Life Sciences Commons^™

Protocol For An Agent-Based Model Of Recombination In Bacteria Playing A Public Goods Game, Isaiah Paolo A. Lee, Omar T. Eldakar, J. Peter Gogarten, Cheryl P. Andam

Biology Faculty Articles

Agent-based models are composed of individual agents coded for traits, such as cooperation and cheating, that interact in a virtual world based on defined rules. Here, we describe the use of an agent-based model of homologous recombination in bacteria playing a public goods game. We describe steps for software installation, setting model parameters, running and testing models, and visualization and statistical analysis. This protocol is useful in analyses of horizontal gene transfer, bacterial sociobiology, and game theory.

For complete details on the use and execution of this protocol, please refer to Lee et al.¹

A Forensic Assessment Of Current Water Quality Using Idexx Techniques In The Himmarshee Canal And New River In Fort Lauderdale, Florida, Catherine Margaret Bilodeau

All HCAS Student Capstones, Theses, and Dissertations

Fecal contamination has continued to plague local communities around the United States and the world, especially in highly populated areas like Southern Florida. Newer techniques are beginning to be used to track microbes properly as they are found in waterways. Some more recent techniques in Microbial Source Tracking (MST) use IDEXX kits to detect fecal indicator bacteria (FIB). Two IDEXX kits Colilert-18^® which detects total coliforms and Escherichia coli in water or fecal coliforms in wastewater, and Enterolert^®, which targets enterococci, were used in the waterways of the Himmarshee Canal and South Fork of the New River …

Effects Of Cyanobacteria Harmful Algal Blooms On The Microbial Community Within Lake Okeechobee, Fl, Usa, Paisley S. Samuel

All HCAS Student Capstones, Theses, and Dissertations

The Lake Okeechobee (Lake O) watershed is a Floridian freshwater ecosystem that has been affected by the increased frequency and intensity of harmful cyanobacterial bloom (cyanoHAB) events occurring over recent decades. Lake O has several ecological and economic purposes such as providing habitats for various organisms and providing drinking water to urban communities surrounding the lake. Toxic cyanoHAB events are posing a threat to the ecosystem and economy of the lake due to the degradation of water quality. This study investigates how the microbial community structure within Lake O is affected by annual cyanobacterial harmful algal blooms over several years …

Change And Stasis Of Distinct Sediment Microbiomes Across Port Everglades Inlet (Pei) And The Adjacent Coral Reefs, Lauren E. Krausfeldt, Jose V. Lopez, Catherine Bilodeau, Hyo Won Lee, Shelby L. Casali

Biology Faculty Articles

Deep water ports are human built coastal structures that by definition welcome ship traffic and disturbance. Evidence is accumulating that enhanced port activities such as dredging or deepening have negatively affected nearby natural habitats. Port Everglades Inlet (PEI) is a large active South Florida cargo port for over two million people and lies adjacent to coral reefs, dwindling mangroves, and recreational beaches. In this study, the microbial communities of PEI and adjacent reef sediments were characterized to serve as indicators for change due to dredging and assess anthropogenic influence on these sensitive ecosystems by sequencing the V4 region of 16S …

The Microbiome Characterization And Comparison Of Loggerhead (Caretta Caretta) And Green (Chelonia Mydas) Sea Turtle Eggs In Broward County, Florida, Colleen Mcmaken

All HCAS Student Capstones, Theses, and Dissertations

All seven extant species of sea turtles are classified as threatened or endangered due to natural and anthropogenic impacts. Sea turtle conservation efforts are largely dedicated to ensuring healthy nesting activity. Hatching success of sea turtle eggs can be affected by several factors including the proliferation of pathogenic microbes. However, it is unclear as to which microbes have the most impact on hatching success and how they are transmitted into the eggs. The primary aims of this study characterized and compared the bacterial communities (microbiomes) from the i) cloaca of nesting sea turtles; ii) sand within and surrounding the nests …

Periodic Spatial Disturbances Alter The Expression Of Quorum Sensing Virulence Factors In Pseudomonas Aeruginosa, Laura García-Diéguez

