We investigated the modifying effects of ultraviolet (UV) light and chemical dispersant (Corexit 9500A) on crude oil toxicity in juvenile mysids (≤ 24 h) (Americamysis bahia) and larval red drum (24-72 hpf) (Sciaenops ocellatus). These results demonstrate that crude oil toxicity significantly increases with co-exposure to environmentally relevant UV levels in both species, indicating photo-induced toxicity. This toxicity was further exacerbated by the application of chemical dispersants which increased the dissolution and concentration of oil-derived polycyclic aromatic hydrocarbons (PAHs) in test solutions. To better understand the mechanisms and initiating events of this observed photo-induced toxicity, the incidence of apoptotic cell death and global transcriptomic changes were assessed in larval red drum (24-72 hpf) following co-exposure to crude oil and UV. These results showed that co-exposure to UV and low concentrations of crude oil (<1 µg/L ∑PAH50) induced apoptotic cell death in skin and eye tissue and altered transcriptomic pathways related to visual processing and dermatological disease. To link these cellular and molecular impacts of photo-induced toxicity to apical endpoints of ecological performance, sublethal impacts to growth, metabolic rate, and visually mediated behaviors were explored in larval red drum at 2 developmental stages. These results suggested that earlier life stages may …

Oil spills are well-known adverse anthropogenic events, as they can induce severe impacts on the environment and negative economic consequences. Still, much remains to be learned regarding the effects of crude oil exposure to aquatic organisms. The objectives of this dissertation were to fill some of those knowledge gaps by examining the effects of Deepwater Horizon (DWH) crude oil exposure on teleost kidney development and function. To this end, I analyzed how these effects translate into potential osmoregulatory impairments and investigated the interactive effects of ubiquitous natural factors, such as dissolved organic carbon (DOC) and ultraviolet (UV) light, on acute crude oil toxicity. Results demonstrated that acute early life stage (ELS) crude oil exposure induces developmental defects to the primordial kidney in teleost fish (i.e., the pronephros) as evident by alterations in: (1) transcriptional responses of key genes involved in pronephros development and function and (2) alterations in pronephros morphology. Crude oil-exposed zebrafish (Danio rerio) larvae presented defective pronephric function characterized by reduced renal clearance capacity and altered filtration selectivity, factors that likely contributed to the formation of edema. Latent osmoregulatory implications of crude oil exposure during ELS were observed in red drum (Sciaenops ocellatus) larvae, which manifested reduced survival …

The bristlenose pleco (Ancistrus cirrhosus) is a species of armored catfish in the Loricariidae family that breathes air facultatively when the aquatic environment becomes hypoxic. The bristlenose pleco uses its highly vascularized stomach as an air breathing organ. The two main goals of this developmental study were to determine the size of onset of air breathing and to determine the frequency of air breathing behavior in bristlenose plecos from juveniles to adults. Developing juveniles reach functional maturity within four to six months of hatching and grow to an adult size of eight to ten cm in length. To examine the developmental timing for the onset of air breathing, we tested different sized juveniles beginning at one cm up until 8 cm in length. The developmental timing for the onset of air breathing was measured by exposing each fish to a slowly decreasing aquatic oxygen content from 100% air saturation down to 8% air saturation. Fish were first able to breathe air at just over 2 cm and 1 gram in mass. There was a weak negative correlation between fish length and % air saturation at which air breathing began. When exposed to 15% air saturation, frequency of air breathing was …

Recent epidemiological studies have demonstrated a correlation between the manifestation of non-alcoholic fatty liver disease (NAFLD) and ambient air pollution levels, which is exacerbated by the presence of other risk factors, such as diabetes, dyslipidemia, obesity, and hypertension. We investigated the hypothesis that exposure to a mixture of gasoline and diesel engine emissions (MVE) coupled with the concurrent consumption of a high-fat (HF) diet promotes the development of a NAFLD phenotype within the liver. Three-month-old male C57Bl/6 mice were placed on either a low fat or HF diet and exposed via whole-body inhalation to either filtered (FA) air or MVE (30 µg PM/m3 gasoline engine emissions + 70 µg PM/m3 diesel engine emissions) 6 hr/day for 30 days. Histology revealed mild microvesicular steatosis and hepatocyte hypertrophy in response to MVE exposure alone, compared to FA controls, yielding a classification of "borderline NASH" under the criteria of the modified NAFLD active score (NAS) system. As anticipated, animals on a HF diet exhibited moderate steatosis; however, we also observed inflammatory infiltrates, hepatocyte hypertrophy, and increased lipid accumulation, with the combined effect of HF diet and MVE exposure. Immunofluorescence staining and RT-qPCR of the liver revealed the presence of lipid peroxidation, altered expression …

