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[Leptospirosis in swine; observations on the serological diagnosis of Leptospira interrogans serotype bratislava].

Leptospirosis is considered to be an important cause of porcine reproductive failure. In this respect recently attention is paid to the possible role of Leptospira interrogans serotype bratislava (bratislava). In the present paper the results of serotype bratislava serology conducted on three farms are presented and discussed. On two of the farms no reproductive failure was observed. On the third farm with a high percentage of return to service a longitudinal search was done. No relation could be demonstrated between bratislava titers and reproductive failure.

Animals↗

A new prepatory method of thermically inactivated Leptospira Patoc antigen for rapid slide agglutination used as serosurvey test for human leptospiroses.

830 sera from leptospirosis-suspect patients and 133 sera from investigations were analysed by the new test--rapid slide agglutination with Patoc antigen, inactivated at 60 degrees C for 15 minutes, in parallel with two standard tests--CFT with merthiolated Patoc antigen and MAR with 17-23 live pathogenic antigens. The new serotest proved efficient for selecting the cases of disease or leptospira infection, acute or recent form, not older than 2 years, both by its sensitivity and specificity and also by its rapid and economical character. In the present paper, the technology of preparation and control of Patoc antigen, inactivated at 60 degrees C for 15 minutes, is described.

Agglutination Tests↗

Isolation of Leptospira interrogans serovar bratislava from stillborn and weak pigs in Iowa.

Leptospira interrogans serovar bratislava was isolated from a herd of swine in Iowa with a history of stillborn and weak neonatal pigs. Placentas, kidneys, and lungs of stillborn and weak pigs from 3 litters were processed to detect leptospires by use of bacteriologic culture and fluorescent antibody testing. Sera from stillborn and weak pigs were tested to detect agglutinating antibody against leptospires. A low antibody titer against L interrogans serovar bratislava was detected in the sera of stillborn and weak pigs. Small numbers of leptospires were sometimes detected in tissues by use of the fluorescent antibody test. Serovar bratislava was isolated from placentas, stillborn pigs or weak pigs from each of the 3 litters.

Animals↗

[Study on the characteristics of agglutination reaction of McAb with Leptospira interrogans outer envelope].

Three McAb were produced against an outer envelope preparation from Leptospira, interrogans, serovar Lai by fusion of SP2/0 myeloma cells with immune BALB/c mice spleen cells. The fusion rate was 96% and the antibody positive rate was 50%. One of the hybridomas, E4B11C9, reacted with 13 of the 13 serovars of the Icterohaemorrhagiae serogroup in microscopic agglutination test (MAT) but did not react with the 18 representative serovars of L. interrogans and L. biflexa serovar patoc and Leptonema illini. For all non-reactive serovars the MAT titres were greater than 1:25. The McAb, E4B7G5, reacted similarly with all serovars except smithi and tonkini. E4B7D4 reacted also similarly with all serovars except serovars birkini, ndambari, bogvere, smithi and tonkini. Therefore, 3 McAb showed serogroup specificity and partial serogroup specificity by agglutination. The agglutination titres were high and hybridomas were stable, so it might be useful in providing a simple, rapid method for the classification and identification of clinical isolates such as pathogenic L. interrogans in place of the complicated and time-consuming conventional methods.

Agglutination Tests↗

Antibody response to genus- and serovar-specific leptospiral antigens in Leptospira-infected cows.

In cows inoculated with Leptospira interrogans serovar pomona or hardjo, the 2-mercaptoethanol-sensitive microscopic agglutination test (MAT) antibody to the serovar appeared 3 to 8 days after inoculation and peaked at 10 to 20 days, whereas the 2-mercaptoethanol-resistant MAT antibody was predominant at 35 to 80 days. A persistent antibody response, probably associated with serovar-specific leptospiral antigens, was detected in the cows inoculated with serovar pomona, using a sonicated or an alkaline-extracted antigen derived from serovar pomona in the enzyme-linked immunosorbent assay (ELISA). In contrast, a short-lived antibody response to the same antigens was demonstrated in cows inoculated with serovar hardjo, probably more typical of the response to genus-specific leptospiral antigens. Antigens derived from L biflexa serovar patoc only detected the latter type of antibody response in cows inoculated with serovar pomona or hardjo. Correlative studies revealed that the antigens derived from serovar patoc seem to be genus specific and serologically closely related, but not identical. The antigens derived from serovar pomona were genus specific on the basis of the early antibody response to leptospiral inoculation in the cows, but serovar specific based on the subsequent more persistent response to leptospiral inoculation. These antigens were also serologically closely related, but not identical. Examination of sera from cows that aborted and were MAT-positive for serovar pomona or hardjo revealed a more serovar-specific antibody response, indicating that there may have been a less recent leptospiral antigenic stimulus, thus emphasizing the caution with which results of the ELISA and other serologic assays for the detection of bovine leptospirosis must be interpreted.