All HCAS Student Capstones, Theses, and Dissertations

Pseudomonas aeruginosa is an opportunistic pathogen associated with severe acute and chronic illnesses. Current antibiotic-based approaches fail to effectively treat P. aeruginosa infections due to the effectiveness and robustness of the quorum sensing signaling system (QS). Pathogenic bacteria, such as P. aeruginosa, employ this population density-dependent communication mechanism to confer antimicrobial resistance, propagate infection, and coordinate the expression of virulence factors, through the production and detection of autoinducing signaling molecules (AI). As such, there is a growing interest in developing novel non-antibiotic-based techniques to attenuate the pathogenicity of P. aeruginosa by disrupting the functionality of its QS system. Previous …

Phage Hunters: The Discovery Of Two Novel Bacteriophages That Infect Gordonia Rubripertincta, Sarah Ballarin, Neel Balusa, Melissa Bell, Samia Caballero, Joshua Chan, Maria Farez, Ashley J. Guillen-Tapia, Nashrah Pierre-Louis, Victoria Polishuk, Bhavya Soni, Julie Torruellas Garcia, Katie E. Crump

Sea-Phages

Antibiotic resistance is a global healthcare concern because it poses a threat to effectively treating bacterial infections and the need for alternative therapeutics are urgently required. Phage therapy, which uses viruses to treat bacterial infections, is a promising alternative to conventional antibiotics. Phages have also been widely used in the food industry to prevent microbial growth on certain foods and are currently being explored as a method for bioremediation of oil spills and wastewater treatment. Although phages are ubiquitous in nature, many have yet to be discovered. Through the SEA-PHAGES (Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science) program …

A Temporal Analysis Of The Microbiota And Biofouling Development On Artificial Substrates In The Port Everglades Inlet, Florida, Denise Swack

All HCAS Student Capstones, Theses, and Dissertations

A pilot project was deployed in Port Everglades Inlet, Florida that aimed to evaluate the biofilm composing the microbiome on ecologically engineered artificial substrates used to build Coastal Marine Infrastructure. In April of 2017, an Articulated Concrete Block Mattress comprised of an ecological engineered concrete substrate and a standard smooth surface control substrates were compared. This study will provide a profile on the microbiome community on artificial substrates within Port Everglades Inlet on bio-enhancing concrete-based solutions in our Coastal Marine Infrastructure. To study the microbial community, the 16s rRNA technology was used in Illumina’s high-throughput DNA sequencing. Samples were collected …

Increasing The Frequency Of Periodic Spatial Disturbance Decreases Surface Attachment Protein Expression In Staphylococcus Aureus, Ivana M. Barraza

All HCAS Student Capstones, Theses, and Dissertations

Staphylococcus aureus is an opportunistic pathogen that can cause a variety of acute and chronic illnesses. The severity of these illnesses such as sepsis, necrotizing pneumonia, and toxic shock syndrome is measured through the virulence that S. aureus inflicts on its host. Methicillin-resistant S. aureus (MRSA) is commonly associated with secondary infections and is challenging to treat given the limited selection of antibiotics that are effective against it. Accordingly, novel approaches to reduce S. aureus pathogenicity are required. S. aureus regulates pathogenesis through a cell-to-cell communication system referred to as quorum sensing. Effective communication determines the production of two broad …

Investigation Of Sulfur Cycling In Marine Sponge Cinachyrella Spp. From A South Florida Reef, Shelby K. Cain

All HCAS Student Capstones, Theses, and Dissertations

Symbionts within marine sponges are actively participating in the biogeochemical cycles. Among them, the role of symbiont microbes in the sulfur cycle remains a mystery. This study measured the abundance of microbes within the genus Cinachyrella before and after exposure to hydrogen sulfide. A four-part study was conducted: a) five-hour drop experiments, b) vertical distribution experiments, c) five-hour uptake experiments, and d) long-term exposure experiments. The five-hour drop experiment utilized a microsensor to measure sulfide levels, which was lowered 1.0 mm every thirty minutes for a total of 5 hours. Three trials were performed, each with one sponge and a …