In this work, we have performed comparative evolutionary analysis, functional genomics analysis, and machine learning analysis to identify the molecular factors that discriminate between multipartite and unipartite bacteria, with the goal to decipher taxon-specific factors and those that are prevalent across the taxa underlying the these traits. We assessed the roles of evolutionary mechanisms, namely, horizontal gene transfer and gene gain, in driving the divergence of bacteria with single and multiple chromosomes. In addition, we performed functional genomic analysis to garner support for our findings from comparative evolutionary analysis. We found genes such as those encoding conserved hypothetical protein DR_A0179 and hypothetical protein DR_A0109 in Deinococcus radiodurans R1, and Putative phage phi-C31 gp36 major capsid-like protein and hypothetical protein RSP_3729 in Rhodobacter sphaeroides 2.4.1, which are located on accessory chromosomes in both bacteria and were not found in the inferred ancestral sequences, and on the primary chromosomes, as well as were not found in their closest relatives with single chromosome within the same clade. These genes emphasize the important potential roles of the secondary chromosomes in helping multipartite bacteria to adapt to specialized environments or conditions. In addition, we applied machine learning algorithms to predict multipartite genomes based on gene …

To better understand the nature and function of N-acylethanolamines (NAEs) in Camelina sativa, we used mass spectrometry analysis to identify and quantify NAE types in developing seeds, desiccated seeds and seedlings. Developing seeds showed a differential increase in individual NAE species and an overall increase in NAE content with seed development and maturation. The NAE composition in mature, desiccated seeds mostly reflected the total fatty acid composition in the seed tissues, except for a noted absence of 11-eicosenoic (20C monounsaturated) fatty acid in the NAE pool. During seed stratification and seedling growth, individual NAE species were depleted at similar rates. Simulated drought treatments during seedling development resulted in a significant rise in NAE levels for the major 18C NAE types compared with untreated seedlings. Arabidopsis and Camelina mutants with reported altered fatty acid profiles were analyzed for their NAE compositions; both Arabidopsis and Camelina had relatively similar changes between compositions of total seed fatty acids and NAEs. Furthermore, seeds were analyzed from transgenic Arabidopsis and Camelina with engineered, non-native, long-chain polyunsaturated fatty acids (18C, 20C and 22C), and the results showed the production of novel N-acylphosphatidylethanolamines (presumed precursors of NAEs) and NAEs with the same long acyl chains. These results …

The present studies determine the accumulation of persistent organic pollutants (POPs) in three pinniped species, evaluate the relationship with relevant biomarkers of exposure, and calculate toxic effect thresholds. Stranded harp and hooded seals were found to be accumulating PBDEs at levels which could pose a based on threshold levels determined in this study. Northern fur seals are accumulating all three classes of POPs (PCBs, PBDEs, and OCPs) with significant relationships being seen with blubber percent lipid. Correlations between contaminant concentrations and expression levels of relevant biomarkers were seen potentially indicating an effect on multiple pathways. Overall risk can be hard to determine due to factors such as sex and age. Broad threshold response values and hazard quotients were calculated for toxic effect endpoints in pinnipeds. Overall these results suggest that certain populations of pinnipeds are at high risk of experiencing toxic effects due to POP exposure, but it is important to understand effects even at lower concentrations. The relationship between exposure, toxic effects, and other stressors, both environmental and physiological, can impact the overall fitness and survival of pinnipeds.

