The closely related killifishes Fundulus heteroclitus and F. grandis hybridize in a small region where their ranges overlap in coastal northeastern Florida. Hybrids of these species are rare in frequency within the contact zone, suggesting the presence of relatively strong reproductive isolation between these species. The objective of this study was to elucidate barriers to reproduction between F. heteroclitus and F. grandis in the lab, as well as to quantify the relative strengths and contributions of various isolating barriers. Pre-zygotic (mating and fertilization) and post-zygotic (hatching) barriers were investigated by performing a variety of choice and no-choice laboratory mating experiments. The results revealed that under no-choice conditions, barriers to mating had the biggest influence on hybrid production in F. grandis, whereas hatching barriers contributed to the majority of reproductive isolation in F. heteroclitus. However, under choice conditions pre-zygotic barriers had the greatest influence on both species’ ability to produce hybrids. The total relative reproductive isolation that was observed in females of each species was stronger in F. heteroclitus than in F. grandis overall, and was nearly complete in F. heteroclitus females under choice conditions while moderate in F. grandis females. These results reveal an asymmetry in the potential gene flow between these two species, with F. grandis being more likely to hybridize than F. heteroclitus in the absence of environmental influences.

Scleractinian coral populations are declining worldwide in response to numerous stressors operating on both global and regional scales. Rising sea surface temperatures associated with global climate change and the increasing frequency of coral-macroalgae competitive interactions are two of the gravest ecological drivers facing coral reef ecosystems. However, little is known about how these stressors interact to impact corals, their health, and potential modes of population recovery. These threats also highlight the need to develop reliable techniques that detect stress in multiple life-history stages of hermatypic corals prior to the degradation of coral reef habitats. To address these concerns we evaluated the effects of elevated sea surface temperatures (+3.5°C), Dictyota menstrualis competition, and their combined impacts on three life-history stages of the reef-building coral Porites astreoides. Elevated temperature induced sub-lethal stress yet had varied responses that were contingent on the life-history stage being examined. Hyperthermal stress did not consistently effect the transcriptional expression of heat shock proteins (Hsp) 16 or 60, but was readily detected utilizing biomarkers of the oxidative stress pathway. The presence of D. menstrualis significantly reduced coral survival and recruitment beyond simple space occupation in every coral life-history stage examined.While macroalgal exposure and elevated temperature had distinct effects on coral survival and physiological condition, the combination of both stressors induced a synergistic impact on biomarkers of oxidative stress in coral larvae. The results highlight the potential of biomarkers of oxidative stress for detecting hyperthermal stress in scleractinian corals. They also support the accepted notion that benthic macroalgae compete with reef-building corals via direct contact for space on coral reefs and that elevated temperatures can reduce the health of the coral holobiont. In addition, the results indicate that larvae from P. astreoides are more susceptible to the impacts of hyperthermal stress compared to established corals and that multiple perturbations can interact to exacerbate coral health.

Reduced reproduction has been shown to increase lifespan in many animals, yet the mechanisms behind this trade-off are mostly unknown. A previous study has shown that in the lubber grasshopper, Romalea microptera, ovariectomized (OVX) individuals have a 30% increase in lifespan relative to controls (Sham). In a separate study, an increase in fat body mass and a halting of ovarian growth were seen upon reduction of vitellogenin transcript via RNAi (VgRNAi). These data suggest that VgRNAi increases lifespan through the trade-off between reproduction and longevity and animals with combined ovariectomy and VgRNAi, might show additive physiological responses. In this study, we used two injection control groups for the VgRNAi treatment, namely buffer injection or injection with RNAi against a 90kDa hexamerin storage protein (Hex90RNAi). We have combined these manipulations to test lifespans upon: OVX & VgRNAi, OVX & Hex90RNAi, OVX & Buffer, Sham & VgRNAi, Sham & Hex90RNAi, and Sham & Buffer. Ovariectomy and VgRNAi exhibited similar reductions in feeding (~40%) and extensions in lifespan (13-21%) but showed differences in vitellogenin protein levels. This study also observed the effects of reduced reproduction on hexamerin storage proteins. We observed that upon ovariectomy and VgRNAi, hexamerins were increased, emphasizing the importance of protein in insect life extension. When methods to reduce reproduction were combined (OVX VgRNAi), no additive physiological responses were observed, suggesting ovariectomy and VgRNAi each extend lifespan by overlapping or convergent pathways.

