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Articles 421 - 450 of 507
Full-Text Articles in Microbiology
Essential Oils And Their Antimicrobial Effect On Serratia Marcescens, Seren C. Davies-Hughes, Patricia I. Marquez, Henna N. Schulz, Laura S. Sanchez
Essential Oils And Their Antimicrobial Effect On Serratia Marcescens, Seren C. Davies-Hughes, Patricia I. Marquez, Henna N. Schulz, Laura S. Sanchez
Humboldt Journal of Microbiology
S. marcescens is a gram-negative, facultatively anaerobic bacterium that is particularly aggressive in its growing capability. It is a concern in medical fields as well as home environments due to its resistance to most antibiotics, high mortality rate, and easy ability to colonize on soap and other fatty substances. Although testing of natural cleansers such as lemon juice, vinegar, and ammonia has been done on S. marcescens and essential oil efficacy has been tested on Escherichia coli and Staphylococcus aureus, no literature has been published on the effect of essential oils on S. marcescens growth. The purpose of this experiment …
Trends In Antibiotic Susceptibility Of Staphylococcus Aureus Isolates In A Pediatric Hospital: An Analysis Of The Impact Of The Sars-Cov-2 Pandemic, Juan Miguel S. Gonzalez Rivero
Trends In Antibiotic Susceptibility Of Staphylococcus Aureus Isolates In A Pediatric Hospital: An Analysis Of The Impact Of The Sars-Cov-2 Pandemic, Juan Miguel S. Gonzalez Rivero
Honors Undergraduate Theses
Infections caused by the organism Staphylococcus aureus are one of the most common causes for community-associated and healthcare-acquired infections (HAI). Isolates of this bacterium found within the healthcare setting often demonstrate a higher prevalence of antibiotic resistance making these infections difficult to treat. Historically, considerable focus has been placed on methicillin-resistant S. aureus (MRSA), which are strains resistant to β-lactam antibiotics like penicillin, oxacillin and cephalosporins; however, methicillin-sensitive (MSSA) strains may also possess resistance to several first-line antibiotics. Resistance to antibiotics can be acquired through horizontal gene transfer (HGT) by means of mobile genetic elements or by random DNA mutations …
Surveillance And Monitoring Of Amphibian Pathogens And Evaluating The Impacts Of An Associated Course-Based Undergraduate Research Experience (Cure), Jennifer Perez
EWU Masters Thesis Collection
Amphibians are a highly diverse class of vertebrates and crucial for natural ecosystems, inhabiting both aquatic and terrestrial environments at different life stages. However, amphibians are facing devastating declines largely due to the disease chytridiomycosis, caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). This disease can lead to population declines, biodiversity loss, and species extinction. However, resources to study and mitigate this disease are limited and an opportunity to assist in these efforts has been created in the form of a course-based undergraduate research experience (CURE). Chapter 1 of my thesis used Roger's Diffusion of Innovations …
Comparison Of Groundwater Chemistry And Associated Biodiversity Of Sulfidic Karst Habitats In Southcentral Kentucky, Kendall Wheeler
Comparison Of Groundwater Chemistry And Associated Biodiversity Of Sulfidic Karst Habitats In Southcentral Kentucky, Kendall Wheeler
Mahurin Honors College Capstone Experience/Thesis Projects
In most ecosystems, primary production is conducted by plants or photosynthetic microbes; however, extremophilic sulfur-oxidizing bacteria can use hydrogen sulfide gas (H2S) as an electron donor to produce metabolic energy in a type of metabolism called chemoautotrophy. These ecosystems are a prominent feature of some karst landscapes, and two have been observed in the Mammoth Cave system, Sulphur River in Parker Cave and Marianne’s Pass in Mammoth Cave National Park respectively.