Alteration in diet and knockdown of detoxification genes impacts the response of C. elegans to oxygen deprivation stress. I hypothesized that feeding worms a vitamin D3-supplementation diet would result in differential oxygen deprivation stress response. We used a combination of wet lab and transcriptomics approach to investigate the effect of a vitamin-D3 supplemented diet on the global gene expression changes and the anoxia response phenotype of C. elegans (Chapter 2). C. elegans genome consists of 143 detoxification genes (cyp and ugt). The presence of a significant number of genes in these detoxification families was a challenge with identifying and selecting specific cyp and ugt genes for detailed analysis. Our goal was to understand the evolution, phylogenetic, and expression of the detoxification enzymes CYPs and UGTs in C. elegans (Chapter 3). We undertook a phylogenetic and bioinformatics approach to analyze the C. elegans, detoxification family. Phylogenetic analysis provided insight into the association of the human and C. elegans xenobiotic/endobiotic detoxification system. Protein coding genes in C. elegans have been predicted to be human orthologs. The results of this work demonstrate the role of C. elegans in the identification and characterization of vitamin D3 induced alterations in gene expression profile and anoxia …

The anti-S2 peptides, the stabilizer and destabilizer, were designed to target myosin sub-fragment 2 (S2) in muscle. When the peptides are coupled to a heart-targeting molecule, they can reach the cardiomyocytes and interfere with cardiac muscle contraction. Monoclonal antibodies, MF20 and MF30, are also known to interact with light meromyosin and S2 respectively. The MF30 antibody compared to anti-S2 peptides and the MF20 antibody is used as a control to test the central hypothesis that: Both the anti-S2 peptides and antibodies bind to myosin S2 with high affinity, compete with MyBPC, and possibly interact with titin, in which case the anti-S2 peptides have further impact on myosin helicity and reach the heart with the aid of tannic acid to modulate cardiomyocytes' contraction in live mice. In this research, the effects of anti-S2 peptides and antibodies on myosin S2 were studied at the molecular and tissue levels. The anti-myosin binding mechanism to whole myosin was determined based on total internal reflectance fluorescence spectroscopy (TIRFS), and a modified cuvette was utilized to accommodate this experiment. The binding graphs indicated the cooperative binding of the peptides and antibodies with high affinity to myosin. Anti-myosin peptides and antibodies competition with Myosin Binding Protein C …

This study examined students' perception between short research experiences (SRE) courses and full-semester course research experiences (CRE) using the Persistence in the Sciences (PITS) survey and the interview questionnaire. The study also aimed to correlate the influence of student's demographic as a predictive indicator for Project Ownership Scores (POS) and Quantitative Literacy (QL) score means. The three courses studied at the University of North Texas were Biology for Science Majors Laboratory (BIOL 1760 SRE), Microbiology with Tiny Earth (BIOL 2042 Tiny Earth SRE), and Introductory Biology Research Laboratory I (BIOL 1750 SEA-PHAGES CRE). The mean scores for the PITS categories leaned favorably towards the research component of each laboratory course assessed in this study. The interview questionnaire showed 66% of the students in the SRE courses and 90% of the students in the CRE course preferred the research component of the lab. Paired survey demographic analysis for BIOL 1760 SRE showed significance for the Science Community Values with associate/bachelor's degree. BIOL 1750 SEA-PHAGES CRE showed significance in three of the six categories when comparing means for Project Ownership Emotion, Self-Efficacy, and Science Identity with Gender. Binary logistics was used to build a regression model to predict demographics with approximately 65% …

Anthropogenic activities have caused many wildlife spices to decline in populations worldwide. The grassland bird communities are especially being impacted by these land use changes. Breeding success is closely tied to functional habitats for most grassland bird species in North Texas. Restoring these degraded habitats is an important component to aid in conserving wildlife biodiversity. We surveyed the bird population at Riverby Ranch Mitigation site by conducting point count sampling. This site consists of recently restored grassland, wetland, and forested habitat. This research was focused on conducting post restoration monitoring of the bird community in the early succession of the restored habitats. We set out to use the bird community as biomonitors to help assess if the restoration practices could be considered successful. We found that density estimates were more than double at the restored site when comparing to three different references sites under different management practices. This included an unrestored working ranch, a wildlife management area, and a conservation managed prairie site. We found that biodiversity metrics were as high or higher than the reference sites. In addition, we also found that there were more observations of species of high conservation concern present at the restored stie. This research …

