It has been known that the ability of Shiga toxinproducing Escherichia coli (STEC) to produce Stx2e in culture media plays a role in the diagnosis of edema disease and determination of subunit vaccine candidates in STEC isolates. To examine the efficiency of Stx2e production in several commercial media, a Stx2e-producing strain (KEFS1302) was grown in four different media: ISO-Sensitest broth (ISB), E. coli broth (ECB), trypticase soy broth (TSB), and Mueller Hinton broth (MHB), with or without mitomycin C at 37°C (250 rpm) for 6 h. Toxin production was measured by enzyme-linked immunosorbent assay. In the presence of mitomycin C, ECB was found to be the most suitable medium, reaching a production peak (OD600 = 1.2) at 1 h; Stx2e was mostly produced during the logarithmic phase (within 3 h). On the other hand, toxin production in ISB reached a peak at 3 h after incubation in the absence of mitomycin C. Stx2e was purified by fast protein liquid chromatography (FPLC) using anion-exchange chromatography. The 43 kDa band of Stx2e was confirmed by western blot using the ECB supernatant. Our results showed that ECB and ISB media would be a suitable medium for mass production of Stx2e even if the toxin production is dependent on time.

A female wild raccoon dog was referred with a history of generalized seizure. Mild leukocytosis was noted on laboratory tests. Gross lesions included nasal hemorrhage, hemothorax, and hemorrhage in the urinary bladder with hematuria. Microscopically, interstitial and purulent bacterial pneumonia was observed in the lungs. In the cerebellum, characteristic eosinophilic intracytoplasmic and intranuclear inclusion bodies were found in Purkinje cells, and severe demyelination was observed in the cerebellar white matter. Canine distemper virus (CDV) infection was suspected and confirmed after detection of CDV nucleoprotein RNA in the cerebrum and the lungs by nested reverse transcription polymerase chain reaction (RT-PCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP). Based on the histopathological and molecular diagnostic findings, it was concluded that the raccoon dog was infected with CDV.

초 록 본 실험은 Corynebacterium속 식물병원세균에 관한 연구의 일환으로서 Corynebacterium속 식물병원세균의 종 및 속으로서의 특성을 구명하기 위하여 행하여진 것으로서 균체구성단백의 정량은 Lang의 Messier 변법을 사용하였으며 균채 구성단백의 아미노산정량은 미생물 정량에 의하였다. Corynebacterium 속 식물병원세균은 타속의 병원세균에 비하여 다소 적은양의 균체 구성단백을 가지고 있었으며 균체구성단백의 아미노산, Cystein, Trypthophane, Histidine, Phenylalanine 및 Isoleucin에 있어서도 같은 결과를 나타내었다. 허나 이 현상은 종적 혹은 개체의 균주의 특성으로서 분류에는 관계를 갖지 않고 있음이 밝혀졌다

본 실험은 Corynebacterium속 식물병원세균에 관한 연구의 일환으로서 Corynebacterium속 식물병원세균의 종 및 속으로서의 특성을 구명함과 동시에 C. sepedonicum이 생성하는 Rumiflabin에 기초를 둔 ultraviolet method의 재검토를 위하여 행하여 진 것으로 vitamin B군의 정량은 미생물 정량에 의하였다. 대다수의 식물병원세균은 Thiamine (vitamin ), Nicotinic acid, Biotin 및 안식향산을 생장물질로서 사용하였다. Riboflavin은 corynebacterium속 식물 병원세균 (C. rathay 및 C. fascians는 제외)은 물론 타속의 식문병원세균도 생성하는 점으로 Corynebacterium속 식물병원 세균의 특성이 아니라는 점이 구명되었으며 sepedonicum에 의하여 발생되는 감자의 윤부병과 청고병과의 감별에 사용되어온 Ultra-violet light법은 재검토되어야 한다.