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Intramuscular Injection of Plant Derived Antimicrobials: a Potenial Use Against M.Bovis in Cattle University of Connecticut OpenCommons@UConn Master's Theses University of Connecticut Graduate School 12-2-2019 Intramuscular Injection of Plant Derived Antimicrobials: A Potenial Use Against M.Bovis in Cattle Elizabeth Johnson [email protected] Follow this and additional works at: https://opencommons.uconn.edu/gs_theses Recommended Citation Johnson, Elizabeth, "Intramuscular Injection of Plant Derived Antimicrobials: A Potenial Use Against M.Bovis in Cattle" (2019). Master's Theses. 1448. https://opencommons.uconn.edu/gs_theses/1448 This work is brought to you for free and open access by the University of Connecticut Graduate School at OpenCommons@UConn. It has been accepted for inclusion in Master's Theses by an authorized administrator of OpenCommons@UConn. For more information, please contact [email protected]. Intramuscular Injection of Plant Derived Antimicrobials: A Potenial Use Against M.Bovis in Cattle Elizabeth Joy Johnson B.S. Biology, Elmira College, 2017 A Masters Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Masters of Science at the University of Connecticut 2019 i Copyright by Elizabeth Joy Johnson 2019 ii APPROVAL PAGE Masters of Science Intramuscular Injection of Plant Derived Antimicrobials: A Potenial Use Against M.Bovis in Cattle Presented by Elizabeth Joy Johnson, B.S. Major Advisor……………………………………………………………….. Dr. Xiuchun (Cindy) Tian Associate Advisor………………………………………………………………. Dr. Mary Anne Roshni Amalaradjou Associate Advisor………………………………………………………………. Dr. Guillermo Risatti University of Connecticut 2019 iii Table of contents Approval page……………………………………………………………………………………………………………………… i Table of contents………………………………………………………………………………………………………………… ii List of figures………………………………………………………………………………………………………………………. iv List of tables………………………………………………………………………………………………………………………… iv List of abbreviations……………………………………………………………………………………………………………. v Chapter I: Mycoplasma I. Mycoplasma General information…………………………………………………………………………… 1 II. Morphology: Physical and Chemical………………………………………………………………………… 1 III. Mycoplasma membranes, transport systems, and metabolism……………………………….. 2 IV. Mycoplasma bovis General information…………………………………………………………………… 7 V. Prevalence……………………………………………………………………………………………………………….. 9 VI. Acquiring infection…………………………………………………………………………………………………… 10 VII. Laboratory diagnosis………………………………………………………………………………………………… 11 VIII. Antibiotic resistance…………………………………………………………………………………………………. 12 IX. Mycoplasma Genome……………………………………………………………………………………………….. 14 X. Virulence factors……………………………………………………………………………………………………….. 16 XI. Mycoplasma bovis immune function…………………………………………………………………………. 22 XII. Environmental survival………………………………………………………………………………………………. 28 XIII. Bovine Respiratory airway…………………………………………………………………………………………. 29 XIV. Mucociliary apparatus……………………………………………………………………………………………….. 31 XV. Pathogenesis and localization of antigen…………………………………………………………………… 34 XVI. Co-infection……………………………………………………………………………………………………………….. 34 XVII. Mycoplasma bovis arthritis……………………………………………………………………………………….. 35 XVIII. Mycoplasma bovis pneumonia………………………………………………………………………………….. 35 XIX. Mycoplasma bovis mastitis………………………………………………………………………………………... 39 Chapter 2: Essential Oils- Plant derived antimicrobials I. Background………………………………………………………………………………………………………………… 46 II. Historical use of essential oils…………………………………………………………………………………….. 48 III. Composition of essential oils……………………………………………………………………………………… 50 IV. Testing essential oils………………………………………………………………………………………………….. 51 V. The growth medium………………………………………………………………………………………………….. 53 VI. Tests of antibacterial activity of essential oils in food systems………………………………….. 54 VII. Antioxidant and anti-inflammatory activities of essential oils…………………………………… 57 VIII. Essential oils on ruminal function……………………………………………………………………………… 62 iv IX. Mode of antibacterial action…………………………………………………………………………………….. 69 a. Carvacrol…………………………………………………………………………………………………….. 72 b. Eugenol………………………………………………………………………………………………………. 74 c. Trans Cinnamaldehyde……………………………………………………………………………….. 74 X. Susceptibility of Gram-negative and Gram-positive organisms……………………………….. 77 XI. Synergism and antagonism between components of essential oils…………………………. 81 XII. Toxicity……………………………………………………………………………………………………………………. 83 a. Liver…………………………………………………………………………………………………………….. 90 b. Kidney…………………………………………………………………………………………………………. 93 XIII. Safety Data……………………………………………………………………………………………………………… 95 XIV. Neurological effects of essential oils……………………………………………………………………….. 96 XV. Part I and II References…………………………………………………………………………………………… 107 Chapter 3: Effects of intramuscular injected plant-derived antimicrobials in the mouse model i. Effects of intramuscular injected plant-derived antimicrobials in the mouse model…. 