Date of Award:


Document Type:


Degree Name:

Doctor of Philosophy (PhD)


Nutrition, Dietetics, and Food Sciences

Department name when degree awarded

Nutrition and Food Sciences

Committee Chair(s)

Bart C. Weimer


Bart C. Weimer


The initial aim of this work was to evaluate processing conditions for extended shelf life (ESL) milk to have a shelf life at refrigeration temperature of 60 d. Milk was processed on a pilot-scale ultra-high-temperature processing plant and evaluated for microbial and sensory quality over 60 d at 7°C storage. Results of this study showed that lower process temperatures were preferable to minimize cooked flavors and that the minimum safe processing temperature was 134°C for 4 s as determined by the destruction of bacterial spores in the processed milk.

Consumer preference panel results indicated that consumers preferred milk processed at 134°C for 4 s (those recommended in this study for ESL processing) to commercial UHT milk although there was a slight preference for pasteurized milk. The critical sensory characteristic of the processed milk was a cooked flavor, which decreased with lower processing temperature and shorter storage time; however, a significant increase in flavors that could be associated with lipolytic activity was also noted.

This study highlighted deficiencies in existing methods for determining heat-stable bacterial products in thermal-processed foods. No rapid, sensitive assay for detection of heat-stable spores or lipases in milk exists. If such assays were available, it would allow processors to determine Lipase activity and bacterial spore counts before processing and direct raw milk with low spore counts and low lipolytic activity into long-shelf-life products. To this end, assays to rapidly quantitate spores and lipolytic activity in milk were developed.

The lipase assay relies on the hydrolysis of p-nitrophenyl caprylate liberating a yellow color that is detected using reflectance colorimetry. The assay is sensitive to 5 mUnits/ml and is linearly correlated to spectrophotometry (r2 = 0.93) and release of titratable free fatty acids (r2 = 0.92 to 0.97).

An immunocapture, enzyme-linked immunoassay coupled with a fluorescent detection system was developed for and resulted in a prototype spore assay using Bacillus stearothermophilus spores. This organism was selected because it is extremely heat resistant, is commonly found in milk, and is associated with spoilage of milk and milk products. The assay was able to quantitate spores down to 103 cfu/ml in milk and other products in about 1.5 h. Other detection limits could be set if needed.