Agglutination Tests↗

[In vitro sensitivity of 3 strains of Leptospira interrogans to 3 different antibiotics].

Leptospirosis, anthropo-zoonosis ubiquitously widespread, is a social and economic problem still to be solved. The experimental and therapeutic employment of many antibiotics has largely been tested "in vitro" and "in vivo". In the following research we tried to evaluate, by experimental "in vitro" method, the sensitivity difference of three serovar strains of Leptospira interrogans to two macrolides, Erythromycin and Josamycin, compared with Penicillin. From standard cultures, previously treated with serial dilution of these antibiotics, the MIC and MSC, as quantitative parameters, have been stated. For the qualitative evaluation of the damages induced at ultrastructural level by the drug activity. Electron Microscopy investigations were performed on specimens from cultures treated for 6 hr with twice and tenfold the MSC. The present research confirms the good sterilizing efficaciousness "in vitro" of the tested macrolides (MSC less than 1 mcg/ml) and their different activity pathway.

Drug Resistance, Microbial↗

Effect of vaccination with a pentavalent leptospiral vaccine containing Leptospira interrogans serovar hardjo type hardjo-bovis on type hardjo-bovis infection of cattle.

Effectiveness of 2 pentavalent leptospiral vaccines containing Leptospira interrogans serovar hardjo was evaluated for protection of steers from infection with serovar hardjo type hardjo-bovis. The hardjo component of 1 vaccine was prepared from serovar hardjo type hardjoprajitno. The hardjo component of the other vaccine was prepared from serovar hardjo type hardjo-bovis. Two steers were vaccinated once and 4 steers were vaccinated twice with the pentavalent vaccine containing type hardjoprajitno. Four steers were vaccinated once and 4 steers were vaccinated twice with the pentavalent vaccine containing type hardjo-bovis. Four steers were maintained as non-vaccinated controls. Steers given vaccine containing type hardjo-bovis developed higher mean serum microscopic agglutination titers against serovar hardjo than steers given vaccine containing hardjoprajitno. Six months after the first vaccination, all steers were challenge-exposed on 3 occasions by conjunctival instillation of 10(7) serovar hardjo type hardjo-bovis organisms, and on 1 occasion by conjunctival instillation of urine from a steer shedding hardjo-bovis. All control and all vaccinated steers became infected and shed serovar hardjo type hardjo-bovis in the urine. Lesions were detected in kidneys of 3 of 4 nonvaccinated control steers, 5 of 6 steers given hardjoprajitno vaccine, and 6 of 8 steers given hardjo-bovis vaccine. Leptospires were detected in kidneys of 4 of 4 control steers and 13 of 14 vaccinated steers.

Animals↗

Effect of vaccination with a pentavalent leptospiral vaccine on Leptospira interrogans serovar hardjo type hardjo-bovis infection of pregnant cattle.

Effectiveness of a pentavalent leptospiral vaccine to protect cattle from infection and reproductive problems caused by Leptospira interrogans serovar hardjo type hardjo-bovis was evaluated. Seven cows were vaccinated once and 8 cows were vaccinated twice with a USDA-licensed pentavalent leptospiral vaccine. Five cows were maintained as nonvaccinated controls. Cows were bred 1 to 2 months after the last vaccination. During the 4th to 6th month of gestation, all cows were challenge exposed on 4 occasions by conjunctival instillation of 10(8) serovar hardjo type hardjo-bovis organisms and on 3 occasions by conjunctival instillation of urine from a cow shedding hardjo-bovis. All control cows and 13 of 15 vaccinated cows became infected and shed leptospires in the urine. Leptospires were detected in fewer urine samples collected from vaccinated cows, compared with those collected from control cows. Four stillborn calves and 3 weak calves were born to control and vaccinated cows. Leptospires were detected in the kidneys of 11 apparently healthy calves born to vaccinated and control cows. Agglutinating antibodies were not detected in the precolostral serum of these calves.

