45.000 On the contrary though, from site 2 did show this pattern where when they were transplanted to site 2 they had the lowest variation. 00:55:45.000 --> 00:55:51.000 Via interactions and the highest. At the 2 sites that you were transmitted to away. 00:55:51.000 --> 00:56:10.000 Finally, Judith Pipes from site 3 express the same variance in genotype by environment interactions, which could potentially be due to the fact that these unit types originated from the site for this from shore, which is likely the most stable under which variation of plasticity might be less beneficial. 00:56:10.000 --> 00:56:24.000 In summary, genotype, genotype origin significantly affected the variance in G by E. And so populations, each of these 3 sites were evolving independently from other from one another due to isolation. 00:56:24.000 --> 00:56:33.000 We could include the genotypes from site 3. Again, maybe expressing this decrease in variance due to having their home site then. 00:56:33.000 --> 00:56:40.000 The site with the least amount of variance. We could also potentially conclude that Gina types from site 2. 00:56:40.000 --> 00:56:49.000 Are showing an increase in variance when transplanted to a way sites as we're seeing. A release of hidden genetic variation or hidden reaction. 00:56:49.000 --> 00:56:59.000 However, because we found no evidence for genetic divergence in my 1st chapter, The pot these populations are likely connected by gene flow. 00:56:59.000 --> 00:57:14.000 And so the differences that we see between origins in the G by E variation. Good potentially or likely you to carry over effects or internal effects that they're experiencing at their home site. 00:57:14.000 --> 00:57:22.000 And so future work is necessary to far support the drivers of these patterns. Invariants that we're seeing here. 00:57:22.000 --> 00:57:28.000 I'll briefly summarize my findings from the Gulf of Mexico before I move into my findings from the Caribbean. 00:57:28.000 --> 00:57:43.000 We found that branching in Alberta. Was plastic in response to an offshore environments, similar to findings previously documented in symbiotic However, we did not find differences in public density or square right size. 00:57:43.000 --> 00:57:52.000 And therefore, it is likely that differences in branching are essentially more. Due to changes in water, than differences in light available. 00:57:52.000 --> 00:58:00.000 Additionally, we found differences in the variation in GBI interactions driven by both colony origin and transplant destination. 00:58:00.000 --> 00:58:14.000 Providing insights into the complex dynamics, underlying evolution. Of plasticity and system. However, it's important to note that in this study we were unable to replicate the exact requirements. 00:58:14.000 --> 00:58:23.000 And so these findings related to environmental variation in G by interaction variation are correlated. 00:58:23.000 --> 00:58:30.000 Additionally, while we uncovered variance in G by interactions, we're unable to determine the underlying mechanisms driving these interactions. 00:58:30.000 --> 00:58:38.000 Again, they could either be due to independent evolution if there is some sort of isolation occurring at these sites. 00:58:38.000 --> 00:58:48.000 However, the genetic analysis that we did doesn't suggest that and so it's likely that these are due to carry over effects for maternal effects experience other sites. 00:58:48.000 --> 00:58:57.000 And so the further I look into that, we would need to do some sort of population genetic studies. On to determine the differences. 00:58:57.000 --> 00:59:00.000 Between these results. 00:59:00.000 --> 00:59:05.000 Alright, so. 00:59:05.000 --> 00:59:15.000 Having looked at interest specific variation in morphology. I wanted to investigate whether intra-specific variation would be morality could be linked to trophic ecology. 00:59:15.000 --> 00:59:23.000 And for this I use 2 species of pseudoplexor, a mixatrophic octopural species or genus in the group. 00:59:23.000 --> 00:59:32.000 And so I talk up until this point has focused entirely on asymptotic species or species that are only obtaining nutrition through heterotrophic feeding. 00:59:32.000 --> 00:59:38.000 However, many Caribbean octoporal species, especially those in fellows are actually symbiotic. 