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PubMed2000

Monoclonal antibodies suitable for incorporation into a competitive enzyme-linked immunosorbent assay (ELISA) for detection of specific antibodies to Leptospira interrogans serovar pomona.

Monoclonal antibodies (mAb) were produced by fusing Sp2/0-Ag14 myeloma cells with spleen cells from BALB/c and ND4 mice that were immunized with killed Leptospira interrogans serovar pomona whole cells. Thirty hybridomas which produced antibodies (of the IgG1, IgG2a, IgG2b, or IgG3 isotype) that bound to epitopes on the serovar pomona whole cell antigen were identified by an indirect enzyme-linked immunosorbent assay (ELISA). Twenty-eight of these 30 mAbs cross-reacted in the indirect ELISA with at least one whole cell antigen prepared from 12 other pathogenic Leptospira serovars, and/or with whole cell antigen from the non-pathogenic Leptospira biflexa serovar patoc. The two serovar pomona-specific mAbs, which were designated M897 and M898, were obtained from the ND4 mouse and were both of the IgG1 isotype. In competitive ELISAs, M897 and M898 were inhibited from binding to the pomona antigen by bovine sera with anti-serovar pomona microscopic agglutination test (MAT) titres ranging from 100 to 6400. No significant inhibition was observed with pomona MAT-negative sera or with sera from animals experimentally infected with serovars canicola, copenhageni, grippotyphosa, hardjo type hardjobovis or sejroe. The epitopes recognized by M897 and M898 were both highly susceptible to sodium meta-periodate oxidation, indicating a carbohydrate composition. Neither of these mAbs reacted in immunoblots with the separated components of the serovar pomona whole cell antigen.

Animals↗
PubMed1994

[Leptospirosis in Pomurje and Slovenia].

In Pomurje leptospirosis is endemic. During the years 1964-1985 there were 407 leptospiral cases reported in Slovenia, 366 of which were in Pomurje. Inclusion in this study required indication of leptospirae with a fourfold increase of the titer of antibodies or the isolation of leptospirae from hemocultures. In Pomurje in this period 214 men (58.47%) and 152 women (41.53%) were infected. The average morbidity rate was 12.75/100,000 inhabitants. The highest rate was in the country of Lendava (23.14/100,000). Ten serotypes were confirmed: grippotyphosa in 122 (33.33%) of the patients, sejroe in 91 (24.86%), icterohaemorrhagie in 50 (13.66%), saxkoebing in 48 (13.11%), australis in 47 (12.84%), canicola and pomona in two cases, tarassovi and autumnalis in one patient. In one case, the serotype could not be differentiated. The disease occurs throughout the years. In contrast to Western Europe in Pomurje there exists a much larger percent of infected women and a high percentage among the elderly. Two patients have died (mortality rate is 0.55%). One of them succumbed on the 5th day of infection due to massive bleeding in the heart and other organs. The other died on the 12th day under the influence of a severe Weil's syndrome.

Female↗
PubMed1977

[In vitro and in vivo activity of tiamulin against leptospires (author's transl)].

The minimal growth inhibiting concentration of tiamulin, a derivative of the diterpen antibiotic pleuromutilin, was evaluated in vitro against 11 different serogroups of leptospira interrogans by twofold serial dilution technique, in comparison to tetracyclin, dihydrostreptomycin and tylosin. The range of the MIC values of tiamulin is between 0.07 and 2.5 microgram/ml and thus comparable to the activities of the standard antibiotics tested (see table 1). The chemotherapeutic efficacy (ED50) of the compound was examined in two experimental leptospiral infections of the Syrian hamster, in comparison to tetracyclin. Both compounds were administered orally for 3 days. In the L. canicola infection, the ED50 values were 103.8 mg/kg and 306.3 mg/kg body-weight for tiamulin and tetracyclin, respectively. In the L. grippotyphosa infection, the ED50 values amounted to 35.16 and 277.5 mg/kg bodyweight for tiamulin and tetracyclin, respectively. Based on these values, tiamulin in comparison to tetracyclin showed 3-8 fold higher efficacy in vivo after oral administration.

Animals↗
PubMed2004

[Estimation of transmission hazard of Leptospira Sp. infections in 2 groups of people].

