Microbiology Commons^™

Integrated Periphyton Biofilters In Marine Recirculating Aquaculture Systems, Adam N. Bell

USF Tampa Graduate Theses and Dissertations

Marine aquaculture is a growing industry that has the potential to reduce eutrophication, overfishing, and loss of biodiversity. Recirculating aquaculture systems (RAS) are a type of aquaculture system that decreases discharge, raises biological control, and conserves water; however, RAS are challenged due to high energy cost, low biodiversity, water quality control, and waste nutrients. Periphyton biofilters are a new technology that allows for feed production, water quality control, microbial diversity, and resource recovery in marine aquaculture systems. The overarching question that this dissertation investigates is, how can periphyton biofilters be integrated into RAS to address the challenges of modern sustainable …

A Novel Application Of Spinning Disk Technology To Collect Plasma From Whole Blood Prior To Quantifying Plasma Rna, Naomi Rapier-Sharman, Mae-Lynn L. Hutchinson, Carlos M. Moreno, Abraham Quaye, Brian D. Poole, K. Scott Weber, William G. Pitt, Brett E. Pickett

Faculty Publications

The spinning disk technology has previously been utilized to isolate bacterial components from blood in hours instead of days. We hypothesized that this platform could be applied as an alternative approach to isolating plasma RNA from a whole blood sample. We consequently tested the efficacy of the spinning disk technology to extract plasma from whole blood upstream of RNA isolation and analysis. To do so, we collected plasma using either the spinning disk or the typical two-spin centrifuge method. We found that the spinning disk method results in significantly more hemolysis during collection than the conventional two-spin centrifuge method. However, …

Towards Better Detection And Treatment Of Mycobacterial Disease., Laura O'Connell

ORBioM (Open Research BioSciences Meeting)

Background

Mycobacterial disease is a major cause of fatality, with 1.5 million deaths per 10 million infections annually. Due to the emergence of multi- and extensively-drug resistant clinical isolates, mycobacteriophage (MP) are being investigated as an alternative to antibiotics.

Objectives

The objectives of this study were: to characterise an emerging cohort of MP, to determine suitable in vivo modes of delivery for MP including encapsulation, to examine the application of a MP-based viability assay, and to examine the functionality of MP in raw milk.

Methods

The fast-growing Mycobacterium smegmatis was used as a host in all assays to demonstrate phage …

Investigating The Ability Of Saltwater Isolates To Inhibit Biofilm Formation By Clinically Significant Pathogens, Monica Murphy

ORBioM (Open Research BioSciences Meeting)

Background

Nosocomial pathogens are responsible for chronic and persistent infections partly due to their ability to form protective biofilms which enhance their antibiotic tolerance. Biofilm disruption combats antimicrobial resistance as dispersed cells regain antibiotic susceptibility. The goal of this study was to evaluate the antibiofilm potential of isolates from marine environments against a pathogen panel consisting of MRSA and clinical Staphylococcus aureus strains, Acinetobacter baumannii and Escherichia coli pathogens.

Method

A large bank of saltwater bacterial isolates was cultured in glucose-supplemented trypticase soy broth (TSBg) and resulting colonies were screened against the pathogen panel. Isolates of interest were further examined …

Short Chain Fatty Acid Combination Treatment Protects Against 6-Ohda And Wt Α-Synuclein Induced Decreases In Neurite Growth In In Vitro Models Of Parkinson’S Disease., Alex Morris

ORBioM (Open Research BioSciences Meeting)

Background

Parkinson’s disease (PD) is a neurodegenerative disorder characterized by dopaminergic neuron degeneration. This degeneration is partly driven by over expression of α-synuclein (α-syn) and development of α-syn aggregates known as Lewy bodies throughout the substantia nigra. As well as motor dysfunction, PD presents with several chronic gastrointestinal comorbidities, which cause a decline of gut microbial diversity and microbially derived short chain fatty acids (SCFAs). Recent in vivo studies have shown SCFAs to be neuroprotective in various degenerative disease states, suggesting that SCFAs may protect against dopaminergic degeneration.

Methods

Human neuroblastoma SH-SY5Y cells were used as a model of human …

Urobiome Screening For Antimicrobial Peptides Against Uropathogenic Escherichia Coli, Jennifer Jones

ORBioM (Open Research BioSciences Meeting)

BACKGROUND: Urinary tract infections (UTIs) are one of the most common bacterial infections worldwide. As global incidences of UTIs caused by multidrug resistant bacteria continue to increase, so too does the demand for novel antimicrobial therapies.