00:59:38.000 --> 00:59:44.000 Meaning that they're obtaining nutrition through both. Better, trophy heating, as well as. 00:59:44.000 --> 00:59:59.000 You go photosynthetic. Alger. However, even within symbiotic species, the amount of reliance on hetero trophy versus autotrophy just exist on a 00:59:59.000 --> 01:00:12.000 Speaker at all. 2015. Down using stable carbon and nitrogen isotopes that the average polyp size of this octic world species was correlated with the amount of heterotroph. 01:00:12.000 --> 01:00:20.000 So they measured polyp size here using Kaelic step or that depth of the opening of which all of its coming out of. 01:00:20.000 --> 01:00:30.000 And so they found through this decrease in Delta C 13 of the host. That colonies with larger polyps like these are more heterotrophic than collies. 01:00:30.000 --> 01:00:41.000 It's fine. However, the study was conducted at the species. And we know from previous studies that Polymer Quality can vary in trust within. 01:00:41.000 --> 01:00:51.000 So for in, for example, in 30 have shown that colonies tend to have larger polyps at deeper depths. 01:00:51.000 --> 01:01:04.000 Then they do at shallower environments. And so because polypsides at the species level has been associated with Perfect strategy and because follow-up sides can vary at interest specifically. 01:01:04.000 --> 01:01:16.000 I wanted to investigate whether interest specific variation of qualified was correlated to trophic strategy, flexibility or differences. 01:01:16.000 --> 01:01:22.000 Good to do this. First, st I, over 250 colonies and pseudo-plexora genus. 01:01:22.000 --> 01:01:31.000 Over 2 years in the US for violence. And the survey was, these surveys were conducted from between one and a half to 13. 01:01:31.000 --> 01:01:41.000 Do this, I photograph colony branches using a scale bar and using image day I measured the length of the tentacles for 10 different 10 different 10 from Polls. 01:01:41.000 --> 01:01:53.000 On each call. We completed the survey at the genus level because species within pseudoplexor can be quite difficult to delineate. 01:01:53.000 --> 01:01:59.000 You surveys were conducted, at all sites shown on this figure. So 3 sites off the north side of St. 01:01:59.000 --> 01:02:04.000 Thomas and wide sites off of the south side of St. John. 01:02:04.000 --> 01:02:14.000 And through the survey, I found that There is interest specific variation or there is variation at the DNF level in technical length. 01:02:14.000 --> 01:02:21.000 We're at deeper depths. We see an increase in technical length. 01:02:21.000 --> 01:02:34.000 However, to investigate differences in trophic strategy, I wanted to look at the species level. And so to do this, I use tutoxora perosa and pseudoplex or a wagner eye, which again, we're very, as you can see, difficult to 28 in the field. 01:02:34.000 --> 01:02:40.000 And so I used sclerite morphology in the lab with selected tissue. 01:02:40.000 --> 01:02:48.000 I conducted these sample collections. From 2022 to 23. From 4 sites, 3 on the north side of St. 01:02:48.000 --> 01:02:52.000 Thomas and one on the south side of Saint John. 01:02:52.000 --> 01:03:00.000 And so again, I took I photographed these colonies and measured their tentacle length and then I collected a tissue sample and preserved it for chlorophyll. 01:03:00.000 --> 01:03:08.000 We can use it the property for. Thank the sets. As well as for Delta C, 13 and Delta N, 15 stable isotope analyses. 01:03:08.000 --> 01:03:20.000 Prior to isopopanalysis, I partitioned the host and symbiotic. Samples, any 2 separate fraction to be able to analyze and dependence. 01:03:20.000 --> 01:03:26.000 In terms of technical length, what we find in both of these species, it's similar to what we found at the penis level. 01:03:26.000 --> 01:03:38.000 We're shallow colonies, which are denoted or which are designated as less than 5 meters deep tended to have significantly shorter tentacles than colonies in deeper sites or greater than 5. 01:03:38.000 --> 01:03:44.000 And this was not dependent on speeches and there was no interaction. 01:03:44.000 --> 01:03:51.000 For choral aid concentration, again, at proxy for photosynthesis. We see in terms of depth that for P. 01:03:51.000 --> 01:03:58.000 There's no change in floral concentration. There's no change in that photosynthesis between shallow and deep environments. 01:03:58.000 --> 01:04:06.000 However, for P Wagner I we see this significant decrease for a deep deep in photosynthesis. 