Benthic Foraminiferal Distributions in a Shallow Subtropical Estuary and the Influence of Environmental Factors

Maria Angelica Zamora-Durán

A thesis submitted to the College of Engineering at Florida Institute of Technology in partial fulfillment of the requirements for the degree of

Masters of Science in Biological Oceanography

Melbourne, Florida December, 2016

We the undersigned committee hereby approve the attached thesis, “Benthic Foraminiferal Distributions in a Shallow Subtropical Estuary and the Influence of Environmental Factors” by Maria Angelica Zamora-Durán.

______Kevin B. Johnson, Ph.D., Major Advisor Associate Professor Department of Ocean Engineering and Sciences

______John Trefry, Ph.D. Professor Department of Ocean Engineering and Sciences

______Ralph Turingan, Ph.D. Professor Department of Biological Sciences

______Stephen Wood, Ph.D. Associate Professor and Department Head Department of Ocean Engineering and Sciences

Abstract

Benthic Foraminiferal Distributions in a Shallow Subtropical Estuary and the Influence of Environmental Factors

By Maria Angelica Zamora-Durán

Advisor: Kevin B. Johnson, Ph. D.

Benthic foraminiferal communities are potentially useful tools for assessing and monitoring estuarine ecosystems and as bioindicators of water quality and pollution. To evaluate this indicator for the Indian River Lagoon (IRL) benthic ecosystem, the densities and distributions of Foraminifera were measured in Turkey

Creek (Palm Bay, Florida), Crane Creek (Melbourne, Florida), and the adjacent

IRL. Spatial population patterns were compared with parameters characterizing benthic sediments, including silt and clay content, organic matter (OM), and associated boundary water column conditions including salinity, temperature, and dissolved oxygen (DO. Samples (n=145) were collected from October 2015 to

December 2015 at 12 locations. Four species, Ammonia tepida, Ammonia parkinsoniana, Elphidium excavatum, and Quinqueloculina seminula made up

>90% of the foraminiferal assemblages, with A. tepida dominating at all sites. The highest average densities were found in the intermediate sediments of Turkey

iii

Creek. The average foraminiferal density was between 124 (±35 SE) and 291 (±91

SE) individuals ml-1 in the low (OM <2%) organic matter sediments of the IRL near Crane Creek and Turkey Creek, respectively. This was significantly lower

(p<0.01) than the 750 (± 180 SE) individuals ml-1 in the intermediate organic sediments in Turkey Creek, and significantly higher (p<0.01) than the 20 (±19 SE) individuals ml-1 in the deep channels containing the highest amounts of fine- grained, organic-rich sediments (OM >10%, “muck”). In general, foraminiferal densities correspond strongly to OM and DO, with the highest densities occurring between 2-4 mgl-1 DO and 1-5% OM.

Paleoecological data from foraminiferal tests provide insight into how foraminiferal communities may respond to the changing estuary over decades. No foraminiferal community shifts were observed in the intermediate organic sediments (TC). There is, however, an increasing abundance of Quinqueloculina seminula in shallower, more recent TC sediments, possibly due to recent increases in sediment organic content.

Table of Contents

Table of Contents ...... iii List of Figures ...... vii List of Tables ...... viii Acknowledgement ...... viii Dedication ...... ix Introduction ...... 1 Methods ...... 13 Results ...... 18 Discussion ...... 35 Conclusion ...... 44 References ...... 46 Appendix ...... 55

List of Figures

Figure 1 — Foraminiferal Life Cycle...... 7 Figure 2 — Pictures of Key Species...... 9 Figure 3 — Map of Study Sites...... 15 Figure 4 — Scanning Electron Microscope Pictures...... 19 Figure 5 — Spatial Comparisions of Average Foraminiferal Densities...... 21 Figure 6 — Average Foraminiferal Densities per Species...... 23 Figure 7 — Live and Dead Foraminiferal Densities per Species...... 24 Figure 8 — Average Live/Dead Ratio for Each Species...... 26 Figure 9 — Canonical Correspondence Analysis Ordination Plot...... 28 Figure 10 — Correlation of Dissolved Oxygen and Organic Matter...... 29 Figure 11 — Correlations of Density and Dissolved Oxygen/Organic Matter...... 30 Figure 12 — Ammonia Correlations with Dissolved Oxygen and Organic Matter...... 32 Figure 13 — Temporal Shifts in Community Structure based on Deep Cores...... 34 Figure 14 — Conceptual Model of Sediment and Community Composition...... 41

List of Tables

Table 1 — Locations of Sampling Areas ...... 16 Table 2 — Sediment and Dissolved Oxygen Parameters of Study Sites ...... 20

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Acknowledgement

I thoroughly appreciate the expertise and support provide by my major advisor, Dr. Kevin Johnson, as well as the other members of my thesis committee, Dr. John Trefry and Dr. Ralph Turingan. Monthly sampling and laboratory work was facilitated by the efforts of Anthony Cox and Daniel Hope. Without the engineering assistance provided by William Battin and Daniel Fernández, I would not have essential materials used during this research. I am also thankful for the financial support offered to me by assistantships from the Department of Ocean Engineering and Sciences, the Edward W. and Lee Hill Snowdon Foundation, as well as the State Legislature through Brevard County, without which I would not have been able to conduct this research.

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Dedication

I dedicate this work to my parents, whose love and support have been unyielding, through this process and throughout my life. I am eternally grateful and promise to keep making you both proud while living out my dreams. To all of the friends, family, and teachers that have offered me guidance and assistance throughout the years in the form of words, lessons, and/or embraces – your encouragement and efforts have truly helped me get to where I am today and will never be forgotten.

Introduction

Estuaries are dynamic and productive aquatic ecosystems that are subject to negative impacts due to coastal development and anthropogenic nutrient inputs. Resulting eutrophication can lead to ecosystem changes in nutrient biogeochemical feedbacks, such as increasing nitrogen and phosphorus which can lead to the formation and persistence of algal blooms by providing nutrients for phytoplankton and, in turn, more food for zooplankton (Schlesinger, 1991; Vitousek et al., 1997; Howarth et al., 2011). Decomposition of the dead algae and other organic matter subsequently result in hypoxic/anoxic zones that can limit population sizes and overall biodiversity (Chapin et al., 1997; Matson et al., 1997; Howarth et al., 2011). To counter estuarine degradation, the US Congress established the National Estuary Program (NEP) in 1987. The goals of this program include protecting and restoring the water quality and ecological integrity of key estuaries. In 1990, the Indian River Lagoon (IRL) was adopted by the NEP as an estuary of national significance. The IRL is a diverse shallow estuary spanning 250 kilometers along the east coast of Florida from Ponce de León Inlet (Volusia County) to Jupiter Inlet (West Palm Beach). This commercially and recreationally valuable ecosystem is suffering from anthropogenic influences including eutrophication. Increased nutrient and organic matter input, as well as higher silt and clay content from soil erosion, has led to an accumulation of fine-grained, organic-rich sediments in >