Disruption Of Pyoverdine Synthesis In Pseudomonas Aeruginosa Through Periodic Disturbance Of Biofilm Structure, Rebecca J. Quinn

All HCAS Student Capstones, Theses, and Dissertations

Infections due to bacteria were once easily treated using antibiotics. However, the effective shelf life of antibiotics is diminishing due to the rise and spread of antibiotic resistant bacteria. Accordingly, novel approaches to treating infections are required. Pseudomonas aeruginosa is a multi-drug resistant, biofilm forming opportunistic pathogen. A determinant of P. aeruginosa’s pathogenicity is pyoverdine, a siderophore that is used to sequester iron from the environment. After being produced by a bacterium, pyoverdine is secreted into the environment where it complexes with iron. The pyoverdine-iron complex is then returned to the bacteria through diffusion, where it drives the synthesis of …

Characterization Of Bacterial Communities In Biscayne Bay Through Genomic Analysis, Eric Fortman

HCNSO Student Theses and Dissertations

Biscayne Bay is a shallow oligotrophic estuary in Southeast Florida. Channelization of rivers, and dredging of canals has greatly altered the historical flow of fresh water into the bay. This, coupled with the rise of a sprawling urban & suburban development, has greatly increased the nutrient load in the bay. This study examined the bacterial community at 14 stations throughout Biscayne Bay —6 stations were located at the mouths of canals; 1 upstream-canal station; 6 stations in the center of the bay; and one ocean influenced station, located near the entrance to the bay. One liter, surface water samples were …

Inferred Function And Dynamics Of Microbial Communities From The Northern Gulf Of Mexico, Deepesh Tourani

HCNSO Student Theses and Dissertations

Microbial communities, or microbiomes, are the major drivers of global biogeochemical cycles, acting as primary producers and decomposers across the water column in the oceans. Thus, they reflect changes in physicochemical properties and nutrient composition of the ocean. However, this correlation between ecological changes and the function of marine microbiomes is poorly understood. Large-scale oceanic events such as the bottom-water oxygen-depleted zone (i.e., “dead zone”) and the Deepwater Horizon oil spill in the Gulf of Mexico (GoM) render the ecosystem fragile. These events decrease survival rates of pelagic and coastal macrofauna and affect the biodiversity of the region. As part …

Next Generation Sequencing Methods For Coastal Zone Water Quality Monitoring, Catherine Iskrenko

HCNSO Student Capstones

When analyzing the water quality of the coastal zone, culture-based techniques have been utilized most often to identify Fecal Indicator Bacteria in samples. Since the advent of the Sanger Method for DNA sequencing, other techniques have arisen that provide significantly more information on the microorganisms in sample, but they are still not the mainstream for water quality analysis. This capstone reviews and compares culture-based techniques, DNA sequencing, RNA sequencing, qPCR for biomarker, and 16S rDNA sequencing to highlight their merits and shortcomings for analyzing environmental water samples. The technique presented that provides the broadest range of information (including the identification …

Microbial Ecology Of South Florida Surface Waters: Examining The Potential For Anthropogenic Influences, Chase P. Donnelly

HCNSO Student Theses and Dissertations

South Florida contains one of the largest subtropical wetlands in the world, and yet not much is known about the microbes that live in these surface waters. These microbes play an important role in chemical cycling and maintaining good water quality for both human and ecosystem health. The hydrology of Florida’s surface waters is tightly regulated with the use of canal and levee systems run by the US Army Corps of Engineers and The South Florida Water Management District. These canals run through the Everglades, agriculture, and urban environments to control water levels in Lake Okeechobee, the Water Conservation Areas, …

Differential Preservation Of Endogenous Human And Microbial Dna In Dental Calculus And Dentin, Allison E. Mann, Susanna Sabin, Kirsten Ziesemer, Ashild J. Vagene, Hannes Schroeder, Andrew T. Ozga, Krithivasan Sankaranarayanan, Courtney A. Hofman, James A. Fellows Yates, Domingo C. Salazar-Garcia, Bruno Frohlich, Mark Aldenderfer, Menno Hoogland, Christopher Read, George R. Milner, Anne C. Stone, Cecil M. Lewis Jr., Johannes Krause, Corinne Hofman, Kirsten I. Bos, Christina Warinner