The aim of the study was to determine the possible risk of the leptospiral transmission from animals to humans. 457 humans (both men and women) were divided into 2 groups: 1) of possible high risk infection consisted of people dealing as farm workers with cattle and pigs 2) of possible low risk infection consisted of people selected randomly. The animals on the farms were previously tested and found positive. All sera were examined using micro-agglutination test (MAT) with a battery of 18 serovars. The statistical evaluation of the results was performed using chi 2 test. The infection rate in I group was 13.79% and in II group was 1.5%. (p < or = 0.001). Sera of I group reacted with 7 serovars (sejroe, bratislava, canicola, tarassovi, bataviae, celledoni, patoc) and of II with 3 (hebdomadis, sejroe, hardjo). The infection rate in the group I was over 9 times higher than in group II. Thus dealing with infected animals is a high risk factor.

Agglutination Tests↗
PubMedDate not supplied

[Microbiological characterization of candidate vaccine strains of Ballum serogroup Leptospira interrogans].

Two candidate vaccines of Ballum serogroup Leptospira interrogans were microbiologically characterized as part of the work directed to the obtention of new antileptospirosis vaccine formulations for human use. The growth kynetics of both strains was evaluated in EMJH protein medium and in 3 protein free media. The virulence was estimated in hamsters by the calculation of the mean lethal dose. The cellular and extracellular antigenic profiles were analyzed by unidimensional SDS-PAGE and compared with those from strains of Canicola, Icterohaemorrhagiae and Pomona serogroups. The antigenic homology among heterologous groups was analyzed by western blotting with serun from hasterms vaccinated with vax-SPIRAL. The results obtained showed a fastidious growth of both strains of Ballum in the studied media, a high virulence in the animal model and a large antigenic homology with strains from other serogroups of Lepstospira prevailing in Cuba.

Animals↗
PubMed1998

[Standardization of the lethal dose 50% of Leptospira interrogans strains used in the control of the leptospirosis cuban vaccine for human use].

The LD50 of strains of L. interrogans used in the potency assay of the Cuban trivalent leptospiral vaccine intended for human use was standardized. The control of the leptospires content of the cultures by means of the adjusting dilution (10 to 12 leptospires per field) was introduced. That was the key of the standardization of the assay, since it guaranteed the reproducibility of the estimate of the LD50 value for each serogroup: 10(5.98) for canicola, 10(5.60) for icterohaemorrhagiae and 10(6.60) for pomona, with the following limits for an interval of 1 log10: 10(4.98)-10(6.98), 10(4.60)-10(6.60) and 10(5.49)-10(7.49), respectively; whereas for a 95% (+/- 2 SD) confidence interval the values were: 10(5.54)-10(6.19); 10(5.33)-10(5.75) and 10(6.35)-10(6.60), respectively. These values were lower than the previous ones, which means ever more reproducibility. The methodology described proved to be satisfactory for the standardization of the LD50 of the challenge strains and it could also be used for evaluating the virulence of other strains of L. interrogans, including those used for the vaccine production.

Cuba↗
PubMed1997

Development of a monoclonal antibody-based competitive enzyme-linked immunosorbent assay for the detection of Leptospira borgpetersenii serovar hardjo type hardjobovis antibodies in bovine sera.

A murine monoclonal antibody (designated M553) that binds to an epitope on whole cell antigens prepared from Leptospira borgpetersenii serovar hardjo type hardjobovis and Leptospira interrogans serovar hardjo type hardjoprajitno, was produced and incorporated into a competitive enzyme-linked immunosorbent assay for the detection of bovine antibodies to serovar hardjo. The epitope recognized by M553 was susceptible to periodate oxidation. The M553 antibody was characterized by western blot with hardjobovis whole cell antigen. This antibody does not cross-react with whole cell antigens prepared from 11 other pathogenic Leptospira serovars, or, Leptospira biflexa serovar patoc. The sensitivity estimate of the competitive ELISA was 100% with field sera (n = 165) with serovar hardjo microscopic agglutination test (MAT) titres of > or = 100. The specificity estimate was 100% with sera (n = 128) obtained from a specific pathogen free herd of cattle that were negative in the MAT at a dilution of 1:100 for serovars hardjo, pomona, sejroe, copenhageni, canicola, and grippotyphosa. The specificity estimate with field sera (n = 301) with serovar hardjo MAT titres of < 100, was 98% (95% confidence interval = +/- 1.58%). There was no cross-reactivity with field sera (n = 306) with serovar pomona titres > or = 100 and serovar hardjo titres < 100. The specificity estimate with the combined populations of sera with serovar hardjo MAT titres of < 100 (n = 735) was 99.18% (95% confidence interval = +/- 0.65%). There was a high level of agreement (kappa = 0.977) between the results of the competitive ELISA and those of the MAT.