01:04:06.000 --> 01:04:16.000 We see the pretty much the same trend for. When you look at it for tentacles, so short tentacles in the Wagner eyes tended to be significantly. 01:04:16.000 --> 01:04:23.000 Higher in chloro than they did for long tentacles but there was no difference for pivorosa 01:04:23.000 --> 01:04:38.000 So now looking at stable isotopes, we expect in Caribbean and oxidoxic worlds that as you become more heterotrophic that you should decrease in Delta C 13 and increase in Delta N 15.th 01:04:38.000 --> 01:04:46.000 And the reason for this is because Plankton in the Caribbean has a delta C, 13 signature of around negative 20. 01:04:46.000 --> 01:04:57.000 And so they are approaching their food source as uptake more. And as they're increasing, heterotrophy, they should be increasing in their trophic level as well. 01:04:57.000 --> 01:05:03.000 However, it's important to note that in symbiotic species There's, cycling occurring between. 01:05:03.000 --> 01:05:09.000 And so this relationship can be quite complex. 01:05:09.000 --> 01:05:18.000 So in terms of how to evaluate our results. In 2020, on teacher pay at all published a study using the our package cyber. 01:05:18.000 --> 01:05:30.000 Were they estimated the trophic strategies of hard worlds shown? I wanna find standard ellipses for host shown in purple and symbiote shown in. 01:05:30.000 --> 01:05:39.000 And they proposed these cut offs of different percentages of overlap of these ellipses were greater than 70% overlap. 01:05:39.000 --> 01:05:52.000 Is indicative of auto trophy. So shown here in an acrobat. And so the more overlap you are, the more the more cycling that is in theory occurring between hosts and. 01:05:52.000 --> 01:06:00.000 On the opposite end of that spectrum, And so we expect that individuals that are more heterotrophic. 01:06:00.000 --> 01:06:09.000 Should have less than 10% overlap between hosts and symbiot because the host fraction is getting external nutrients from heterotrophic feeding. 01:06:09.000 --> 01:06:25.000 And so you see shifts in that. Terrific level. And then mix of trophy, they proposed should be between 10 and 70% where you have something in the middle where you're starting to see that divergence of the host but there's still quite a bit of overlap between hosts. 01:06:25.000 --> 01:06:32.000 Additionally, we can look for ships in trophic niche as well. Using hotelings t squared test to calculate a p-value. 01:06:32.000 --> 01:06:39.000 Where we can compare the centroids of these standard ellipses. And look for significant differences. 01:06:39.000 --> 01:06:51.000 And so this from this week in gain whether or not host and semi are occurring and significantly different trophic. And so by using both of these 2 methods, we can gather. 01:06:51.000 --> 01:06:58.000 Differences in trophic strategy as well as differences in trophic. 01:06:58.000 --> 01:07:04.000 Okay, so looking at this, what we found is that for They are mixatropic and approaching autotropy. 01:07:04.000 --> 01:07:09.000 They have 69% overlap, which is right at that cusp of the cutoff for auditorium. 01:07:09.000 --> 01:07:17.000 Additionally, they have a d value of point one, indicating no difference between host and symbiotic. 01:07:17.000 --> 01:07:26.000 However, for peat, we see something different. Or we see that they're heterotropic as there's no overlap between host and and by out fractions. 01:07:26.000 --> 01:07:33.000 And there's a significant difference in centroid of these ellipses indicating separate trophic. 01:07:33.000 --> 01:07:39.000 Additionally, when comparing 2 species. We see using hoteling c squared test again. 01:07:39.000 --> 01:07:49.000 That the host and the symbiote have significantly different, or in significantly different trophic. And so therefore not only are these 2 species. 01:07:49.000 --> 01:08:00.000 Utilizing different perfect strategies, but they're also existing in different prophic niches despite living in synth battery and having almost identical morphology. 01:08:00.000 --> 01:08:07.000 Now looking at intro specific variation, I want to 1st orient you to this figure, and show you a couple like this. 01:08:07.000 --> 01:08:17.000 For those that are either deep colonies or long tentacled colonies, they'll be displayed in triangles and dashed lines and those that are shallow. 01:08:17.000 --> 01:08:23.000 Or have short tentacles will be circles and solid ones. 01:08:23.000 --> 01:08:28.000 And so for P Wagner, right? We see it with changes in depth. We see no change in that overlap. 