• Korean Journal of Veterinary Research
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• v.15 no.1
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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.

• Journal of Veterinary Clinics
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• v.25 no.4
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• pp.241-244
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• 2008

This study examined the prevalence of Brucella spp. and Leptospira interrogans in 360 clinically healthy dogs using multiplex nested PCR. Four dogs (1.1%, 2 females and 2 males) tested positive to Brucella spp. by multiplex nested PCR. Fifty nine (16.4%, 31 females and 28 males) of 360 dogs tested positive L. interrogans. In 1 and 2 of the samples that tested positive to Brucella spp. and L. interrogans, the partial sequences of the virB1 and 16S rRNA genes were identified by direct sequence analysis, respectively. In conclusion, prevalence of Brucella spp. and L. interrogans by multiplex nested PCR revealed low and high, respectively. Multiplex nested PCR is can be useful for early detection of Brucella spp. and L. interrogans in the canine blood from asymptomatic dogs.

• Genomics & Informatics
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• v.19 no.3
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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.

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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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.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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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.

• Korean Journal of Veterinary Research
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• v.32 no.2
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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.

• Korean Journal of Veterinary Research
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• v.36 no.2
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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.

• The Journal of the Korean Society for Microbiology
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• v.21 no.3
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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.

• Korean Journal of Veterinary Research
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• v.52 no.2
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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.

• Korean Journal of Microbiology
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• v.23 no.3
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• pp.223-229
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• 1985

We have previously isolated seven strains of spiral form bacteria, which have shown several similar characteristics to Leptospira from patient with epidemic pulmonary hemorrhagic fever and matural paddy water. Further studies have performed to characterize them. All of the isolated bacteria were aerobic, gram negative and spiral form and their growth were completely inhibited at temperature below $13^{\circ}C\;and\;50^{\circ}C$. In sensitivity test to drugs, the bacteria were resistant to the bile salts, 5-Fu and Amphotericine B, but sensitive to the 8-Azaguanine. The isolated bacteria as well as 5 serotypes of Leptospira interrogans reacted the patient sera and it was also shown that antisera from rabbit immunized with the isolated bacteria reacted with 5 serotypes of Leptospira interrogans. These results suggest that the isolated bacteria may belong to the genus Leptospira.