Environmental Microbiology and Microbial Ecology Commons^™

Biomass Estimation Of Marine Biofilms On Plastic Surfaces, Kian Banihashemi, Fernando Javier Gil

Biological Sciences

Plastics have become a major source of marine pollution, which threatens food safety and quality, human health, and marine ecosystems. Due to the drastic negative effects of plastics in a marine environment, alternative biodegradable plastics are being generated that are more eco-friendly and have less environmental impact. Though some of these plastics are known to biodegrade, the process of degradation for bioplastics has not been heavily studied in a marine environment. This pilot project sought to both quantify the process of biodegradation and compare across different methods for effectiveness of biomass estimation, which serves as an indicator of biodegradation. Plastics …

The Response Of Marine Synechococcus To A Landscape Of Environmental Stressors: A Proteomic Exploration, Dana E. Michels

Master's Theses

In the field of marine microbial ecology, many questions remain unanswered with regards to the physiological trade-offs made by phytoplankton to maximize growth (e.g., nutrient acquisition) and minimize loss (e.g., predation defenses). These tradeoffs, which occur at the cellular level, have wide reaching impacts on food web dynamics and global biogeochemical cycles. In the first chapter, we explored the use of a non-canonical amino acid (NCAA) technique, bioorthogonal non-canonical amino-acid tagging (BONCAT), in phytoplankton model systems. This technique has potential to work well in natural systems by enabling isolation of only newly synthesized proteins during an incubation period with the …

Analysis Of Biofilm Remediation Capacity For Octenyl Succinic Anhydride (Osa), A Bioactive Food Starch Modifier Compound, Matthew R. Borglin

Master's Theses

Matthew R. Borglin

This thesis demonstrates efficacy of Octenyl Succinic Anhydride (OSA), as a biofilm sanitizer. Biofilms allow bacteria to adhere to solid surfaces with the use of excreted polymeric compounds. For example, surfaces found in food production or processing facilities such as the interior of a raw milk holding tank, are some of the most susceptible to biofilm contamination. When present, biofilms can cause a variety of negative effects, which include; reduction of product shelf life, corrosion, and outbreaks of foodborne illnesses. The close association of biofilms with the majority of foodborne illness cases led the US Environmental Protection …

Glycolic Acid Utilization In Two Species Of Marine Bacteria, Erik S. Timsak

STAR Program Research Presentations

Interactions between bacteria and phytoplankton are crucial for the cycling of organic matter in marine environments. Around 50% of organic carbon taken up by marine bacteria is converted into inorganic carbon. The uptake of organic carbon by marine bacteria exuded from phytoplankton is a key factor in regulating the marine carbon cycle. One such molecule that is exuded by phytoplankton and then uptaken by marine bacteria is called glycolate - the anion of glycolic acid, a two caron molecule. Glycolate is exuded by phytoplankton during photorespiration and 10-50% of dissolved organic carbon in marine environments is comprised of glycolate. Additionally, …

Effectiveness Of Windrow Composting Methodology In Killing A Thermo-Tolerant Species Of Salmonella During Mortality Composting, Spencer Gabriel Myers

Master's Theses

In a large agricultural operation, such as the one at Cal Poly San Luis Obispo, disposal of deceased animals is an immense issue. The cost of transporting and rendering every dead animal is inhibitory to the general function of the agricultural operations and their thin budget. Therefore, we propose that composting mortalities could be an economical alternative. Composting is a recognized method for taking animal waste products along with carbon waste and turning it into a pathogen-free, nutrient-rich topsoil. Carcass composting is in fact performed in other countries and states to varying degrees of success. However, the California EPA limits …

Influence Of A Guanidine Riboswitch In Bacterial Cells, Erik S. Timsak, Jefferey A. Kimbrel, Chris S. Ward, Rhona K. Stuart, Michael P. Thelen

STAR Program Research Presentations

Bacteria live in environments containing complex ecologies of other microbes that communicate and survive through the action of a variety of small metabolic compounds. One common yet relatively unstudied metabolite is guanidine. Although it can be toxic to cells, recent studies have revealed that guanidine may function as a cellular metabolite through a specialized RNA sequence known as a riboswitch. Within our overall project on improving algal productivity, the focus of this study is to: (a) describe various guanidine riboswitch sequences in bacteria that interact with biofuel producing algae; and (b) determine if guanidine has a positive or negative influence …

