Microbiology Commons^™

Expression, Characterization, And Comparative Analysis Of Bacillus Bacteriophage Endolysins, Stephen D. Zdgiebloski

Research and Creativity Symposium

Bacteriophages present a promising alternative for combating antibiotic-resistant bacteria without disrupting the normal microbiome or harming human cells. The antibacterial properties of lytic bacteriophages are largely attributed to endolysins, enzymes active in the lytic cycle. Endolysins recognize bacterial cell walls and cleave the peptidoglycan matrix, leading to cell wall disruption. Their specificity and effectiveness against bacterial hosts are determined by two functional domains, including the cell wall binding domain. Our research focuses on characterizing Bacillus phage endolysins. We analyzed the protein sequences of endolysins from six Bacillus phages isolated at the University of Mary Washington. Although these phages were isolated …

Genome Analysis Of Aeromonas Hydrophila Strain S14-452, Zarin Tasnim Raya

Undergraduate Research Conference

The Aeromonas hydrophila are gram-negative, opportunistic bacteria that can infect many groups of organisms, mainly fish and amphibians. In channel catfish, A. hydrophila causes fatal bacterial septicemia, leading to massive economic losses for catfish farmers. Therefore, an understanding of its genome will aid in identifying its virulence factors, metabolic pathways and survival mechanisms within the host. This study details the draft genome sequence of A. hydrophila strain S14-452, isolated from channel catfish exhibiting bacterial septicemia during a disease outbreak at a catfish farm.

A. hydrophila strain S14-452 was sequenced using the Illumina iSeq 100 System. Assembly was performed with SPAdes, …

Improved Protein Splicing Through Viral Passaging, John Smetana

Scholars Week

Intervening proteins (inteins) are translated as subdomains within host proteins and removed through an intein-driven splicing reaction where the flanking sequences (exteins) are joined with a peptide bond. Previously, we developed a self-removing translation reporter for labeling Ebola virus (EBOV). In this reporter, an intein (RadA) containing the fluorescent protein ZsGreen (ZsG) is inserted within the EBOV protein VP30. Upon VP30-RadA-ZsG expression from the viral genome, RadA-ZsG is removed from VP30 through the protein splicing activity of RadA, generating functional, non-tagged VP30 and functional ZsGreen. While incorporation of our VP30-RadA-ZsG fusion reporter into recombinant EBOV (rEBOV- RadA-ZsG) resulted in an …

Rainwater Harvesting Systems Metagenomics, Jade Riddle, Julia Parsons

Spring Showcase for Research and Creative Inquiry

Climate change induced water scarcity has led to an increasing interest in non-traditional water sources such as rainwater. However, there are lingering health and safety concerns due to the lack of research into the microbial communities contained within collection systems. To address this gap, water samples were collected from rain barrels around northern Virginia for microbial analysis. Each microbe within the sample has a unique genome subject to DNA testing. Environmental DNA was prepared from the collected water samples. To characterize the microbial community, both targeted sequencing and whole genome sequencing approaches were used. Using targeted sequencing of the 16s …

“Pinkalicious": Outer Membrane Vesicles Isolated From Pseudoalteromonas Rubra Carry Prodigiosin Antibiotics As Cargo, Margaret Hill

Senior Honors Projects

Astrangia poculata, the state coral of Rhode Island, is susceptible to infection by Vibrio coralliilyticus RE22, which is a common marine pathogen affecting various invertebrate species. The settlement of coral larvae prior to the establishment of colonies is mediated by crustose coralline algae (CCA). This process may be induced by CCA-associated protective bacteria that colonize corals in early metamorphosis; furthermore, such bacteria potentially have downstream effects in the susceptibility of corals to pathogens. Strains of Pseudoalteromonas rubra isolated from CCA have been shown to induce larval settlement of tropical corals. We hypothesize that P. rubra induces the settlement of …