Results from the present study found for the first time that the use of bacterial strains of Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium thermophilum, and Enterococcus faecium from the commercial product PrimaLac® had the potential to act as a possible probiotic for juvenile red drum. The addition of PrimaLac® probiotics [whether as a water-soluble probiotic (WSP) or in a probiotic enhanced starter feed (PESF)] reduced mortality (%M), enhanced growth rates (MW, LT, SGR, and DGR), improved feed conversion efficiencies (FCEs), and physical condition factors in the juvenile red drum. Improvement was quantified using external morphological condition indices (MCIs) and blood physiological condition indices (PCIs). Results showed a strong positive relationship between MCIs and PCIs, which suggested that probiotics treated fish were in better health (lower MCIs) with lower fasting blood glucose and lactate levels than control fish. Addition of probiotics also resulted in improved water quality (lower nitrate, nitrite, and ammonia) in the treatment tanks compared to controls. Application of PrimaLac® probiotics on the morphology and histology of three different regions of the intestine (proximal, mid-and distal) improved intestinal length (Li), mass (Mi), and digesta mass (Md). In 5μm histological sections examined for differences among treatments (probiotics vs. controls), five variables …

The Dallas-Fort Worth International (DFW) Airport is located in a densely urbanized area with one of the fastest-growing populations in the U.S.A. The airport property includes a large tract of "protected" riparian forest that is unique to the urban surroundings. This dissertation explores variables that influence the benthic macroinvertebrate community structure found in urbanized prairie streams that were initially assessed by the University of North Texas (UNT) Benthic Ecology Lab during four, non-consecutive biomonitoring studies (2004, 2005, 2008, and 2014) funded by the DFW Airport. Additionally, land use analysis was performed using 5-meter resolution satellite imagery and eCognition to characterize the imperviousness of the study area watersheds at multiple scales. Overall, flow conditions and imperviousness at the watershed scale explained the most variability in the benthic stream community. Chironomidae taxa made up 20-50% of stream communities and outperformed all other taxa groups in discriminating between sites of similar flows and urban impairments. This finding highlights the need for genus level identifications of the chironomid family, especially as the dominant taxa in urban prairie streams. Over the course of these biomonitoring survey events, normal flow conditions and flows associated with supra-seasonal drought were experienced. Prevailing drought conditions of 2014 did not …

In my thesis, I examined impacts of nesting and floral resources on ground-nesting bee assemblages at nine ranch sites with differing grazing histories. Because ecological disturbances can strongly affect the availability of foraging and nesting resources over time, I also examined the impacts of grazing history on nesting and floral resources for ground-nesting bees. I sampled bee assemblages, a comprehensive array of vegetation and soil measures, and floral abundance and richness. I used these data to determine (1) the importance of different nesting habitat resources and livestock grazing history for ground-nesting bees and (2) how livestock grazing history influenced the availability of nesting and floral resources. I found that sites with sandier soils had greater abundance and richness of ground-nesting bees, and sites with less compacted soils had greater bee abundance. Contrary to many other studies, the availability of bare ground was not important for ground-nesting bee abundance or richness. Grazing history did not affect any measures of nesting or floral habitat, and had negligible effects on ground-nesting bee abundance, richness, and community composition. My results suggest that the availability of sandier or less compacted soils may be limiting for ground-nesting bee abundance or richness, especially in areas with predominately …

In the extensive and remote sub-Antarctic forests of South America, birds are the dominant terrestrial vertebrates. Despite considerable efforts to understand the ecology of birds breeding in these forests, our current knowledge for many species is still incomplete. During three breeding seasons (2014 – 2017), I studied the breeding ecology of the five most abundant open-cup forest-dwelling passerines in the sub-Antarctic forest of Navarino Island, Chile (55°04′S, 67°40′W). There were differences in some of the breeding strategies used by birds breeding on Navarino Island versus conspecific populations breeding at lower latitudes. Milvago chimango was the main nest predator of open-cup nesting forest passerines, and the main cause of nest failure. In addition, I found that species built their nests in sites with higher density and taller understory; however, these two factors decreased their nest survival. This mismatch could be due to a change in depredation risk on Navarino Island, and thus, passerines breeding there may be in an ecological trap. In addition, using light-level geolocators, I determined that the migratory connectivity of Elaenia albiceps is weak as a result of the large spatial spread of individuals on the wintering ground, and that the distances among individuals on the breeding grounds …

