Oceanography Commons^™

Growth Phase Proteomics Of The Heterotrophic Marine Bacterium Ruegeria Pomeroyi, Dasha Krayushkina, Emma Timmins-Schiffman, Jessica Faux, Damon H. May, Michael Riffle, H. Rodger Harvey, Brook L. Nunn

OES Faculty Publications

The heterotrophic marine bacterium, Ruegeria pomeroyi, was experimentally cultured under environmentally realistic carbon conditions and with a tracer-level addition of ¹³C-labeled leucine to track bacterial protein biosynthesis through growth phases. A combination of methods allowed observation of real-time bacterial protein production to understand metabolic priorities through the different growth phases. Over 2000 proteins were identified in each experimental culture from exponential and stationary growth phases. Within two hours of the ¹³C-labeled leucine addition, R. pomeroyi significantly assimilated the newly encountered substrate into new proteins. This dataset provides a fundamental baseline for understanding growth phase differences in molecular …

Habitat Associations And Reproduction Of Fishes On The Northwestern Gulf Of Mexico Shelf Edge, Elizabeth Marie Keller

LSU Doctoral Dissertations

Several of the northwestern Gulf of Mexico (GOM) shelf-edge banks provide critical hard bottom habitat for coral and fish communities, supporting a wide diversity of ecologically and economically important species. These sites may be fish aggregation and spawning sites and provide important habitat for fish growth and reproduction. Already designated as habitat areas of particular concern, many of these banks are also under consideration for inclusion in the expansion of the Flower Garden Banks National Marine Sanctuary. This project aimed to gain a more comprehensive understanding of the communities and fish species on shelf-edge banks by way of gonad histology, …

Seaflow Data V1, High-Resolution Abundance, Size And Biomass Of Small Phytoplankton In The North Pacific, François Ribalet, Chris Berthiaume, Annette Hynes, Jarred Swalwell, Michael Carlson, Sophie Clayton, Gwenn Hennon, Camille Poirier, Eric Shimabukuro, Angelicque White, E. Virginia Armhurst

OES Faculty Publications

SeaFlow is an underway flow cytometer that provides continuous shipboard observations of the abundance and optical properties of small phytoplankton (μm in equivalent spherical diameter, ESD). Here we present data sets consisting of SeaFlow-based cell abundance, forward light scatter, and pigment fluorescence of individual cells, as well as derived estimates of ESD and cellular carbon content of picophytoplankton, which includes the cyanobacteria Prochlorococcus, Synechococcus and small-sized Crocosphaera (μm ESD), and picophytoplankton and nanophytoplankton (2–5 μm ESD). Data were collected in surface waters (≈5 m depth) from 27 oceanographic cruises carried out in the Northeast Pacific Ocean between 2010 and 2018. …

Closing The Oxygen Mass Balance In Shallow Coastal Ecosystems, Matthew H. Long, Jennie E. Rheuban, Daniel C. Mccorkle, David J. Burdige, Richard C. Zimmerman

OES Faculty Publications

The oxygen concentration in marine ecosystems is influenced by production and consumption in the water column and fluxes across both the atmosphere-water and benthic-water boundaries. Each of these fluxes has the potential to be significant in shallow ecosystems due to high fluxes and low water volumes. This study evaluated the contributions of these three fluxes to the oxygen budget in two contrasting ecosystems, a Zostera marina (eelgrass) meadow in Virginia, U.S.A., and a coral reef in Bermuda. Benthic oxygen fluxes were evaluated by eddy covariance. Water column oxygen production and consumption were measured using an automated water incubation system. Atmosphere-water …

The Influence Of Location, Positioning, And Seasonality On Feeding Behavior Of The Sydney Oyster (Saccostrea Glomerata) In New South Wales, Australia, Nathaniel Hess

Independent Study Project (ISP) Collection

Oysters are hard shelled bivalves that aggregate over time to create structures in estuarine systems, known as oyster reefs. These reefs are important for the promotion of estuarine biodiversity by supporting many species of fish, invertebrates, and plants. They also act as a key contributor to water health by using active suspension feeding mechanisms and selective feeding to remove nutrients and water borne pollutants from estuarine systems. They have been touted as possible bio-remediators. They also effect rates of sedimentation in estuaries.

