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Analysis of Leptospira antigens by crossed immunoelectrophoresis.

Serovars icterohaemorrhagiae, canicola, and arboreae of pathogenic Leptospira interrogans and serovar patoc of saprophytic L. biflexa were examined by crossed immunoelectrophoresis. A close antigenic relationship was found between the interrogans serovars, particularly between icterohaemorrhagiae and canicola. To a much lesser extent cross-reactions were found between interrogans serovars and patoc. Comparison of three different antigenic preparations of icterohaemorrhagiae by tandem crossed immunoelectrophoresis and by absorption experiments with the patoc reference system showed the presence of several common broadly reactive antigens. Antigen 1 of the reference system, a genus-specific heat-stable protein antigen, was found in all antigenic preparations. It is a diagnostically important antigen.

Animals↗

[Occurrence of leptospiral antibodies in dogs in Brno and the surrounding area].

A serological examination by the microagglutination reaction of lysis (RMAL) was performed with 12 leptospira strains to investigate 1272 blood serums of dogs coming for various kinds of treatment to the outpatient ward of the veterinary hospital. The higher infestation rate was recorded in the dogs from outside the city (14.08%) than in the city dogs (7.96%). A considerably high infestation was found in the dogs of the service breeds (13.47%). The males were infested more frequently (10.99%) than the bitches (7.22%). As for age, young dogs until the age of one year were found to be infested more frequently. Separate significant titres were demonstrated with the leptospira of the Grippotyphosa, Icterohaemorrhagiae, Sejroe, and Australis (L. bratislava) serological groups. Reactions with the leptospira of other groups were recorded only at low titres: they were as coagglutinations with parallel high-titre reactions with the leptospira of the Grippotyphosa and Icterohaemorrhagiae groups.

Animals↗

Broth microdilution susceptibility testing for Leptospira spp.

Leptospirosis in humans has traditionally been treated with penicillin or doxycycline. The choice of therapy offered at the time of initial patient presentation is often empirical, as definitive diagnosis can take weeks. Determining the activity of numerous antimicrobial agents against a wide range of Leptospira serovars may broaden empirical therapeutic options. Various antimicrobials have been shown to be active against a limited number of serovars in in vitro studies, chiefly by the use of broth macrodilution techniques. We developed a broth microdilution technique using the commercially available growth indicator alamarBlue. MICs produced by this technique were compared to MICs and minimal bactericidal concentrations produced by the traditional broth macrodilution technique. The internal validity of our methods was assessed with 11 runs over numerous days with a single isolate of Leptospira interrogans serovar Icterohaemorrhagiae. By either method, the MICs for these internal-validity runs fell within 2 dilutions of each other for more than 90% of antimicrobials. A broader application of these two techniques included 12 serovars (including seven species) of Leptospira and six antimicrobials (penicillin G, doxycycline, chloramphenicol, erythromycin, cefotaxime, and ciprofloxacin). Observed reproducibility fell within 2 dilutions for 99% of the duplicate result sets for the MIC microdilution method, compared to 89% for the MIC macrodilution method. The macrodilution method tended to have a higher MIC at which 90% of the isolates were inhibited (MIC(90)) than did the microdilution method, but the MIC(90)s of both methods were within 2 dilutions of each other for all six drugs. The macrodilution and microdilution techniques produced similar results, with microdilution allowing a faster, more streamlined method of producing MIC results.

Anti-Bacterial Agents↗

Proposal to list ATCC 43642 as the type strain of Leptospira interrogans in the Approved Lists of Bacterial Names. Request for an opinion.

ATCC 23581(T) has been cited as the type strain of Leptospira interrogans since 1980. In 1986, the Centers for Disease Control informed the ATCC that this strain was Leptospira interrogans serovar budapest, not serovar icterohaemorrhagiae as deposited originally. An authentic culture of the Leptospira interrogans serovar icterohaemorrhagiae RGA(T) strain was then submitted and assigned ATCC 43642, which was designated as the type strain of Leptospira interrogans in an article by Yasuda et al. [Yasuda et al. (1987). Int J Syst Bacteriol 37, 407-415]. In this Request for an Opinion to the Judicial Commission, it is proposed that a correction be made in the Approved Lists of Bacterial Names to acknowledge ATCC 43642 as the type strain of Leptospira interrogans.

Leptospira interrogans serovar icterohaemorrhagiae↗

What is the correct specific epithet for pathogenic Leptospirae?