139 a. Abstract………………………………………………………………………………………………………… 140 b. Introduction………………………………………………………………………………………………….. 141 c. Methods……………………………………………………………………………………………………….. 143 d. Results………………………………………………………………………………………………………….. 148 e. Discussion…………………………………………………………………………………………………….. 154 f. Conclusion……………………………………………………………………………………………………. 159 ii. References………………………………………………………………………………………………………………. 161 Chapter 4: VigorSential I. University support……………………………………………………………………………………………………. 164 II. Executive Summary………………………………………………………………………………………………….. 167 III. Company description………………………………………………………………………………………………. 168 IV. Products and Services………………………………………………………………………………………………. 168 V. Markey analysis……………………………………………………………………………………………………….. 170 VI. Strategy of implementation…………………………………………………………………………………….. 171 VII. Financial plan and projections/ Organization and management team……………………… 172 v Lists of Figures Title Page Chapter III Figure 1 Animal treatment and observation schemes. Mice arrived on Day -2 144 and were sexed. The mice then had a 48h acclimation time. On the morning of Day 0 the mice were weighed and injected. Mice were observed hourly for 2 h, and twice daily post-injection until Day 4 when mice were weighed, euthanized for dissection. Red vertical bars indicate the times of observation. Figure 2 The averaged foot/leg scores (means+SD; upper panel) and number 151 of mice with scores of >0 (lower panel) in controls and TC treated mice. H and D: hour and day post-injection. Scoring system: 0=normal; 1=closed digits while walking well; 2=closed digits, no grip; 3=closed digits, no grip, dragging leg, foot flipped. Figure 3 The averaged foot/leg scores (means+SD; upper panel) and number 152 of mice with scores of >0 (lower panel) in controls and CAR treated mice. H and D: hour and day post-injection. Scoring system: 0=normal; 1=closed digits while walking well; 2=closed digits, no grip; 3 Lists of Tables Title Page Chapter III Table 1 Levels of PDA treatments. DMSO was used as the vehicle control. 145 Table 2 Scoring systems for eating/drinking, physical activities, physical 146 appearance, and foot/leg function/appearance Table 3 Mortality from CAR treatments (dead/total). No mortality was found 148 in the corresponding DMSO controls. Table 4 The averaged behavior scores and the number of animals receiving a 149 score of >0 during the first 12 h of TC and CAR treatments Table 5 Weights (g) and relative weights (%) of the liver and kidney in 153 control and PDA treated mice. vi List of Abbreviations BRD Bovine Respiratory Disease LPS Lipopolysaccharide PBP Periplasmaic binding proteins TCA Tricarboxylic acid cycle VSP Variable Surface Lipoprotein MALP Macrophage activating lipoprotein E2 Prostaglandin estrogen NO Nitric oxide TACA Serum antioxidant capacity Mb-LIP M.bovis lympho-inhibitory peptide PIMs Pulmonary intravascular macrophage ASL Air-surface liquid TAP Tracheal antimicrobial peptide LAP Lingual antimicrobial peptide SP-A Surfactant proteins A SP-D Surfactant proteins D UBRD Undifferentiated bovine respiratory disease EO Essential Oils MLC Minimal lethal concentration ICAM Intracellular adhesion molecule VCAM Vascular cell adhesion molecule LTA Lipoteichoic acid PPARs Peroxisome Proliferator Activated Receptor TC Transcinnamaldehyde OM Outer membrane HHP High Hydrostatic Pressure MIC Minimum inhibitory concentration Ip Intraperitoneal Iv Intravenous PDAs Plant Derived Antimicrobials vii Chapter I: Mycoplasma Mycoplasma General Information Mycoplasmas are among the smallest free-living microorganisms, comprising over 100 species. The class Mollicutes was founded in 1956 by Edward and Freundt. The class Mollicutes is now used to classify organisms of the pleuropneumonia group and the order of Mollicutes. Mollicutes are part of the Gram-positive lineage originally evolving from Clostridium-like bacteria via gene deletion. Most Mycoplasmas are facultative anaerobes, have a tri-layered cell membrane and cannot form a cell wall. Due to the lack of a cell wall, they have a pleomorphic shape, and antibiotic resistance. One family, Mycoplasmataceae, is composed of two generas, Mycoplasma and Acholeplasma. Mycoplasma bovis (M. bovis) was initially called M.agalactiae var. (later subsp) bovis because of both clinical and biochemical similarities to M.agalactiae, the cause of contagious agalactia in small ruminants (Nicholas, 1998). Many species of Mycoplasma are important veterinary pathogens that cause Epizooties list A diseases. It is generally believed that Mycoplasmas have a secondary role in infections by exacerbating pre-existing conditions. However, Mycoplasma bovis has been shown in recent years to have a primary role in infections such
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