01:08:28.000 --> 01:08:38.000 Between host and SIMBIANT and hosting SIMBI, indicating that regardless of depth, They're a header. 01:08:38.000 --> 01:08:51.000 However, now we can use hotelings t squared test instead of comparing the centroids of host and symbiot, we can compare the centroids of host within shallow and host with deep to look for differences in trophic niche. 01:08:51.000 --> 01:09:00.000 And we see that trophic niche does significantly shift with depth despite the fact that they're still heterotrophic in both areas. 01:09:00.000 --> 01:09:14.000 We found the same pattern. People, in both shallow and deep environments. However, the host did shift. 01:09:14.000 --> 01:09:25.000 Now looking at technical length. We see in P Wagner that we don't see a shift in trophic strategy between long short colonies with short pinnacles and colonies with long pinnacles. 01:09:25.000 --> 01:09:34.000 Regardless of their clinical morphology, their heterotrophic. And we do not see a shift in trophic niche up the host. 01:09:34.000 --> 01:09:41.000 However, for people, so we see something different. We see that in colonies with short tentacles. 01:09:41.000 --> 01:09:50.000 They have 77% overlap, putting them in that autotrophic zone. And colonies with long tentacles have only 41% overlap. 01:09:50.000 --> 01:09:57.000 Putting them in the mix. And so we also see a significant change in host trophic niche. 01:09:57.000 --> 01:10:07.000 And so these findings that we found in paper roads are similar to what's been found in TUR specifically in previous studies, where species with different technical length exist at different. 01:10:07.000 --> 01:10:11.000 Under different perfect shops. 01:10:11.000 --> 01:10:17.000 So in summary, I found interest specific variation in tenant point in both K. Wagner I and P. 01:10:17.000 --> 01:10:29.000 Rosa. This difference in tentacle length. Could be the result of differences in morphology. It could also be the result of differences in behavior where under a shallow environment with higher light, higher water movement. 01:10:29.000 --> 01:10:39.000 They might partially contract those chemicals for under deeper environments with lower of both of those on environmental variables, they might expand those kind of more. 01:10:39.000 --> 01:10:45.000 But regardless, we're seeing this phenotype across the genus as well as the species level. 01:10:45.000 --> 01:10:53.000 All these 2 species of pseudoplexor exist in the same habitat and have really similar, We see that they have different trophic strategies. 01:10:53.000 --> 01:11:02.000 And that these terrific strategies differ with or that these traffic strategies. Our their flexibility differs between the species. 01:11:02.000 --> 01:11:09.000 And so with, we see that with increasing pinnacle length. An increase in heterotrophic input. 01:11:09.000 --> 01:11:19.000 Okay, and a shift between autotrophy and pederrophy while maintaining that same level of photosynthesis from But with the Wagner I, we see that they. 01:11:19.000 --> 01:11:25.000 Do not ship in their strategy or their trophic niche and their fluorophiles significantly decreases. 01:11:25.000 --> 01:11:32.000 And so even though they're experiencing a significant decline in photosynthesis, that might not actually have as much of an effect on the column. 01:11:32.000 --> 01:11:36.000 Because they're predominantly reliant on. 01:11:36.000 --> 01:11:46.000 Therefore we found these trophic niche and strategy differences not only between species like previously documented but within species as well. 01:11:46.000 --> 01:11:57.000 So the return to my research objectives and remind you of the goals of my dissertation, I wanted to 1st start by documenting the color mark distribution in Alberta and the northeastern Gulf of Mexico. 01:11:57.000 --> 01:12:09.000 And look for potential barriers to reproduction as well as potential genetic differentiation. Then I wanted to look at peanut plasticity and GPI interactions in this species. 01:12:09.000 --> 01:12:18.000 And then finally, I looked at the association between morephological variation and 01:12:18.000 --> 01:12:25.000 So in conclusion, I found that Albert, a lot of color morphs are differentially distributed between inshore and offward sites. 