Increasing global occurrences of highly infectious, easily transmissible diseases unfamiliar to nurses affect the learning environment and the required skill set for professional nurses. The global threat of Ebola Virus Disease and other high-risk diseases requiring biocontainment necessitates competency in the management of complex patient needs, while ensuring safety measures that prevent spread of the potentially fatal disease. Guided by Bandura's social cognitive theory, this quantitative correlational study addressed the relationships between nurses' professional characteristics and their perceived self-efficacy when providing care to highly infectious patients requiring biocontainment. A full census of 92 nurses was used to recruit participants from eligible nurses for this study. Participants anonymously completed a cross-sectional electronic survey consisting of the Nursing Care Self-Efficacy Scale (NCSES) and questions related to the nurses' professional practice characteristics. Data analysis included descriptive statistics, correlations, and multiple linear regression. Results showed that the number of biocontainment drills and a higher level of formal education were significantly correlated with a higher total NCSES score. Years of nursing significantly predicted a higher total NCSES score. Results support the establishment of prerequisites criteria for learner participation in biocontainment training and the inclusion of multiple drill within the education design. Findings from this study may inform positive social change through educational enhancements that support the development of professional self-efficacy and competency in skill performance for nurses who care for patients with highly contagious diseases requiring biocontainment.

Genetic engineering is important to enhance crop characteristics and certain traits. Genetically engineered crop cultivation brings environmental and ecological concerns with the potential of unwanted transgene escape and introgression. Transgene escape has been considered as a major environmental and regulatory concern. This concern could be alleviated by appropriate biocontainment strategies. Therefore, it is important to develop efficient and reliable biocontainment strategies. Removing transgenes from pollen has been known to be the most environmentally friendly biocontainment strategy. A transgene excision vector containing a codon optimized serine resolvase CinH recombinase (CinH) and its recognition sites RS2 were constructed and transformed into tobacco (Nicotiana tabacum cv. Xanthi). In this system, the pollen-specific LAT52 promoter from tomato was employed to control the expression of CinH recombinase. Loss of expression of a green fluorescent protein (GFP) gene under the control of the LAT59 promoter from tomato was used as an indicator of transgene excision. Efficiency of transgene excision from pollen was determined by flow cytometry (FCM)-based pollen screening. While a transgenic event in the absence of CinH recombinase contained about 70% of GFP-synthesizing pollen, three single-copy transgene events contained less than 1% of GFP-synthesizing pollen based on 30,000 pollen grains analyzed per event. This suggests that CinH-RS2 recombination system could be effectively utilized for transgene biocontainment. A novel approach for selective male sterility in pollen was developed and evaluated as a biocontainment strategy. Overexpression of the EcoRI restriction endonuclease caused pollen ablation and/or infertility in tobacco, but exhibited normal phenotypes when compared to non-transgenic tobacco. Three EcoRI contained 0% GFP positive pollen, while GFP control plants contained 64% GFP positive pollen based on 9,000 pollen grains analyzed by flow cytometry-based transgenic pollen screening method. However, seven EcoRI events appeared to have 100% efficiency on selective male sterility based on the test-crosses. The results suggested that this selective male sterility could be used as a highly efficient and reliable biocontainment strategy for genetically engineered crop cultivation.

Master of Science / Department of Clinical Sciences / Michael Sanderson / Biosecurity, biocontainment and security practices are important in production animal agriculture. Procuring cattle from multiple sources and commingling them into a single confinement operation increases risk of disease introduction. The large concentration of animals makes a feedyard a more likely target of a domestic or international terror group. Controlling or eradicating an intentionally introduced pathogen or toxin would be costly. The aim of these surveys was to gather information from experts about perceived risks and mitigation strategies and to assess current practices of biosecurity, biocontainment and security in Central Plains feedyards. Consulting veterinarians and feedyard managers shared similar views on the likelihood of disease caused by terrorism, natural introduction or accidental introduction, and on the importance of on-site security. They disagreed on the importance of preventative products, disease transmission control, and environmental control. Generally speaking, feedyard managers believed environmental control to be more important than consulting veterinarians. In reference to a survey of current practices, some feedyards use equipment for both manure and feed handling. Many feedyards are not cleaning and disinfecting oral treatment equipment, treatment facilities, or unloading facilities on a regular basis which may increases their risk for indirect disease transmission of endemic agents such as Salmonella or BVDV. Most feedyards in this survey import some cattle directly from an auction market, do not require clean boots or foot covering to be worn by visitors, and do not require trailers to be cleaned. Smaller feedyards were more likely to require trailers to be cleaned before loading incoming cattle. Less than half of the feedyards reported having a fence that will stop humans or kept protein supplements or micro-nutrients secured from access. Some feedyards enforced a visitor log or employed a night watchman. Most feedyards learned about a future employee by calling references listed in resume, but some performed a criminal background check. A cost-benefit analysis should be done on all management practices to determine economic benefits. More research is needed to better understand which practices are most beneficial.

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Genetically modified organisms (GMOs) are confined physically in order to contain their spread in nature and to minimise chances of horizontal gene transfer. However, with the potential that GMOs hold as cheap, reliable and efficient micro-machines, their eventual uncontrolled release into the wider space is becoming more likely. Indeed, their application as environmental sensors is largely increasing. Nevertheless, the field of synthetic biology may also afford solutions to the problem. A major potential application of GMOs is the delivery of antigens to human and animal hosts, through the utilization of live, engineered microbes. Recombinant technology is promising for several reasons including their capacity to be less reactogenic, more potent, safer and genetically definable. Also, they have the potential to provide protection against multiple targets simultaneously, are relatively inexpensive and can be eradicated with antibiotics, as the need arises. Besides, delivery of vaccines to mucosal surfaces is more efficient. Mutant Salmonella expressing heterologous antigens have been shown to induce protection against a variety of pathogens. Nevertheless, limited containment systems are available that can be applicable for bacterial antigen carriers. This project aims to design safeguards for the bacterial antigen delivery systems that limit ORF translatability and self-inactivates/destructs upon exit from the host. In this work, double quadruplet codons were suppressed by orthogonal tRNAs, providing a barrier for gene translation in the recipient cells when antigen is horizontally transferred. Furthermore, three kill switches were designed that are activated by a decrease in temperature from 37 °C. First, Sau3AI endonuclease was activated by protein self-splicing at low temperature mediated by Mtu recA intein. The activation of the endonuclease led to three-fold logarithmic decrease in the number of viable cells within two hours of gene expression. Second, RNA-dependent activation of RNase 7 showed a reduction in the number of viable cells at low temperature of three logarithmic folds. RNase 7 was controlled by the cspA 5’UTR, which sequesters ribosome binding site at 37 °C and allows translation at low temperature. Third, CspA 5’UTR was shown to regulate expression of TEV protease at 37 °C and low temperature. This led to bacterial cellular inhibition within two hours of TEV induction and five-fold logarithmic reduction in the number of viable cells at low temperature. In addition, for the first time and contrary to previous studies, the TEV protease was shown to inhibit cellular growth. It was also shown that biofilm formation was drastically impaired by the TEV activity. The three killing switches and the quadruplet translation system are poised to function as robust safeguards for bacterial antigen delivery systems.