Detecting A Probiotic Product Within The Gut Of Broiler Chickens, Anneka Pisula

Master's Theses

As of January 2017, the U.S. poultry industry banned the use of antibiotics and now relies on alternatives such as probiotics to help protect animal health. Although probiotic use is not a new concept in the poultry industry, identifying the best combination of bacterial strains to generate an effective probiotic formula requires further investigation. This study aimed to detect a probiotic product of four bacterial strains (Pedioccoccus acidilactici, Pediococcus pentosaceus, Lactobacillus plantarum, and Bacillus subtilis) in a feeding trial with broiler chickens. Birds given the probiotic were predicted to show an improved growth performance with …

Spatial And Temporal Heterogeneity Of Microbial Life In Artificial Landscapes, Roopkamal Kaur, Aditi Sengupta, Peter A. Troch

STAR Program Research Presentations

The Landscape Evolution Observatory (LEO) project at Biosphere 2 consists of three replicated artificial landscapes which are sealed within a climate-controlled glass house. LEO is composed of basaltic soil material with low organic matter, nutrients, and microbes. The landscapes are built to resemble zero-order basins and enables researchers to observe hydrological, biological, and geochemical evolution of landscapes in a controlled environment. This study is focused on capturing microbial community dynamics in LEO soil, pre- and post-controlled rainfall episodes. Soil samples were collected from six different positions and at five depths in each of the three slopes followed by DNA extraction …

An Assessment Of Potential False Positive E.Coli Pyroprints In The Cplop Database, Skyler A. Gordon

Master's Theses

The genetic information found in each species of organism is unique, and can be used as a tool to differentiate at the molecular level. This has caused rapid genotyping methods to become the cornerstone of a new area of research dependent on reading the genome as a form of identification. One of these specific identification methods, known as pyroprinting, relies on the small variation of DNA sequences within the same species to develop a unique, reproducible fingerprint. By simultaneously pyrosequencing multiple polymorphic loci within the ribosomal operons known as the intergenic transcribed spacers, a reproducible output is obtained, known as …

Production And Harvest Of Microalgae In Wastewater Raceways With Resource Recycling, Alexander Colin Roberts

Master's Theses

Microalgae can be grown on municipal wastewater media to both treat the wastewater and produce feedstock for algae biofuel production. However the reliability of treatment must be demonstrated, as well as high areal algae productivity on recycled wastewater media and efficient sedimentation harvesting. This processes was studied at pilot scale in the present research.

A pilot facility was operated with nine CO₂-supplemented raceway ponds, each with a 33-m² surface area and a 0.3-m depth, continuously from March 6, 2013 through September 24, 2014. The ponds were operated as three sets of triplicates with two sets continuously fed …

Demographics And Transfer Of Escherichia Coli Within Bos Taurus Populations, Joshua Ryan Dillard

Master's Theses

In the United States, symptoms caused by pathogenic strains of Escherichia coli are on the rise. A major source of these pathogenic strains is the E. coli in the digestive tract of cattle. The purpose of this project was to determine if E. coli are transferred between individuals of the same species and if interspecies transmission is possible. Proximity of cattle was also studied as a contributing factor to the transfer of E. coli. To accomplish this goal, E. coli isolates from cattle and cohabitating ground squirrels were compared through a new method of bacterial strain typing called pyroprinting. …

How Many Particles Are Present In The Air? Bioaerosol Detection Using An Air Particle Counter, Angie Pamela Rivera, Parag Vaishampayan

STAR Program Research Presentations

Relative cleanliness in terms of particle abundance in spacecraft assembly facilities is determined by particle counts carried out in clean rooms during resting conditions. Particle counters assess total particles and particle size distribution, but do not distinguish inert particles from biological particles, which may include bacterial spores that are resistant to standard cleanroom sterilization procedures. Current cleanroom certifications do not fully assess the effects of human presence on spacecraft contamination since humans are known symbionts to enumerate microorganisms and assessments are performed at rest when there is no human presence. In this study, contamination risks and bioburden in spacecraft assembly …

Collecting Diverse Microorganisms From Rover Spacecraft, Jennifer I. Jacobs, Arianna Jefferson, Heidi Aronson, James Tan, Wayne Schubert, Parag Vaishampayan