Animals↗
PubMed2001

Competitive enzyme-linked immunosorbent assay for detection of Leptospira interrogans serovar pomona antibodies in bovine sera.

A competitive enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody (M898) was developed for detection of bovine antibodies to Leptospira interrogans serovar pomona. This assay was evaluated using field sera (n = 190) with serovar pomona microscopic agglutination test (MAT) titers of > or =100 as the positive population (group A); field sera (n = 1,445) which were negative in the MAT (1:100 dilution) for serovar pomona (group B); and sera (from a specific-pathogen-free cattle herd [n = 210]) which were negative in the MAT (1:100 dilution) for serovars canicola, copenhageni, grippotyphosa, hardjo, pomona, and sejroe (group C). At the cutoff point recommended by receiver operating characteristic (ROC) curve analysis of the combined ELISA results of serum groups A, B, and C, the sensitivity and specificity values were 93.7 and 96.3%, respectively. The value for the area under this ROC curve was 0.977, indicating a high level of accuracy for the ELISA. Similar results were obtained from the analysis of the combined results of serum groups A and B and from the analysis of the combined results of serum groups A and C.

Animals↗
PubMed1984

[The Pavia Center for the study and diagnosis of leptospirosis].

According to his deep experience in this field, the Author greatly appreciates the program agreed during this congress. He thinks that Leptospirosis is a still widely diffused zoonosis, for which accurate diagnostic work-up is requested, coupled to an adequate epidemiologic study. Among his own contributions to this argument, the A. remarks the studies on the experimental infection of the guineapig with L. icterohaemorrhagiae, providing evidence that jaundice and lethal course of the illness are due to the Leptospira and not, as it was currently supposed, to an overcoming Staphylococcal septicemia. Furthermore the A. observed a striking epidemic by L. pomona affecting 90 out of 120 rice-workers in the neighbourhood of Pavia. This Leptospira, previously unreported in Europe, was found to be carried by Norvegian rats captured in that country. These data were in contrast with the more frequent incidence, in this work environment, of L. bataviae, carried by Micromis minutus sorcinus. Following the amount of observations and researches in this field "Center for study and diagnosis of Leptospirosis" was founded in 1954, and is still working in Pavia. The necessity of isolation of the infective agent, in addition to the simple serologic study, was claimed by the observation of five cases of meningitis in young pigherds, in which L. canicola (instead of the more common L. pomona) was demonstrated and îelded from the CSF of a patient. In the field of veterinary medicine, the A. demonstrated the L. australis A as responsible of some cases of miscarriages in openair breeded cattle. In those cases the hedgehog (Erinaceus europeus) was the natural carrier.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Diseases↗
PubMed1999

Leptospiral infection among primitive tribes of Andaman and Nicobar Islands.

The Andaman islands were known to be endemic for leptospirosis during the early part of the century. Later, for about six decades no information about the status of the disease in these islands was available. In the late 1980s leptospirosis reappeared among the settler population and several outbreaks have been reported with high case fatality rates. Besides settlers, these islands are the home of six primitive tribes of which two are still hostile. These tribes have ample exposure to environment conducive for transmission of leptospirosis. Since no information about the level of endemicity of the disease among the tribes is available, a seroprevalence study was carried out among all the accessible tribes of the islands. A total of 1557 serum samples from four of the tribes were collected and examined for presence of antileptospiral antibodies using Microscopic Agglutination Test (MAT) employing 10 serogroups as antigens. An overall seropositivity rate of 191% was observed with the highest rate of 53.5% among the Shompens. The seropositivity rates in the other tribes were 16.4% among Nicobarese, 222% among the Onges and 14.8% among the Great Andamanese. All of the tribes except the Onges showed a similar pattern of change in the seroprevalence rates with age. The prevalence rates were rising from low values among children to reach a peak in those aged 2140 years and then declined. Among Onges the seroprevalence rates continued to rise beyond 40 years. In all the tribes, seroprevalence rates were found to be significantly higher among the males. The commonest serogroups encountered were Australis followed by Grippotyphosa, Icterohaemorrhagiae, Pomona and Canicola.