Oyster reefs have historically been threatened by anthropogenic influences such as overharvesting, destructive fishing practices, water pollution, CO2 …

36 Years After The Species' Mass-Mortality: Diadema Antillarum Test Sizes, Population Densities, And Substrate Preferences In Three Guna Yala Reefs Near Wichub Wala Island, Bimini Horstmann

Independent Study Project (ISP) Collection

In January 1983, a waterborne pathogen beginning near the Atlantic Panama Canal entrance caused history's largest recorded marine animal die-off, wiping out 95-99% of Diadema antillarum populations throughout the Caribbean. D. antillarum, the long-spined sea urchin, is a keystone herbivore in coral reefs and its decreased densities have caused many reefs to suffer macroalgal phase shifts. Modest recovery of this species has been documented in some Caribbean locations, but reefs in Guna Yala, Panama continued to experience population decline. This study investigates density, test size, and substrate preferences of D. antillarum in three shallow coral reef areas to update the …

Nutrient Controls Over Cyanobacterial Synthesis Of The Neurotoxin Β-N-Methylamino-L-Alanine (Bmaa) And Its Potential Accumulation In The Blue Crab (Callinectes Sapidus), Madeline M. Hummel

OES Theses and Dissertations

Cyanobacteria are known to produce a variety of toxins that negatively impact both aquatic and terrestrial organisms. One putative neurotoxic compound is the non-protein amino acid β-N-methylamino-L-alanine (BMAA), which has epidemiological linkages to the development of several human neurological diseases. Three cyanobacterial species thought to produce BMAA —Microcystis aeruginosa, Synechococcus bacillaris, and Nostoc sp. —were grown in nutrient replete cultures to examine its synthesis and cellular distribution over a growth cycle. Production of BMAA was also examined in nutrient (nitrogen and phosphorus) deplete cultures of Microcystis aeruginosa. In addition, natural assemblages of phytoplankton dominated by …

Lipid Biomarkers Of Bering And Chukchi Sea Euphausiids And Their Application To Diet History, Rachel L. Pleuthner

OES Theses and Dissertations

In the eastern Bering Sea, Thysanoessa raschii are the most abundant krill species and a keystone trophic member that serve as both an important grazer and link to upper level consumers. In this system krill experience large annual variation in food resources, especially during ice advance and retreat; multiple lipid classes are used to temper the effects of those fluctuations, as well as to fuel reproduction and growth. Two shipboard feeding experiments that occurred during late spring and early summer of 2010, respectively, monitored the lipid retention in adult T. raschii and examined the fluctuation of specific lipid biomarkers under …

Carbonate Chemistry Characterization In A Low-Inflow Estuary With Recent Seagrass Loss, Jolie Higgins

Master's Theses

Estuaries are dynamic environments that are strongly affected by natural variability, as well as direct and indirect anthropogenic impacts. A better understanding of the drivers of carbon fluxes and biogeochemical variability in estuarine systems is needed, particularly with the increasing threat of ocean acidification. Morro Bay in Central California is a small nationally protected estuary, with seasonally low freshwater inputs. Since 2007, the bay has experienced a significant loss of native seagrass, Zostera marina, which is an important component of the marine ecosystem. Because seagrass photosynthesis decreases carbon dioxide and increases oxygen in the water column, the loss of seagrass …

A Fishy Problem: Effects Of Atlantic Salmon Farming In The Pacific Ocean, Madeleine A. Griffith

Student Theses 2015-Present

In this report, I explore the historical, climatological, economic, and ethical issues created by the contemporary industrial salmon farming practices off Pacific coast of the United States and Canada. Chapter 1 utilizes a variety of sources from Stephen Hume’s A Stain upon the Sea to Miller’s Living in the Environment, to examine the integral part salmon plays in both freshwater and marine ecosystems, the ecosystem services salmon contribute in wild and farmed settings, and the trends in salmon consumption around the world. Chapter 2 examines the historically relevant role salmon held among indigenous societies and how that role has changed …

Coupling Metaproteomics With Taxonomy To Determine Responses Of Bacterioplankton To Organic Perturbations In The Western Arctic Ocean, Molly P. Mikan