STAR Program Research Presentations

. The Planetary Protection discipline at NASA’s Jet Propulsion Laboratory develops and implements procedures to prevent both forward and backward contamination between the Earth and solar system bodies. However, there will always be some microorganisms that will be resistant to the strictest of sterilization methods. In order understand the microorganisms found on spacecraft during assembly, and to rapidly identify them, a mass spectrometry approach was developed. As an experimental approach, a custom database was created for a subset of microorganisms in the Planetary Protection Archive. In order to make the database as accurate and efficient as possible, several different procedures …

Exploring The Physiological Role Of Vibrio Fischeri Pepn, Sally L. Cello

Master's Theses

The primary contributor to Vibrio fischeri aminopeptidase activity is aminopeptidase N, PepN. Colonization assays revealed the pepN mutant strain to be deficient at forming dense aggregates and populating the host’s light organ compared to wildtype within the first 12 hours of colonization; however the mutant competed normally at 24 hours. To address the role of PepN in colonization initiation and establish additional phenotypes for the pepN mutant strain, stress response and other physiological assays were employed. Marked differences were found between pepN mutant and wildtype strain in response to salinity, acidity, and antibiotic tolerance. This study has provided a foundation …

Methane Production By A Packed-Bed Anaerobic Digester Fed Dairy Barn Flush Water, Sean Richard Thomson

Master's Theses

Packed-bed digesters are an alternative to covered lagoon digesters for methane production and anaerobic treatment of dilute wastewaters such as dairy barn flush water. The physical media of packed-beds retain biofilms, often allowing increased treatment rates. Previous studies have evaluated several types of media for digestion of dilute wastewaters, but cost and media fouling have setback commercial development. A major operational cost has been effluent recirculation pumping.

In the present effort, a novel approach to anaerobic digestion of flush dairy water was developed at pilot-scale: broken walnut shells were used as a low-cost packed-bed medium and effluent recirculation was replaced …

Phylogenetic Diversity Of Microbial Isolates From The Mars Pathfinder, Kyla Bradylong, Adriana Blachowicz, Parag Vaishampayan, James N. Benardini, Wayne Schubert

STAR Program Research Presentations

As spacecraft are sent to different planets, they take with them microscopic pieces of life from Earth. It is the task of the Biotechnology and Planetary Protection Group to keep as much of this life off other planets as possible as well as document any life that may have been sent. During the construction of the Mars Pathfinder, samples were collected from various locations on the spacecraft to test for contamination. These samples were then isolated, grown, documented, preserved and their 16S rRNA genes were sequenced for identification. The 16S rRNA gene sequence is utilized because it is a highly …

How Clean Is The Clean Room?, Erik J. Mcfarland

STAR Program Research Presentations

In order to limit contamination and protect the integrity of search-for-life missions, spacecraft assembly clean rooms should be free of microorganisms to the extent that planetary protection guidelines require of the mission. This study monitored the bioburden of the clean room in which the Mars-destined InSight lander was being assembled. Samples of ten locations inside the clean room and one location outside were taken at monthly intervals, starting in March 2014 and scheduled to end in September 2014.

Bioburden was determined using qPCR and ATP analyses with qPCR targating 16S rRNA gene copy number and ATP targating metabolic activity of …

The Influence Of Hydrogeomorphology, Soil Redox Conditions, And Salinity On The Spatial Zoning Of Saltgrass, Salt Rush, And Cattails In Scotts Creek Marsh, Swanton Pacific Ranch, Ca, Mark D. Gormley

Master's Theses

Scotts Creek Marsh (SCM) is a small coastal wetland ecosystem in Davenport, CA. The vegetation of SCM is dominated by three halophytic zones comprised of saltgrass, salt rush, cattails. The objectives of the study were (i) to investigate the variables that influence the zoning of the three dominant halophyte communities in SCM and (ii) to the test the effectiveness of Indicator of Reduction in Soil (IRIS) tubes to indicate the reduction of S. The study examined the following parameters from April 6 to July 21, 2013: (i) the HGM of Scotts Creek Marsh, (ii) soil oxidation and reduction (redox) conditions, …

Life Support: Long Term Storage Of Solid Waste In An Enclosed Membrane System, Emmy O. Trieu, Michael Flynn, Rocco Mancinelli