Biology Faculty Articles

Dental calculus (calcified dental plaque) is prevalent in archaeological skeletal collections and is a rich source of oral microbiome and host-derived ancient biomolecules. Recently, it has been proposed that dental calculus may provide a more robust environment for DNA preservation than other skeletal remains, but this has not been systematically tested. In this study, shotgun-sequenced data from paired dental calculus and dentin samples from 48 globally distributed individuals are compared using a metagenomic approach. Overall, we find DNA from dental calculus is consistently more abundant and less contaminated than DNA from dentin. The majority of DNA in dental calculus is …

Fine Grained Compositional Analysis Of Port Everglades Inlet Microbiome Using High Throughput Dna Sequencing, Lauren M. O'Connell, Song Gao, Donald S. Mccorquodale Jr., Jay M. Fleisher, Jose Lopez

Marine & Environmental Sciences Faculty Articles

Background

Similar to natural rivers, manmade inlets connect inland runoff to the ocean. Port Everglades Inlet (PEI) is a busy cargo and cruise ship port in South Florida, which can act as a source of pollution to surrounding beaches and offshore coral reefs. Understanding the composition and fluctuations of bacterioplankton communities (“microbiomes”) in major port inlets is important due to potential impacts on surrounding environments. We hypothesize seasonal microbial fluctuations, which were profiled by high throughput 16S rRNA amplicon sequencing and analysis.

Methods & Results

Surface water samples were collected every week for one year. A total of four samples …

Microbiome And Bacterioplankton Rrna Gene Sequence Data Collected From Gulf Of Mexico Seawater Samples, Cruises Dp03 And Dp04 From April - August 2016, Cole Easson, Lindsey Freed

DEEPEND Datasets

Seawater was collected and filtered for microbiome and bacterioplankton sequencing and analyses at various depths during planned DEEPEND cruise expeditions to the GOM in 2016. Filters were stored and then processed for total environmental genomic DNA according to standard methods (see earthmicrobiome.org). 16S rRNA amplicon libraries covering the V4 hypervariable regions were generated with universal PCR primers and then sequenced on an Illumina MiSeq DNA sequencing platform. Raw paired-end sequences were joined and quality filtered in the bioinformatics program, QIIME. Vertical baseline characterizations will track alpha and beta diversity at different depths ranging from 0 – 1500 m, assess seasonal …

Is There A Link Between Aging And Microbiome Diversity In Exceptional Mammalian Longevity?, Graham M. Hughes, John Leech, Sebastien J. Puechmaille, Jose V. Lopez, Emma C. Teeling

Biology Faculty Articles

A changing microbiome has been linked to biological aging in mice and humans, suggesting a possible role of gut flora in pathogenic aging phenotypes. Many bat species have exceptional longevity given their body size and some can live up to ten times longer than expected with little signs of aging. This study explores the anal microbiome of the exceptionally long-lived Myotis myotis bat, investigating bacterial composition in both adult and juvenile bats to determine if the microbiome changes with age in a wild, long-lived non-model organism, using non-lethal sampling. The anal microbiome was sequenced using metabarcoding in more than 50 …

Pulmonary Alveolar Proteinosis- A Case Report, Renee M. Horner, Douglas S. Gardenhire, Ralph D. Zimmerman

Internet Journal of Allied Health Sciences and Practice

ABSTRACT

Background: Pulmonary Alveolar Proteinosis (PAP) is a rare pulmonary disorder caused by a congregation of excessive lipoproteinaceous material in the alveolar spaces due to impaired surfactant metabolism. The congregation of the protein in the alveolar space leads to difficulty in breathing, impaired pulmonary immunity, and susceptibility to both opportunistic and acquired pulmonary infections. Although Pulmonary Alveolar Proteinosis is rare, there are potential treatments. Whole-lung lavage is the most widely accepted therapy and course of treatment. An additional form of therapy, GM-CSF stimulating therapy, uses recombinant deoxyribonucleic acid (DNA) technology to increase white cell production.