Adolescent↗
PubMed1984

Leptospirosis in Israel: a report of 14 cases caused by icterohaemorrhagiae serogroup (1968-82).

Fourteen cases of leptospirosis due to the Leptospira interrogans serogroup Icterohaemorrhagiae (Weil's disease) were serologically confirmed in the last 15 years. In two cases, Icterohaemorrhagiae was also isolated from the patients' blood. Both positive cultures were obtained in Korthof's medium by indirect passage through brain-heart infusion medium. One of the isolates was identified as serovar budapest, which has never been reported in Israel. In most of the cases, paradoxical cross-reactions with serovars of the Autumnalis, Hebdomadis and Canicola serogroups were observed in the early stages of the illness. In all cases, we suspected that the source of the infection was serovars of the Icterohaemorrhagiae serogroup shed in the urine of Rattus norvegicus rats. Thirteen of the affected persons were working and living in the Tel Aviv area, all in places not known to be Icterohaemorrhagiae endemic before 1968. One patient was working and living in the Hadera region. All patients were jaundiced, and one died. The illness was usually accompanied by pronounced and long-lasting elevation of blood bilirubin and urea levels.

Adolescent↗
PubMed1984

[Diagnostic research on leptospirosis in the Milanese territory].

168 serum specimens have been tested for Leptospirosis in the years from 1980 to 1983. The identification procedure was a macroscopic plate agglutination screening test with formalin fied antigens and a confirmatory microscopic agglutination-lysis test of live antigens of actively growing cultures of different pathogenic serotypes propagated in a fluid medium. 34 serum specimens (23.8%) relevant to 18 cases were positive. L. icterohaemorrhagiae was positive in 12 cases (63.1%), L. pomona in 2 cases (10.5%), L. canicola in 1 case (5.3%), 1 case was positive for both L. grippotyphosa and L. bataviae and 1 case was positive for L. poi and L. sejroe. During the summer-fall of 1981 40 specimens of surface water from 4 pre-alpin lakes of the Provincia of Como were tested for Leptospiras with the cultural and biological examinations, following the detection of several leptospirosis cases on the relevant area. All water specimens gave negative results.

Antibodies, Bacterial↗
PubMed1984

Characterization of monoclonal antibodies against etiological agents of Weil's disease.

Monoclonal antibodies against etiological agents of Weil's disease were produced by cell fusion technology. Twenty hybridomas were produced through the fusion of P3X63Ag8 .653 cells with spleen cells from BALB/c mice immunized against Leptospira interrogans serovar icterohaemorrhagiae RGA strain and serovar copenhageni Shiromizu and M20 strains. Reactivities of the antibodies produced by the hybridomas were determined by the microscopic agglutination test. Among the five hybridoma antibodies to the RGA strain, two reacted specifically to serovar icterohaemorrhagiae, two reacted to serovar icterohaemorrhagiae at a high titer and serovar copenhageni at a low titer, and one reacted to serovars icterohaemorrhagiae, copenhageni, pyrogens, and canicola. Of the ten hybridoma antibodies to the Shiromizu strain, one reacted specifically to serovar copenhageni, seven reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, and two exhibited intermediate properties. Of the five hybridoma antibodies to the M20 strain, three reacted to both serovars copenhageni and icterohaemorrhagiae at almost the same titer, one reacted to serovar copenhageni at a low titer and serovar icterohaemorrhagiae at a high titer, and one reacted to serovars copenhageni, icterohaemorrhagiae, and pyrogens. The results revealed that each serovar has its own antigen(s) and their common antigens. In addition, 20 strains of leptospires were recently isolated and tested with three monoclonal antibodies characterized by different reactivities. Twenty strains were clearly identified by their antibodies, i.e., 16 strains were identified as serovar icterohaemorrhagiae and three strains were identified as serovar copenhageni. The remaining strain, which was not agglutinated by three antibodies, was identified as serovar autumnalis by an agglutination test with immune rabbit sera.

Agglutination↗
PubMed1992

Epidemiology of an outbreak of leptospirosis in man and dog.