OES Theses and Dissertations

Understanding how the functionality of marine microbial communities change over time and space, and which taxonomic groups dominate distinct metabolic pathways, are essential to understanding the ecology of these microbiomes and the factors contributing to their regulation of elemental cycles in the oceans. The primary goal of this dissertation was to investigate the community metabolic and taxonomic responses and the degradation potential of two compositionally distinct marine microbiomes within the shallow shelf ecosystem of the Chukchi Sea after rapid fluctuations in algal organic matter availability. Novel bioinformatics tools were collaboratively developed and used together with community proteomics (metaproteomics) to characterize …

Beach Composition Preferences For Nesting Populations Of Leatherback Sea Turtles (Dermochelys Coriacea), Armila Beach, Guna Yala Comarca, Scott Campbell

Independent Study Project (ISP) Collection

Sea turtles play a critical role in marine ecosystems all over the world, including the Caribbean Sea. However, many sea turtle species are under threat due to anthropogenic impacts, such as habitat destruction and fisheries bycatch. This has caused significant declines in sea turtle populations around the world, which in turn has impacted marine ecosystems where sea turtles play critical roles in proper ecosystem functioning. A crucial part of the sea turtle life cycle that has been threatened by anthropogenic factors is nesting. Sea turtles rely on unspoiled beaches with particular physical characteristics for laying their eggs. One of the …

Sponge Community Biocomplexity, Competition, And Functional Significance In Hard-Bottom Habitats Of The Florida Keys, Fl (Usa), Marla Maxine Valentine

Biological Sciences Theses & Dissertations

Sponges can have powerful effects on ecosystem processes in shallow, tropical marine ecosystems and are an integral component of the bentho-pelagic cycle of nutrients, via filtering of dissolved and particulate organic matter from the water column. The diversity of marine communities is thought to play a determining role in intensity of ecosystem processes; thus the loss of taxa alters community function and by extension ecosystem processes. Coastal sponge populations worldwide are increasingly exposed to declining water quality that in several regions has resulted in mass sponge mortalities and reduced sponge diversity. In the Florida Keys (Florida, USA), for example, frequent …

The House Of The Fish: Collaborative Coral Reef Awareness Project On Nosy Be, Meghan Hurley

Independent Study Project (ISP) Collection

The purpose of this project was to initiate a place-based coral reef education project, with the ultimate goal of improved localized conservation efforts in the face of growing global threats. Three principal groups that interact with coral reefs on Nosy Be were interviewed: fishers, tourist guides, and conservationists. Research was conducted on the biology of coral reefs, and on the most prominent threats facing coral reefs in the Indian Ocean and around Nosy Be. Information and direct quotes from interviews, and the results of research, were incorporated into a photographic journal entitled "In the House of the Fish" or "An-tragno …

Impacts Of Anthropogenic Waste On Tasmanian Pacific Gull (Larus Pacificus) Diet, Anne Bartlett

Independent Study Project (ISP) Collection

Plastic pollution is one of the foremost environmental issues that the world is currently experiencing. The effects of plastic pollution are great and range from leaching of hazardous chemicals into the environment to ingestion of plastic waste by wildlife, including seabirds. Due to the high rates of plastic consumption by seabirds, many recent studies have been performed to determine the biological impacts of plastic consumption on various seabird species. The Pacific Gull (Larus pacificus) is the world’s largest sea gull and the only gull endemic to Australia, yet very limited information exists on this species. This study examines the impacts …

Species Abundance, Spatial And Vertical Distributionsof Large Heteropods (Pterotracheidae And Carinariidae)In The Northern Gulf Of Mexico, Kristine A. Clark

USF Tampa Graduate Theses and Dissertations

A description of species abundance, richness and distribution, and eye size of heteropod molluscs from the families Pterotracheidae and Carinariidae in the oligotrophic ecosystem of the northern Gulf of Mexico (GOM) is described based on discrete-depth sampling protocols. The collections were comprised from two midwater sampling programs conducted sequentially after the 2010 Deepwater Horizon oil spill (DWHOS): the Offshore Nekton Sampling and Analysis Program (ONSAP, 2011) and the Deep Pelagic Nekton Dynamics of the Gulf of Mexico (DEEPEND, 2015-2018). Study materials from DEEPEND were collected during the initial five cruises of 2015-2017. These programs collected over 3,495 heteropods in two …

Ecological Response Of Phytoplankton To The Oil Spills In The Oceans, Danling Tang, Jing Sun, Li Zhou, Sufen Wang, Ramesh P. Singh, Gang Pan