10% of the Indian River Lagoon (Trefry et al., 1990). Extreme organic sediments are referred to as “muck”, which is operationally defined as being comprised of > 10% organic matter, > 60% silt and clay, and > 75% water by weight (Trefry and Trocine, 2011). Muck in the IRL is detrimental to the overall quality of the ecosystem because it serves as an internal source of nutrients (ammonium and phosphate) to the lagoon (Gu et al., 1987; Trefry et al., 2007), compounding eutrophication and fueling algal blooms. When blooms die off, they fall to the sediment and are subsequently decomposed by bacteria leading to hypoxic and even anoxic conditions in the bottom water. Areas of low dissolved oxygen also foster anaerobic, sulfate-reducing bacteria that release toxic hydrogen sulfide into the water column. If dissolved oxygen levels become too low, or hydrogen sulfide too high, the environment becomes uninhabitable to aerobic organisms and even the most resilient species of benthic infauna, seagrasses, and fish may die. Mitigation and restoration projects attempting to correct nutrient problems have been implemented in the IRL. These include the implementation of Total Maximum Daily Load limits (TMDLs), restoration of seagrass and oyster reef habitat, and environmental dredging of muck. Some restoration techniques are unproven, while others have a history of facilitating positive change. In either case, monitoring is necessary to document the degree of success. Monitoring allows managers to evaluate restoration methods and improve techniques. One efficient approach to evaluating an ecosystem and determining the success of restoration efforts is to monitor the species’ populations. Foraminifera have been used as environmental bioindicators of water quality and pollution in estuarine ecosystems (Ishman et al., 1997; Gapotchenko et al., 2000; Gibson et al., 2000; Karlsen et al., 2000; Alves Martins et al., 2015). They are an effective tool due to their pervasive distributions, high abundances, 2

short generation times, reliable fossil record, and sensitivity to certain environmental factors (Schönfeld et al., 2012). There are spatial and temporal advantages in using small invertebrates or protozoans, such as Foraminifera, in bioassessments. For example, a foraminifer may be 50 μm in size compared to a macroinvertebrate which may be 5 cm in size. In addition, Foraminifera have several generations per year, whereas macroinvertebrates exhibit a few or just one generation per year (Buzas et al., 2002). Indices have been proposed whereby resource managers can monitor the condition of benthic environments and evaluate restoration efforts by using the presence and abundance of living foraminiferal assemblages (Sen Gupta et al., 1996; Hallock et al., 2003; Bouchet, 2012). In addition to monitoring living populations, paleoecological data from fossilized foraminiferal tests provide valuable historical perspectives to ecologists and restoration planners (Gooday, 2003; Willard and Cronin, 2007). The United States Environmental Protection Agency has published a Bioassessment and Biocriteria Technical Guidance for estuarine and coastal marine waters which are used by federal, state, and local government agencies to design biosurveys for assessment of water quality, which highlights the advantages of using Foraminifera in ecological assessment (Gibson et al., 2000). Foraminifera are among the most abundant protists in the marine environment, occurring in both planktonic and benthic habitats of coastal waters and the deep sea (Sen Gupta et al., 2002). They are characterized by their ability to secrete a protective shell or test and by their reticulopodia. The test is either composed of a single chamber (unilocular), or multiple chambers that are separated by walls called foramen (multilocular). During growth, the unilocular forms will increase the size of their single chamber by adding material to the inner organic lining of the test (Goldstein and Barker, 1988). Those species with multiple chambers, most commonly the calcareous species, will intermittently add a new 3

chamber by forming a cyst made of sediment or other foreign material which will be filled in with an organic lining, ultimately resulting in calcification of the chamber and the whole test (Goldstein, 2002). There are four major groups of Foraminifera, classified by the mechanism in which they build their shells (Sen Gupta, 2002). The agglutinated species construct their tests by cementing particles from their environment, such as organic matter or sand grains. Agglutinated forms of Foraminifera can be found in most marine environments; however, they dominate only in areas where calcareous forms are stressed by reduced levels of calcium carbonate (Murray, 2006). The other groups of Foraminifera use a form of calcite to build their test. These calcareous taxa include species that secrete porcelaneous imperforate shells made of rods of calcite that are randomly arranged into inner and outer surface layers and do not contain pores. The other group is comprised of hyaline perforate species whose shells are made up of interlocking crystals of calcite which contain pores. Studies on the phylogeny of these taxa support their distinction, indicating that the porcelaneous taxa and the hyaline taxa diverged early in the evolutionary history of these lineages (Longet and Pawlowski, 2007). Lee (1990) elevated Foraminifera from Order to Class and currently 16 orders are recognized (Loeblich and Tappan, 1992; Sen Gupta, 2002). Foraminifera also produce reticulopodia, fine pseudopodia that they use for motility, attachment, feeding, building and structuring tests, protection, excretion, and some aspects of respiration and reproduction. Foraminifera extrude their reticulopodia through apertures in the test. Particles can be moved bidirectionally along these reticulopodia (Allen, 1964). Most benthic Foraminifera live in fine- grained sediments through which they actively migrate and use their reticulopodia to collect a variety of materials that form mounds around tests, creating cysts for growth or feeding (Goldstein, 2002). 4

Only one suborder of the order Rotaliida is planktonic, Globigerinida, whereas all other extant orders of Foraminifera are benthic, yet can have planktonic reproductive stages (Goldstein, 2002). Benthic species of Foraminifera are widely influenced by a variety of environmental factors and can live epifaunally, epiphytically, or infaunally. Studies have shown that food supply determines the population abundances of benthic foraminifers as long as physical and chemical parameters are suitable (Loubere and Faiduddin, 2002). In areas of high organic matter and decomposition, increased hypoxia/anoxia and sulfidic conditions limit the depth in the sediments to which Foraminifera can live. In oligotrophic environments, the availability of food is the dominant factor that limits the depth of Foraminifera (Hallock, 2003). Benthic foraminifers exhibit a variety of feeding strategies including grazing, suspension feeding, deposit feeding, carnivory, parasitism, direct uptake of DOC, and symbiosis (Goldstein, 2002). Heterotrophic forms feed on detritus, bacteria, algae, or a combination thereof. Some taxa have symbiotic relationships with algae allowing them to feed and photosynthesize. Symbiosis drastically increases the amount of fixed carbon available to these individuals compared to the purely heterotrophic organisms that share the same environment (Hallock, 2002; Lee, 2006). Species that live in muddy sediments, within and below the photic zone, can use their reticulopodia to construct feeding cysts (Hofker, 1927; Nyholm, 1957; Goldstein and Corliss, 1994). In this process, the reticulopodia gather material including sediment, algal cells, bacteria, and organic detritus and partition the material into small bundles which the foraminifer then ingests by phagotrophy from the terminal chamber (Goldstein and Corliss 1994). Most foraminifers are microscopic but can grow to be several centimeters in diameter, with the extinct Lepidocyclina elephatina being the largest ever reported at 14 cm (Grell, 1973). Size is proportional to life span in Foraminifera, with small 5

taxa living for days and larger taxa for years. Benthic Foraminifera undergo alternation of generations (Figure 1) (Goldstein, 2002). Foraminifera can have one or more asexual generations that typically reproduce by multiple fission, producing broods of tens to thousands of juveniles from the protoplasm of the parent cell (Cushman, 1928). These juveniles are not necessarily clones of the parent cell since meiosis can precede asexual reproduction. Most species of Foraminifera also exhibit sexual reproduction where they produce biflagellated gametes released directly into the water (Goldstein, 1997). Unlike most animals, meiosis in the Foraminifera serves as a separate part of the life cycle, occurring as an integral part of multiple fission. The young produced by meiosis are haploid and grow to become uninucleate adults, which subsequently produce gametes by mitosis (Le Calvez, 1946, 1950). For the Foraminifera whose life cycles are known (~30 species), most have life cycles along this general theme of alternation of generations, some exhibiting obligatory alternation of generations while others show facultative alternation of generations (Lee et al., 1991; Goldstein, 1997). The obligatory forms always undergo meiosis before multiple fission, while the facultative forms include asexual cycles that reproduce by multiple fission (Bradshaw, 1957; Schnitker, 1974). The life cycle in Foraminifera is more varied than any other group of protists, with other protist groups exhibiting binary fission, various forms of budding, and nuclear/test dimorphism (Goldstein, 2002).