An outbreak of leptospirosis in man and dog occurred during the monsoon in Madras city, India. 48 (50.5%) of the 95 human sera tested were positive for leptospirosis. Of these, 32 showed the presence of agglutinin to serovar icterohaemorrhagiae. 20 (21.3%) of the 94 canine sera tested were positive for leptospirosis. Of these, 10 had agglutinins to serovar icterohaemorrhagiae and 9 to serovar canicola. Among murine reservoirs, 8 (25%) of 32 rat sera and 10 (41.6%) of 24 bandicoot sera were positive. Bandicoot and rat sera contained 70 and 50% of agglutinins to serovar icterohaemorrhagiae, respectively. Urine culture lead to isolate a icterohaemorrhagiae strain from a patient and a canicola one from a dog. The changing trends of epidemiology in the transmission of leptospirosis to man and dog has been discussed.

Animals↗
PubMed1997

Serological survey of leptospiral infections in sheep, goats and dogs in Cordillera province, Bolivta.

A serological survey for antibodies to Leptospira spp. was conducted on sheep, goat and dog serum samples collected in three localities in Cordillera province in the southern part of the Santa Cruz Department (Bolivia) in 1992. A total of 98 sheep, 218 goats and 43 dogs were tested against 29 leptospiral serovars using the microscopic agglutination test. At the time of blood collection all of the examined animals appeared healthy and presented no clinical sign suggestive of leptospirosis. Antibody prevalences, as determined by positive results at a 1:100 dilution or higher, was 14.3% in sheep, 19.7% in goats, and 14.0% in dogs. Agglutinins against six serovars (poi. shermani, pomona, canicola, javanica, djasiman) were found in positive animals. The highest serological prevalence in sheep and goats was recorded for serovar poi, followed by pomona in sheep and shermani in goats. Titres to shermani were the commonest in dogs. The results of this survey indicate that leptospiral infection is common in south-east Bolivia and that serovars of several serogroups concur in the etiology.

Agglutination Tests↗
PubMed1966

Recent observations on leptospirosis in Northern Ireland and their bearing on current diagnostic methods.

An investigation was undertaken to assess the present importance of leptospiral infections in Northern Ireland, and in particular to look for evidence of infection by leptospiral serotypes other than L. icterohaemorrhagiae and L. canicola. Blood samples from 335 patients, sent to the laboratory for a variety of tests, were examined. After initial screening with five groups of pooled antigens, tests for leptospiral agglutinins were completed with formolized antigens prepared from 13 different serotypes. In seven patients a diagnosis of acute leptospirosis was made while nine others showed serological evidence of previous leptospiral infection. Attempts to isolate leptospirae by culture from 29 blood samples were unsuccessful. The serological results indicate that two additional leptospiral serotypes, namely L. ballum and L. bratislava, are causing human infection in Northern Ireland, and presumably also in other parts of the British Isles. Some clinical and epidemiological features associated with different types of leptospiral infection are described. It is stressed that leptospirosis is essentially a febrile illness, that meningeal symptoms are common, and that (contrary to popular belief) jaundice is by no means a constant occurrence. The implications of these findings are discussed, with special reference to the diagnosis of leptospiral infections. Laboratory diagnostic procedures are briefly reviewed, and the possible deficiencies of the agglutination test commonly used in Britain are pointed out. Some suggestions are made concerning both clinical and laboratory aspects of diagnosis, and the need for a reliable screening test for all forms of leptospiral infection is emphasized.

Adolescent↗
PubMed1989

Serological investigations for leptospirosis in humans in Columbia.

The authors examined serologically 353 clinically healthy persons in 5 localities in South Columbia for leptospirosis using the MAL-reaction with 15 serovars of 15 serogroups. The positivity was high - 18.4% namely with the serovars of the serogroups Icterohaemorrhagiae, Canicola, Pyrogenes, Australis, Pomona, Grippotyphosa, Sejroe, Hebdomadis, Bataviae, Panama and Shermani. Sera reacted positive in titer levels of 1:400 and higher only with the serovars of the serogroups Icterohaemorrhagiae, Canicola, Australis, Grippotyphosa, Sejroe, Hebdomadis, Panama and Shermani. In the total positivity were the serovars of the serogroups Icterohaemorrhagiae with 30.8%, Grippotyphosa with 23.1%, Australis with 15.4%, Panama and Pyrogenes with 10.8% each, Pomona with 6.2%, Shermani with 4.6% and Canicola, Sejroe, Hebdomadis, Bataviae with 3.1% each. The high positivity of the sera from the Mocoa for L.grippotyphosa was surprising - 20.7%.

Adolescent↗