Mathematics, Physics, and Computer Science Faculty Articles and Research

Oil spills in oceans have substantial influence on marine ecosystems. This study investigates 21 oil spills in the world. Analyzing Chlorophyll-a (Chl-a) from Moderate Resolution Imaging Spectroradiomerer (MODIS) data after Penglai oil spills on 4 June 2011, found a bloom with peak value of Chl-a (13.66 mg m⁻³) spread over an area of 800 km² during 18–25 June 2011, and a pronounced increase in the monthly Chl-a concentration (6.40 mg m⁻³) on June 2012 in the Bohai Sea. Out of the 21 oil spills, 14 blooms were observed, while 11 …

Habitat Usage Patterns Of Bottlenose Dolphins (Tursiops Truncatus) In Terrebonne And Timbalier Bays, Louisiana, Mary Allison Manning

LSU Master's Theses

I coupled fine-scale environmental data with observed behavior and group composition data to examine overall distribution within the bay system and to characterize the habitat associated with foraging and the presence of calves. Semi-isolated populations of bottlenose dolphins (Tursiops truncatus) along Louisiana’s coast are undergoing increased risks from boat traffic, oil spills, land subsidence, and planned water diversions (CPRA 2017). Characterizing the habitat use of bottlenose dolphins in Terrebonne and Timbalier bays, Louisiana, is important given the likely high site fidelity, small home ranges, and low exchange of individuals with nearby coastal populations (Lane et al. 2015, McDonald …

Dinitrogen Fixation Across Physico-Chemical Gradients Of The Eastern Tropical North Pacific Oxygen Deficient Zone, C. R. Selden, M. R. Mulholland, P.W. Bernhardt, B. Widner, A. Macías-Tapia, Q. Ji, A. Jayakumar

OES Faculty Publications

The Eastern Tropical North Pacific Ocean hosts one of the world's largest oceanic oxygen deficient zones (ODZs). Hot spots for reactive nitrogen (N_r) removal processes, ODZs generate conditions proposed to promote N_r inputs via dinitrogen (N₂) fixation. In this study, we quantified N₂ fixation rates by ¹⁵N tracer bioassay across oxygen, nutrient, and light gradients within and adjacent to the ODZ. Within subeuphotic oxygen‐deplete waters, N₂ fixation was largely undetectable; however, addition of dissolved organic carbon stimulated N₂ fixation in suboxic (μmol/kg O₂) waters, suggesting that diazotroph communities are …

Ebullition Of Oxygen From Seagrasses Under Supersaturated Conditions, Matthew H. Long, Kevin Sutherland, Scott D. Wankel, David J. Burdige, Richard C. Zimmerman

OES Faculty Publications

Gas ebullition from aquatic systems to the atmosphere represents a potentially important fraction of primary production that goes unquantified by measurements of dissolved gas concentrations. Although gas ebullition from photosynthetic surfaces has often been observed, it is rarely quantified. The resulting underestimation of photosynthetic activity may significantly bias the determination of ecosystem trophic status and estimated rates of biogeochemical cycling from in situ measures of dissolved oxygen. Here, we quantified gas ebullition rates in Zostera marina meadows in Virginia, U.S.A. using simple funnel traps and analyzed the oxygen concentration and isotopic composition of the captured gas. Maximum hourly rates of …

Metagenomic Sequencing Identifies Highly Diverse Assemblages Of Dinoflagellate Cysts In Sediments From Ships' Ballast Tanks, Lixia Shang, Zhangxi Hu, Yunyan Deng, Yuyang Liu, Xinyu Zhai, Zhaoyang Chai, Xiaohan Liu, Zifeng Zhan, Fred C. Dobbs, Ying Zhong Tang

OES Faculty Publications

Ships' ballast tanks have long been known as vectors for the introduction of organisms. We applied next-generation sequencing to detect dinoflagellates (mainly as cysts) in 32 ballast tank sediments collected during 2001-2003 from ships entering the Great Lakes or Chesapeake Bay and subsequently archived. Seventy-three dinoflagellates were fully identified to species level by this metagenomic approach and single-cell polymerase chain reaction (PCR)-based sequencing, including 19 toxic species, 36 harmful algal bloom (HAB) forming species, 22 previously unreported as producing cysts, and 55 reported from ballast tank sediments for the first time (including 13 freshwater species), plus 545 operational taxonomic units …