Figure 1. The foraminiferal life cycle, showing alternation of generations. The life cycle alternates between a haploid, uninucleate, megalospheric gamont (produces gametes), and a diploid, multinucleate, microspheric agamont (produces haploid individuals through multiple fission). Some species also include a schizont form in their life cycles which is produced by the agamont and reproduces asexually (from Goldstein, 2002).

Antoine van Leuwenhoek and Carl von Linné first described the complexity of Foraminifera, initially classifying them as mollusks. Dujardin aided in the classification in 1835 by concluding that Foraminifera are unicellular. Most of the studies completed in the 19th century on Foraminifera do not focus on their biology, but instead investigate their geology, with over 50,000 fossil species having been described (Pawlowski and Holzmann, 2008). Conversely, only 3,000-5,000 species are estimated to be living today (Goldstein, 2002).

Rotaliida is a dominant calcareous benthic foraminiferal order that includes some of the most stress-tolerant generalists and specialized species found among the Foraminifera, such as the members of the genus Ammonia. Ammonia species are often the only eukaryotes to be found in hypoxic or highly polluted areas (Schafer, 2000). Ammonia was the first genus assigned to Foraminifera in 1772 (Holzmann, 2000), and since then has been a focus of research due to its wide global distribution and variable morphology. This variability has led to complications when classifying species, especially since morphology has been shown to vary due to environmental characteristics and pollution (Boltovskoy, et al. 1991; Alve, 1995; Colburn, 1998; Frontalini, 2015). Over 46 species, subspecies and varieties have been described (Holzmann, 2000). As a genus, Ammonia is common in benthic, nearshore environments, with Ammonia parkinsoniana (Figure 2) and Ammonia tepida being found most commonly in shallower waters (Jorissen, 1988). These species are deposit feeders that can ingest large amounts of sediment, algal cells, bacteria, and organic detritus (Goldstein and Corliss, 1994). Both are tolerant of wide ranges of temperature (0°C-35°C) and salinity (0.5 up to 50 ppt) (Sen Gupta, 2002) and, due to their being facultative anaerobes, hypoxic and anoxic conditions (Cushman, 1928; Karlsen et al., 2000). The main distinction between the two species is their morphology, with A. parkinsoniana containing an umbilical plug while A. tepida does not (Figure 2).

A) B)

C) D)

Figure 2. The benthic rotaliids Ammonia parkinsoniana (diameter 500 μm) and Ammonia tepida (300μm) A. A. parkinsoniana spiral side B. A. parkinsoniana umbilical side. C. A. tepida spiral side D. A tepida ventral side (A. tepida specimen stained with Rose Bengal).

Studies by Buzas et al. (1982, 2000, and 2002) have focused on the distribution of Foraminifera in the IRL. A study done from 1975-1976 documented 94 species in the IRL and showed that species richness increased from the north (Haulover Canal) to the south (St. Lucie). Canonical variate analysis of the 15 most abundant species, with Ammonia parkinsoniana (previously Ammonia beccarii, Holzmann, 2000) being the most abundant, distinguished inlets and Haulover from other areas in the IRL. This distinction is likely to the conditions associated with the tidal changes at the inlets vs. Haulover, where tidal influence is almost non- 9

existent. Though species diversity generally increased southward, it was not a function of inlet proximity. Caging experiments indicate that foraminiferal densities in the IRL are regulated by predation (Buzas, 1978) by a wide variety of invertebrate and vertebrate animals (Buzas and Carle, 1979). The increase in densities towards the south, was positively correlated with decapod densities (Young et al., 1976), but negatively correlated with polychaetes and amphipods (Young et al., 1976). Alternatively, the physical lagoon environment is less variable toward the southern end of the IRL near the St. Lucie Inlet (Young et al., 1976; Young and Young, 1977; Nelson et al., 1982). In addition to characterizing the spatial distributions of Foraminifera, Buzas et al. (2000, 2002) conducted long-term monitoring to investigate the temporal variability of foraminiferal densities. The researchers concluded that there are large variations through seasons, years, and even decades. However, over the 20-year study, there was no general increase or decrease of foraminiferal densities leading to an overall long-term equilibrium. Nonetheless, species densities were shown to exhibit maximum densities at particular times during the year. For instance, Ammonia parkinsoniana reached maximum densities in the IRL during the summer similar to other studies (Boltovskoy, 1964; Jones and Ross 1979); however, other researchers recorded a maximum for this species in fall and spring (Murray and Alve, 2000). These inconsistencies were due to the fact that Foraminifera exhibit neither the same pattern nor density from year to year. As explained by the long- term studies: “We propose a model wherein individual foraminifers are spatially distributed as a heterogeneous continuum forming patches with different densities that are only meters apart; reproduction is asynchronous causing pulsating patches that vary in space and time. Thus we would expect significant differences among stations, years, seasons and their interaction. At the same time, no long-term increase or decrease in density for any of 10

the taxa is observed. Evidently, long-term stability is achieved through considerable short-term variability in space and time. Although observations at a single station are not indicative of a larger area at any particular time, the concept of pulsating patches indicates that observations at a station will in the long-term give an assessment of a larger area” (Buzas et al., 2002, p. 68)

Although Foraminifera within the IRL exhibit tremendous complexities both spatially and temporally, Buzas et al. (2002) argue that they are still valuable bioindicators due to their high densities and short generation times. Their high densities allow for a relatively small sampling area. Additionally, Foraminifera pass through several generations annually, unlike larger organisms which require decades of observation. Studies on distribution and diversity of foraminiferal assemblages in the Indian River Lagoon (Buzas et al., 2000) encourage long-term monitoring to clarify the relationship between seagrass and infaunal organisms, as well as follow long-term population trends. Foraminifera have not been used, however, as bioindicators in the IRL. This study seeks to examine foraminiferal populations of the IRL and characterize their environmental and sediment tolerances. It may then be possible to utilize foraminiferal population data as bioindicators of habitat quality. The objectives of this study are to (1) characterize the populations of benthic Foraminifera within Turkey Creek, Crane Creek, and surrounding areas of the IRL; (2) relate foraminiferal presence to various environmental parameters (salinity, temperature, dissolved oxygen, and sediment characteristics), and (3) document a pre-anthropogenic impact baseline of the diversity and abundance of Foraminifera in Turkey Creek and the IRL for comparison with the modern community. It is expected that overall foraminiferal densities will be inversely correlated with the presence of fine-grained organic-rich sediments (FGORS) and that abundance of the FGORS-tolerant Ammonia tepida species will increase

with the presence of FGORS up to a certain threshold. In addition, it is expected that the diversity of modern foraminiferal assemblages will be lower than that of historical assemblages due to FGORS accumulation.