Sulfidic water is ubiquitous in southcentral Kentucky and is strongly associated with hydrocarbon deposits. This oil boom of the 1880s in this region predates most environmental regulation and records …
Staff Education To Promote Probiotic Breastfeeding Therapy, José Diaz
Staff Education To Promote Probiotic Breastfeeding Therapy, José Diaz
Walden Dissertations and Doctoral Studies
Preterm birth is the leading cause of death in children under 5. In addition, there are 22 million babies born underweight each year which also contributes to the infant mortality rate. Some reasons for the inability to thrive and survive are the infants' immature system, allostatic load inadaptability, and undiversified and underdeveloped microbiota. Human breast milk (HBM) provides lifesaving nutrients, which prevents necrotizing enterocolitis and mortality. In addition, probiotic supplementation helps reduce late-onset sepsis, morbidities, and mortality. A literature review provided substantial evidence that HBM and probiotics can improve an infant's microbiota, thus improving thrivability and survivability. Probiotics offset the …
Investigation Of Key Mechanisms Of Contact Dependent Growth Inhibition Systems In Burkholderia Cepacia Complex Species, Zaria K. Elery
Investigation Of Key Mechanisms Of Contact Dependent Growth Inhibition Systems In Burkholderia Cepacia Complex Species, Zaria K. Elery
Theses and Dissertations--Microbiology, Immunology, and Molecular Genetics
Burkholderia cepacia complex is a group of closely related environmental bacteria that can exacerbate disease in immunocompromised individuals. B. cepacia bacteria are equipped with an arsenal of mechanisms to cooperate and compete for nutrients in various polymicrobial environments. Contact-dependent growth inhibition (CDI) systems are antagonistic mechanisms widespread among proteobacteria. CDI systems consist of Two Partner Secretion pathway proteins that function to deliver the toxic C-terminal portion of the large effector exoprotein (termed ‘BcpA’ in Burkholderia-type CDI systems) to the cytoplasm of a compatible recipient bacterial cell upon direct cell to cell contact. The translocation of BcpA out of the …
Substrate Selection And Effector Gating In The Helicobacter Pylori Cag Type Iv Secretion System, Mackenzie E. Ryan
Substrate Selection And Effector Gating In The Helicobacter Pylori Cag Type Iv Secretion System, Mackenzie E. Ryan
Theses and Dissertations--Microbiology, Immunology, and Molecular Genetics
Helicobacter pylori is a recognized carcinogen and gastric colonization by strains that harbor the cag type IV secretion system (1) is the strongest known risk factor for stomach malignancy. Gastric adenocarcinoma is the fourth leading cause of cancer-related deaths world-wide ( > 700,000 deaths annually), with H. pylori directly contributing to the development of more than one million new cases of cancer per year accounting for 5.5% of all malignancies. H. pylori exploits cag T4SS activity to alter the mucosal microenvironment by delivering diverse immunostimulatory cargo into target gastric epithelial cells. Currently, the mechanisms by which the cag T4SS …
Receptor Usage And Infection Biology Of Influenza D Virus, Tirth Uprety
Receptor Usage And Infection Biology Of Influenza D Virus, Tirth Uprety
Theses and Dissertations--Veterinary Science
Influenza D virus (IDV) utilizes bovines as a primary reservoir with periodical spillover to other hosts. We have previously demonstrated by the traditional hemagglutination assay coupled with sialoglycan microarray and functional assays that IDV can recognize and bind both 9-O-acetylated N-acetylneuraminic acid (Neu5,9Ac2) and 9-O-acetylated N-glycolylneuraminic acid (Neu5Gc9Ac). Agricultural animals such as cattle can produce both Neu5,9Ac2 and Neu5Gc9Ac, while humans are genetically unable to synthesize Neu5Gc9Ac. The bifunctional enzyme, glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase (GNE) is a master regulator of de-novo sialic acid synthesis while 9-O-Acetylation of sialic acids is catalyzed by …
