• 대한수의학회지
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• 제15권1호
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• pp.75-81
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• 1975

The rate of growth of Leptospires were studied by culturing in Korthof's media and Cox's liquid media which were treated at four different temperatures. The effect of thiamine hydrochloride in the culture of leptospires were also studied. The results obtained were summerized as followings: 1. The best growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in Korthof's media with the rabbit serum heated for 60 minutes at $56^{\circ}C$. 2. A good degree of growth of Leptospira icterohaemorrhagiae and Leptospira canicola was observed in both Cox's liquid media with rabbit serum heated for 30 minutes at $100^{\circ}C$ and with fresh rabbit serum. 3. The growth of Leptospira was enhanced by adding small amount of thiamine to the both media which were shown to be unsatisfactory for the growth of the organisms. 4. Culture of leptospires were attained by simply heating for 30 minutes at $100^{\circ}C$ both Korthof's media and Cox's liquid media containing rabbit serum obtained without aspetic procedures.

• 한국임상수의학회지
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• 제25권4호
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• pp.241-244
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• 2008

본 연구는 특이적인 임상증상이 없는 360두의 개를 대상으로 multiplex nested PCR 기법을 이용하여 Brucella spp. 및 Leptospira interrogans를 검출하였다. Brucella spp.는 총4두 (1.1%, 암컷 2두, 수컷 2두)가 검출되었고, Leptospira interrogans는 59두 (16.4%, 암컷31두, 수컷28두)에서 검출되었다. 결론적으로 Brucella spp.의 검출률은 낮은 반면 Leptospira interrogans는 높았다. 또한 multiplex nested PCR기법은 무증상의 개 혈액에서 Brucella spp. 및 Leptospira interrogans를 조기에 검출하는데 빠르고 편리한 기법으로 판단된다.

• Genomics & Informatics
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• 제19권3호
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• pp.31.1-31.9
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• 2021

Leptospirosis is a zoonotic disease caused by spirochetes from the genus Leptospira. In Thailand, Leptospira interrogans is a major cause of leptospirosis. Leptospirosis patients present with a wide range of clinical manifestations from asymptomatic, mild infections to severe illness involving organ failure. For better understanding the difference between Leptospira isolates causing mild and severe leptospirosis, illumina sequencing was used to sequence genomic DNA in both serotypes. DNA of Leptospira isolated from two patients, one with mild and another with severe symptoms, were included in this study. The paired-end reads were removed adapters and trimmed with Q30 score using Trimmomatic. Trimmed reads were constructed to contigs and scaffolds using SPAdes. Cross-contamination of scaffolds was evaluated by ContEst16s. Prokka tool for bacterial annotation was used to annotate sequences from both Leptospira isolates. Predicted amino acid sequences from Prokka were searched in EggNOG and David gene ontology database to characterize gene ontology. In addition, Leptospira from mild and severe patients, that passed the criteria e-value < 10e^-5 from blastP against virulence factor database, were used to analyze with Venn diagram. From this study, we found 13 and 12 genes that were unique in the isolates from mild and severe patients, respectively. The 12 genes in the severe isolate might be virulence factor genes that affect disease severity. However, these genes should be validated in further study.

• 대한미생물학회지
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• 제21권2호
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• pp.191-204
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• 1986

Leptospira isolated from the patients with so called epidemic pulmonary hemorrhagic fever were comparatively studied with standard strains obtained from ATCC. The specific aim of this study was to clarify the morphologic heterogeneity of the isolates, i. e., coexistence of spiral forms in both handness, right and left, rod and spherical forms in their population by comparing them with those of the ATCC strains. No differences between our strains and ATCC strains were noticed in their growth characteristics, responses to the culture media, temperature, antibiotics and antifungal agents. Furthermore, the morphologic heterogeneity had been repeatedly observed even in cultures of standard ATCC strains, which had been noticed in the cultures of bacteria isolated in this laboratory. The serologic analysis of our isolates demonstrated that the bacteria reacted with L. icterohaemorrhagiae and L. australis regardless their differences in time of isolation(1984, 1985). Thus, it was concluded that the spiral bacteria isolated in this laboratory are Leptospira of a new serovar which still remained to be determined. And the previous reports on the morphology of the Leptospira, which described that the Leprospira population has only spiral forms with right. handed coils might be reconsidered.

