Palumbo S A
Appl Environ Microbiol. 1984 Sep;48(3):477-80. doi: 10.1128/aem.48.3.477-480.1984.
A procedure for detecting and quantitating heat injury in Campylobacter jejuni was developed. Washed cells of C. jejuni A7455 were heated in potassium phosphate buffer (0.1 M, pH 7.3) at 46 degrees C. Samples were plated on brucella agar supplemented with Na2S2O3, FeSO4 X 7H2O, and sodium pyruvate and on a medium containing brilliant green, bile, Na2S2O3, FeSO4 X 7H2O, and sodium pyruvate. Colonies were counted after 5 days of incubation at 37 degrees C in an atmosphere containing 5% O2, 10% CO2, and 85% N2. After 45 min at 46 degrees C, there was virtually no killing and ca. two log cycles of injury. Cells grown at 42 degrees C were more susceptible to injury than cells grown at 37 degrees C. The addition to brucella agar supplemented with Na2S2O3, FeSO4 X 7H2O, and sodium pyruvate of three different antibiotic mixtures used in the isolation of C. jejuni from foods or clinical specimens did not prevent recovery of heat-injured C. jejuni. Cells lost 260 nm of absorbing materials during heat injury. The addition of 5% NaCl or 40% sucrose to the heating buffer prevented leakage but did not prevent injury. Of the additional salts, sugars, and amino acids tested for protection, only NH4Cl, KCl, and LiCl2 prevented injury. Heat-injured C. jejuni repaired (regained dye and bile tolerance) in brucella broth supplemented with Na2S2O3, FeSO4 X 7H2O, and sodium pyruvate within 4 h. Increasing the NaCl in this medium to 1.25% inhibited repair, and increasing it to 2% was lethal. Heat-injured C. jejuni will repair at 42 degrees C but not at 5 degrees C.
开发了一种检测和定量空肠弯曲菌热损伤的方法。将空肠弯曲菌A7455的洗涤细胞在磷酸钾缓冲液(0.1M,pH7.3)中于46℃加热。将样品接种在补充有Na2S2O3、FeSO4·7H2O和丙酮酸钠的布鲁氏菌琼脂上,以及含有亮绿、胆汁、Na2S2O3、FeSO4·7H2O和丙酮酸钠的培养基上。在含有5%O2、10%CO2和85%N2的气氛中于37℃孵育5天后计数菌落。在46℃处理45分钟后,几乎没有杀菌作用,约有两个对数周期的损伤。在42℃生长的细胞比在37℃生长的细胞更容易受到损伤。在从食品或临床标本中分离空肠弯曲菌时使用的三种不同抗生素混合物添加到补充有Na2S2O3、FeSO4·7H2O和丙酮酸钠的布鲁氏菌琼脂中,不能阻止热损伤的空肠弯曲菌的恢复。细胞在热损伤过程中损失了260nm的吸收物质。在加热缓冲液中添加5%NaCl或40%蔗糖可防止渗漏,但不能防止损伤。在测试的用于保护的其他盐、糖和氨基酸中,只有NH4Cl、KCl和LiCl2可防止损伤。热损伤的空肠弯曲菌在补充有Na2S2O3、FeSO4·7H2O和丙酮酸钠的布鲁氏菌肉汤中4小时内修复(恢复对染料和胆汁的耐受性)。将该培养基中的NaCl增加到1.25%会抑制修复,增加到2%则具有致死性。热损伤的空肠弯曲菌在42℃时会修复,但在5℃时不会。