Spatial Structuring And Life History Connectivity Of Antarctic Silverfish Along The Southern Continental Shelf Of The Weddell Sea, Jilda Alicia Caccavo, Julian R. Ashford, Svenja Ryan, Chiara Papetti, Michael Schröder, Lorenzo Zane

OES Faculty Publications

A multi-disciplinary approach was employed to examine a physical-biological population hypothesis for a critical forage species, the Antarctic silverfish Pleuragramma antarctica. Caccavo et al. (2018; Sci Rep 8:17856) had shown strong gene flow along the westward Antarctic Slope Current, in addition to spatially recurring length modes that provided evidence for episodic connectivity. In this paper, otolith nucleus chemistry from a subset of fish collected in the southern Weddell Sea as part of a hydrographic survey of the Filchner Trough system was used to test between connectivity scenarios. Nucleus chemistry, which reflects environmental exposure during early life, showed significant …

A Full-Ocean-Depth Rated Modular Lander And Pressure-Retaining Sampler Capable Of Collecting Hadal-Endemic Microbes Under In Situ Conditions, Logan M. Peoples, Matthew Norenberg, David Price, Madeline Mcgoldrick, Mark Novotny, Alexander Bochdansky, Douglas H. Bartlett

OES Faculty Publications

The hadal zone remains one of the least studied environments because of its inaccessibility, in part because of hydrostatic pressures extending to 110 MPa. Few instruments are capable of sampling from such great depths. We have developed a full-ocean-depth-capable lander that can be fit with sampling packages for the collection of still images, video, motile megafauna, and hadal seawater. One payload includes a pressure-retaining sampler (PRS) able to maintain seawater samples under in situ pressure during recovery. We describe the technical specifications of the lander and the PRS and preliminary results from three deployments at depths in excess of 10,700 …

Metaproteomics Reveal That Rapid Perturbations In Organic Matter Prioritize Functional Restructuring Over Taxonomy In Western Arctic Ocean Microbiomes, Molly P. Mikan, H. Rodger Harvey, Emma Timmins-Schiffman, Michael Riffle, Damon H. May, Ian Salter, William S. Noble, Brook L. Nunn

OES Faculty Publications

We examined metaproteome profiles from two Arctic microbiomes during 10-day shipboard incubations to directly track early functional and taxonomic responses to a simulated algal bloom and an oligotrophic control. Using a novel peptide-based enrichment analysis, significant changes (p-value < 0.01) in biological and molecular functions associated with carbon and nitrogen recycling were observed. Within the first day under both organic matter conditions, Bering Strait surface microbiomes increased protein synthesis, carbohydrate degradation, and cellular redox processes while decreasing C1 metabolism. Taxonomic assignments revealed that the core microbiome collectively responded to algal substrates by assimilating carbon before select taxa utilize and metabolize nitrogen intracellularly. Incubations of Chukchi Sea bottom water microbiomes showed similar, but delayed functional responses to identical treatments. Although 24 functional terms were shared between experimental treatments, the timing, and degree of the remaining responses were highly variable, showing that organic matter perturbation directs community functionality prior to alterations to the taxonomic distribution at the microbiome class level. The dynamic responses of these two oceanic microbial communities have important implications for timing and magnitude of responses to organic perturbations within the Arctic Ocean and how community-level functions may forecast biogeochemical gradients in oceans.

Nitrogen Assimilation In Picocyanobacteria Inhabiting The Oxygen-Deficient Waters Of The Eastern Tropical North And South Pacific, Montserrat Aldunate, Carlos Henriquez-Castillo, Qixing Ji, Jessica Lueders-Dumont, Margaret R. Mulholland, Bess B. Ward, Peter Von Dassow, Osvaldo Ulloa

OES Faculty Publications

Prochlorococcus and Synechococcus are the most abundant free-living photosynthetic microorganisms in the ocean. Uncultivated lineages of these picocyanobacteria also thrive in the dimly illuminated upper part of oxygen-deficient zones (ODZs), where an important portion of ocean nitrogen (N) loss takes place via denitrification and anaerobic ammonium oxidation. Recent metagenomic studies revealed that ODZ Prochlorococcus have the genetic potential for using different N forms, including nitrate and nitrite, uncommon N sources for Prochlorococcus, but common for Synechococcus. To determine which N sources ODZ picocyanobacteria are actually using in nature, the cellular 15N natural abundance (δ15N) and assimilation rates of …