• 대한미생물학회지
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• 제34권6호
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• pp.591-594
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• 1999

Leptospirosis has been known as endemic disease in Korea since 1984. Wild rodent, mostly Apodemus agrarius, has been known as an important source of leptospiral infection especially in rainy circumstances in harvest reason of rural area. The infection rates of Leptospira interrogans in field rodents, Apodemus agrarius, was investigated by culture and PCR detection of leptospiral DNA, and compared with previous data. Furthermore, the serogroup and serovar were investigated. Two hundred twenty two Apodemus agrarius were captured during October to December 1996. Spirohaetes were isolated from 22 (9.9%) and leptospiral DNA was detected in an additional six rodents (12.6%). Subsequent cross-agglutinin absorption test, monoclonal antibody reactivity classified 21 cultures among 22 isolates as Leptospira interrogans serogroup Icterohemorrhagiae serovar lai. The above data did not differ from previous survey in 1984 to 1987. There was no significant change of Leptospira interrogans infection in field rodents in Korea.

• 대한수의학회지
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• 제32권2호
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• pp.195-205
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• 1992

The soluble antigen profiles and antigenic specificities of Leptospira interrogans serovars icterhaemorrhagiae, canicola, pomona and hardjo were examined by SDS-polyacrylamide gel electrophoresis, crossed immunoeletrophoresis and immunoblotting. The profiles of protein, glycoprotein and fraction containing N-acetylglucosamine of 4 serovars were compared. The protein profiles of 4 serovars were very similar except the range of 14,400 to 30,000 daltons. Molecular weight of glycoprotein of L, pomona was lower than other serovars. L canicola showed extra N-acetylglucosamine bands having molecular weight of 82,000 and 90,000 daltons. In crossed immunoelectrophoresis, a close antigenic relationship was found between L icterohaemorrhagiae and L canicola. In immunoblottings conducted with soluble antigens and rabbit antisera of 4 serovars, Leptospira interrogans serovars possessed cross-reactive antigens and serovar-specific antigens. The molecular weights of serovar-specific antigens were 45,000, 82,000 and 90,000, 31,000 and 24,000 daltons in L icterohaemorrhagiae, L canicola, L pomona, and L hardjo, respectively.

• 대한수의학회지
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• 제36권2호
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• pp.349-358
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• 1996

The present study was done to detect Leptospira canicola antigens in paraffin sections from experimentally infected hamsters with PAP method. The anti-Leptospira canicola serum used as first antibody was prepared by immunizing rabbits with Leptospira canicola. Positive reactions were detected in interstitium, tubular epithelial cells and glomerulus of the kidney, and in the hepatic sinusoid. With PAP method, leptospiral antigens were detected at the 48 hours after bacterial infection, but isolation, silver stain, and dark field microscopic examination of the samples did 48 hours later than PAP method. The results suggested that PAP method is considered as a reliable tool for confirmative diagnosis of this disease.

• 대한미생물학회지
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• 제21권3호
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• pp.331-336
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• 1986

A microbiological study for the isolation and characterization of Leptospira interrogans was performed in an attempt to define the characteristics of Leptospirosis in Korea. The results are summerised as follows. Thirty-five cultures were isolated from 11 patients with leptospirosis and 24 wild rodents captured in Paju area. The isolation rate of Leptospira from wild rodents reached 23.1%. All 35 cultures were identified as Leptospira interrogans by their characteristic morphology and motility in dark field microscopy, pathogenecity in Guinea pig and sensitivity to 8-Azaguanine. Cultures formed diffuse subsurface colonies with hazy margin and were catalase, peroxidase and oxdase positive. The fifty percent lethal dose of isolate HM 4 in Guinea pig was $8.9{\times}10^4$ org. Mouse passage method was sucecssfully applied in maintenance of fresh isolates without any loss of their original virulency.

• 대한수의학회지
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• 제52권2호
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• pp.75-82
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• 2012

We have developed and optimized a multiplex polymerase chain reaction (mPCR) for simultaneous detection of Brucella, Salmonella and Leptospira with high sensitivity and specificity. Three pairs of oligonucleotide primers were designed to specifically amplify the targeted genes of Salmonella, Leptospira and Brucella species with sizes of 521, 408 and 223 bp, respectively. The mPCR did not produce any nonspecific amplification products when tested against 15 related species of bacteria. The sensitivity of the mPCR was 100 fg for Brucella and 1 pg for both Salmonella and Leptospira species. In the field application, kidney, liver and spleen were collected from wild rats and stray cats and examined by mPCR. The high specificity and sensitivity of this mPCR assay provide a valuable tool for diagnosis and for the simultaneous and rapid detection of three zoonotic bacteria that cause disease in both humans and animals. Therefore, this assay could be a useful alternative to the conventional method of culture and single PCR for the detection of each pathogen.

• 미생물학회지
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• 제23권3호
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• pp.223-229
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• 1985

유행성 출혈형 폐염양 질환의 환자 및 논물에서 분리한 병원체의 세균학적 특성을 조사한바, 형태적 및 물리생화학적 특성이 L.inteerogans와 동일하였으며 또한 형태학적 검사에서도 분류균과 L. interrogans는 항원적으로 같은 반응을 보였다.