Due to its relatively understudied nature, the urinary microbiome represents a niche with an untapped source of potentially novel antimicrobials (e.g., bacteriocins). Improvements to bacterial culturing and sequencing techniques have highlighted these potential alternative treatments and control strategies to target antibiotic resistant uropathogenic E. coli (UPEC).

METHODS: Expanded quantitative urine culture (EQUC) was used to culture bacterial isolates from mid-stream urine samples. Urinary isolates …

Examining The Absorbance Maximum, Protein Concentration And Antimicrobial Activity Of Seaweed Extracts, Claudia Kenneally

ORBioM (Open Research BioSciences Meeting)

Background

The emergence of antimicrobial resistance (AMR) has highlighted the need for novel antimicrobials. Antimicrobial compounds can be isolated from various marine sources, including seaweed, which contain pigments and proteins. This study focuses on screening seaweeds for potential antimicrobial compounds.

Methods

Four seaweed types (A, B, C, D) were collected from beaches in Cork, Ireland, and extracts of each were prepared using both methanol (-M) and water (-H₂O). The absorbance maximum (λ_max)of seaweed extracts was determined from 220 – 800 nm to investigate potential pigment interference at 600 nm. The bicinchoninic acid (BCA) assay was preformed …

Bactericidal Effects Of Chlorine And Bacteriophages On Mycobacteria In Conventionally Treated Water., Max Kevane-Campbell

ORBioM (Open Research BioSciences Meeting)

Introduction

Mycobacteria are members of the order Actinomycetales and the only genus in the family Mycobacteriaceae. The distinguishing characteristics that are found in mycobacteria include acid-fastness and the presence of mycolic acids. Mycobacteria can be commonly classified as non-spore-forming, aerobic, slender rod-shaped, and are slow-growers. Natural reservoirs that mycobacteria can be found in are aquatic and terrestrial environments. Recent studies have shown that mycobacteria that can cause skin lesions, immune and/or pulmonary dysfunctions and chronic diseases, i.e., Mycobacterium avium subsp. Paratuberculosis, M. kansasii, and M. xenopi, can be isolated from common household tap water. Tap water can originate from …

Development Of A Protocol For The Extraction Of Genomic Material From Fecal Matter For Metagenome And Virome Sequencing, Keith Coughlan

ORBioM (Open Research BioSciences Meeting)

With the advent of increasingly more accurate and reliable sequencing methods, the requirement for extraction methods yielding large volumes of high-quality genomic material has become progressively more significant. While the analysis of genomic material through bioinformatics can offer some methods of “cleaning” and standardizing data, it is essential that the “wet lab” aspects of data produce abundant amounts of high-quality genetic material. Therefore, the purpose of this project is to develop an optimized method of DNA extraction for fecal material sequencing in microbiome and virome investigations. The samples used for the optimized protocol are infant fecal samples collected and stored …

Mycobacteriophage: A Demonstration Of The Reduction Of M. Smegmatis In Various Matrices., Laura O'Connell

ORBioM (Open Research BioSciences Meeting)

Background

Mycobacterial disease is a major cause of fatality worldwide, with approximately 1.5 million deaths for every 10 million infections. These infections are difficult to treat due to the intrinsic resistance of its mycolic acid rich cell wall to many antibiotics. There is potential for mycobacteriophage (MP) to be used therapeutically for multidrug- and extensively-drug resistant infections. Mycobacterium smegmatis mc² 155 is a useful substitute for slow-growing pathogenic mycobacteria, as it propagates quickly under lab conditions. This feature of M. smegmatis increases the pace of analysis, by ensuring the quick isolation and characterisation of MP and acting as a …

Bioengineered Nisin A Derivatives Display Enhanced Activity Against Clinical Neonatal Pathogens, Anna Desmond

ORBioM (Open Research BioSciences Meeting)

Neonatal infection is a significant cause of mortality and morbidity in infants. Globally, neonatal pathogens are increasingly becoming multi-drug resistant, indicating the need to discover novel alternative treatment strategies. Nisin is an antimicrobial peptide that exhibits broad-spectrum activity against Gram-positive and Gram-negative bacteria including a wide variety of clinical pathogens. Nisin can be used in combination with antibiotics to improve their efficacy. This study examined the activity of nisin and bioengineered derivatives against multi-drug resistant Streptococcus agalactiae and Staphylococcus capitis isolates and investigated the potential synergy between nisin peptides and selected antibiotics. Whole genome sequence analysis of the strains revealed …