STAR Program Research Presentations

In deep space missions, maintaining life support is of the utmost priority. In such a closed system, human waste must be stored and treated. Simulated feces (ersatz) will be inoculated with microbes representing normal fecal flora, mixed with urine brine and shredded refuse of products typically used in space missions. Composting methods often use alternating layers of waste with scraps of carboniferous materials (finely shredded refuse). By preparing membrane bags with a homogenized ersatz and carboniferous refuse mixture and membrane bags with alternating layers of ersatz and carboniferous refuse, it may be possible to monitor anaerobic thermophillic digestion of the …

Escherichia Coli Strain Diversity In Humans: Effects Of Sampling Effort And Methodology, Emily R. Neal

Master's Theses

Studies investigating Escherichia coli strain diversity and demographics in human hosts are frequently inconsistent regarding sampling effort and methodology while current strain typing methods are often expensive or laborious. To rectify these inconsistencies, sampling effort was investigated by comparing the diversity of 15-isolate collections to 100-isolate collections from 3 human subjects. Temporal variation in E. coli strain diversity was also studied by collecting 15 isolates once every 6 months. Additionally, strain identification and diversity collected by different sampling methods (fecal swabs vs. anal swabs collected at different times around defecation) were compared to identify any inherent biases in sampling method. …

Simplified Microbial Mats: Unraveling A Complex Community One Alga At A Time., Donna A. Dela Calzada, Brad M. Bebout, Angela Detweiler

STAR Program Research Presentations

Microbial mats are the oldest known ecosystems with an over 3.4 billion-year fossil record. Understanding their structure and function will help us better understand the biological evolution of life on early earth. They are rich in diversity (phylogenetically and physiologically) ranking amongst the most complex systems on earth. In this investigation, we aim to develop an effective method to study microbial mat organisms in relatively uncomplicated ecosystems that mimic the way they grow in nature. By growing “simplified microbial mats” on polypropylene cloth, we are able to observe the range of biogeochemical processes of a single species in a biogeochemical …

Enhanced Hydrocarbon Biodegradation Using Bioaugmentation With Biowish™-Aqua Fog, Michael Robert Lehrer

Master's Theses

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSH^TM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSH^TM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSH^TM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The …

Investigation Of Bacillus Subtilis As A Biopesticide Against Botrytis Cinerea, Kenneth K. Ng

Master's Theses

The objective of this thesis was to investigate BiOWiSH^TM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSH^TM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSH^TM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed …

Growth And Hydrogen Production Of Rubrivivax Gelatinosus Cbs, Shannon A. Swanson, Karen Wawrousek

STAR Program Research Presentations

Rubrivivax gelatinosus CBS is a purple non-sulfur photosynthetic bacterium that has been found to produce substantial amounts of Hydrogen (H2) which could be used as a clean burning biofuel. CBS has been known to grow on a variety of carbon sources, but it is unknown as to which carbon sources it has the greatest potential to use, and this understanding could be helpful in engineering future strains of the bacteria. Hydrogen production in CBS occurs when carbon monoxide (CO) is present and is presumed to be under the control of the RcoM transcription factor. When CBS wild-type strains were grown …

Pcr Primers For The Detection Of Propane And Butane-Oxidizing Microorganisms, Brian Jeremy Chan

Master's Theses

In an increasingly energy-hungry world, our capacity to meet the heightened energy demands of the future has become a pressing matter. The most urgent of these concerns are tied to the accessibility of petroleum. Various experts have proselytized both the imminent arrival of peak oil production rates and the ensuing decline of those rates thereafter. And to that end, the development of novel and advanced oil exploration methodologies has become almost as important as finding the sources of oil themselves.

The soils above petroleum reservoirs play host to various communities of alkane- oxidizing bacteria that can utilize the natural gas …

Temperature Influence And Heat Management Requirements Of Microalgae Cultivation In Photobioreactors, Thomas Hagen Mehlitz

Master's Theses

Microalgae are considered one of the most promising feedstocks for biofuel production for the future. The most efficient way to produce vast amounts of algal biomass is the use of closed tubular photobioreactors (PBR). The heat requirement for a given system is a major concern since the best algae growth rates are obtained between 25-30 °C, depending on the specific strain. A procedure to determine temperature influence on algal growth rates was developed for a lab-scale PBR system using the species Chlorella. A maximum growth rate of 1.44 doublings per day at 29 °C (optimal temperature) was determined. In addition, …