Purpose: The purpose …

Microbial Community Richness Distinguishes Shark Species Microbiomes In South Florida, Rachael Cassandra Karns

HCNSO Student Theses and Dissertations

The microbiome (microbial community) of individuals is crucial when characterizing and understanding processes that are required for organism function and survival. Microbial organisms, which make up an individual’s microbiome, can be linked to disease or function of the host organism. In humans, individuals differ substantially in their microbiome compositions in various areas of the body. The cause of much of the composition diversity is yet unexplained, however, it is speculated that habitat, diet, and early exposure to microbes could be altering the microbiomes of individuals (Human Microbiome Project Consortium, 2012b, 2012a). To date, only one study has reported on microbiome …

Bacterial Communities Associated With Healthy And Diseased Acropora Cervicornis (Staghorn Coral) Using High-Throughput Sequencing, Charles Walton

HCNSO Student Theses and Dissertations

Coral diseases were first noted in the 1960s and 1970s and have had major impacts globally on coral reef community structures. In the Caribbean, a major outbreak of white band disease has been considered responsible for the drastic decline of Caribbean Acroporids since the 1970s. In addition to white band disease, another more recently described condition known as rapid tissue loss (RTL) has had major impacts on Acropora cervicornis populations, specifically offshore Broward County Southeast Florida. While these diseases have contributed to the population decline, determining their etiologies has been elusive.

Coral diseases have been characterized by shifts in their …

The Human Microbiome: An Emerging Tool In Forensics, Jarrad T. Hampton-Marcell, Jose V. Lopez, Jack A. Gilbert

Biology Faculty Articles

No abstract provided.

Microbiome And Bacterioplankton Rrna Gene Sequence Data Collected From Gulf Of Mexico Seawater Samples, Cruises Dp01 And Dp02 From May 2015 - August 2015, Cole Easson, Jose V. Lopez

DEEPEND Datasets

Seawater was collected and filtered for microbiome and bacterioplankton sequencing and analyses at various depths during two planned DEEPEND cruise expeditions to the GOM. Filters were stored and then processed for total environmental genomic DNA according to standard methods (see earthmicrobiome.org). 16S rRNA amplicon libraries covering the V4 hypervariable region was generated with universal PCR primers and then sequenced on an Illumina MiSeq DNA sequencing platform. Bioinformatics analyses in QIIME and R was used to quality filter and assess microbial community diversity and taxonomic composition. These baseline characterizations track alpha and beta diversity at different depths ranging from 0 – …

Otopathogenic Pseudomonas Aeruginosa Enters And Survives Inside Macrophages, Rahul Mittal, Christopher V. Lisi, Hansi Kumari, M'Hamed Grati, Patricia Blackwelder, Denise Yan, Chaitanya Jain, Kalai Mathee, Paulo H. Weckwerth, Xue Zhong Liu

Biology Faculty Articles

Otitis media (OM) is a broad term describing a group of infectious and inflammatory disorders of the middle ear. Despite antibiotic therapy, acute OM can progress to chronic suppurative otitis media (CSOM) characterized by ear drum perforation and purulent discharge. Pseudomonas aeruginosa is the most common pathogen associated with CSOM. Although, macrophages play an important role in innate immune responses but their role in the pathogenesis of P. aeruginosa-induced CSOM is not known. The objective of this study is to examine the interaction of P. aeruginosa with primary macrophages. We observed that P. aeruginosa enters and multiplies inside human …

Characterization Of The Marine Sponge Amphimedon Compressa Microbiome Across A Spatial Gradient, Renee Michelle Potens

HCNSO Student Theses and Dissertations

Diverse and ecologically important microbial communities (microbiomes) are symbiotic within marine sponges. In this study, the microbiome of Amphimedon compressa from three sample locations (Broward and Dade Counties, Southeast Florida, USA and the Southern Caribbean, Bocas del Toro, Panama) is characterized using 16S rRNA Illumina sequencing. The predominant taxa are Proteobacteria and Cyanobacteria, as expected for Low Microbial Abundance sponges, accounting for over 53% of the total microbiome community. The numbers of Operational Taxonomic Units (OTUs) decrease from Broward County (2,900) to Dade County (2,300) and then Bocas del Toro (1,200). The correlates to a decreasing north-south gradient of …