Investigating Alternative Induction Of The S. Aureus 80a Prophage, Jody Caretti

Student Scholar Showcase

Widespread antibiotic resistance has quickly become one of the most concerning crises affecting modern medicine, especially in cases of Staphylococcal infections, which have become resistant to all ��-lactam antibiotics. As a result, research into alternative forms of treatment for bacterial infections is a top priority. Bacteriophage therapy is a popular candidate for replacing antibiotics, due to the way the viruses interact with the bacteria. The 80�� prophage integrates its DNA into Staphylococcus aureus strain 10616 via the lysogenic cycle, and replicates with the bacterial cell. When stressed, the bacteria stop replicating and the prophage stimulates the lytic cycle, activating transcription …

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 …

Insights Into Viral Genome Function Through Comparative Structural Analysis, Lydia Phillips

UNO Student Research and Creative Activity Fair

Enteroviruses are single stranded RNA viruses which have caused many public health concerns, particularly in children. These viruses are responsible for polio, hand, foot, and mouth disease, many polio-like neurological diseases, and the common cold. The enterovirus called Coxsackievirus B3 (CVB3), a close relative to poliovirus, has been shown responsible for severe human disease, including pancreatitis, myocarditis, and type 1 diabetes. A unique advantage of studying CVB3 is the existence of a naturally occurring strain (CVB3/GA) which displays no known pathogenicity. Earlier comparative genomic studies have shown that the primary difference between avirulent CVB3/GA and the virulent strains of CVB3 …

Lipoarabinomannan From Mycobacterium Smegmatis Stimulates An Inflammatory Response In Raw 264.7 Murine Macrophages, Aysha Zaman

Student Scholar Showcase

Macrophages are a key leukocyte in defense against Mycobacterium tuberculosis infection. The precise mechanism by which M. tuberculosis evades host macrophage defenses remains unknown, so understanding how macrophages interact with cell wall components of mycobacteria is critical. Lipoarabinomannan (LAM) is a glycolipid Toll-like receptor-2 (TLR-2) ligand found on the cell wall of mycobacteria, and is thought to contribute to the cell wall structural integrity. When LAM binds to TLR on host leukocytes, this activates cellular responses and phagocytosis by host macrophages. Nitric oxide production can be used as a measure of inflammatory response and can be evaluated via nitrite response …

Co-Occurrence Of Manganese Oxidase Genes Indicates Lateral Transfer Between Classes Of Proteobacteria, Jacob Olichney

Student Scholar Showcase

Pseudomonas putida GB-1 is a model organism for the study of manganese oxidation in bacteria, however, the frequency of co-localization of multiple known and suspected manganese oxidizing proteins, as well as their occurrence between species, is unknown. Eight different genes isolated from known manganese oxidizing bacteria (MnxG, MopA, McoA, PputGB1_2552, PputGB1_2553, MoxA, MofA, and Bacillus MnxG) were tested individually using BioPython and BLAST (Basic Local Alignment Search Tool) on multiple genomic databases. BLAST searches had an expect value cutoff of 1e-50, limiting gene homologs to those with high sequence similarity. The abundance of homologous genes across classes of proteobacteria point …

Targeted Therapy For The Future: The Use Of Novel Antimicrobial Peptides Against P. Aeurginosa, Matthew Froid

UNO Student Research and Creative Activity Fair

Pathogenic bacteria, such as the gram-negative bacterium Pseudomonas aeruginosa, are becoming resistant to our current arsenal of antibiotics at an alarming rate. P. aeruginosa is a leading cause of nosocomial acquired infections and is a primary co-morbidity in patients with compromised immune systems. One potential source of new antibiotic agents is antimicrobial peptides. Antimicrobial peptides (AMPs) are small proteins, and some have shown a high degree of efficacy and broad-spectrum activity against both Gram-positive and Gram-negative bacteria. An experimental AMP that has been developed by Dr. Wang at UNMC, DASamp2, has shown to be effective against virulent bacteria, including P. …

Investigating The Impact Of Oxidative Stress On Tetrahymena Thermophila Sirtuin,Thd 18, Emmanuel Dubuisson

Publications and Research

Sirtuins are a family of enzymes that fulfill various important biological functions. Investigators have looked for the implication of Sirtuin genes in cell signaling mechanisms, in the formation and silencing of heterochromatin, in the regulation of ion channels, and in the modulation of the cellular redox reactions[2]. Different model organisms have been previously used to conduct these studies; among them, there are yeasts, nematodes, and fruit flies. Each one has made some valuable contribution into the vast body of knowledge related to this field.

