Reddy N Rukma, Patazca Eduardo, Morrissey Travis R, Skinner Guy E, Loeza Viviana, Schill Kristin M, Larkin John W
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Division of Food Processing Science and Technology, 6502 South Archer Road, Bedford Park, Illinois 60501 USA.
Institute for Food Safety and Health, Illinois Institute of Technology, 6502 South Archer Road, Bedford Park, Illinois 60501 USA.
J Food Prot. 2016 Feb;79(2):253-62. doi: 10.4315/0362-028X.JFP-15-310.
The purpose of this study was to determine the inactivation kinetics of the spores of the most resistant proteolytic Clostridium botulinum strains (Giorgio-A and 69-A, as determined from an earlier screening study) and of Clostridium sporogenes PA3679 and to compare the thermal and pressure-assisted thermal resistance of these spores. Spores of these strains were prepared using a biphasic medium method. C. sporogenes PA3679 spores were heat treated before spore preparation. Using laboratory-scale and pilot-scale pressure test systems, spores of Giorgio-A, 69-A, and PA3679 suspended in ACES [N-(2-acetamido)-2-aminoethanesulfonic acid] buffer (pH 7.0) were exposed to various combinations of temperature (93 to 121°C) and pressure (0.1 to 750 MPa) to determine their resistance. More than a 5-log reduction occurred after 3 min at 113°C for spores of Giorgio-A and 69-A and after 5 min at 117°C for spores of PA3679. A combination of high temperatures (93 to 121°C) and pressures yielded greater log reductions of spores of Giorgio-A, 69-A, and PA3679 compared with reduction obtained with high temperatures alone. No survivors from initial levels (>5.0 log CFU) of Giorgio-A and 69-A were detected when processed at a combination of high temperature (117 and 121°C) and high pressure (600 and 750 MPa) for <1 min in a pilot-scale pressure test system. Increasing pressure from 600 to 750 MPa at 117°C decreased the time from 2.7 to 1 min for a >4.5-log reduction of PA3679 spores. Thermal D-values of Giorgio-A, 69-A, and PA3679 spores decreased (i.e., 29.1 to 0.33 min for Giorgio-A, 40.5 to 0.27 min for 69-A, and 335.2 to 2.16 min for PA3679) as the temperature increased from 97 to 117°C. Pressure-assisted thermal D-values of Giorgio-A, 69-A, and PA3679 also decreased as temperature increased from 97 to 121°C at both pressures (600 and 750 MPa) (i.e., 17.19 to 0.15 min for Giorgio-A, 9.58 to 0.15 min for 69-A, and 12.93 to 0.33 min for PA3679 at 600 MPa). At higher temperatures (117 or 121°C), increasing pressure from 600 to 750 MPa had an effect on pressure-assisted thermal D-values of PA3679 (i.e., at 117°C, pressure-assisted thermal D-value decreased from 0.55 to 0.28 min as pressure increased from 600 to 750 MPa), but pressure had no effect on pressure-assisted thermal D-values of Giorgio-A and 69-A. When compared with Giorgio-A and 69-A, PA3679 had higher thermal and pressure-assisted thermal D-values. C. sporogenes PA3679 spores were generally more resistant to combinations of high pressure and high temperature than were the spores of the C. botulinum strains tested in this study.
本研究的目的是确定最具抗性的蛋白水解型肉毒梭菌菌株(根据早期筛选研究确定的乔治亚 - A和69 - A)以及生孢梭菌PA3679孢子的失活动力学,并比较这些孢子的热抗性和压力辅助热抗性。这些菌株的孢子采用双相培养基法制备。生孢梭菌PA3679孢子在制备孢子前进行了热处理。使用实验室规模和中试规模的压力测试系统,将悬浮在ACES [N -(2 - 乙酰氨基)-2 - 氨基乙烷磺酸]缓冲液(pH 7.0)中的乔治亚 - A、69 - A和PA3679孢子暴露于不同的温度(93至121°C)和压力(0.1至750 MPa)组合下,以确定它们的抗性。对于乔治亚 - A和69 - A的孢子,在113°C下处理3分钟后,对数减少超过5个对数级;对于PA3679的孢子,在117°C下处理5分钟后,对数减少超过5个对数级。与仅高温处理相比,高温(93至121°C)和压力组合使乔治亚 - A、69 - A和PA3679的孢子对数减少更多。在中试规模的压力测试系统中,当在高温(117和121°C)和高压(600和750 MPa)组合下处理不到1分钟时,未检测到乔治亚 - A和69 - A初始水平(>5.0 log CFU)的存活菌。在117°C下,将压力从600 MPa增加到750 MPa,使PA3679孢子对数减少>4.5个对数级的时间从2.7分钟减少到1分钟。随着温度从97°C升高到117°C,乔治亚 - A、69 - A和PA3679孢子的热D值降低(即乔治亚 - A从29.1分钟降至0.33分钟,69 - A从40.5分钟降至0.27分钟,PA3679从335.2分钟降至2.16分钟)。在600和750 MPa这两种压力下,随着温度从97°C升高到121°C,乔治亚 - A、69 - A和PA3679的压力辅助热D值也降低(即在600 MPa下,乔治亚 - A从17.19分钟降至0.15分钟,69 - A从9.58分钟降至0.15分钟,PA3679从12.93分钟降至0.33分钟)。在较高温度(117或121°C)下,将压力从600 MPa增加到750 MPa对PA3679的压力辅助热D值有影响(即在117°C下,随着压力从600 MPa增加到750 MPa,压力辅助热D值从0.55分钟降至0.28分钟),但压力对乔治亚 - A和69 - A的压力辅助热D值没有影响。与乔治亚 - A和69 - A相比,PA3679具有更高的热抗性和压力辅助热抗性。一般来说,生孢梭菌PA3679孢子比本研究中测试的肉毒梭菌菌株的孢子对高压和高温组合更具抗性。