The Deepwater Horizon Oil Spill (DWH) released about 4.4 million barrels of crude oil into the Gulf of Mexico (GOM), making it one of the largest oil spills in U.S. history. Additionally, the depth of the spill (i.e., 1500 meters) created a unique research opportunity because most oil spills occur at the surface and affect coastal rather than deepwater habitats. Polycyclic aromatic hydrocarbons (PAHs) are the most toxic components of oil, and are often the focus of oil exposure studies. PAHs are quickly metabolized by vertebrates; therefore, indicators of biological responses to PAH exposure (PAH “biomarkers”) such as the levels of PAH detoxification enzymes and the resulting metabolites are commonly used to examine oil exposure. This study measured multiple PAH biomarkers including hepatic activity of the PAH detoxification enzymes cytochrome P4501a1 (CYP1A) and glutathione-S-transferase (GST), as well as biliary PAH metabolites in deep sea sharks and bony fishes from areas affected by the Deepwater Horizon Oil Spill. Samples were collected from 2011-2013 from seven species of sharks, with special focus on the four most abundant deep sea species: Centrophorus niakang, Centrophorus cf granulosus, Squalus cubensis and Squalus cf mitsikurii. Overall enzyme activity was low in these sharks, yet it was higher in oiled sites compared to reference locations. Additionally some species showed declining CYP1A activity since the time of the oil spill, suggesting exposure to CYP-inducing compounds during the beginning of the survey period. Last, PAHs of a petrogenic nature were more abundant in oiled sites compared to reference locations. Overall, this project provides the much need biomarker data for sharks as well as insight on exposure and metabolism of PAHs in deep sea sharks after the DWH.

Sindbis virus (SINV) is an arthropod-borne Alphavirus in the family Togaviridae. Sindbis virus has a broad host range that includes avian, mammalian, and human hosts; therefore, its receptor(s) is/are highly conserved. When the mosquito imbibes a viremic blood meal, the virus infects the midgut cells, disseminates into the hemolymph, and eventually infects the salivary glands. The midgut is an organ of transmission and the virus must overcome the midgut epithelia infection- and escape-barriers. Sindbis virus infection is determined by the chance collision of the glycoproteins with a compatible receptor. Research has supported the involvement of high-affinity laminin receptor and heparan sulfate in SINV binding to host cells. However, it has been suggested that not all strains of SINV are dependent on heparan sulfate for attachment/entry and that SINV could be utilizing multiple receptors. A study using Drosophila demonstrated that, of the nine genes that encode for proteins that enhance SINV infection, only natural resistance-associated macrophage protein (NRAMP) was conserved. A symporter of divalent metals and hydrogen ions, NRAMP is ubiquitously expressed. Overexpression of NRAMP led to an increase in SINV infection of human cells while deletion of NRAMP in mouse and Drosophila decreased SINV infection. Sindbis virus could be utilizing this protein to overcome the infection barriers of mosquito midgut epithelia. In this study, NRAMP was localized to Aedes aegypti and Anopheles quadrimaculatus tissues via immunofluorescence assay and TR339-TaV-eGFP was detected in the midgut epithelia and visceral muscles. We suspect that NRAMP was detected on midguts and/or Malpighian tubules of Aedes aegypti and Anopheles quadrimaculatus. The similarities between the pattern of NRAMP labeling and TR339-TaV-eGFP infection of the midgut suggest that SINV infection is influenced by NRAMP in the midgut epithelia. Because NRAMP is ubiquitously expressed, this research provides insight into the attachment and entry phase of the arbovirus lifecycle.

