Differentiation of Pathogenic and Saprophytic Leptospires With 8-Azaguanine

Johnson, Russell C.; Rogers, Palmer

doi:10.1128/jb.88.6.1618-1623.1964

Cited by 102 publications

(56 citation statements)

References 12 publications

(11 reference statements)

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“…Growth inhibition studies. Growth inhibition of leptospires by CuS04 (7), 8-azaguanine (15), and normal rabbit serum (14) were performed as described previously except that SM-7 and SM-4 with 0.01% Tween 80 and SM-4 and 0.2% tryptose-phosphate broth with 10.0% rabbit serum were used as growth media.…”

Section: Methodsmentioning

confidence: 99%

Enzymatic degradation of H2O2 by Leptospira

Corin¹,

Boggs²,

Cox³

1978

Infect Immun

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The enzymes responsible for reducing H202 were surveyed in 49 strains of Leptospira by using semiquantitative assays for catalase and peroxidase. The survey revealed a differential distribution of catalase and peroxidase activities between the two leptospiral complexes. The pathogenic Leptospira interrogans strains gave strong catalase and weak or negative peroxidase reactions. Conversely, the nonpathogenic Leptospira biflexa strains gave strong peroxidase and negative or weak catalase reactions. An intermediate group of four L. biflexa strains, which were isolated from mammals, fell into the high peroxidase, low or negative catalase group. One water isolate, H-23, gave strong reactions for both enzymes and was examined for virulence and in vitro growth parameters. Results indicate metabolic differences between pathogens and water forms in their abilities to reduce H202.The genus Leptospira is composed of two complexes, the pathogenic Leptospira interrogans and the free-living Leptospira biftexa (13),

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Section: Methodsmentioning

confidence: 99%

Enzymatic degradation of H2O2 by Leptospira

Corin¹,

Boggs²,

Cox³

1978

Infect Immun

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“…Leptospires were grown in 5-mL homemade Korthof 's liquid medium (Korthof, 1932) and EMJH liquid medium (Ellinghausen & McCullough, 1965;Johnson & Harris, 1967) at 30 1C to obtain a density suitable for agglutination reactions.…”

Section: Bacterial Strainsmentioning

confidence: 99%

Molecular analysis ofLeptospiraspp. isolated from humans by restriction fragment length polymorphism, real-time PCR and pulsed-field gel electrophoresis

Turk¹,

Milas²,

Mojčec³

et al. 2009

FEMS Microbiology Letters

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A total of 17 Leptospira clinical strains isolated from humans in Croatia were serologically and genetically analysed. For serovar identification, the microscopic agglutination test (MAT) and pulsed-field gel electrophoresis (PFGE) were used. To identify isolates on genomic species level, PCR-based restriction fragment length polymorphism (RFLP) and real-time PCR were performed. MAT revealed the following serogroup affinities: Grippotyphosa (seven isolates), Icterohaemorrhagiae (eight isolates) and Javanica (two isolates). RFLP of PCR products from a 331-bp-long fragment of rrs (16S rRNA gene) digested with endonucleases MnlI and DdeI and real-time PCR revealed three Leptospira genomic species. Grippotyphosa isolates belonged to Leptospira kirschneri, Icterohaemorrhagiae isolates to Leptospira interrogans and Javanica isolates to Leptospira borgpetersenii. Genomic DNA from 17 leptospiral isolates was digested with NotI and SgrAI restriction enzymes and analysed by PFGE. Results showed that seven isolates have the same binding pattern to serovar Grippotyphosa, eight isolates to serovar Icterohaemorrhagiae and two isolates to serovar Poi. Results demonstrate the diversity of leptospires circulating in Croatia. We point out the usefulness of a combination of PFGE, RFLP and real-time PCR as appropriate molecular methods in molecular analysis of leptospires.

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“…The agglutination was read by dark-¢eld microscopy. Those strains with no agglutination were submitted to other phenotypic tests, namely, the growth at 13³C [15] and the resistance to 8azaguanine [16], and ¢nally to PCR detection of signatures from both saprophytic and pathogenic leptospires on rrs (16rRNA) genes (data not shown) to con¢rm a saprophytic nature [17].…”

Section: Bacterial Strains and Agglutinationsmentioning

confidence: 99%

Analysis of Leptospira isolates from mainland Portugal and the Azores islands

Collares-Pereira¹

2000

FEMS Microbiology Letters

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From 228 recent Leptospira isolates from mainland Portugal and Azorean wild mammals, 149 were characterized at the serovar level by monoclonal antibodies (MAbs), a quick serological method in epidemiological studies. In order to compare this antigenic information with that from new genetic techniques, a sample of isolates was analyzed through pulsed-field agarose gel electrophoresis (PFGE) (n = 71), mapped restriction site polymorphisms (MRSPs) in PCR-amplified rRNA genes (n = 45, including 13 saprophytes) and arbitrarily primed polymerase chain reaction (AP-PCR) fingerprinting (n = 32). MRSP and AP-PCR lead to species identification of the studied 32 pathogenic isolates: Leptospira interrogans (n = 3), Leptospira kirschneri (n = 8) and Leptospira borgpetersenii (n = 21). MAbs and PFGE characterized pathogenic isolates at the serovar level and resulted mainly in agreement (64%) although many discrepancies (35%) were observed. ß

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Differentiation of Pathogenic and Saprophytic Leptospires With 8-Azaguanine

Abstract: The use of the purine analogue, 8-azaguanine, as a differential agent for the separation of pathogenic and saprophytic leptospires was investigated. Growth of strains of the saprophyte Leptospira biflexa was almost insensitive to the

Cited by 102 publications

References 12 publications

Enzymatic degradation of H2O2 by Leptospira

Enzymatic degradation of H2O2 by Leptospira

Molecular analysis ofLeptospiraspp. isolated from humans by restriction fragment length polymorphism, real-time PCR and pulsed-field gel electrophoresis

Analysis of Leptospira isolates from mainland Portugal and the Azores islands

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