Effect Of Soil Ph And Indigenous Bacterial Populations In Selecting Rhizobial Endophytes Within Soybean Root Nodules, Brianne Danielle Edwards
Effect Of Soil Ph And Indigenous Bacterial Populations In Selecting Rhizobial Endophytes Within Soybean Root Nodules, Brianne Danielle Edwards
Graduate Theses/Dissertations
Soybean plants fulfill most of their nitrogen requirement by developing symbiotic associations with four different rhizobial genera, including Bradyrhizobium, Sinorhizobium, Mesorhizobium, and genus Rhizobium. In general, members of Bradyrhizobium and Sinorhizobium have been reported as dominant root nodule endophytes under acidic soils and alkaline soil conditions, respectively. So far, it is unknown whether the selection of rhizobial endophytes is regulated by their ability to survive under different soil pH or driven by host plants regardless of their relative abundance in soil. This study was focused on the assessment of the potential role of soil pH in …
Location Of The Fibronectin Binding Domain On The Fibronectin Attachment Protein Of Mycobacterium Avium Subsp. Paratuberculosis, Chrissy Rose Hattery
Location Of The Fibronectin Binding Domain On The Fibronectin Attachment Protein Of Mycobacterium Avium Subsp. Paratuberculosis, Chrissy Rose Hattery
All Graduate Theses, Dissertations, and Other Capstone Projects
Mycobacterium avium paratuberculosis (MAP) is an acid-fast bacillus that causes a fatal granulomatous ileocolitis called Johne’s Disease that affects primarily ruminants. The impact of MAP infection in the US dairy industry is particularly severe, with up to 70.4% of the herds being infected and annual losses as high as $1.5 billion. MAP is capable of persisting for months in the livestock environment and surviving the pasteurization process. While several vaccines are available, none can prevent infection and treatment with antimycobacterial chemotherapy does not result in a cure. Management and control of MAP infection requires characterization of host-pathogen interaction at the …
Novel Amino Acids Dictate Differences In Sivmac239 And Hiv-2 Rod9 Vpx Cellular Localization, Hunter D. Mayhew
Novel Amino Acids Dictate Differences In Sivmac239 And Hiv-2 Rod9 Vpx Cellular Localization, Hunter D. Mayhew
All Graduate Theses, Dissertations, and Other Capstone Projects
Vpx is a protein encoded by Human Immunodeficiency Virus Type 2 (HIV-2) and specific Simian Immunodeficiency Viruses (SIVs) that is essential for successful infection of non-dividing myeloid cells. Vpx facilitates viral replication by trafficking the viral genome into the nucleus of infected cells and targeting the primarily nuclear innate immune protein SAMHD1 for proteasomal degradation. Considering that these activities occur at the nucleus, we hypothesized that Vpx nuclear localization would be tightly correlated with these functions. To assess this, site directed mutagenesis in combination with confocal microscopy or Western Blotting analyses were utilized to evaluate the determinants of nuclear accumulation …
Odd Bedfellows & Marriage Of Necessity: Public Health And Politics In American Federalism, Gabriella Victoria Chianese
Odd Bedfellows & Marriage Of Necessity: Public Health And Politics In American Federalism, Gabriella Victoria Chianese
Honors Theses and Capstones
In the wake of the coronavirus disease 2019 (COVID-19) pandemic, while the public disagrees over mask mandates, required vaccinations, and social distancing, it seems like one common sentiment exists – a distaste for the odd bedfellows of politics and public health. There are those who cry for the compartmentalization of the science of public health and the art of politics to rectify this situation. In the wake of so much confusion and chaos, it is not unjustified to demand the depoliticization of public health; however, this response is unrealistic given the modern political climate, demonstrates a narrow understanding of the …
Understanding Human Gut Bacterial Interactions And Their Role In Colonization Resistance Against Clostridioides Difficile Using Mini-Bioreactor Array And Caenorhabditis Elegans As Model Systems, Achuthan Ambat
Electronic Theses and Dissertations
No abstract provided.
