Mori T, Kohsaka K
National Institute for Leprosy Research.
Nihon Rai Gakkai Zasshi. 1990 Apr-Jun;59(2):113-9. doi: 10.5025/hansen1977.59.113.
When leprosy bacilli grown in nude mouse foot pad were used for culture experiments, cultivable acid-fast bacillus was sometimes isolated as a contaminant. Whenever bacilli were inoculated to nude mice, the same leprosy bacilli were killed by autoclaving and were inoculated in to foot pads of 5 nude mice for examination of this cause of the contamination. Acid-fast bacillus was cultivated on 3% Ogawa egg medium at 33 degrees C from homogenates of foot pads of nude mice infected with M. leprae after one year and a while of infection. Foot pad of nude mouse injected with leprosy bacilli was cut off, ground in mortar and passed through sterile absorbent cotton and the filtrate was centrifuged at 10,000 rpm for 30 minutes. The sediment was inoculated on 3% Ogawa egg medium after treating with a small amount of sterile 1 N sodium hydroxide. Acid-fast bacilli were isolated from 3 out of 41 mice inoculoted with heat killed bacilli. The isolated acid-fast bacillus did not be observed in the same experimental group inocudated with live bacilli, positive cases were scattered in another groups. Four out of 16 tubes were positive for acid-fast bacilli in mice infected with Kurume-naha and 5 out of 7 tubes in the Amami-KM infected mouse group. The two negative tubes were discarded due to contamination. Kurume-Oki strain which has yellow colonial morphology was isolated from one out of 6 culture tubes. Strains Kurume-naha and Amami-KM have the same characteristics as follows: slow grower with pale yellow smooth colonial morphology, strongly positive for niacin production and ureas; positive for nicotinamidase, pyradinamidase and 68 degrees C catalase; no growth at 45 degrees C, negative for nitrate reduction, hydrolysis of Tween 80, diamine oxidase, heat stable acid-phosphatase and arylsulphatase; resistant to streptomycin, isoniazid, rifampicin and B 663. Two isolates were identified as Mycobacterium simiae from these characteristics. Characteristics of a Kurume-Oki isolate was as follows: slow grower with yellow smooth colonial morphology, positive for urease, 68 degrees C catalase, hydrolysis of Tween 80 and arylsulfatase; no growth at 45 degrees C, negative for niacin production, nicotinamidase, pyradinamidase, nitrate reduction, daimine oxidase and heat stable acid-phosphatase; resistant to streptomycin, isoniazid, rifampicin and B. 663. This bacillus was identified as Mycobacterium gordonae from these characteristics.(ABSTRACT TRUNCATED AT 400 WORDS)
将在裸鼠足垫中培养的麻风杆菌用于培养实验时,有时会分离出可培养的抗酸杆菌作为污染物。每当将杆菌接种到裸鼠体内时,同样的麻风杆菌经高压灭菌后被接种到5只裸鼠的足垫中,以检查这种污染的原因。在感染麻风分枝杆菌一年及一段时间后,从感染裸鼠的足垫匀浆中,于33℃在3%小川鸡蛋培养基上培养出抗酸杆菌。将注射了麻风杆菌的裸鼠足垫切断,在研钵中研磨,通过无菌脱脂棉过滤,滤液以10000转/分钟离心30分钟。沉淀物用少量无菌1N氢氧化钠处理后接种到3%小川鸡蛋培养基上。在接种热灭活杆菌的41只小鼠中,有3只分离出抗酸杆菌。在接种活杆菌的同一实验组中未观察到分离出的抗酸杆菌,阳性病例分散在其他组中。在感染久留米 - 那霸株的小鼠中,16管中有4管抗酸杆菌呈阳性,在奄美 - KM感染小鼠组的7管中有5管呈阳性。由于污染,另外两管阴性样本被丢弃。从6管培养物中的1管中分离出具有黄色菌落形态的久留米 - 冲株。久留米 - 那霸株和奄美 - KM株具有以下相同特征:生长缓慢,菌落形态为淡黄色光滑,烟酸产生和尿素反应呈强阳性;烟酰胺酶、吡哆酰胺酶和68℃过氧化氢酶呈阳性;45℃不生长,硝酸盐还原、吐温80水解、二胺氧化酶、热稳定酸性磷酸酶和芳基硫酸酯酶呈阴性;对链霉素、异烟肼、利福平和B663耐药。根据这些特征,有两株分离菌被鉴定为猿猴分枝杆菌。久留米 - 冲株分离菌的特征如下:生长缓慢,菌落形态为黄色光滑,脲酶、68℃过氧化氢酶、吐温80水解和芳基硫酸酯酶呈阳性;45℃不生长,烟酸产生、烟酰胺酶、吡哆酰胺酶、硝酸盐还原、二胺氧化酶和热稳定酸性磷酸酶呈阴性;对链霉素、异烟肼、利福平和B663耐药。根据这些特征,该杆菌被鉴定为戈登分枝杆菌。(摘要截取自400字)