请问：亚硫酸还原梭状芽胞杆菌的“呼吸试验”怎么做？

tangyinming381

ISO 15213：2003 厌氧亚硫酸还原梭状芽胞杆菌计数  9.5  注2中：有一个证实试验："respiratory test ",不知如何做？
为方便看标准，标准我放在附件了。

pushful

亚硫酸盐还原菌是能利用硫酸盐或者其它氧化态硫化物来异化有机物质的微生物,在其生长繁殖代谢活动中，除CO2和水外，还产生高浓度的H2S为呼吸终产物。用含有亚硫酸铁的培养基，该菌生长繁殖过程中会将亚硫酸铁还原成为硫化铁，因而形成黑色菌落。

tangyinming381

pushful 发表于 2014-10-13 08:59
亚硫酸盐还原菌是能利用硫酸盐或者其它氧化态硫化物来异化有机物质的微生物,在其生长繁殖代谢活动中，除CO2 ...

respiratory test  呼吸试验
  这几天我查阅了资料，有如下认识：
微生物呼吸方式有数种，好氧、微需氧、兼性厌氧、厌氧。对厌氧亚硫酸还原梭状芽胞杆菌，第一步是厌氧培养，有可疑菌生长，说明是厌氧条件下可生长；取这可疑菌划线于亚硫酸铁琼脂平板，有氧条件下培养，如果生长，说明是兼性厌氧菌，不是目标菌；如果不生长，哪就是目标菌。
   如此做试验，是否可行？

pushful

tangyinming381 发表于 2014-10-13 09:29
respiratory test  呼吸试验
  这几天我查阅了资料，有如下认识：
微生物呼吸方式有数种，好氧、微需 ...

这么做完全可以

小伙5

请问ISO15213亚硫酸盐还原梭菌中，孢子形成实验怎么做呀谢谢啦！

威爱丽

真的可以这样？

tangyinming381

新版标准 ISO 15213-1:2023(E)    确认了这个方法，见下面原文：
9.6 Confirmation of sulfite-reducing Clostridium spp.
9.6.1 For confirmation, take five presumptive colonies from each dish retained for enumeration (see 9.5.2). If more than one morphology is present among the colonies, select one of each morphology for subculture and confirmation.
9.6.2 Streak each of the selected colonies with a sterile loop (6.8) onto two non-selective blood agar plates, e.g. Columbia blood agar (Clause B.3). If blood is not available, Columbia agar base or another nutrient-rich medium (e.g. tryptone soya agar or brain heart infusion agar) can be used.Allow the plates to equilibrate at room temperature if they were stored at a lower temperature. If necessary, dry the surface of the plates before use (see ISO 11133).Several isolates can be streaked onto identified sectors of each of the two non-selective agar plates. Streaks should obtain well-isolated colonies.From each pair of plates, one is incubated in an aerobic atmosphere and the other in an anaerobic atmosphere (6.1) at 37 °C for 20 h ± 2 h (6.3). After incubation, the plates can be refrigerated at 5 °C
(6.5) for a maximum of 48 h before reading. For plates which were incubated anaerobically, maintain the anaerobic atmosphere.
9.6.3 Typical colonies are confirmed as follows:
— If growth from one typical colony occurs on the anaerobically incubated (blood) agar plate and not on the aerobically incubated (blood) agar plate, the colony belongs to the genus Clostridia. This colony and other colonies with the same morphology on ISA medium are counted as sulfite-reducing Clostridium spp.
— If growth occurs from one typical colony on both the anaerobically and aerobically incubated blood agar plates, the colony does not belong to the genus Clostridia. Therefore, this colony and other colonies with the same morphology on ISA medium cannot be counted as sulfite-reducing Clostridium spp.
NOTE Alternative procedures (see ISO 7218) can be used to confirm the isolate as sulfite-reducing Clostridium spp., provided that the suitability of the alternative procedure has been validated (see ISO 16140-4 or ISO 16140-6).