In Vitro Antimicrobial Activity Of Cell-Free Supernatants From Asymptomatic Bacteriuria (Abu) Isolates Against Multi-Drug Resistant Uropathogenic Escherichia Coli (Upec), Ciara Kenneally

ORBioM (Open Research BioSciences Meeting)

Background

Multi-drug resistant uropathogenic Escherichia coli (UPEC) cause > 80% of community acquired urinary tract infections (UTIs) and > 40% of nosocomial UTIs. Preliminary evidence suggests a beneficial role for UPEC’s commensal counterparts, asymptomatic bacteriuria (ABU), in establishing bacterial interference against UPEC. The aim of this study was to analyse the antimicrobial activity of cell free supernatants (CFS) from ABU isolates against UPEC.

Methods

In vitro antagonistic effects of nine uncharacterised ABU isolates were compared against the inhibitory affect induced by the prototype ABU isolate, E. coli 83972. Turbidimetric growth assays were performed using the CFS of the ABU isolates against eight …

Investigating The Thermotolerance Of Cronobacter Sakazakii In Powdered Infant Formula, Fiona Ryan O'Connor

ORBioM (Open Research BioSciences Meeting)

Background

The powdered infant formula (PIF) sector is a growing global market, with Ireland at the forefront of the European exports market; reaching €1.5bn and accounting for 35% of all dairy exports in 2015. However, a major concern associated with PIF is the risk of contamination with the neonatal pathogen Cronobacter sakazakii. Although PIF undergoes thermal treatment during manufacture, it is possible that re-contamination of the product may occur via later processing measures. We investigated the resistance of C. sakazakii to thermal inactivation following inoculation in commercial PIF as a potential treatment for contaminated powder.

Method

Five strains of C. …

Tracing Evolution Of Gene Transfer Agents Using Comparative Genomics, Roman Kogay

Dartmouth College Ph.D Dissertations

The accumulating evidence suggest that viruses and their components can be domesticated by their hosts, equipping them with convenient molecular toolkits for various functions. One of such domesticated system is Gene Transfer Agents (GTAs) that are produced by some bacteria and archaea. GTAs morphologically resemble small phage-like particles and contain random fragments of their host genome. They are produced only by a small fraction of the microbial population and are released through a lysis of the host cell. Bioinformatic analyses suggest that GTAs are especially abundant in the taxonomic class of Alphaproteobacteria, where they are vertically inherited and evolve …

Cell Surface And Plasma Biomarkers Of Kaposi Sarcoma, Sara R. Privatt

Dissertations and Doctoral Documents, University of Nebraska-Lincoln, 2023–

Kaposi sarcoma-associated herpesvirus is the etiological agent of several pathologies including Kaposi sarcoma, primary effusion lymphoma, and multicentric Castleman’s disease. Here, we investigated both the tumor microenvironment (TME) in KS lesions as well as plasma from KSHV-infected individuals and how metabolic dysfunction and dysregulated cytokines might have a systemic effect on KS tumorigenesis. Based on previously published KS tumor transcriptomics, one aspect of our investigation involved characterizing cell surface glycoproteins with potential to serve as therapeutic targets against KSHV-infected cells or KS tumors or as biomarkers for disease. Expression patterns of such markers may also suggest the cellular origin of …

Connecting The Nebraska Water Quality Index To The Aquatic Microbial Community Of The North Platte River Basin, Nebraska, Paula R. Guastello

School of Natural Resources: Dissertations, Theses, and Student Research

The Nebraska Water Quality Index, under development by the Nebraska Department of Environment and Energy, summarizes in a single value eight environmental parameters that have been monitored in Nebraska for nearly 20 years. Water quality parameters including those used in the Nebraska Water Quality Index have been shown in previous studies to impact bacterial growth. As such, this index has the potential to correlate with the freshwater microbial community. Here, I relate the Nebraska Water Quality Index to microbial community composition and structure using 16S rRNA gene amplicon sequence data collected from the North Platte River Basin, Nebraska. This index …

The Pseudomonas Syringae Pv. Tomato Dc3000 Effector Hopd1 Interferes With Cellular Dynamics Associated With The Function Of The Plant Immune Protein Atnhr2b, Luis Francisco Marin-Ponce, Catalina Rodriguez-Puerto, Perla Rocha-Loyola, Clemencia M. Rojas