Reptiles can often experience perturbations that greatly influence their metabolic status (e.g., temperature, exercise, digestion, and ontogeny). The most common cause of fluctuations in metabolic status in post-embryonic reptiles is arguably digestion and physical activity (which will be further referred to as exercise). The objective of this thesis is to determine the mechanisms involved in CO2 transport during digestion, determine the mechanisms that allow for the maintenance of acid-base homeostasis during digestion, and observing the effect of an understudied form of exercise in semi-aquatic reptiles on the regulation of metabolic acidosis and base deficit. This dissertation provided evidence for potentially novel and under investigated mechanisms for acid-base homeostasis (e.g., small intestine and tissue buffering capacity; Chapters 3 & 4), while also debunking a proposed hypothesis for the function of an anatomical feature that still remains a mystery to comparative physiologist (Chapter 2). This thesis is far from systematic and exhaustive in its approach, however, the work accomplished in this dissertation has become the foundation for multiple distinct paths for ecologically relevant investigations of the regulation of metabolic acidosis/alkalosis in reptiles.

Crude oil commonly enters freshwater aquatic ecosystems as thin sheens forming on the water surface. Oil contains mixtures of toxic compounds called polycyclic aromatic hydrocarbons (PAHs), some of which are known to be photodynamic, increasing toxicity when combined with ultraviolet radiation. Benthic macroinvertebrate communities are commonly utilized as bioindicators, and as such rely on abundant data in literature concerning benthic macroinvertebrates' relative tolerances to a wide range of pollutants. A series of 10 plastic traps, half of which were filtered from UV radiation, were deployed in an urban pond for 27 days to determine colonization preferences of benthic macroinvertebrates to UV exposure. Results of this in situ experiment indicated that the majority of aquatic insects collected from traps inhabited the UV exposed treatment group, particularly the nonbiting midge, Chironomidae. A series of bioassays were then completed to investigate the sensitivities of a Chironomidae species to thin sheens of crude oil in the presence and absence of UV radiation. All bioassays were conducted using 10 day old Chironomus dilutus larvae cultured in the lab. The series of C. dilutus bioassays were all conducted under the same water quality parameters, temperatures, and oil sheen dosing methods, under a 16:8 photoperiod and exposed …

Increased exposure to antibiotics has led to the dissemination of genes conferring resistance to antimicrobial metabolites throughout human microbiomes globally via horizontal gene transfer (HGT). This has resulted in the emergence of new resistant strains leading to a rising epidemic of deaths from previously treatable infections. Evidence suggests that before the age of anthropogenic antibiotic use, microbes living within a community produced antibiotic metabolites and, subsequently, maintained such genes for several useful functions and a balance of diversity in nature. The question of the origin of these resistant genes is difficult to answer, but with continued advancements in ancient genomic analysis, researchers have developed methods of acquiring a more accurate representation of the microbiome associated with our human ancestors by extracting fossilized microbial specimens from dental calculus and directly sequencing the metagenomes. This thesis outlines the production of taxonomic and functional profiles of 20 different human and non-human oral microbiome samples using metagenomics tools originally developed for living individuals, altered for use with ancient microbial specimens. Putative antimicrobial resistant (AMR) genes derived from these profiles were reconstructed and conserved functional regions were identified. From the data that is available regarding the human microbiome from a range of time points throughout …

The field of vector-borne disease research uses multidisciplinary approaches to help understand complicated interactions. This dissertation, covers three different aspects of tick-borne disease research which all focus on exploring tick-borne diseases in the non-endemic areas of Denton, County Texas and the state of Texas with a focus on Rickettsia spp. These aspects include tick sampling, testing ticks for the presence of Rickettsia spp., and creating species distribution maps of the Rickettsia spp. Rickettsia amblyommatis and tick species Amblyomma americanum.

Current evidence suggests that thyroid hormones (THs) may impact development of the immune system. However, studies that explore the role of THs in immune development are limited, and the mechanisms leading to alterations in immune function are poorly understood. It is important to elucidate the role of THs in immune development given that many environmental contaminants have been shown to disrupt TH homeostasis and may also have negative impacts on the immune system. As such, the main goal of this study was to determine the long-term consequences of early life stage (ELS) hypothyroidism on immune function. To achieve this goal, it was first necessary to further characterize basic immune function in the selected model species, the fathead minnow (FHM, Pimephales promelas). Preliminary studies were conducted to describe the transcriptomic response to Yersinia ruckeri and adapt assays for the assessment of respiratory burst and phagocytic cell activity. To determine the long-term effects of ELS hypothyroidism, FHMs were exposed to the model thyroid suppressant propylthiouracil (PTU) from <1 to 30 days post hatch and reared under normal conditions. Upon reaching adulthood, ex vivo immune cell function and the in vivo immune response to Y. ruckeri were assessed. Fish exposed to PTU experienced …