Modeling The Seasonal Cycle Of Iron And Carbon Fluxes In The Amundsen Sea Polynya, Antarctica, P. St-Laurent, P. L. Yager, R. M. Sherrell, H. Oliver, M. S. Dinniman, S. E. Stammerjohn

CCPO Publications

The Amundsen Sea Polynya (ASP) is distinguished by having the highest net primary production per unit area in the coastal Antarctic. Recent studies have related this high productivity to the presence of fast-melting ice shelves, but the mechanisms involved are not well understood. In this study we describe the first numerical model of the ASP to represent explicitly the ocean-ice interactions, nitrogen and iron cycles, and the coastal circulation at high resolution. The study focuses on the seasonal cycle of iron and carbon, and the results are broadly consistent with field observations collected during the summer of 2010–2011. The simulated …

Sponges Structure Water-Column Characteristics In Shallow Tropical Coastal Ecosystems, Marla M. Valentine, Mark J. Butler Iv

Biological Sciences Faculty Publications

Sponges can have powerful effects on ecosystem processes in shallow tropical marine ecosystems. They drive benthic-pelagic coupling by filtering dissolved and particulate organic matter from the water column, alter water chemistry in association with their symbiotic microorganisms, and increase habitat structural complexity. Anthropogenic degradation of coastal waters is widespread and can reduce the density and diversity of foundation species such as sponges, potentially diminishing their contributions to ecosystem processes. We used a novel mesocosm design that minimized artifacts associated with traditional single-species and closed-system filtration experiments to examine the effects of water turnover and sponge biomass on water-column properties. Using …

Spatial Variability In Recruitment Of Chilipepper Rockfish (Sebastes Goodei) In The California Current System, Laura K. Solinger

Cal Poly Humboldt theses and projects

Properly describing variability in population dynamics (e.g., growth, fecundity, recruitment) is expected to improve management of important fisheries stocks (Maunder & Piner, 2014). As recruitment to most fish stocks is determined during early life history stages (Houde, 1997; Iles & Beverton, 2000), and early life history stages are influenced by oceanographic conditions (Bjorkstedt et al. 2002; Laidig, 2010), understanding how environmental stochasticity influences recruitment deviations has potential to improve estimates of stock productivity and how productivity might change over time to support more effective management. Chilipepper Rockfish (Sebastes goodei) are an important commercial species that is managed as …

Along-Shelf Connectivity And Circumpolar Gene Flow In Antarctic Silverfish (Pleuragramma Antarctica), Jilda Alicia Caccavo, Chiara Papetti, Maj Wetjen, Rainer Knust, Julian R. Ashford, Lorenzo Zane

OES Faculty Publications

The Antarctic silverfish (Pleuragramma antarctica) is a critically important forage species with a circumpolar distribution and is unique among other notothenioid species for its wholly pelagic life cycle. Previous studies have provided mixed evidence of population structure over regional and circumpolar scales. The aim of the present study was to test the recent population hypothesis for Antarctic silverfish, which emphasizes the interplay between life history and hydrography in shaping connectivity. A total of 1067 individuals were collected over 25 years from different locations on a circumpolar scale. Samples were genotyped at fifteen microsatellites to assess population differentiation and …

Dynamics Of Prochlorococcus Diversity And Photoacclimation During Short-Term Shifts In Water Column Stratification At Station Aloha, Anne W. Thompson, Ger Van Den Engh, Nathan A. Ahlgren, Kathleen Kouba, Samantha Ward, Samuel T. Wilson, David M. Karl

Biology Faculty Publications and Presentations

The cyanobacterium Prochlorococcus is the dominant phototroph in surface waters of the vast oligotrophic oceans, the foundation of marine food webs, and an important component of global biogeochemical cycles. The prominence of Prochlorococcus across the environmental gradients of the open ocean is attributed to its extensive genetic diversity and flexible chlorophyll physiology, enabling light capture over a wide range of intensities. What remains unknown is the balance between temporal dynamics of genetic diversity and chlorophyll physiology in the ability of Prochlorococcus to respond to a variety of short (approximately 1 day) and longer (months to year) changes in the environment. …