A Tale Of Two Theories: Using An Engineered Strain Of E. Coli To Bridge The Gap Between Quorum Sensing And Diffusion Sensing, Cortney E. Wilson

HCNSO Student Theses and Dissertations

Cooperation is a trait that is found at all levels of biological organization. Interestingly, cooperation appears to occur in bacteria that produce small, easily diffusible molecules called autoinducers. To understand why bacteria produce these autoinducers, the scientific community has focused on one predominant theory called quorum sensing. Under this theory, bacteria produce autoinducers so they can sense the density of the population. Once a sufficiently high population density is reached, autoinducers initiate the production of a costly gene product that serves to benefit the population. In contrast, a competing theory called diffusion sensing suggests that autoinducers are used by the …

Intrinsic Challenges In Ancient Microbiome Reconstruction Using 16s Rrna Gene Amplification, Kirsten Ziesemer, Allison Mann, Krithivasan Sankaranarayanan, Hannes Schroeder, Andrew T. Ozga, Bernd W. Brandt, Egija Zaura, Andrea Waters-Rist, Menno Hoogland, Domingo C. Salazar-Garcia, Mark Aldenderfer, Camilla Speller, Jessica Hendy, Darlene A. Weston, Sandy J. Macdonald, Gavin H. Thomas, Matthew J. Collins, Cecil M. Lewis Jr., Corinne Hofman, Christina Warinner

Biology Faculty Articles

To date, characterization of ancient oral (dental calculus) and gut (coprolite) microbiota has been primarily accomplished through a metataxonomic approach involving targeted amplification of one or more variable regions in the 16S rRNA gene. Specifically, the V3 region (E. coli341–534) of this gene has been suggested as an excellent candidate for ancient DNA amplification and microbial community reconstruction. However, in practice this metataxonomic approach often produces highly skewed taxonomic frequency data. In this study, we use non-targeted (shotgun metagenomics) sequencing methods to better understand skewed microbial profiles observed in four ancient dental calculus specimens previously analyzed by amplicon …

Toll-Like Receptor 9-Mediated Inflammation Triggers Alveolar Bone Loss In Experimental Murine Periodontitis, Paul D. Kim, Xia Xia-Juan, Katie E. Crump, Toshiharu Abe, George Hajishengallis, Sinem E. Sahingur

Biology Faculty Articles

Chronic periodontitis is a local inflammatory disease induced by a dysbiotic microbiota and leading to destruction of the tooth-supporting structures. Microbial nucleic acids are abundantly present in the periodontium, derived through release after phagocytic uptake of microbes and/or from biofilm-associated extracellular DNA. Binding of microbial DNA to its cognate receptors, such as Toll-like receptor 9 (TLR9), can trigger inflammation. In this study, we utilized TLR9 knockout (TLR9^−/−) mice and wild-type (WT) controls in a murine model of Porphyromonas gingivalis-induced periodontitis and report the first in vivo evidence that TLR9 signaling mediates the induction of periodontal bone loss.P. gingivalis-infected …

Two Distinct Microbial Communities Revealed In The Sponge Cinachyrella, Marie L. Cuvelier, Emily Blake, Rebecca Mulheron, Peter J. Mccarthy, Patricia Blackwelder, Rebecca Vega-Thurber, Jose V. Lopez

Marine & Environmental Sciences Faculty Articles

Marine sponges are vital components of benthic and coral reef ecosystems, providing shelter and nutrition for many organisms. In addition, sponges act as an essential carbon and nutrient link between the pelagic and benthic environment by filtering large quantities of seawater. Many sponge species harbor a diverse microbial community (including Archaea, Bacteria and Eukaryotes), which can constitute up to 50% of the sponge biomass. Sponges of the genus Cinachyrella are common in Caribbean and Floridian reefs and their archaeal and bacterial microbiomes were explored here using 16S rRNA gene tag pyrosequencing. Cinachyrella specimens and seawater samples were collected from the …