Murano A, Miyake M, Kato J, Tanzawa H, Takeo K, Noda M
Second Department of Microbiology, Chiba University School of Medicine, Japan.
Microbiol Immunol. 1999;43(11):1009-15. doi: 10.1111/j.1348-0421.1999.tb01230.x.
We found that a sub-lethal concentration of hydrogen peroxide (HPOx) enhanced the growth of Helicobacter pylori in Brucella broth supplemented with 10% fetal bovine serum (BB/FBS). The enhancement was evident at 0.1 mM HPOx and reached a maximun at 3.5 mM. The growth stimulation was dependent on the basal media used; when brain heart infusion broth (BHIB) was used instead of BB, the growth was not altered regardless of the presence or absence of HPOx. Furthermore, the growth in BHIB/FBS was comparable to that in BB/FBS plus 3.5 mM HPOx. This suggested that the enhancement of growth by HPOx resulted from the derepression of the inhibitory factor existing in BB by HPOx. The inhibitory substance seemed to be bisulfite salt since the bacteria grew to a similar extent in bisulfite-less Brucella broth (BLBB0)/FBS compared to the bacterial growth in BHIB/FBS and BB/FBS plus HPOx. These results indicate that the detoxification of bisulfite in BB can be easily achieved by simply adding HPOx to the medium, which causes the oxidation of bisulfite to bisulfate, a less-toxic compound to the bacterial growth. Since we also found that the morphology and cellular protein profile of BB/FBS-cultured bacteria were apparently different from those cultured in BLBB/FBS, we propose that the use of BB for primary isolation and cultivation of H. pylori should be limited on certain occasions, or if necessary, BB can be used after detoxification of the bisulfite by the addition of a low concentration of HPOx.
我们发现,在补充有10%胎牛血清的布鲁氏菌肉汤(BB/FBS)中,亚致死浓度的过氧化氢(HPOx)可促进幽门螺杆菌的生长。在0.1 mM HPOx时这种促进作用就很明显,在3.5 mM时达到最大值。生长刺激作用取决于所使用的基础培养基;当使用脑心浸液肉汤(BHIB)代替BB时,无论有无HPOx,生长情况都不会改变。此外,BHIB/FBS中的生长情况与BB/FBS加3.5 mM HPOx中的生长情况相当。这表明HPOx对生长的促进作用是由于HPOx解除了BB中存在的抑制因子的抑制作用。这种抑制物质似乎是亚硫酸盐,因为与BHIB/FBS以及BB/FBS加HPOx中的细菌生长情况相比,细菌在无亚硫酸盐的布鲁氏菌肉汤(BLBB0)/FBS中生长到了相似的程度。这些结果表明,只需向培养基中添加HPOx就能轻松实现BB中亚硫酸盐的解毒,这会使亚硫酸盐氧化为硫酸氢盐,而硫酸氢盐对细菌生长的毒性较小。由于我们还发现,BB/FBS培养的细菌的形态和细胞蛋白质谱与BLBB/FBS培养的细菌明显不同,我们建议在某些情况下应限制使用BB进行幽门螺杆菌的初次分离和培养,或者如有必要,可在添加低浓度HPOx对亚硫酸盐进行解毒后再使用BB。