The underlying documentation of the first descriptions of members of the genus Leptospira is carefully revised. Stimson's publication of? Spirochaeta interrogans (1907) is to be inadequate and not suitable to be recognized as the first valid description of a new genus being only a description of a spirochete-like microorganism without any documentation. The naming of the species of pathogenic leptospira interrogans is therefore not in confirmity with the requirements of the International Code of Nomenclature. The first valid description of saprophytic leptospira is given by Wolbach and Binger (1914) and that of pathogenic ones by Inada and Ido (1915). The correct name for the species of pathogenic leptospira is therefore icterohaemorrhagiae and for the saprophytic ones biflexa.

Humans↗

Identification of some Japanese leptospiral strains as serotypes copenhageni and icterohaemorrhagiae by preciptin-absorption test in gel.

Nine strains of Leptospira belonging to the Icterohaemorrhagiae serogroups isolated in Japan were identified by the precipitin-absorption test in gel. The results of the precipitin-absorption test matched closely with those of the agglutinin-absorption test. Two strains were identified as serotype copenhageni and 7 were serotype icterohaemorrhagiae. Strain Shibaura, which was thought to belong to serotype icterohaemorrhagiae and used in many laboratories in Japan, was corrected as belonging to serotype copenhageni.

Agglutination Tests↗

Experimental leptospirosis (L. interrogans serovar icterohaemorrhagiae) of the guinea pig: leptospiral antigen, gamma globulin and complement C3 detection in the kidney.

Morphofunctional and immunofluorescent studies were done in guinea pigs experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae to determine the role of leptospiral antigen, gamma-globulin, and complement C3 deposits in the pathogenesis of the renal lesions. A sharp increase in leptospiral antigen deposits was observed in the late phase of the experimental infection. Immunoglobulin and complement C3 were small compared with the heavy leptospiral antigen deposits. Moreover, leptospiral antigen was observed mainly in the interstitium whereas immunoglobulin and complement C3 deposits were seen in glomeruli and small blood vessel walls. In our experimental model bacterial migration and local liberation of factors causing virulence seems more likely to determine the renal damage.

Animals↗

[Populations of pathogenic Leptospira of varying virulence in nature].

The study of geographically remote populations of Norway rats (Rattus norvegicus) revealed that in one of these populations a highly virulent population of Leptospira copenhageni, serogroup icterohaemorrhagiae, and in another population of rats a faintly virulent population of these microorganisms circulated simultaneously. At the same time in vitro experiments with Leptospira cultures showed the absence of the constant probability of sharp changes in the level of their virulence in time.

Animals↗

[Diagnosis and surveillance of leptospirosis in the Venetia Region].

The Microbiological Laboratory of Verona Hospital is the leptospirosis reference Centre since 1980. It uses routinely the lysis/agglutination test with leptospires strains in order to confirm the clinical diagnosis and to define the typing of strains. Although this Centre is not yet officially recognized, it is in fact, at the service of the Veneto region and greater part of the Trentino Alto Adige. It examines 600 samples a year (about 3000 determinations) from these two regions, therefore it can construct a biregional epidemiology although the data is neither referred to superior Committees nor, perhaps, utilized in the requesting territory. The aspects of this service to be improved are the incompleteness of data with samples and their not perfect condition on arrival for testing. The most interesting point is that the epidemiology in the Veneto region has not changed since 1980: the prevailing leptospira is the icterohaemorrhagiae and then the canicola and pomona.

Humans↗

[Leptospirosis in slaughter cattle--serologic and bacterial study].

Antibodies to leptospiras were demonstrated in 91 cases (7.4%) out of 1,239 animals examined by serological assays of blood sera of cattle slaughtered in slaughter-houses and coming from 21 farms of one district. The antibodies were detected in animals coming from eleven out of the farms investigated (52.4%). The most frequent reactions were proved with leptospiras of the serovar L. grippotyphosa (61.8%) and with leptospiras of the serological group Sejroe (18.9%), rarely with leptospiras of the serovar L. icterohaemorrhagiae or copenhageni (5.7%). Antibodies to leptospiras of other serovars (L. canicola, L. bulgarica and L. hardjo) were demonstrated only as coagglutionations with the above-mentioned leptospira serovars. We failed to demonstrate leptospiras in the animals examined by culture examinations as well as by bacterioscopy. The results of examinations have shown that the cattle on Czech farms is also exposed to infections by various serovars of leptospiras. The infections are not accompanied by manifest clinical symptoms in many cases nor do they cause significant elimination of leptospiras in urine in the serologically positive animals. On the Czech cattle farms the occurrence of antibodies to leptospiras of the serovar L. grippotyphosa is prevailing; this serovar seems not to be expressly pathogenic to cattle and does not cause the formation of anthropurgic foci. Reduction in the occurrence of antibodies to leptospiras of this type is possible to achieve by preventing the contact with natural foci of this type. Hence the prognosis of epizootologic situation in bovine leptospirosis in the conditions of this country is relatively favorable. But special attention should be paid to animals bought and imported from foreign countries. These animals could be a source of some highly pathogenic serovars of leptospiras to cattle.