In the past century, seagrasses have experienced massive die off episodes in what has been collectively referred to as “wasting disease.” Researchers generally agree that wasting disease is caused by a protist of the Labyrinthula genus, and that environmental stressors can make some populations of seagrasses more susceptible to infection. The purpose of this study was to examine the combined effects of elevated salinity, elevated temperature, elevated sulfide and night-time hypoxia on Thalassia testudinum health and its response to Labyrinthula sp. infection under controlled conditions. To test these effects, microcosms were utilized and individual seagrass shoots were randomly assigned to treatment groups consisting of various combinations of abiotic stressors. They were then infected with Labyrinthula sp. and monitored for lesion formation and a reduction in photosynthetic efficiency. It was hypothesized that seagrasses incubated under the application of a given stressor would show evidence of declining health, and, in turn, would be more prone to infection, as quantified by lesion size and reduced photosynthetic capacity. Results indicated that abiotic stressors have little effect on T. testudium’s ability to resist infection from Labyrinthula sp. However, the Labyrinthula sp. was highly sensitive to abiotic stressors, specifically salinity, indicating that the health of the pathogen greatly contributed to the severity of the disease. Therefore, the stress thresholds of both the host seagrass and the pathogen need to be considered. Indeed, the interaction(s) among T. testudinum, Labyrinthula spp. and the environment are complex and not as linear as previously thought.

Carnivorous plants perform as both producers and consumers. Botanical carnivory has evolved in sunny, moist, nutrient-poor environments, and the primary nutrient supplied by prey is proposed to be nitrogen. There is a trade-off between carnivorous and photosynthetic structures which corresponds to degree of carnivory expression and available nutrients. This study was conducted on the hooded pitcher plant, Sarracenia minor, which is a facultative wetland plant and Florida-threatened species. Sarracenia minor is considered a specialist myrmecophage and ants characterize the majority of attracted and captured prey. Ants not only provide nutritional benefit, but also protection against herbivory. A natural population of S. minor in northeast Florida was selected to test response to prey and fertilizer nitrogen in a press-experimental design. Introduced fire ants (Solenopsis invicta) and ammonium nitrate (NH4NO3) were used as prey and fertilizer nitrogen sources, respectively. Treatments included: 1) ant addition; 2) fertilizer addition; 3) ant addition/fertilizer addition; 4) no ants/no fertilizer; 5) control. Treatments were administered biweekly and morphological characteristics and herbivory were measured monthly from April-November 2012. Results indicated no significant treatment effects on plant performance and morphological characteristics, except for a significantly greater number of flowers displayed by the nutrient-deprived group (p < .005). Herbivory by Exyra semicrocea also showed a marginally significant negative effect on the tallest pitchers per ramet. Since nitrogen is primarily stored by pitchers and allocated to new growth in the following growing season, the predictive power of this study may be limited. However, increased flowering rate in the nutrient-deprived group suggests that plants were induced to flower from nutrient stress. Also, a burn at the beginning of the study likely influenced nutrient availability and plant response to experimental treatments. In conclusion, stress may have occurred from both fire and nutrients, and S. minor showed resistance and poor response to nitrogen addition.