Effectiveness Of Atmospheric Cold Plasma In Inactivating Microorganisms, Nadee Shanika Kaluwahandi
Effectiveness Of Atmospheric Cold Plasma In Inactivating Microorganisms, Nadee Shanika Kaluwahandi
Electronic Theses and Dissertations
In the food processing industry, Cold Plasma (CP) is an emerging green process with a number of potential applications. Cold plasma has mostly been used to reduce microbial counts in foodstuffs and biological materials as well as in different levels of packaging, particularly in cases where there is thermal sensitivity. Recent studies have demonstrated that CP technology is being developed for use in the food and agriculture industries. The primary focus is on the interactions between reactive species and food-borne microbes to inactivate them. The literature review discusses both proven and potential applications for cold plasma in food processing, as …
Marineepi: A Gui-Based Matlab Toolbox To Simulate Marine Pathogen Transmission, Gorka Bidegain, Tal Ben-Horin, Eric N. Powell, John M. Klinck, Eileen E. Hofmann
Marineepi: A Gui-Based Matlab Toolbox To Simulate Marine Pathogen Transmission, Gorka Bidegain, Tal Ben-Horin, Eric N. Powell, John M. Klinck, Eileen E. Hofmann
CCPO Publications
The Graphical User Interface (GUI) MarineEpi is presented as a Matlab toolbox for easily (i) constructing disease transmission models for different marine host-pathogen systems, (ii) running simulations by specifying initial conditions and model parameters, and (iii) interpreting the resulting time series of the host and pathogen population dynamics. MarineEpi users can generate models for systems in which pathogen transmission occurs through contact with infected individuals (SI), contact with dead infected individuals (SID), contact with environmental pathogens released by infected individuals (SIP), and contact with environmental pathogens released by dead infected individuals (SIPD). MarineEpi is a freely available GUI that provides …
Soil And Compost Tea: A New Restoration Technique?, Carol E. Day
Soil And Compost Tea: A New Restoration Technique?, Carol E. Day
Master's Theses and Doctoral Dissertations
Historically, prairie restorations have lacked the plant diversity seen in remnant prairies. Most restoration practices focus on reestablishing the plant community but overlook the soil microbial community even though microbes are critical to habitat functioning. Developing techniques that increase soil microbes in prairie restorations is critical to ensuring diverse restored habitats. We compared how microbial communities differed between remnant and restored prairie sites. We also investigated if soil and compost teas could be used to reintroduce microbes to restored prairie soil and if the teas affected native plant establishment. We found significant differences in the levels of bacterial taxa between …
Antimicrobial Resistance Profile Of Escherichia Coli Isolated From Poultry Litter, M. J. Khong, A. M. Snyder, M. M. Young, N. L. Barbieri, S. L. Weimer
Antimicrobial Resistance Profile Of Escherichia Coli Isolated From Poultry Litter, M. J. Khong, A. M. Snyder, M. M. Young, N. L. Barbieri, S. L. Weimer
Poultry Science Faculty Publications and Presentations
Antimicrobial resistance is a threat to animal and human health. As a commensal and zoonotic bacterium, Escherichia coli has the potential to be a pathogenic source of antimicrobial resistance. The purpose of this study aimed to investigate the antimicrobial resistance profile of E. coli isolated from litter collected from pens in a broiler chicken experiment. E. coli was isolated from litter samples (n = 68 isolates) of 16 pens housing broilers to d 53 of age. Resistance to 10 antimicrobials was observed by disc diffusion. The presence of 23 antimicrobial and heavy metal resistance genes, O serogroups, and avian pathogenic …
College Of Natural Sciences 2022 Year-End Publication, College Of Natural Sciences
College Of Natural Sciences 2022 Year-End Publication, College Of Natural Sciences
College of Natural Sciences Newsletters and Reports
This is the 2022 issue of the annual College of Natural Sciences year-end publication.
Contents:
[Page] 2 Dean's message
[Page] 3 Department highlights
[Page] 4 Overview of Bold & Blue Campaign
[Page] 5 Dr. Edward Hogan recognition & endowment
[Page] 6 Career milestones
[Page] 7 Student travel and research
[Page] 8 $11 million COBRE grant
[Page] 9 Professional Science Masters & Research highlights
[Page]10 Outreach highlights throughout the state
[Page] 11 2022 events recap – join us in 2023!
