Enzymatic and microbiological inhibitory activity in eggshell membranes as influenced by layer strains and age and storage variables

Eggshell membranes (ESM) have been shown to exhibit antibacterial activity. The purpose of this study was to evaluate the enzymatic and biological [decimal reduction times (D-values)] activities of ESM as a function of bird breed, age, and ESM stabilization treatments. Younger White Leghorn (WL) hens produced ESM with 28% higher lysozyme activity than Rhode Island Red (RIR) layers. In contrast, older WL layers produced ESM with 17% less lysozyme activity than ESM from RIR layers. Similarly, beta-N-acetylglucosaminidase (beta-NAGase) ESM activities differed by hen age within breeds with younger hens yielding 14 to 16% more enzyme activity. D54 degrees C-values of Salmonella Typhimurium cells preexposed to WL ESM did not differ as a function of bird age (33, 50, and 81 wk). The ESM Lysozyme and beta-NAGase activities varied somewhat over a 6-mo storage study after treatment with 1 of 5 stabilization methods [i.e., storage at 4 degrees C, -20 degrees C, or ambient air storage after freeze drying, air drying (23 degrees C), or forced-air drying (50 degrees C)]. Both air and forced-air drying yielded significant reductions in beta-NAGase and lysozyme ESM activity (ca 12 to 30%) after the initial 24 h and then remained fairly stable during the extended storage. Freeze-dried samples retained the most enzymatic activity (95%) throughout the 6-mo trial, whereas refrigerated ESM lost 20 and 18% of the beta-NAGase and lysozyme activities, respectively. Frozen ESM lost 22% of the beta-NAGase activity, whereas lysozyme was nearly unaffected after 6 mo. The ESM biological activities against S. Typhimurium were not adversely impacted by layer breed or age. No significant loss in biological activity of ESM was detected 24 h after processing or after 6 mo of storage for refrigerated, frozen, and freeze-dried membranes, whereas significant reductions were observed for air- and heat-dried ESM. These findings demonstrate that ESM enzyme and biological activities are relatively constant across layer breeds and over extended storage. Based on these and other findings, ESM may have potential commercial value as a processing adjuvant in food and pharmaceutical product applications.