01:12:25.000 --> 01:12:33.000 But this is unlikely a result of barriers to reproduction. Or genetic divergence, indicative of. 01:12:33.000 --> 01:12:41.000 Using the Magenta Warps, I documented that growth and branching are under scientific plasticity. 01:12:41.000 --> 01:12:48.000 And that those that were transplanted closer to shore tended to grow and branch more than they transmitted away from shore. 01:12:48.000 --> 01:12:57.000 And this is occurring in a species. Again, that's ace antibiotic. So in the absence of those symbiosis, we're seeing similar patterns to what's been found in previous symbiotics. 01:12:57.000 --> 01:13:04.000 And therefore, growth and branching may be strongly influenced by water. 01:13:04.000 --> 01:13:11.000 I also found that both colony and origin affected transplant. Plasticity and UI interactions. 01:13:11.000 --> 01:13:27.000 And so I was able to uncover some of the complex dynamics that might underlie the evolution of My work therefore contributes to our understanding of interest specific variation and phenotypic plasticity in sessile invertebrates. 01:13:27.000 --> 01:13:40.000 And it exemplifies the interplay between HOAZ and symbiote. And how that might. 01:13:40.000 --> 01:13:46.000 Then building off of my findings from the asymptotic species, I uncovered trophic niche differentiation. 01:13:46.000 --> 01:13:55.000 At the species level as well as flexibility in its species. I found that Peter Rosa and P. Wagner utilize different perfect strategies. 01:13:55.000 --> 01:14:07.000 Despite more logical similarity. Was the trophy and had flexibility in their trophic strategy in relation to Well, P. 01:14:07.000 --> 01:14:17.000 Wagner, I rely heavily on autotrophy. And did not vary in their trophic strategy in relation. 01:14:17.000 --> 01:14:25.000 In the future, I plan to build off of my dissertation work by investigating the interplay between coast and symbiotic plasticity. 01:14:25.000 --> 01:14:34.000 And how plastic changes in one member can influence the dynamics between partners and symbiotic relationships. Not only in, but in other systems as well. 01:14:34.000 --> 01:14:45.000 I want to examine the full amount and how scientific plasticity among community member members can influence unity dynamics, nutrients, cycling and. 01:14:45.000 --> 01:14:57.000 So, that I have a ton of people to think. 1st I, I want to thank my advisor, Don, for all the opportunities and experiences that. 01:14:57.000 --> 01:15:13.000 And that's Like awesome place and time to learn. I also wanna thank my lab members, especially Melanie, who's been my lab m since the beginning and who truly showed me, what it means to be a strong woman in science. 01:15:13.000 --> 01:15:21.000 I want to thank you, Ling How, for her expertise in molecular techniques and for teaching me everything I know about that. 01:15:21.000 --> 01:15:29.000 It was always a pleasure working with you in the lab. I wanna thank my committee for all of your feedback and guidance throughout this process. 01:15:29.000 --> 01:15:37.000 Wanna thank the Marine Lab as well as the Cornell University stable isotope laboratory for helping me processing my isotope. 01:15:37.000 --> 01:15:48.000 I have a huge list of love and field help that I need to think for these projects. I think we typically say this work has been done with these people, but in this case that's really true. 01:15:48.000 --> 01:15:55.000 I could not have go and done all this work about these people, especially in the months that I was unable to dive. 01:15:55.000 --> 01:16:02.000 So thank you to everyone that built in when I needed things. Without the field brought back into the from the field while I was out. 01:16:02.000 --> 01:16:11.000 Thank you to all of you. Additional help I received from faculty both in the department as well as at external universities. 01:16:11.000 --> 01:16:23.000 And also community members. I wanna especially thank the late Bob Brooks. They gave me a lot of the sites where I found, my octopus in shore up here just based off of where he was from fishing. 01:16:23.000 --> 01:16:29.000 Probably like which was really good. I wanna thank my funding sources. 01:16:29.000 --> 01:16:39.000 From remote endowment, the Board of Trustees from Marine Lab, the Grambling Scholarship, the Roman Divers Hall, Same, International Fishing Association as well as And it's up grants. 01:16:39.000 --> 01:16:49.000 And then I want to thank my parents. For always supporting me from. All like whatever interests I had, they were on board. 