Methods

To characterize the modern foraminiferal assemblages, surface samples (0-1 cm) were collected using a polyvinyl chloride (PVC) corer (inner diameter 8 cm), except for in deeper waters (>3m) where a petite ponar grab sampler was used. The corer area (50 cm2) meets that recommended for proper statistical sampling of foraminifers (Schönfeld et al., 2012). Sampling the upper 1cm of the sediment is an approach used in other monitoring studies (Cearreta et al., 2000; Murray and Alve, 2000; Leorri et al., 2008), and provides the best evaluation of foraminiferal populations (Bouchet et al., 2012). Standardized foraminiferal monitoring methodologies were adapted from Schönfeld et al. (2012). Sampling was conducted monthly from October-December 2015 at 16 stations in Turkey Creek, Crane Creek, and the surrounding IRL (Figure 3) with three replicate cores (or grabs) per station. Samples were preserved in 50 mL of 95% ethanol with 10 mg rose Bengal following collection. The total volume of the sample was taken to determine foraminiferal densities. Samples were sieved (63 μm) to remove fine grained material and then split. The split samples were sieved again (125 μm) and the coarser part of the assemblage was enumerated. Living foraminifers and skeletons were counted via stereomicroscopy. The densities from replicate cores were averaged from each station in order to characterize the patchiness of foraminiferal populations. Environmental conditions (temperature, salinity, and dissolved oxygen) were measured during each month and at each

station, both near the surface and benthos (within 2 cm on top of the benthic sediment), using a YSI Model 85 Probe. One deeper core (between 40-74 cm) at each station in Turkey Creek (Figure 3A, TC-1 through TC-4) was taken to determine if fossil foraminiferal densities change through time. A narrower core (2-cm diameter) was used to sample coarser sediments underneath muck at each of the four Turkey Creek stations. Sediment was subsampled at the top, middle, and bottom of each core. Each subsample was dried and sieved (125 μm) and the coarser portion used for identification and enumeration of organisms. Relative proportions (out of 300 total individuals) of different foraminiferal species were used to determine temporal shifts in the community (Buzas, 1970; Buzas, 1990; Schönfeld et al., 2012).

Figure 3 Primary study sites, including A. Turkey Creek (“Palm Bay”). B. Comparison study site in the Indian River Lagoon near the mouth of Turkey Creek (TCL). C. Comparison study site in the Indian River Lagoon near the mouth of Crane Creek (CCL). Yellow dots indicate locations of surface cores (triplicates of monthly sampling at each marked location) Yellow dots with no red line indicate deep sites where grab sampler was used instead of corer, and correspond to the muck sites TCM (A) and CCM (C).

Table 1 Location, region and site name of sampling stations. Site Region Name Latitude (N) Longitude (W) Palm Bay, Mouth of Turkey Creek TC 1 28° 2'20.09"N 80°34'50.65" Palm Bay, Mouth of Turkey Creek TC 2 28° 2'18.39"N 80°34'54.03" Palm Bay, Mouth of Turkey Creek TC 3 28° 2'16.07"N 80°34'54.41" Palm Bay, Mouth of Turkey Creek TC 4 28° 2'13.25"N 80°34'53.82" Shelf of Turkey Creek in IRL TCL 1 28° 2'33.97"N 80°34'54.21" Shelf of Turkey Creek in IRL TCL 2 28° 2'26.47"N 80°34'48.16" Shelf of Turkey Creek in IRL TCL 3 28° 1'49.46"N 80°34'33.28"W Shelf of Turkey Creek in IRL TCL 4 28° 1'38.55"N 80°34'28.94" Channel in Palm Bay TCM 1 28° 2'13.01"N 80°34'48.23" Channel in Palm Bay TCM 2 28° 2'8.51"N 80°34'49.64" Shelf of Crane Creek in IRL CCL 1 28° 4'35.89"N 80°35'52.77" Shelf of Crane Creek in IRL CCL 2 28° '24.25"N 80°35'54.38" Shelf of Crane Creek in IRL CCL 3 28° 4'15.29"N 80°35'51.48"W Shelf of Crane Creek in IRL CCL 4 28° 4'7.11"N 80°35'46.42" Crane Creek Muck #1 CCM 1 28° 4'39.16"N 80°36'4.52" Crane Creek Muck #2 CCM 2 28° 4'35.77"N 80°36'11.31"

Sediment from each station was analyzed for organic matter, silt and clay, and grain size. A sediment sample was heated at 105° C, weighed before and after to determine dry weight. The sample was then rehydrated, sieved (63 μm) to remove silt and clay, and re-baked, allowing dry weight of determination of the removed silt-clay portion. Grain size distributions (>63 μm) were determined using a shaker sieve and dried sediment. Since the distribution was unimodal and means and medians tended to match the mode of the sediment distributions, the mode (particle size most commonly found) was used as a measurement of the grain size. 16

Sediment organic content was determined by the loss-on-ignition (LOI) method (contrasting weight before and after heating at 550° C for 4 hours) (Heiri et al., 2001). Densities of total, living (stained), and dead (unstained) Foraminifera were calculated by dividing the number of individuals by the milliliters of wet sediment and compared using Analysis of Variance (ANOVA). Canonical Correspondence Analysis (CCA) was used to determine dominant relationships between community data and the 7 measured environmental variables (temperature, salinity, percent dissolved oxygen, oxygen saturation, silt and clay, organic content, and grain size).

Results

In the 144 samples, 4 species dominate: Ammonia parkinsoniana, Ammonia tepida, Elphidium excavatum, and Quinqueloculina seminula (Figure 4). Because species richness was low at all sites, with only 4 species making up >90% of all samples, the primary focus of this study is on foraminiferal densities and distributions. Total average foraminiferal density (Figure 5a) is lowest in the sites where the sediment fits the operational definition of muck (> 10% organic matter, > 60% silt and clay, and > 75% water by weight Trefry and Trocine, 2011), namely the sites in the channels of Crane Creek (CCM) and Turkey Creek (TCM) (Table 2). Total foraminiferal density is greatest in the transitional zone within Palm Bay at the mouth of Turkey Creek (TC), which exhibits intermediate quantities of percent water by weight, organic and silt and clay content, as well as percent dissolved oxygen (Table 2). Finally, in the sandy sediments within the shelves of Turkey Creek and Crane Creek (TCL and CCL, respectively) the average foraminiferal density is between 120-300 individuals per ml of wet sediment, significantly lower (p<0.01) than the average of about 750 individuals/ml in TC and significantly higher (p<0.01) than the average of ~50 individuals/ml in the muck sites. The average dead (unstained) and living (stained) populations of Foraminifera (Figures 5b and 5c, respectively) follow the same general pattern as the total average populations, with TC exhibiting the highest densities (p<0.001) of live and dead Foraminifera, TCL and CCL exhibiting intermediate densities of both 18

live and dead Foraminifera, and the muck sites showing the lowest densities (p<0.001) of both live and Foraminifera. In addition, all sites exhibit significantly lower densities of living individuals than dead individuals (TC, CCL, TCM/CCM p<0.001, TCL p<0.05).

A) B)

C) D)

Figure 4 4 major species of Foraminifera found in the study. (A) Ammonia parkinsoniana, (B) Ammonia tepida, (C) Elphidium excavatum, (D) Quinqueloculina seminula.

To determine the average proportions of living and dead individuals at each site without the influence of density, the living to dead ratios were calculated (Figure 8). The living to dead ratios were significantly higher (p<0.001) within intermediate and sandy sediments (TC, TCL and CCL) than in the muck sites, indicating that even though there are more dead individuals overall at each site,

there are a greater number of living individuals in TC, TCL, and CCL when compared to the muck sites. The graph also shows that the ratio of living to dead Foraminifera within the intermediate sediment (TC) is higher (P<0.001) than within the sandier areas (CCL and TCL).