[Page] 12 Updates on our VR initiative
[Page] 14 Overview of awards and recognitions from 2022
Investigating The Effectiveness Of Microplastic Removal And The Relationship Between Microplastics, Bacterial Communities, And Nitrogen Cycling In The Natural Treatment Systems At The Arcata Wastewater Treatment Facility, Fiona Connor
Cal Poly Humboldt theses and projects
This thesis addressed two challenging aspects of microplastic research, including the quantification of microplastics in organic-rich water, and the investigation of how microplastics impact microorganisms and related nutrient-cycling. Two projects were designed and executed to address specific research objectives relating to these broad topics. The first objective was to develop and implement methods to determine the effectiveness of microplastic removal across the natural treatment systems at the Arcata Wastewater Treatment Facility (AWTF). The second objective was to observe how microplastics found in domestic wastewater impact the bacterial communities and nitrogen cycling in freshwater treatment wetland sediment through a bench-scale experiment, …
Host-Microbe Interactions In Non-Native Estuarine Anemones: Biogeography And Temperature, Parker K. Lund
Host-Microbe Interactions In Non-Native Estuarine Anemones: Biogeography And Temperature, Parker K. Lund
Cal Poly Humboldt theses and projects
Non-native species are increasing in prevalence around the world, resulting in negative economic and ecological impacts. However, the broad distributions of non-native species also offer a system for investigating the response of host-associated microbial communities to environmental factors across a range of ecological scales. At the broadest scale, I investigated the geography of microbial communities in the non-native estuarine anemone Diadumene lineata on the west coast of the United States of America. Across latitudes, microbial community composition was very similar and displayed a high percentage of Klebsiella spp. at all sites. However, the communities in California tended to exhibit higher …
Community Ecology And Disease Dynamics In Pacific Oysters: Unraveling Microbiome-Pathogen Interactions In The Wild, Victoria E. Cifelli
Community Ecology And Disease Dynamics In Pacific Oysters: Unraveling Microbiome-Pathogen Interactions In The Wild, Victoria E. Cifelli
Cal Poly Humboldt theses and projects
In the context of multispecies microbial assemblages, disruptions can occur when there are alterations in host conditions, such as the onset of a disease. Notably, viruses have the potential to reshape a host's microbial community. However, the role of the host's habitat and environment, which could be pivotal in communities with shifting niche space and habitat filters, is often overlooked in host-microbe-pathogen interactions. Recognizing the importance of these factors, I employed a field-based approach to understand microbial community dynamics in the presence of disease. To address the influence of geographical location, I conducted an analysis involving healthy and infected oysters …
Probing The Porphyromonas Gingivalis Nitrosative Stress Response Network, Alexandra Galina
Probing The Porphyromonas Gingivalis Nitrosative Stress Response Network, Alexandra Galina
Theses and Dissertations
Objective: To persist in the oral cavity, the perio-pathogen Porphyromonas gingivalis must have an efficient nitrosative stress response mechanism. To better understand how P. gingivalis responds to nitrite (NO2-) we must determine what genes are involved in the nitrosative stress response network.
Methods: Through genomic screening, a list of genes was generated that were hypothesized to be part of the nitrosative stress response network and believed to be involved in nitrate/nitrite metabolism or redox state of the cells. Protein coding regions were replaced with the ermF sequence by allelic exchange mutagenesis. 15 mutants were created …
Extension Of The Ergot Alkaloid Gene Cluster, Samantha Joy Fabian
Extension Of The Ergot Alkaloid Gene Cluster, Samantha Joy Fabian
Graduate Theses, Dissertations, and Problem Reports (ETD)
Specialized metabolites produced by fungi impact human health. A large portion of the pharmaceuticals currently on the market are derived from metabolites biosynthesized by microbes. Ergot alkaloids are a class of fungal metabolites that are important in the interactions of environmental fungi with insects and mammals and also are used in the production of pharmaceuticals. In animals, ergot alkaloids can act as partial agonists or antagonists at receptors for 5-hydroxytryptamine (serotonin), dopamine, and noradrenaline as ergot alkaloids have chemical structures similar to those neurotransmitters. Therefore, they affect insects and mammals that consume them and can be used to produce drugs …
Effect Of Soil Transplantation On Fungal Communities During Restoration, Thomas Luehl
Effect Of Soil Transplantation On Fungal Communities During Restoration, Thomas Luehl
All Master's Theses
Ectomycorrhizal fungi are integral parts of natural ecological systems that provide access to nutrients that would otherwise be unavailable by forming symbiotic relationships with plants. Relationships with ectomycorrhizal fungi are known to improve plant growth rate, increase plant resistance to pathogens, and improve survival. Current revegetation practices often ignore this relationship between fungi and plants. During the revegetation of the Interstate 90 Keechelus Lake wildlife overcrossing (Easton, WA), soil from forest sites adjacent to the restoration area were collected and used to inoculate a set of experimental plots to determine whether ectomycorrhizal fungi would become established in the plots and …