Animals↗

Comparison of three techniques to detect Leptospira interrogans serovar hardjo type hardjo-bovis in bovine urine.

Nucleic acid hybridization, bacteriologic culture, and a fluorescent antibody test were compared for detection of Leptospira interrogans serovar hardjo type hardjo-bovis in bovine urine. Seventy-five urine samples were collected from pregnant cows challenge exposed with type hardjo-bovis. Twenty samples were collected from steers not exposed to hardjo-bovis. Sediments from each sample were examined, using fluorescent antibodies and a repetitive sequence element nucleic acid probe, to detect the presence of leptospires. Urine samples were processed for bacteriologic culture, using standard techniques. Under laboratory conditions typically used for these techniques, leptospires were detected in 60 of 75 urine samples from challenge exposed cows by nucleic acid hybridization, in 24 samples by fluorescent antibody test, and in 13 samples by bacteriologic culture. Leptospires were not detected in the urine of steers not exposed to hardjo-bovis.

Animals↗

[Leptospirosis caused by Leptospira icterohaemorrhagiae of the pseudo-surgical type: a case].

The authors report a case of fatal leptospirosis due to Leptospira icterohaemorrhagiae revealed by typical signs of acute cholecystitis and associated with pancreatitis in a 73 year old patient presenting with gallstones. The initial clinical findings were highly suggestive of severe but typical cholecystitis and the final diagnosis was only considered when the patient's condition worsened despite surgery, with increasing obstructive jaundice and multiple organ failure. Pancreatitis was an autopsy finding. Misleading, especially gastrointestinal symptoms are frequent in leptospirosis. Hence an early diagnosis is an essential condition for a successful antibiotic management in severe cases of leptospirosis. This possibility should be considered whenever a patient presents with infectious obstructive jaundice. The patient has to be questioned concerning possible contact with contaminated animals and, when in doubt, the presence of specific antibodies should be investigated.

Acute Disease↗

The susceptibility of New Zealand isolates of leptospira to three antibiotics.

The susceptibilities of 14 recent isolates of Leptospira to benzyl penicillin, tetracycline and minocycline, were tested. All strains were highly susceptible to penicillin. The minimum inhibitory concentrations (MIC) of the two tetracyclines were generally low, and no significant difference was found between them.

Anti-Bacterial Agents↗

[Survival rate of Leptospira pomona in the soil at a natural leptospirosis focus].

At the area of a natural focus of leptospirosis (caused by L. pomona) in the Mozdok region of the North Occetic ASSR leptospires were detected by the method of dark field microscopy in 22% of intact soil samples. The presence of pathogenic leptospires in this soil was not confirmed by the method of the biological assay on Syrian golden hamsters. In controlled tests lasting 1-277 days, in 30.4% of cases L. pomona retained their viability, pathogenic and antigenic properties for as long as 74 days, while staying in the soil at the focus of infection with humidity being 15.2-31.4% and pH = 6.7-7.2. 11 Leptospira cultures isolated after staying in the soil retained their pathogenic properties and the death of the animals used in the bioassay from the acute form of leptospiral infection.

Animals↗

Diagnostic specificity, sensitivity and cross-reactivity of an enzyme-linked immunosorbent assay for the detection of antibody against Leptospira interrogans serovars pomona, sejroe and hardjo in cattle.

Outer sheath antigen was prepared from Leptospira interrogans serovars pomona, sejroe and hardjo by treating the organisms with 1.0M NaC1 followed by 0.04% sodium dodecyl sulfate (SDS). Sodium dodecyl sulfate was removed from the SDS-protein complexes by the extraction of dodecyl sulfate anions as ion pairs with triethylammonium cations into an organic solvent. The outer sheath antigen was recovered from the organic solvent as a precipitate and used as the source of leptospiral enzyme-linked immunosorbent assay (ELISA) antigen. Utilizing this antigen, ELISA was adapted to detect bovine serum antibody to L. interrogans serovars pomona, sejroe and hardjo. The specificity of this assay in 344 bovine sera, which were negative in the microscopic agglutination test (MAT) for seven serovars, was 99.4%. In sera from 37 and 87 cattle which revealed MAT titers greater than or equal to 1:50 for L. interrogans serovars pomona and sejroe, the relative sensitivity of the test was 100%. The ELISA also showed a considerable degree of low level cross-reactivity with other serovars. Sixty-six (75.9%) out of 87 bovine sera which were MAT-positive (MAT titer of greater than or equal to 1:50) with serovars sejroe and hardjo only were ELISA positive with heterologous pomona antigen; 16 (43.2%) and six 16.2%) out of 37 bovine sera which were MAT positive MAT titer of greater than or equal to 1:50) with serovar pomona only were ELISA positive with heterologous sejroe and hardjo ELISA antigen respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of vaccination with a bacterin containing Leptospira interrogans serovar bratislava on the breeding performance of swine herds.