01:16:49.000 --> 01:16:54.000 I wanted to see that they're on board when I wanted to be a musician. They were on board. 01:16:54.000 --> 01:17:10.000 And like really excited and so thank you for that and for all I want to think, for not only being a great partner, but also a really awesome. 01:17:10.000 --> 01:17:20.000 And always being willing to dive in those little disability sites with me. Our cat obviously for all of her constant commentary on this. 01:17:20.000 --> 01:17:28.000 And then my family and friends, both people here in Tallahassee. This community has been more than I could have ever imagined. 01:17:28.000 --> 01:17:34.000 As well as my friends from home, especially my good friend Kelly that has supported me through full processing. 01:17:34.000 --> 01:18:04.000 So, with that, I will take questions. 01:18:07.000 --> 01:18:15.000 The obvious question. Okay, I don't know. So the colors are genetically determined. 01:18:15.000 --> 01:18:28.000 And they can either be like In general, they present as these like singular color morphs, but they are comprised of One or 2 colors of square, but never more than that. 01:18:28.000 --> 01:18:32.000 I am not sure what's maintaining the differences in color variation. We went down a lot of, with that. 01:18:32.000 --> 01:18:43.000 We look, there is differences for our differences in morality. Different, as well as where it's with. 01:18:43.000 --> 01:18:53.000 But. When we ended up doing a transplant study with them as well and there were there was no differences in growth or branches. 01:18:53.000 --> 01:18:58.000 So, also looked for different different invertebrates on them and didn't find that either. 01:18:58.000 --> 01:19:04.000 I'm not sure honestly what's maintaining the different colors. Not something I wanna follow up on, but it is curious. 01:19:04.000 --> 01:19:10.000 Okay. 01:19:10.000 --> 01:19:39.000 Oh, Yeah. 01:19:39.000 --> 01:19:47.000 So within the same individual, why the carbon 13 is the same. Like within, or like is it around the same position of carbon 13? 01:19:47.000 --> 01:20:01.000 I think that that's because of the nutrient cycling going on between hosts and And the fact that the carbon 13 signature is really affected by the level of photosynthesis occurring within those individuals is what I think is driving. 01:20:01.000 --> 01:20:10.000 The lack of difference between hosts and S. And then you see that shift with with depth of carbon 13, which can be due to increasing hunter trophy as well as. 01:20:10.000 --> 01:20:40.000 Photosynthesis. Does that answer your question? 01:20:51.000 --> 01:21:04.000 Yeah, so we had that thought in that it would be really cool to either do something like that or to even just get like younger recruits instead of having to fragment them and kind of simulate some sort of. 01:21:04.000 --> 01:21:14.000 Because they're making them smaller, actually few miles. That's why that You could do that. 01:21:14.000 --> 01:21:20.000 Yeah, I guess you could do that. And that would be really cool. We've also talked about that to try to get away. 01:21:20.000 --> 01:21:27.000 1st of all, this study was only to try to get away. 1st of all, the study was only 7 days because of all the study was only 7 days because the hurricane came through and so we like rushed off. 01:21:27.000 --> 01:21:52.000 That's which like, but also we talked about the fact that it potentially would be better to transplant them to like a common garden and let them acclimate to the same location before back transferring them out because they kind of have a similar common history for a period of time out here trying to get rid of those central like The fernal effect or area effects. 01:21:52.000 --> 01:21:56.000 Yeah. 01:21:56.000 --> 01:22:10.000 Okay. 01:22:10.000 --> 01:22:17.000 Yeah, so during the summer time when we did the study, the I'm sure it's probably similar between those 2 areas. 01:22:17.000 --> 01:22:24.000 I think that the temperature in inshore probably more variable because of like the turnover and mixing. 01:22:24.000 --> 01:22:36.000 And just like being but they're probably pretty similar at that time of year. During the winter you end up with like those thermo clients happening, but that's not, looking at them. 01:22:36.000 --> 01:23:06.000 But that's a good point that like if you were to look at it across the, you could definitely have it. 01:23:24.000 --> 01:23:54.000 Thank you.