The Antibacterial Efficacy Of Silver (I) Cyanoximates Against Streptococcus Mutans Ua159 Biofilm Growth And Infection In Galleria Mellonella Larvae, Jill Hughes
Honors Program Theses
Streptococcus mutans is the known etiological agent of dental caries. S. mutans’ virulence factors, namely biofilm formation and lactic acid fermentation, contribute to its pathogenic behavior within the oral cavity. Biofilms are thick, antibiotically resistant communities of bacteria that are much more difficult to treat than planktonic or free-floating bacteria. Continued misuse of antibiotics against dynamic communities, like biofilms, has led to increased research on non-antibiotic alternatives. Amongst these alternatives, there are silver and silver-based compounds. Silver is commonly incorporated into medicine due to its inhibitory and bactericidal effects and antibacterial properties. Silver (I) cyanoximates have demonstrated promising antibacterial activity …
Contrast Of Smokeless Tobacco And It's Microbiological Footprint In Users And Non-Users, Monayjia Brown, Katie Robbins, Elyse Mckinney, Sydney Snell
Contrast Of Smokeless Tobacco And It's Microbiological Footprint In Users And Non-Users, Monayjia Brown, Katie Robbins, Elyse Mckinney, Sydney Snell
Humboldt Journal of Microbiology
Smokeless tobacco products such as vapes have gained significant popularity over the last 20 years (7), and it is clear that the effects of these products are primarily unknown. This study examines how the microbial profiles found in smokeless tobacco products, such as vapes, can be used in forensic science. By linking these profiles to their owners, investigators can identify suspects in cases where traditional evidence is lacking. This information could provide valuable assistance in tracking victims and matching perpetrators or victims to crime scenes. Forensic science involves any evidence found at a crime scene that is examined and analyzed …
Under Ice Photosynthetic Primary Production And Dark Carbon Fixation In A Temperate Freshwater System, Vanessa Cubillos Tellez
Under Ice Photosynthetic Primary Production And Dark Carbon Fixation In A Temperate Freshwater System, Vanessa Cubillos Tellez
Dissertations, Master's Theses and Master's Reports
Ice-covered lakes are vulnerable to environmental change, especially those in the Northern Hemisphere where ice cover is rapidly declining due to global warming. These changes can alter metabolic processes and disrupt carbon cycling driven by primary producers who form the base of the food chain and are key to sustaining ecosystem function. Photosynthetic primary production and dark carbon fixation under the ice in the Keweenaw Waterway, a temperate freshwater system that is ice-covered for ~3 months out of the year, were studied using a carbon isotopic labeling (14C-bicarbonate) technique. Water samples were collected weekly during ice cover and monthly during …
Stem And Pre-Health Students’ Diminishing Online Conversation Volume Regarding Majors And Careers Within The Field: Why Higher Education Should Care, Susan Roh
Electronic Theses and Dissertations
The purpose of this study was to determine the STEM and pre-health online mention volume change regarding STEM and pre-health majors, careers, and universities, with the data collection period encompassing August of 2018 through June of 2021. We evaluated if the mention sentiment shifted between academic year periods. The data gathered for this study was procured in collaboration with Campus Sonar and the social-listening tool of Brandwatch, which allowed for specifications of the data collected. Kruskal-Wallis tests were used to determine the means of the majors, careers, and universities mentions and mean sentiment volumes. Changes in not only online conversation …
Characterisation Of Selected Microbes In Terrestrial And Simulated Extraterrestrial Habitats: Human Health And Environmental Applications, Daniel Joseph Keaney
Characterisation Of Selected Microbes In Terrestrial And Simulated Extraterrestrial Habitats: Human Health And Environmental Applications, Daniel Joseph Keaney
Theses
The rapid development of the Space industry and associated technologies over the past decade is leading to the inevitable consideration of establishing human-based colonies on extra-terrestrial bodies such as the moon and Mars. With Mars being the focus of not only this body of work, but current New Space Age travel (NASA and SpaceX), a significant issue posed by Martian colonists includes establishing a safe and sustainable food source. Owing to its constantly evolving prehistoric landscape, the soils of Mars are contaminated with sodium perchlorate, a water-soluble salt that is toxic to humans, leftover from once inferred brine seeps that …
Evaluation Of Welded And Polished Stainless-Steel Surfaces In Supporting Sporeformer Biofilms In Dairy Plants, Taghreed Almalki
Evaluation Of Welded And Polished Stainless-Steel Surfaces In Supporting Sporeformer Biofilms In Dairy Plants, Taghreed Almalki
Electronic Theses and Dissertations
No abstract provided.