Table 2 Averages of sediment parameters, and dissolved oxygen in bottom water at each site. Average Average Average Water Average Silt and Dissolved by Organic Clay Oxygen Weight Content Content (%) Site (%)(±1SE) (%)(±1SE) (%)(±1SE) (±1SE) 52.2 31.0 TC (±3.8) 6.28 (± 1.3) 29.3 (±5.2) (±7.4) 26.5 91.6 TCL (±1.5) 1.62 (±0.6) 2.55 (±0.7) (±11.6) 30.0 69.8 CCL (±2.2) 1.29 (±0.3) 3.3 (±0.4) (±4.9) 81.1 6.37 TCM/CCM (±1.7) 18.7 (±1.04) 83.3 (±4.24) (±3.31)

A) B)

B B

A A C A A C

C) D)

B B

A A C A A C

Figure 5 Average densities (number of individuals per ml of wet sediment) of quantitatively important benthic Foraminifera species within the shelf of Crane Creek in the Indian River Lagoon (CCL), the mouth of Turkey Creek (TC), the shelf of Turkey Creek in the Lagoon (TCL), and the channels within Turkey Creek and Crane Creek representing the muck sites (TCM/CCM). (A) average total (live plus dead) foraminiferal densities, (B) average total dead (unstained) densities, (C) Average total live (stained) densities of Foraminifera, and (D) the average total ratios of living to dead Foraminifera. Different letters indicate significant differences among sites based on ANOVA, with TC having the highest densities (p<0.01) and TCM/CCM having the lowest (p<0.01).

At all stations, Ammonia species made up >90% of the total foraminiferal population, with E. excavatum having the second highest densities within TC and TCL and Q. seminula having the second highest densities in CCL and the muck sites (Figure 6). Densities of Ammonia parkinsoniana are higher in Turkey Creek than in the sandy areas on the shelves of the lagoon, and at all three locations (TC, TCL, CCL) average total densities for A. parkinsoniana are significantly greater (p<0.001) than within the muck sites. This same pattern is followed by densities of 21

Elphidium excavatum, with significantly greater (p<0.001) densities within the intermediate and sandy sediments than within the muck sediments. For Ammonia tepida, the highest densities are found in the intermediate sediments of Turkey Creek, with the second highest densities (p<0.001) being found in the sandy sediments and the overall lowest densities found in the muck sediments (p<0.001). Densities of Quinqueloculina seminula were very low in all sites but surprisingly they were significantly higher (p<0.05) in the muck sites, and higher in the sediments on the shelf of Crane Creek than within the sandy areas on the shelf of Turkey Creek (TCL).

A) B) 600 B

350 A

300 500

1SE)

Density 250

400

)(+/

1 - 200 1SE) A - 300

150 )(+/

1 - A (individualsDensity 200 A C 100 A B ml

parkinsoniana 100

(individuals ml (individuals

0 tepida A. 0

C) D) 45 30 A A 40

25 A 35

1SE) -

20 30

Density

)(+/

1 - A B 25 15

1SE) 20 - AB A B

10 15

)(+/

Density (individualsDensity 1 - 10

5 ml

E. excavatum E. 5 (individuals ml (individuals

0 0 Q. seminula Q.

Figure 6 Spatial comparisons of mean total densities for each species: (A) total densities of Ammonia parkinsoniana species at each station, (B) total densities of Ammonia tepida at each station, (C) total densities of Elphidium excavatum at each station, and (D) total densities of Quinqueloculina seminula at each station. Densities are higher within the mouth of Turkey Creek (TC) for all foraminiferal species except Q. seminula, which has higher densities within the shelf of Crane Creek (CCL). Letters indicate significant differences based on ANOVA.

Average total living and dead Foraminiferal densities per species A)

250 )

1SE) 200 ±

)( Live 1 - 150 Dead

100 Densities (# of of (# Densities

A. parkinsoniana parkinsoniana A. 50

individuals ml individuals 0

B) 600

)

1 - 400

Live

Densities (# (# Densities

1SE) ±

( 200 Dead of individuals ml individuals of A. tepida tepida A. 0 C)

1SE) 20

± Live

)(

individuals individuals ml

E. excavatum excavatum E. Dead

Densities (# of of (# Densities 10

0 D)

1SE) 30

± Live

)(

1 - 20 Densities of (# Densities Dead

10 individuals ml individuals

Q. seminula seminula Q. 0 CCL TC TCL TCM/CCM Figure 7 Spatial comparisons of mean living and dead species: (A) average living and dead densities of Ammonia parkinsoniana species at each station, (B) average living and dead densities of Ammonia tepida at each station, (C) average living and dead densities of Elphidium excavatum at each station, and (D) average living and dead densities of Quinqueloculina seminula at each station. In all locations and for each species, the mean number of dead individuals is higher than the mean number of living individuals.

Figure 7 compares the average living and dead densities of each species at each site. For all species and sites, there are more dead individuals than living ones. The greater densities of Quinqueloculina seminula in the sediments of the shelf of Crane Creek (CCL) are driven by significantly higher densities of Q. seminula in station CCL-1. The sediment characteristics of CCL 1 are more closely related to those of intermediate sediments found in Palm Bay at the mouth of TC with average silt and clay and organic content averaging 9.4% and 2.8%, respectively. When compared to the other 3 stations within CCL, CCL-1 has significantly higher (p<0.01) levels of silt and clay and organic matter (9.4% vs. 1.3% silt and clay, and 2.8% vs. 0.8% organic content).

A) B) A B A A

B A A A

C) D)

A B B

Figure 8 Spatial comparisons within each species of average ratio of living densities to dead densities: (a) average ratio of living to dead A. parkinsoniana individuals for each station, (b) average ratio of living to dead A. tepida individuals for each station, (c) average ratio of living to dead E. elphidium individuals for each station, and (d) average ratio of living to dead Q. seminula individuals for each station. Letters represent significant differences based on ANOVA.

When looking at the living to dead ratios of each individual species (Figure 8) we see that in all sites for each species, there are more dead individuals on average than living individuals (ratio <1) except for Ammonia tepida within the intermediate sediments (TC). In addition, Elphidium excavatum is the only species that exhibits a higher living/dead ratio in the sites characterized by sandy sediments (TCL and CCL) than those characterized by intermediate or muck sediments (TC and TCM/CCM, respectively). The ratios of living/dead densities of Ammonia

parkinsoniana species are consistent among the sites with sandy and intermediate sediment, and significantly lower in the muck sediments. To relate densities of species to the environmental conditions studied, a Canonical Correspondence Analysis was performed. The results of the CCA (Figure 9) show that silt and clay content, OM and oxygen saturation/percentage of the bottom water are not only the major controlling factors of community composition (lengthiest vectors) but that they are inversely correlated with each other meaning that areas with higher OM tend to have a lower oxygen concentration. This inverse relationship between OM and DO is also shown in Figure 10. Temperature (range 21.2-28.3°C) and salinity (range 18.8-27.8 ppt) of the bottom water have the shortest vectors, suggesting that these environmental variables do not contribute much to the difference in community composition. The points represent the samples, with the colors indicating the location of the sample. Site points are based on linear combinations of the 7 measured environmental variables (raw data in Appendix). The position and separation of sites indicate that the sandier sites (TCL and CCL, excluding CCL-1 and TCL-2 see Appendix) are more positively correlated with increased oxygen concentrations and grain size, while the intermediate and muck sites (TC and TCM/CCM) are correlated with lower oxygen concentrations, and increased silt and clay content and organic matter. There are three main associations of foraminifers that can be identified based on the environmental influences of each site. First, the CCA suggests that Ammonia parkinsoniana and Elphidium excavatum are more closely associated with sandy sediments (TCL and CCL), increased oxygen concentrations (> 4 mg l- 1) and larger grain sizes, as well as decreased silt and clay content (< 2%) and organic matter (< 2%). Ammonia tepida densities are more closely associated with intermediate sediments (TC), with higher levels of silt and clay (> 20%) and organic matter (> 4%) and less dissolved oxygen (< 3 mg l-1). Quinqueloculina 27

seminula are outliers, yet seem to be obliquely associated with higher levels of silt and clay, and plot close to some muck (TCM/CCM) sites.