Swine herds suspected to be infected with Leptospira interrogans serovar bratislava were vaccinated with bacterins containing 5 or 6 leptospiral serovars in which serovar bratislava was the unique component. The principal diagnostic feature indicating an infection by this organism was demonstration of antibody against serovar bratislava in sera from stillborn pigs. For 1 breeding cycle after vaccination of herds on 3 farms, 255 of 266 (95.9%) sows and gilts given the 6-serovar bacterin farrowed. In contrast, 233 of 311 (74.9%) sows and gilts given the 5-serovar bacterin farrowed. These results, as evaluated by analysis of variance techniques, showed a significant improvement (P less than 0.01) in reproductive performance for groups vaccinated against serovar bratislava.

Animals↗

Restriction endonuclease analysis of Leptospira interrogans serovar hardjo isolates from cattle.

The genomes of 253 strains of Leptospira interrogans serovar hardjo were examined by restriction endonuclease analysis of their DNA. The strains had been isolated from cattle at an abattoir (190), milk of agalactic cows (seven) and from aborted bovine fetuses (56). Two distinct genome types, Hardjoprajitno and Hardjobovis, were detected. The majority (91 per cent) of isolates from abattoir cattle were of the Hardjobovis type while most (76 per cent) of the isolates from clinical/pathological material were similar to Hardjoprajitno.

Animals↗

Survival of Leptospira interrogans serovar pomona in an acidic soil under simulated New Zealand field conditions.

Leptospira interrogans serovar pomona was found to survive for at least 42 days in a typical New Zealand soil under simulated winter field conditions. The soil was markedly acidic with a pH of 5.5 and survival times were not reduced even when its water content was only 23%. The values of both these parameters are considerably less than previously recorded for the survival of leptospires in soil. Two methods were used to recover leptospires from the soil microflora. One was the culture of a membrane filtrate in EMJH media with or without contaminant-suppressing additives and the other was the direct inoculation of soil-washings into hamsters. Both techniques proved to equally sensitive. It was estimated that following the addition of 5 X 10(8) leptospires to the soil samples less than 2 X 10(4) were present after six weeks.

Animals↗

[Relation between the incidence of antibodies against Leptospira interrogans in working dogs and in free-living small mammals].

Three localities were chosen in the borderland of the South Bohemian Region to investigate the problems and relations (if there are any) between the incidence of antibodies to Leptospira interrogans (L. i.) in wild ground small mammals and dogs moving in the same environment. In these regions, service dogs work as border guards every day. Blood samples of dogs at these localities were taken at about three-month intervals (from the June 1982 to the October 1984) to investigate the antibodies to L. i. serovars. Parallelly, wild ground small mammals and wild birds were caught at the same localities to be subject to the same examinations. A close relationship between the incidence of L. i. antibodies in ground small mammals and in service dogs was demonstrated at all three localities. At localities no. 1 and 2, the serovar grippotyphosa was found to be dominant in service dogs, which was also found in small rodents. On the other hand, the serovar icterohaemorrhagiae dominated in service dogs at locality no. 3. When changes in the incidence of L. i. antibodies were studied in service dogs and in wild small mammals, the peak of incidence at localities 1 and 2 was in the other half of the year; this relates to the usual maximum incidence in the natural reservoir of the serovar grippotyphosa, i.e. in rodents. On the contrary, at the third locality the peak of incidence of L. i. antibodies was recorded in the winter 1983-1984.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Variation in the hemolytic activity of Leptospira interrogans serovar canicola.

Leptospira interrogans serovar canicola strain moulton was grown to a high cell density in a protein-free medium with hemin. The hemolysin produced in this culture system showed a greatly expanded spectrum of hemolysis as compared to previous reports of leptospiral hemolysin produced in a more traditional culture system containing serum.

Animals↗