Kim A Y, Thayer D W
Eastern Regional Research Center, USDA Agricultural Research Service, Philadelphia, Pennsylvania 19118, USA.
Appl Environ Microbiol. 1996 May;62(5):1759-63. doi: 10.1128/aem.62.5.1759-1763.1996.
Effects of irradiation and heating on survival of Salmonella typhimurium ATCC 14028 were examined by measuring DNA damage and the integrity of the cytoplasmic membrane. S. typhimurium cells fell into two distinct groups following heating: (i) heat-sensitive cells, which were rapidly inactivated at 65 degrees C and (ii) heat-resistant cells, which were only slowly inactivated at 65 degrees C. Radiation sensitivity of S. typhimurium was greater in the presence of air than in the presence of N2 gas (radiation doses required to inactivate 90% of the cells, 0.394 +/- 0.029 in air and 0.561 +/- 0.035 in N2). Recovery of the covalently closed circular form of plasmid pBR322 from S. typhimurium transformants (Ampr Tetr) was decreased by irradiation but not by heating. Heating prior to irradiation significantly decreased the recovery of plasmid DNA without affecting survival of S. typhimurium. Transformability of the recovered plasmid pBR322 was affected by neither irradiation nor heating, and mutation of antibiotic resistance genes was not detected in S. typhimurium. Heating, but not irradiation, caused destabilization of the cytoplasmic membrane, allowing penetration of hydrophobic dye. These results suggest that lethality of heating followed by irradiation for S. typhimurium was additive, reflecting irradiation-induced DNA damage and heat-induced membrane destabilization. When irradiation preceded heating in the absence of air, more cells were inactivated than was expected, because of heat-inactivating radiation-damaged DNA.
通过测量DNA损伤和细胞质膜的完整性,研究了辐照和加热对鼠伤寒沙门氏菌ATCC 14028存活的影响。加热后,鼠伤寒沙门氏菌细胞分为两个不同的组:(i)热敏细胞,在65℃时迅速失活;(ii)耐热细胞,在65℃时仅缓慢失活。在空气中,鼠伤寒沙门氏菌的辐射敏感性高于在N2气体中(使90%的细胞失活所需的辐射剂量,空气中为0.394±0.029,N2中为0.561±0.035)。从鼠伤寒沙门氏菌转化体(Ampr Tetr)中回收共价闭合环状形式的质粒pBR322会因辐照而减少,但不会因加热而减少。辐照前加热显著降低了质粒DNA的回收率,但不影响鼠伤寒沙门氏菌的存活。回收的质粒pBR322的转化能力不受辐照和加热的影响,且在鼠伤寒沙门氏菌中未检测到抗生素抗性基因的突变。加热而非辐照会导致细胞质膜不稳定,使疏水性染料能够渗透。这些结果表明,加热后辐照对鼠伤寒沙门氏菌的致死作用是累加的,反映了辐照诱导的DNA损伤和加热诱导的膜不稳定。在没有空气的情况下,当辐照先于加热时,由于热使辐射损伤的DNA失活,更多的细胞被灭活。