Raw oyster shellstock was subjected to abuse conditions (7, 13, and 21°C) and sampled over a ten day storage period to gather scientific data to aid in determining whether spoilage occurred in the raw product over time before proliferation of pathogenic flora (Vibrio vulnificus) made the product unsafe. Spoilage was evaluated through pH measurements of a homogenate of the shucked meat and liquor. The olfactory acceptability of the raw oysters was evaluated in concert with the microbial and chemical evaluations. At all storage conditions, halophilic bacteria outgrew V. vulnificus by a minimum of 1 log CFU/g oyster (Colony Forming Units per gram) (p < 0.05). Olfactory acceptability was below 40% when V. vulnificus growth was at its highest (p < 0.05). Refrigerated storage should be considered a CCP for raw shellstock since even moderate temperature control kept V. vulnificus below 10⁴ approximately 1-2 Logs below the estimated infective dose for the majority of the population. / Master of Science

Many nonferrous industries such as mining and surface treatment plants produce co-products that are high in heavy metals and therefore toxic to the environment. A less obvious producer of heavy metal containing co-products is the whisky industry. Current methods of copper removal from such co-products include electrolysis and membrane filtration which are impractical and costly. When copper is found as a salt, current methods of removal include settlement, filtration and precipitation. Alternatives such as biological copper ion removal from effluents has also been shown to be effective. This study aimed to develop a biological method for the synthesis of stable copper nanoparticles. Morganella psychrotolerans was used to reduce Cu2+ to insoluble Cu0 nanoparticles. The nanoparticles were purified and characterised using X-Ray Photoelectron Spectroscopy (XPS) and High-Resolution Transmission Electron Microscopy (HR-TEM). Whisky distillery co-products were tested as a growth medium for M. psychrotolerans with concomitant copper nanoparticle synthesis. The copper nanoparticles were also studied for their application in electronics in order to make conductive circuits. Genomics studies combined with proteomics, helped develop possible models for copper nanoparticle synthesis by M. psychrotolerans, as well as identify proteins and genes not previously thought to be related to this pathway. The genome sequence of M. psychrotolerans obtained in this work allowed for a far more detailed study on the mechanism of copper nanoparticle synthesis than previously possible. This thesis also focused on understanding this mechanism better through proteomics and qRT-PCR. In order to study the identified copper ion reduction pathway in the future, a genetic modification toolkit was developed for M. psychrotolerans.

Changes in histamine, putrescine, and cadaverine concentrations in fresh and stored bluefish (Pomatomus saltatrix) were determined using a new HPLC method. The HPLC method utilized a 5.0% (w/v) trichloroacetic acid (TCA) extraction, pre-column fluorescamine derivitization, and fluorescence detection. The derivatives were stable over 24 h. The 5% TCA extraction produced percent recoveries of 98.6%, 98.7, and 100.0% for histamine, cadaverine, and putrescine respectively. The HPLC process including extraction, derivatization, and HPLC analyses was conducted in less than 45 minutes. Fresh bluefish was found to contain between <1 ppm and 99 ppm histamine, and no cadaverine or putrescine. Fresh bluefish fillets were stored at 5, 10, and 15 degrees C until sensory rejection. Fresh bluefish fillets inoculated with Morganella morganii were also stored at the same conditions. Histamine levels as high as 2200 ppm were observed in the inoculated fish stored at 15 degrees C. Overall, histamine achieved higher levels in the bluefish pieces inoculated with Morganella morganii. Histamine was present in greater amounts than putrescine and cadaverine in the bluefish samples. Histamine levels at each temperature exceeded the 50 ppm advisory level established by the FDA before 100% sensory rejection. Putrescine levels increased at each temperature during storage. Cadaverine was present only in uninoculated bluefish stored at 15 degrees C. Consumer risk from histamine poisoning seems to be the greatest in those fish stored at 5 degrees C where acceptance levels were higher and histamine levels above 100 ppm were observed. The presence of histamine-forming bacteria in fish-processing facilities was studied. Environmental sampling techniques were conducted in the Hampton Roads area of Virginia in fish-processing facilities that regularly handle scombroid fish or other fish which are known to accumulate histamine levels greater than 50 ppm. Surfaces that come into contact with the fish were swabbed and the histamine-forming bacteria from these areas were identified. One isolate each of Klebsiella ozaenae and Vibrio alginolyticus, and two isolates of Aeromonas sp. were found in the processing facilities. The study concluded that histamine-forming bacteria do not make up a large part of the microflora associated with fish-processing facilities. Fishing vessels were also sampled and no histamine-forming bacteria were identified. / Master of Science