The role of Mycobacterium avium subsp. paratuberculosis (MAP) in Crohn's disease in humans has been debated for many years. Milk and milk products have been suggested as possible vectors for transmission since the beginning of this debate, whereas recent publications show that slaughtered cattle and their carcasses, meat, and organs can also serve as reservoirs for MAP transmission. The objective of this study was to generate heat-inactivation data for MAP during the cooking of hamburger patties. Hamburger patties of lean ground beef weighing 70 and 50 g were cooked for 6, 5, 4, 3, and 2 min, which were sterilized by irradiation and spiked with three different MAP strains at levels between 10² and 10⁶ CFU/ml. Single-sided cooking with one flip was applied, and the temperatures within the patties were recorded by seven thermocouples. Counting of the surviving bacteria was performed by direct plating onto Herrold's egg yolk medium and a three-vial most-probable-number method by using modified Dubos medium. There was considerable variability in temperature throughout the patties during frying. In addition, the log reduction in MAP numbers showed strong variations. In patties weighing 70 g, considerable bacterial reduction of 4 log or larger could only be achieved after 6 min of cooking. For all other cooking times, the bacterial reduction was less than 2 log. Patties weighing 50 g showed a 5-log or larger reduction after cooking times of 5 and 6 min. To determine the inactivation kinetics, a log-linear regression model was used, showing a constant decrease of MAP numbers over cooking time.
Experiments to determine the efficacy of high temperature, short time (HTST) pasteurization of milk in terms of inactivation of pathogenic microorganisms were mainly performed between 1930 and 1960. Among the target organisms were Mycobacterium bovis and Mycobacterium tuberculosis. As a result, the Codex Alimentarius prescribes that HTST treatment of milk should lead to a significant reduction of pathogenic microorganisms during milk pasteurization. Due to the development of improved methods for the detection of survivors and of more advanced heating technology, verification of this requirement seemed to be necessary. To address recent outbreaks of tuberculosis in cattle caused by M. bovis ssp. caprae (M. caprae) in the southern regions of Germany, this organism was tested and compared with M. bovis ssp. bovis (M. bovis). Experiments were performed in a pilot plant for HTST pasteurization of milk with 3 strains of M. caprae and 1 strain of M. bovis. In preliminary trials at a fixed holding time of 25 s, the temperature at which significant inactivation occurred was 62.5°C for all strains. To determine D-values (decimal reduction times) for the inactivation kinetics, the strains were tested at 65, 62.5, and 60°C at holding times of 16.5, 25, and 35 s. At 65°C, the D-values of all strains ranged from 6.8 to 7.8 s, and at 62.5°C, D-values ranged from 14.5 to 18.1 s. Low inactivation was observed at 60°C. When the low slope of the inactivation curve allowed calculation of a D-value, these ranged from 40.8 to 129.9 s. In terms of log10 reductions, the highest values for all strains were 4.1 to 4.9 log at 65°C, with a holding time of 35 s. The tested strains of M. caprae and M. bovis showed similar low resistance to heat. Standard HTST treatment should result in a high reduction of these organisms and thus the requirements of the Codex Alimentarius for inactivation of pathogens by this process are far exceeded.
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