CCL 3.5 TC TCL Q. seminula 3.0 TCM/CCM

2.5

2.0

i E. excavatum

x 1.5 A

1.0

0.5 do osat

0.0 Grain sal silt temp organic

-0.5 A. park A. tepida

-1.0 -2.0 -1.6 -1.2 -0.8 -0.4 0.0 0.4 0.8 1.2 1.6 Axis 1

Figure 9 Canonical Correspondence Analysis (CCA) ordination diagram for living benthic foraminiferal densities and sites (TCL in red, CCL in blue, TC in black, and TCM/CCM in green) based on environmental variables (green vectors) and community composition. The lengthier vectors indicate the environmental variables that contribute the most to the differences in community composition, which include dissolved oxygen, organic matter, and silt and clay.

140

120

y = -21.41ln(x) + 63.945 100 R² = 0.7964 80

40 Oxygen Saturation Oxygen (%) 20

0 0 5 10 15 20 25 30 Organic Matter (%)

Figure 10 Scatter plot showing the inverse logarithmic relationship between OM and percent oxygen saturation.

The relationships between total foraminiferal density and DO as well as OM are shown in Figures 11a and 11b, respectively. The scatter plot (11a) shows low and high amounts of dissolved oxygen correlating with low densities of Foraminifera. Total foraminiferal density peaks between 2-5 mgl-1 DO. foraminiferal density is initially low with low amounts of OM (lowest 0.4%) and increases from 0 individuals ml-1 of sediment to around 750 individuals ml-1 from 0.4-5% OM. Density rapidly declines as OM increases beyond 5%.

A) 800

700

) 600

1 - 500 400 300

(individuals (individuals ml 200 100

Total Live Live ForaminiferalTotal Density 0 0 2 4 6 8 10 12 Dissolved Oxygen (mg l -1)

800

) 1 - 700 600 500 400 300 200

Total Live Live ForaminiferalTotal 100 Density (individuals Density ml 0 0 5 10 15 20 25 30 Organic Matter (%)

Figure 11 Total living foraminiferal density against dissolved oxygen (mg l-1) in bottom water (A) and percent organic matter (B). Scatter plot (A) shows high foraminiferal densities occurring between 2 and 4 mgl-1 of oxygen and low densities occurring at both low and high amounts of dissolved oxygen. Plot (B) shows highest foraminiferal densities occurring between 2-5% organic matter, with densities dropping off almost completely above 7% and below 2% organic matter.

The highly tolerant Ammonia species follow the same general patterns as the total live species in terms of density responses to DO and OM with the highest densities occurring between 2 and 5 mg l-1 DO and 1-7% OM (Figure 12a, b). There are higher densities of A. tepida with very low dissolved oxygen (0.03 mg l- 1) than A. parkinsoniana, suggesting that they are more tolerant of hypoxic conditions than A. parkinsoniana species. Overall, there are lower peak densities of A. tepida (360 individuals ml-1) than A. parkinsoniana (520 individuals ml-1) suggesting that in the right conditions, A. parkinsoniana may out compete A. tepida.

600

)

1 -

500

400

300 A. parkinsoniana

200 A. tepida

Live Densities (individuals ml (individualsDensitiesLive

100

Ammonia 0 0 5 10 15 Dissolved Oxygen (mg l-1)

600

B) )

1 -

500

400

300 A. parkinsoniana

200 A. tepida

Live Densities (individuals ml (individualsDensitiesLive

100

Ammonia 0 0 10 20 30 Organic Matter (%)

Figure 12 Scatter plots relating living densities of Ammonia parkinsoniana (filled in diamonds) and Ammonia tepida (open squares) to DO (mg l-1) (A) and OM (%) (B). Greater densities of Ammonia tepida are found at very low DO (0.09 mg l-1) than Ammonia parkinsoniana, suggesting that they are more tolerant of hypoxia. High densities (up to 100 individuals ml-1) of Ammonia tepida are even present at 25% organic matter, unlike Ammonia parkinsoniana.

To compare historical foraminiferal assemblages to contemporary assemblages, deep cores (> 40 cm) below the muck sediments were taken at the four stations within the intermediate sediments. Figure 12 shows the results of the analysis of the temporal shifts in community structure within TC, with the top 0-2 cm signifying the most recent sediment, and the deep 37-74 cm signifying older sediment which is below the muck layer. The species proportions follow the same density patterns as stated above, with Ammonia species comprising >90% of all samples. Based on the results of ANOVA, there were no significant differences among recent, intermediate, or older sediments for any of the 4 measured species. However, the graphs indicate an increasing trend of Quinqueloculina seminula from older to more recent sediments, which, when compared to the results of the CCA, would correlate to increasing amounts of silt and clay from the accumulation of fine-grained organic-rich sediments throughout the years. In addition, there seems to be a decreasing trend of Ammonia parkinsoniana abundances and an increasing trend of Ammonia tepida abundances which, when analyzed in conjunction with the results of the CCA correlate to decreasing oxygen levels and increased silt/clay and organic matter.

A) B)

C) D)

Figure 13 Results from deep cores (>40 cm) show temporal community shifts within the intermediate sediments of TC. Bar plots show foraminiferal abundances (out of 300 total individuals) of the 4 major species from the top (most recent sediment), middle, and bottom (oldest sediment, below muck layer) of deep cores. P-values of >0.05 for all 4 species indicate that there was no significant difference in the foraminiferal abundances from the top, middle or deep layers of the sediment core.

Discussion

Foraminiferal densities varied among TC, TCL, CCL and the muck sites

(CCM and TCM) and the primary drivers of spatial differences and community composition are dissolved oxygen and oxygen saturation, silt and clay content, and organic matter. This falls in line with views of most authors, that the most important factors controlling the foraminiferal microhabitats are oxygen and food

(Jorissen, 1995; Gooday, 1986; Corliss and Emerson, 1990). The mouth of Turkey

Creek contains sediment with intermediate oxygen concentrations (31.0% mean; ranging from 6% to 90%), silt and clay content (29.3% mean, ranging from 2.55% to 83.3%), and organic matter content (6.28% mean, ranging from 1.27% to 18.7%) and exhibits the highest foraminiferal densities. The muck-filled channels within

Crane Creek and Turkey Creek (TCM/CCM) have the lowest foraminiferal densities. It appears these Foraminifera favor some organic matter and silt and clay content, but only up to a certain point. We hypothesize that dissolved oxygen concentrations eventually become too low.