However, gaps in the understanding of Sirtuins functions still remain to be filled. In this optic, …

Effects Of Evolution On Laboratory Sublines Of Myxococcus Xanthus Dk1622, Mackenzie Ryan, Francesca Scribano, Kimberly Murphy

Celebration of Learning

Microbes have served as effective models for studying evolution because of their ability to be easily replicated, stored, and manipulated. Myxococcus xanthus is a soil bacterium that has served as a model organism in many laboratories. The unique social and motile behaviors exhibited by this bacterium make it ideal for phenotypic assays. A wild-type strain of M. xanthus, DK1622, has been distributed to laboratories across the United States and therefore we now have DK1622 sublines. The genomes of a number of these sublines have been sequenced and their social and motile phenotypes have been analyzed. When nine of these sublines …

Identification Of Microbiota Associated With The Ectosymbiotic Community Structure Of The Appalachian Brook Crayfish (Cambarus Bartonii), Matthew M. Cooke 6268927, Luke T. Fischer, Taylor Griffin, Sherrie Jeffers

Montview Journal of Research & Scholarship

Microbes play a significant role in ecosystems, effecting the vital functions of other organisms, interspecies relationships, and population sizes. Based upon previous research (Skelton et al., 2016), it has been hypothesized that crayfish symbionts (branchiobdellidans) play a significant role in regulating the amount of microbiota associated with the host. A growing body of knowledge is known regarding both the condition of gill fouling and the annelid worm’s symbiotic relationship with the crayfish (Skelton et al., 2016; Skelton et al., 2016). However, there is little to no evidence available concerning the bacteria’s role in the process (Skelton et al., 2016). Microbial …

Discovery Of Geographical Gene Variants In Related Pseudomonas Aeruginosa Bacteriophages, Emilee Carr, Elise Melhado, Emily Loertscher, Trever Thurgood, Ruchira Sharma, Julianne H. Grose

Library/Life Sciences Undergraduate Poster Competition 2019

Antibiotic resistant bacterial strains are a major crisis in the world due to the difficult nature of curing individuals afflicted with them. Phage therapy has been proposed as an alternate treatment for these bacterium. In Dr. Julianne Grose's lab, bacteriophages were against the bacteria, Pseudomonas aeruginosa. To isolate, environmental samples were utilized in enrichment cultures that were ultimately used in serial dilutions, plaque purification, electron microscopy, DNA isolation, sequencing, and genome annotation. The P. aeruginosa phage, TF17, infects a bacterial strain that is highly related to a strain that causes fatalities as an opportunistic infection in patients with cystic fibrosis. …

Microflora In The Cheek Pouches Of Ord's Kangaroo Rat (Dipodomys Ordii) From The Nebraska Sandhills, Devin M. Christensen

UNO Student Research and Creative Activity Fair

The microflora living within the Ord’s kangaroo rat (Dipodymis ordii) cheek pouches has never been extensively studied before. Kangaroo rats use their cheek pouches to transport seeds and grasses in their mouths that they later store. It is predicted that the cheek pouches harbor fungal and bacterial growth that could be either helpful or harmful to the kangaroo rat and its environment. 8 kangaroo rats were humanely captured using Sherman live traps in the Nebraska Sandhills; their cheek pouches were swabbed, and the microorganisms obtained were grown on agar plates and in liquid media until an adequate amount …

The Effects Of The Fungus Beauvaria Sp. On The Cave Cricket, Hadenoecus Subterraneus, Christina N. Walker, Derrick J. Jent, Claire A. Fuller

Scholars Week

The cave cricket, Hadenoecus subterraneus, is a keystone species in cave ecosystems within Mammoth Cave National Park (MCNP). Within MCNP, many cricket cadavers have been found

with a thick, white fungus growing on them; this fungus has previously been identified to be Beauveria bassiana. However, new molecular data suggests that this may actually be the species B. amorpha. Cricket cadavers with Beauveria sp. were collected from MCNP and cultured on potato dextrose agar. Cultures will be sent to the USDA for a genetic analysis and identification of the fungus. The purpose of this study is to examine if the relationship …

How Does The Exploitation And Degradation Of Finite Natural Resources By Industrial Oil Producers Effect Our Future Planetary Environment?, Zachary J. King Mr.