Animals↗

Comparative study between microscopic agglutination and complement fixation tests in experimental canine leptospirosis.

In order to evaluate the antibody response to pathologic leptospira from both serovars icterohaemorrhagiae and canicola, dogs were experimentally inoculated with these leptospiras. Each of the serovar were inoculated in 15 dogs and the antibody response in the acute phase of infected was followed by microscopic agglutination and complement fixation tests. Complement fixing antibodies were detected in both groups at the 4th day post infection and agglutinins appeared in circulation at the 7th day post infection when homologous antigens were used. On the 63rd day post infection with serovar canicola and on 56th day post infection with serovar icterohaemorrhagiae, complement fixing antibodies could no longer be detected but, in both experimental groups, specific agglutinins were presented in significant titers.

Agglutination Tests↗

Characterization of serovars of the genus Leptospira by DNA hybridization with hardjobovis and icterohaemorrhagiae recombinant probes with special attention to serogroup sejroe.

Recombinant DNA probes derived from genomic libraries of serovars hardjobovis and icterohaemorrhagiae were applied for the characterization of leptospires. Differences in hybridization signals in combination with the banding pattern appear to provide good characteristics for strain typing. The banding patterns were easy to distinguish, since the recombinant DNA probes hybridized with a limited number of fragments. They were also indicative of genomic relationships between serovars. The probes suggested the existence of four subgroups with extensive genomic homology within the serogroup Sejroe. A number of serovars outside the serogroup Sejroe showed genomic homology with these subgroups. Amplification with the polymerase chain reaction showed a correlation with the genomic homologies demonstrated by Southern analysis. Knowledge about genomic relationships between leptospiral strains, as revealed by Southern analysis, may lead to a more rational approach for primer selection for polymerase chain reaction or cloning of particular genes.

Blotting, Southern↗

An outbreak of leptospirosis in seals (Phoca vitulina) in captivity.

An outbreak of leptospirosis in seals (Phoca vitulina) in captivity is described. In a zoo in The Netherlands 5 adult seals died within 12 days. At necropsy all animals showed signs of acute septicaemia, consistent with acute leptospirosis. Serological examination of one animal was positive for antibodies against Leptospira interrogans serovar Icterohaemorrhagiae and the serologically closely related serovar Copenhageni. Polymerase chain reaction was positive in one other animal. 8 nutria (Myocastor coypus) were examined, serologically, through bacteriological culture and PCR. 81,8% (9/11) were serologically positive for Leptospira. The seals and nutria were housed in the same water system.

Acute Disease↗

Potency of leptospiral vaccines and protection against chronic infection in golden hamsters.

Seven vaccines prepared from pathogenic strains of different origin of Leptospira interrogans [serovars icterohaemorrhagiae (one strain) and copenhageni (6 strains)] were examined in protection tests on golden hamsters. Two of the copenhageni strains were used for challenge. The organs (kidneys, spleen, liver) in the vaccinated animals surviving challenge were protected to a varying degree. Low rates of survival were associated with a high incidence of Leptospira-positive findings, partly connected with focal lesions of the kidneys. On the other hand, in the groups in which all the animals survived, it was not possible to culture leptospires from their organs or to detect leptospiral antigen in these organs by immunohistochemical investigation. A protection of the organs that prevents vaccinated animals from shedding leptospires after infection clearly depends on the vaccine dose administered and the efficacy of the vaccine which can be measured in potency tests based on the survival rate as the relevant parameter.

Animals↗

Human leptospirosis in a slum area in the city of Rio de Janeiro, Brazil--a serological and epidemiological study.

A serologic survey was carried out on slum dwellers in the city of Rio de Janeiro. A total of 259 serum samples from male and female individuals of different age groups were tested for the presence of antileptospire antibodies by microagglutination. Prevalence data were analyzed in relation to the major risk factors present at the site, mainly represented by the presence of carrier animals and the occurrence of frequent floods. Of the samples tested, 25% reacted with antigens of different serogroups at titres ranging from 1:100 to 1:6400, with a predominance of titres less than or equal to 1:400; 35% of positive sera reacted with leptospirae of the Icterohaemorrhagiae serogroup. Reactions with Djasiman, Panama, Javanica, Canicola, Pyrogenes, Australis, Ballum, Sejroe, Bataviae, Grippotyphosa, Autumnalis and Cynopteri were also detected, though at lower frequencies. There was no statistically significant difference between sexes, but higher prevalence rates were found to be associated with increasing age. A focus of infection was characterized, in which social and economic factors contribute to the persistence of leptospirae by favoring the proliferation of the main reservoir.

Adolescent↗