Conservation corridors, areas of land connecting patches of natural land cover, are frequently cited and implemented as a restorative strategy to counteract fragmentation. Current corridor ecology focuses on experimental corridor systems or designed and built conservation corridors to assess functionality. Such systems and designs are typically short, straight swaths of homogenous land cover with unambiguous transitions between patches. Quantifying the degree to which amorphous landscape configurations, tortuosity, and heterogeneity of land cover and land uses within the corridor has on functional connectedness is a crucial yet overlooked component of corridor efficacy studies. Corridor literature lacks a robust and repeatable methodology for delineating existing landscape elements, recognizing arbitrary edges, and identifying the start and end of ambiguous transitions between the patches and corridor. Using a set of landscapes being studied as part of a global assessment of corridor efficacy, I designed a workflow that standardizes the boundary of corridor-patch interfaces. The proposed method is a quantitative and repeatable approach that minimizes the subjectivity in corridor delineations. This research investigates the degree to which the existence of a corridor modifies the structural and functional connectivity between patches connected by a corridor compared to an intact reference area.

Water quality of a stream or river is influenced by the surrounding landscape and vegetation. The Normalized Difference Vegetation Index (NDVI) is commonly used to characterize landcover and vegetation density. Benthic macroinvertebrates are ubiquitous in freshwater streams and are excellent indicators of the quality of freshwater habitats. Data from one NDVI remote sensing flight and one macroinvertebrate sampling event for eight wadeable stream study sites in the National Ecological Observatory Network (NEON) were acquired. Proportions of high, moderate, and sparse vegetation were calculated for each stream watershed using ArcGIS. Functional feeding groups and tolerance values were assigned to macroinvertebrate taxa. The Fourth-corner and RLQ methods of analysis, available in the ade4 package for R software, were used to evaluate the relationships of macroinvertebrate traits with environmental variables. Hypothesis testing using Model 6 in the ade4 package resulted in p-values of 0.066 and 0.057 for global (overall) significance. Mean NDVI values of moderately vegetated areas and proportion of sparse vegetation were found to be significant to percent shredders at alpha ≤ 0.05. Results of these methods of analysis, when combined with traditional macroinvertebrate sampling metrics, show that NDVI can be a useful, additional tool to characterize a watershed and its effects …

This article is the fourth paper of the Special Issue Memory and Criticality. It bridges the the theoretical debate on the role of memory and criticality discussed in the three earlier manuscripts, with a review of key concepts in biology and focus on cell-to-cell communication in organismal development. The authors suggest that in conjunction with morphogenetic gradients, there exist gradients of information transfer creating cybernetic loops of stability and disorder, setting the stage for adaptive capability. Criticality, therefore, appears to be an important factor in the transmission, transfer and coding of information for complex adaptive system development.

There has been a rapid surge toward "organic" products devoid of GMOs, MSGs, and other common compounds found in processed foods that continue to indicate an association with an increased risk for disease. These consumers seek nutrients and vitamins that are lacking in their diet and lifestyle in the form of nutraceuticals for disease prevention and treatment as well as overall lifestyle enhancement. However, these products generally lack clinical evidence as well as legal definition. Due to this ambiguity, nutraceuticals are neither considered a food product nor a pharmaceutical product. Furthermore, due to their alleged natural properties allowing for safe, therapeutic effects, nutraceuticals are being eagerly sought after by consumers in the place of pharmaceuticals. Additionally, since nutraceutical substances are "naturally" derived, there is a general lack of regulation regarding the manufacturing and distribution process. This mismanagement leads to lack of quality assurance (QA) and quality control (QC) protocols strictly implemented to define appropriate production and storage parameters. Without these critical measures, consumers are subjected to contamination of their products resulting from improper storage conditions and unmanaged production. These contaminants often include heavy metal impurities, pesticides, bacterial activity, and may also be adulterated with illicit drugs, all leading to detrimental …