Symposium of Student Scholars

Zachary King

Dr. Tu

26 March 2015

GEOG 3700

Abstract

The purpose of this presentation is to communicate content of the reports given by credible institution on the degradation and exploitation of finite natural resources by corporate institutions, for short term financial gain, and how it will effect the future planetary environment. Methods used in obtaining this research range from point source observation directly from oil spills in the Gulf or pipe leaks in the Mid-West, to analytical and numerical data quantifications gathered over a period of two decades. This presentation encompasses a wide scope of information concerning the way …

Comparison Of The Promega And Kikkoman Atp Devices, Ryan Hendrickson, James Nick Benardini, Kasthuri Venkateswaran

STAR Program Research Presentations

Planetary Protection (PP) is a scientific discipline that aims to protect other celestial bodies from microorganisms found on Earth by preventing forward and backward contamination. Forward contamination is bringing life from Earth to a given celestial body and backward contamination is preventing contamination of eventual sample return missions from Mars back to Earth. PP verifies that NASA Planetary Protection requirements are met in part via the NASA Standard Assay in order to support the launch approval process and to comply with international law. The NASA Standard Assay verifies these bioburden requirements by quantifying any aerobic, heterotrophic, and heat-shock resistant isolates. …

Analysis Of The Chondroitinase Operon Of Flavobacterium Columnare, Erin L. Sorlien

Senior Honors Projects

Analysis of the chondroitinase operon of Flavobacterium columnare

Erin Sorlien

Major

Cell and Molecular Biology, Chemistry

Advisor

Dr. David R. Nelson

Date

May 2, 2013

Keywords

Flavobacterium columnare, columnaris disease, chondroitin AC lyase, complementation, csl operon

Abstract
Flavobacterium columnare, an opportunistic bacterial pathogen of fish, is the causative agent of columnaris disease (CD). The bacterium is a Gram-negative rod that exhibits gliding motility and avidly forms biofilms. CD affects both wild and cultured freshwater fish, and continues to cause large economic losses to the fish farming industry. According to an investigation conducted by the National Animal Health Monitoring System, CD …

Survival Of B. Horneckiae Spores Under Ground-Simulated, Sneha Tharayil, Parag Vaishampayan

STAR Program Research Presentations

Increasing efforts to find and study habitable conditions and the possibility of life on extraterrestrial cosmic bodies have also deepened the need for planetary protection advancements to prevent possibilities of forward and backward contamination. Despite stringent precautions to ensure spacecraft cleanliness, a number of spore-forming bacteria, including the biofilm forming strain, Bacillus horneckiae have been isolated from spacecraft associated surfaces. B. horneckiae spores have demonstrated unusually high resistance to rigorous sterilization procedures, and extreme conditions, including intense UV radiation. This exceptionally high tolerance of extreme conditions demonstrated by spore-forming bacteria highlighted the need to assess the viability of these microbes …

The Us27 Protein Of Human Cytomegalovirus Enhances Signaling By Human Chemokine Receptor Cxcr4, Kathleen Arnolds, Juliet V. Spencer

Creative Activity and Research Day - CARD

Human cytomegalovirus (HCMV) is a prevalent pathogen that causes little disease except in immune-compromised hosts. HCMV establishes life-long latency by manipulating host immune responses. We have found that the US27 protein profoundly alters the activity of host chemokine receptor CXCR4. Cells expressing US27 and CXCR4 exhibited greater migration and calcium signaling than cells expressing CXCR4 alone. The mechanism for this enhanced signaling appears to be increased expression of CXCR4, and we are continuing to investigate how US27 causes up-regulation of CXCR4. This work is expected to clarify the role of US27 in immune evasion and provide insights into treatment strategies.

Annotation Of The Bacteriophage 933w Genome: An In-Class Interactive Web-Based Exercise, Kai F. Hung

Faculty Research & Creative Activity

No abstract provided.