Shark fisheries are a multimillion dollar industry in the United States and have significant contributions both recreationally and commercially. In order to maintain this industry, fisheries must be properly managed. An understanding of life history and reproduction is crucial in order to adequately manage shark fisheries. The finetooth shark, Carcharhinus isodon, is a member of the small coastal shark (SCS) fishery complex. It is found in Atlantic waters from South Carolina to Florida and throughout the Gulf of Mexico. This species has recently come under increased fishing pressure and has previously been overfished. New data is needed in order to properly assess the reproductive capacity of this species so that healthy populations can be sustained. This study evaluated the overall reproductive cycle of northwest Atlantic populations of C. isodon using composite variables of morphology, histology and endocrinology. Atlantic C. isodon were found to display seasonal reproduction with biennial reproductive periodicity with a 12 month gestation period and litters of 2-6 pups. E2 and T displayed similar patterns and were highest during gametogenesis. P4 did not show any discernible patterns. Estrogen receptors were found in developing follicles of gravid and non-gravid females, in the oviducal glands of non-gravid and early pregnant females and in the uterus of pre-ovulatory females. P4 was found in the oviducal glands in all reproductive stages and in the uterus of pre-ovulatory females. Androgen receptors were found in Sertoli cells and mature spermatozoa as well as epididymal epithelial cells.

Oyster reefs are declining worldwide, as well as the economic and ecological value of oysters to their respective systems. Numerous restoration efforts have been undertaken in hopes of re-establishing these shellfish populations. This study evaluated a restoration project within the Guana Tolomato Matanzas estuary in northeast Florida, U.S.A., to investigate community structure as well as seasonal patterns in species abundance and diversity of juvenile fish and benthic macrofauna within restored and unrestored intertidal habitats along the Guana Peninsula. The first objective was to determine whether the artificially created reefs provided similar quantity and diversity of benthic macrofauna as adjacent unrestored habitats. The second objective was to specifically characterize resident and transient fish assemblages associated with the artificial reef and adjacent unrestored habitats. Benthic macrofauna were quantified using plastic settlement trays deployed in triplicate at each site and sampled monthly for a year. Community structure differed by habitat, confirmed through an analysis of similarity. High abundances of Petrolisthes armatus on the natural reef sites largely contributed to dissimilarity in community composition between the natural reef and the restored sites. Fish assemblages were quantified using monthly seine and gill nets set adjacent to restored and unrestored intertidal habitats. Diversity was similar between the restored and unrestored sites, however, there was very little species overlap between the two sites. The dominance of post-larval and juvenile spot (Leiostomus xanthurus), anchovies (Anchoa mitchelli and A. hepsetus) and mullet (Mugil sp.) led to high density and low diversity in seine collections during the winter months. Overall, the gill net survey did not show any patterns in fish abundance associated with particular habitats in the area, however this is the first assessment in this region using gill netting. Constructed oyster reefs created immediate habitat for resident species and enhanced habitat value compared to unstructured mud bottom.

Lipopolysaccharide (LPS) was shown to serve as a strong elicitor of the early defense response in the subtropical seagrass Thalassia testudinum Banks ex König and was capable of inducing an oxidative burst identified at the single cell level. The formation of reactive oxygen species (ROS) included a diphenylene iodonium sensitive response, suggesting the involvement of an NADPH oxidase. A 900 bp fragment of this enzyme was sequenced and found to encode a NAD binding pocket domain with extensive homology to the Arabidopsis thaliana rbohF (respiratory burst oxidase homolog) gene. Pharmacological dissection of the early events preceding ROS emission revealed that seagrasses contain ROS-generating machinery and signal transduction components that appear to be evolutionarily conserved with the defense response systems of terrestrial plants. It is undetermined whether or not the increased ROS associated with the oxidative burst is simply an antimicrobial agent or a signaling molecule that will initiate programmed cell death (PCD) and lead to the hypersensitive response (HR), a process not yet characterized in seagrasses. ROS accumulation was found to increase around the lesion as the duration of infection increased. The only PCD characteristic observed following infection was a slight increase in caspase-like protease activity around the lesion. Immunohistochemistry revealed inconsistent activity of proteases. Detection of nuclear condensation by TUNEL and Hoechst staining were also inconclusive and showed diffuse genetic material throughout the cytoplasm. It appears as though lesions from Labyrinthula spp. infection are likely to be a direct result of pathogen-based damage as opposed to host PCD.