As oxygen concentrations begin to decrease and intolerant species die back, the total foraminiferal biomass may increase due to decreased predation and 35

competition with the declining macrofauna (Verhallen, 1991). The relationship between live foraminiferal density and dissolved oxygen (Figure 11a) illustrates the population increase that occurs before hypoxic conditions become intolerable. The highest foraminiferal densities occur between 2-6 mg l-1 and the lowest densities occur at either extreme. At a certain critical oxygen level, metazoan meiofauna diminish and subsequently even the most hypoxia tolerant species of Foraminifera decline, leaving only microbial communities behind (Josefson and Widbom, 1988).

Foraminifera most commonly use dissolved organic matter uptake and deposit feeding as their main trophic strategy (Lipps, 1983), feeding on bacterial biofilm, algal mats, and other types of readily available organic matter (Armynot du

Chatelet et al., 2009; Matera and Lee, 1972; Langer et al., 1989; Ozarco et al.,

1997) and this likely contributes to their success at the intermediately organic TC site where the average organic matter content is ~6%, as live foraminiferal densities are shown to be highest between 2-7% organic matter (Figure 11b). Alves Martins et al. (2015a) also recorded increases in foraminiferal density in association with enriched organic matter (4-7% TOC or ~12%-24% OM) in the Aveiro Lagoon,

Portugal. The decreased foraminiferal densities in the sandy locations of the shelves of Crane Creek and Turkey Creek (CCL and TCL) are, on the other hand, most likely a result of the decreased organic matter, or food, at these sites resulting in less reproduction and growth. In addition, increased predation by benthic

macroinvertebrates could be limiting foraminiferal densities in the sandier sites

(TCL and CCL), as predation has been shown to regulate foraminiferal densities within the IRL (Buzas, 1978).

Foraminiferal assemblages in the Indian River Lagoon are typical of estuarine environments, with just one or two species dominating at all sites

(Murray, 1991). The foraminiferal communities at all sites are dominated by

Ammonia tepida, a species known to be tolerant of hypoxic and even anoxic conditions, temperature increases, and high organic sediments (Cushman, 1928;

Karlsen et al,. 2000; Bouchet et al., 2007). The results of the CCA show that

Ammonia tepida are more closely associated with the intermediate sediments, which exhibit higher levels of organic matter content and lower dissolved oxygen percentages when compared to the sandy areas of TCL and CCL. Ammonia tepida even exhibit densities above 100 individuals ml-1 in dissolved oxygen concentrations as low as 0.03 mg l-1. The foraminiferal community within the

Aviero Lagoon, a polyhaline anthropized lagoon, was also dominated by Ammonia tepida and Elphidium excavatum, both of which are associated with high enrichment of organic matter (~12%-24% OM) in that area (Alves Martins et al.,

2015a). The CCA suggests that Ammonia parkinsoniana and Elphidium excavatum are more closely associated with higher oxygen concentrations and lower silt and clay and organic matter concentrations than Ammonia tepida, also correlating with

sandier environments. E. excavatum has been documented as being less tolerant to hypoxic conditions (Sen Gupta, et al., 1996). In the Bizerte Lagoon in Tunisia, which is also influenced organic matter flux, foraminiferal assemblages were shown to be dominated by A. parkinsoniana (Alves Martins et al., 2015b). The more polluted sediments in the Rio de Aveiro (Alves Martins et al., 2015a) could explain A. tepida’s dominance in that region relative to A. parkinsoniana. A. parkinsoniana has been shown to be more sensitive to polluted environments and high heavy metal concentrations (Coccioni et al., 2009). These sensitivities could explain A. parkinsoniana’s correlation with higher oxygen and sandier sediments in this study.

All species except for Q. seminula have higher total average densities within the intermediate sediments. The CCA indicates Quinqueloculina seminula is associated with sediments containing higher silt and clay and organic matter content. Q. seminula exhibits highest mean densities in CCL, within the IRL where one would not expect high silt and clay. However, looking at individual sampling stations, it is apparent that high CCL Q. seminula densities are being driven largely by station CCL-1 alone, where the average sediment silt and clay content (9.4%) and organic matter content (2.8%) more closely resemble intermediate sediments within TC. The average silt and clay content and organic matter content in the other

3 CCL stations are 1.3% and 0.8%, respectively. Though the dissolved oxygen

content is lower at CCL-1 (average 3.5 mgl-1) than the other stations within CCL

(average 4.9 mgl-1), it is still within the range where the highest foraminiferal densities are seen. Therefore, the higher organic matter, along with the higher amounts of oxygen could be providing the optimal conditions for the proliferation of this species. Previous studies have suggested that Quinqueloculina respond to influxes of phytodetritus, and will grow and reproduce until they exhaust that food source (Gooday et al., 2010). This suggests that Quinqueloculina are likely opportunists (Jorissen et al., 1995), possibly reproducing more rapidly than normal or migrating to the surface sediment (Gooday et al., 2010) when there are pulselike inputs of food such as phytodetritus.

Overall, there were more dead than living individuals at all sites; however, the living/dead ratios indicate that there are higher ratios of living/dead in the intermediate sediment (TC). When looking at individual species, however, the ratio of living/dead E. excavatum is higher in the sandy sediments relative to intermediate or muck sediments, most likely due to their limited tolerance of hypoxic conditions (Sen Gupta et al., 1996). In addition, there is a greater density of living Ammonia tepida species (ratio>1) within the intermediate sediments of

Turkey Creek, most likely due to the increased organic matter and their ability to survive in low oxygen conditions (Cushman, 1928; Karlsen et al,. 2000; Bouchet et al., 2007). Numerous studies have shown that hypoxic/anoxic conditions impact

foraminiferal populations, probably through reproductive inhibition due to low oxygen (Alve and Bernhard, 1995; Moodley et al., 1997; Jannik et al., 1998). In general, these results, along with the results of the CCA, indicate a ranking of low oxygen tolerance for the 3 species from the taxonomic Order Rotaliida: A. tepida,

A. parkinsoniana, and E. elphidium (highest to lowest hypoxic tolerance). The higher total living/dead ratios within the intermediate sediments might best be explained by limiting factors in sandy and muck sediments. There may not be enough organic matter in sandy areas, nor any oxygen in muck sediments, for survival or reproduction of Foraminifera. In a model proposed by Jorissen et al.

(1995) based on meiobenthos in bathyal, abyssal and hadal deep sea systems of the western Pacific, benthic foraminiferal distributions were limited by oxygen in eutrophic environments (<0.2ml l-1), but by food in oligotrophic ones. This interpretation was also suggested by Corliss and Emerson (1990) in the northwest

Atlantic Ocean. In the case of this study, the intermediate and muck sediments (TC and TCM/CCM) could be considered eutrophic environments limited by oxygen, while the sandy sediments (TCL and CCL) would be more akin to oligotrophic environments, limited by food. This relationship is shown by the conceptual model in Figure 14. By looking at the foraminiferal density relationships with oxygen and

OM in this study, the critical oxygen requirement appears to be >2 mg/l, while the

OM requirement ranges from 1-10%.

H2S H2S

- Oxygen + + Food -

Muck Intermediate Sandy TCM/CCM TC TCL/CCL

Based on Jorrissen et al., 1995

Figure 14 Conceptual model based on Jorissen et al. (1995) to generally fit the areas of study within the IRL ecosystem. The sizes of the species pictures are relatively based off of their densities within each site. The muck sites are characterized by increasing OM (food) and decreasing DO, as well as lower densities of Foraminifera. The intermediate sites (TC and CCL-1) have the overall highest densities, and are dominated by Ammonia tepida. The sandy areas are characterized by higher levels of DO and lower percent OM, and have lower foraminiferal densities than the intermediate sediment.