Entomology and Plant Pathology Faculty Publications and Presentations

The plant pathogenic bacterium Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) causes disease in tomato, in the model plant Arabidopsis thaliana, and conditionally in Nicotiana benthamiana. The pathogenicity of Pst DC3000 is mostly due to bacterial virulence proteins, known as effectors, that are translocated into the plant cytoplasm through the type III secretion system (T3SS). Bacterial type III secreted effectors (T3SEs) target plants physiological processes and suppress defense responses to enable and support bacterial proliferation. The Pst DC3000 T3SE HopD1 interferes with plant defense responses by targeting the transcription factor NTL9. This work shows that HopD1 also targets the …

Gastrointestinal Infections Modulate The Risk For Insulin Autoantibodies As The First-Appearing Autoantibody In The Teddy Study, Maria Lönnrot, Kristian F Lynch, Marian Rewers, Åke Lernmark, Kendra Vehik, Beena Akolkar, William Hagopian, Jeffrey Krischer, Rickhard A Mcindoe, Jorma Toppari, Anette-G Ziegler, Joseph F Petrosino, Richard Lloyd, Heikki Hyöty, Teddy Study Group

Faculty, Staff and Students Publications

OBJECTIVE: To investigate gastrointestinal infection episodes (GIEs) in relation to the appearance of islet autoantibodies in The Environmental Determinants of Diabetes in the Young (TEDDY) cohort.

RESEARCH DESIGN AND METHODS: GIEs on risk of autoantibodies against either insulin (IAA) or GAD (GADA) as the first-appearing autoantibody were assessed in a 10-year follow-up of 7,867 children. Stool virome was characterized in a nested case-control study.

RESULTS: GIE reports (odds ratio [OR] 2.17 [95% CI 1.39-3.39]) as well as Norwalk viruses found in stool (OR 5.69 [1.36-23.7]) at(OR 0.48 [0.35-0.68]). GIE reports at any other age were associated with an increase in …

Increasing Glutathione Levels By A Novel Posttranslational Mechanism Inhibits Neuronal Hyperexcitability, Ashwini Sri Hari, Rajeswari Banerji, Li-Ping Liang, Ruth E Fulton, Christopher Quoc Huynh, Timothy Fabisiak, Pallavi Bhuyan Mcelroy, James R Roede, Manisha Patel

Faculty, Staff and Students Publications

Glutathione (GSH) depletion, and impaired redox homeostasis have been observed in experimental animal models and patients with epilepsy. Pleiotropic strategies that elevate GSH levels via transcriptional regulation have been shown to significantly decrease oxidative stress and seizure frequency, increase seizure threshold, and rescue certain cognitive deficits. Whether elevation of GSH per se alters neuronal hyperexcitability remains unanswered. We previously showed that thiols such as dimercaprol (DMP) elevate GSH via post-translational activation of glutamate cysteine ligase (GCL), the rate limiting GSH biosynthetic enzyme. Here, we asked if elevation of cellular GSH by DMP altered neuronal hyperexcitability in-vitro and in-vivo. Treatment of …

P2y2 Purinergic Receptor Gene Deletion Protects Mice From Bacterial Endotoxin And Sepsis-Associated Liver Injury And Mortality, Athis R Arunachalam, Sanju S Samuel, Arunmani Mani, Janielle P Maynard, Kelsey M Stayer, Eric Dybbro, Subapradha Narayanan, Aalekhya Biswas, Saliha Pathan, Krishnakant Soni, Abu Hena Mostafa Kamal, Chandra Shekar R Ambati, Nagireddy Putluri, Moreshwar S Desai, Sundararajah Thevananther

Faculty, Staff and Students Publications

The liver plays a significant role in regulating a wide range of metabolic, homeostatic, and host-defense functions. However, the impact of liver injury on the host's ability to control bacteremia and morbidity in sepsis is not well understood. Leukocyte recruitment and activation lead to cytokine and chemokine release, which, in turn, trigger hepatocellular injury and elevate nucleotide levels in the extracellular milieu. P2Y2 purinergic receptors, G protein-coupled and activated by extracellular ATP/UTP, are expressed at the cell surface of hepatocytes and nonparenchymal cells. We sought to determine whether P2Y2 purinergic receptor function is necessary for the maladaptive host response to …