A parameter not measured in this study, but that can limit benthic foraminiferal survival, is hydrogen sulfide (Moodley et al., 1998a, b). In the northern Arabian Sea Oxygen Minimum Zone (OMZ) Jannink et al. (1998) found a flourishing benthic foraminiferal community in hypoxic to anoxic bottom waters.

However, sulfate reduction was absent in the surface sediments of that ecosystem.

The north Arabian Sea species were of the same taxonomic Order (Rotaliida) as

Ammonia and Elphidium, all of which possess perforate hyaline tests. Hydrogen sulfide odors were evident in this study and especially high in the muck sites. This is consistent with what is known about the degradation of polluted organic sediments (Mojtahid et al., 2008). Therefore, it is a fair assumption that sites with the highest sediment organic matter content and lowest dissolved oxygen (e.g.,

TCM and CCM) will also have the highest hydrogen sulfide content, further stressing foraminiferal communities and limiting their densities.

Deep cores penetrating the muck layer were taken to document changes in foraminiferal densities, species, and live/dead ratios over recent time. The accumulation of muck layers in the IRL began around 60-80 years ago (Trefry,

1990). Following foraminiferal populations from deep cores penetrating the muck layer, temporal community shifts were not seen in the intermediate sediments (TC)

(Figure 11). However, there is an overall increasing abundance of Quinqueloculina seminula in more recent sediments within TC. Taking into account the results of the CCA, Q. seminula display an oblique correlation with sediment silt and clay content and organic matter. Therefore, the increased abundances of Q. seminula could be a response to increasing silt and clay content, as fine-grained organic-rich sediment has been increasing at an average rate of 0.7 ±0.6 cm/yr in Crane Creek

(Trefry, 2004). In addition, increasingly frequent algal blooms (Philips et al., 2003) could also influence Q. seminula abundances. Q. seminula populations have been associated with high quality OM (high fresh chlorophyll (Chl a)) (Dessandier et al.,

2016), and increased densities of Q. seminula in response to phytodetritus (Gooday et al., 2010).

Conclusion

Overall, studying the impact of environmental conditions on foraminiferal densities and distributions reveals the importance of OM and DO for foraminiferal abundances in the IRL. The sensitivity of Foraminifera to OM and DO in the IRL, as well as their short generation times (Schönfeld, et al. 2012), yield an ecosystem where the community can respond rapidly to anthropogenic activities, including pollution and mitigation. In general, foraminiferal densities are highest in sediments characterized by intermediate levels of DO and OM. In muck sediments, densities are likely limited by oxygen, whereas in sandier sediment densities organic matter appears to be more important.

In terms of individual species, Ammonia tepida are not only dominant, they are also the most tolerant, surviving in hypoxic conditions. Ammonia parkinsoniana are the second most tolerant species and Elphidium excavatum are the least tolerant of the rotaliids. Quinqueloculina seminula were most successful at one station with a unique combination of high DO and high sediment organic content, and they are obliquely associated with silt and clay and OM. Collectively,

the relative abundances of these species can serve as proxy indicators of sediment and associated environmental conditions.

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Appendix

Environmental Data for Each Station Grain Size Site Month Temp (°C) Salinity (ppt) DO (mg l¯¹) Oxygen Saturation (%) Silt and Clay (%) Organic Matter (%) (microns) TC 1 Oct 27.3 21.4 2.8 62.5 6.5 1.77 125 TC2 Oct 27.4 21.4 0.09 0.8 37.8 6.91 180 TC3 Oct 27.4 19.1 2 20 37.9 3.02 125 TC 4 Oct 27.4 21.4 0.02 0.3 24.5 9.5 125 TC 1 Nov 27.8 21.8 2.85 40.9 7.9 2.4 250 TC2 Nov 27.9 21.8 2.95 43.5 23.3 5.5 180 TC3 Nov 28.1 20.8 0.2 5 42.3 9.3 125 TC 4 Nov 28.3 20.3 2.16 19 13.8 4.1 125 TC 1 Dec 22.5 20.6 4.3 57 8 2.2 335 TC2 Dec 22.8 21.9 4.1 53.5 30 5.5 125 TC3 Dec 22.8 22.4 1.14 13.8 40.6 7.1 125 TC 4 Dec 22.9 19.2 0.03 0.5 87.7 24.5 90

56 TCL 1 Oct 27.8 21.2 6.35 81.6 1.3 0.8 180

TCL 2 Oct 27 21.4 2.99 42 5 1.5 180 TCL 3 Oct 27.3 20.9 5.2 76.2 0.8 10 180 TCL 4 Oct 27.1 20.6 5.44 77 0.7 0.6 180 TCL 1 Nov 27.6 27.8 5.6 74.1 1.8 0.6 180 TCL 2 Nov 27.2 22 2.57 36.4 10.3 2.2 180 TCL 3 Nov 28.3 22.2 6.09 89.5 0.1 0.5 180 TCL 4 Nov 28.1 22.1 5.83 78.8 3.6 0.5 250 TCL 1 Dec 21.8 21.6 7.99 102.5 1 0.1 180 TCL 2 Dec 21.2 19.8 0.007 0.2 3.8 2.5 125 TCL 3 Dec 22 21.5 10.04 120.2 1.1 0.3 250 TCL 4 Dec 22.1 19.3 7.1 89.7 0.7 0.7 180 CCL 1 Oct 27.3 21 2.88 39.5 0.7 3.3 125

Grain Size Site Month Temp (°C) Salinity (ppt) DO (mg l¯¹) Oxygen Saturation (%) Silt and Clay (%) Organic Matter (%) (microns) CCL 3 Oct 27.5 20.9 4.38 62.5 1.6 1.3 180 CCL 4 Oct 27.7 18.8 5.6 76.5 1.3 0.8 180 CCL 1 Nov 26.8 22.3 2.49 31.2 18.7 4.1 125 CCL 2 Nov 27.6 21.9 3.58 50.7 1.2 0.6 180 CCL 3 Nov 28 21.9 3.88 57 1 0.6 125 CCL 4 Nov 27.9 22 5 75.5 1 0.4 180 CCL 1 Dec 21.2 23.6 5.17 66.5 8.8 1 250 CCL 2 Dec 21.7 23.3 5.69 73.5 1.8 1.3 180 CCL 3 Dec 21.7 21.9 5.67 75 1.4 0.8 125 CCL 4 Dec 22 21.7 6.13 76.6 1.1 0.7 125 TCM 1 Oct 27 21.7 1.65 2.5 91.4 16.8 125 TCM 2 Oct 27.3 21.1 0.03 0.1 98.5 18.7 125

57 TCM 1 Nov 27.7 21 0.01 0 96 19.1 125

TCM 2 Nov 26.9 19.8 0.01 0.1 89.9 18.1 125 TCM 1 Dec 22.8 21.9 0.01 0.2 91.2 21.4 90 TCM 2 Dec 22.7 22.4 0.06 0.8 90.9 20.5 90 CCM 1 Oct 27.6 21.4 0.03 0.8 95 19 125 CCM 2 Oct 27.6 21.3 0.3 2.2 63.3 16.4 90 CCM 1 Nov 27 20 0.02 0.2 87 18.7 125 CCM 2 Nov 25.5 21.6 0.02 0.1 49.2 14.1 90 CCM 1 Dec 21.7 22.7 0 0.1 97.2 24.1 125 CCM 2 Dec 21.8 23.3 0.4 10 55.7 11.3 125