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[Seroepidemiologic study of human leptospirosis in Lombardy].

In 1987 a seroepidemiological research was performed to verify the prevalence of leptospira agglutinins on the urban and rural population of Milan, Bergamo and Mantova Provinces in Lombardia Region. Were controlled 150 subjects (75 males and 15 females) in Milan town, 58 (49 males and 9 females) in municipalities of Bergamo Province, and 68 (55 males and 13 females) of Mantova Province. La prevalence of leptospira agglutinins in exposed subjects was 14.3% and in unexposed subjects 8.6%. In the females was noted a significative higher prevalence of agglutinins (14.4%) than in males (9.5%), especially in the females exposed (36.3%) than in females unexposed (9.6%). The leptospira serovars most interested were L. sejroe (15.9%), L. poi (13.6%), L. bratislava, L. canicola and L. Zanoni (9.1%). The sera tested proved reactivity of 56.8% also against two or three different serovars of pathogenic leptospira and prevalence of 13.6% of coagglutinins against nonpathogenic L. patoc serovar.

Agglutinins↗

A serosurvey for antibodies to Leptospira in dogs in the lower North Island of New Zealand.

AIMS: To investigate the prevalence of antibodies to endemic and exotic Leptospira serovars in samples from a serum bank, collected from dogs in the lower North Island of New Zealand. METHODS: Sera (n=466), which had been collected from apparently healthy dogs, were screened using the microscopic agglutination test (MAT) for antibodies to serovars L. borgpeterseni serovar hardjo, L. interrogans serovars pomona, copenhageni and canicola, and L. kirschneri serovar grippotyphosa. RESULTS: Antibody to Leptospiral antigen was found in 14.2% of dogs tested. The highest level of reactivity was with serovar copenhageni, to which 9.5% (41/433) of sera were positive. Antibodies to serovars grippotyphosa and canicola were not detected in this population of dogs. CONCLUSIONS: Leptospira infection is relatively common in dogs in the lower North Island . CLINICAL RELEVANCE: Vaccination of dogs against leptospirosis should be considered using vaccine containing antigen to serovars hardjo, pomona and copenhageni. The presence of moderate levels of copenhageni antibody in dogs in the lower North Island raises the possibility that this serovar has become established in rodent populations in this region.

Journal Article↗

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↗

Evaluation of a macroscopic plate test and an indirect immunofluorescence test to detect leptospiral antibodies in bovine serum.

A macroscopic plate test was found to be reliable and convenient for detection of Leptospira serotype pomona antibodies in bovine sera. However, the procedure was unreliable for detecting L. serotype canicola antibodies because of false positive reactions. An indirect immunofluorescence test and the microscopic agglutination test provided comparable results and they effectively detected serotype pomona antibodies in bovine sera.

Agglutination Tests↗

Effect of heat or chemical treatment on leptospiral antigens.

The use of bacterins is one of the primary methods in the control of leptospirosis in domestic animals, especially cattle, swine, and dogs. Bacterins have been made using chemically treated or heat-inactivated organisms. Many of the initial studies indicated that the bacterins gave complete protection, whereas later reports found that some inactivation methods resulted in bacterins that protect against death but not against kidney infection and renal shedding. This raised the possibility that some of the antigens had been altered. To investigate this possibility, we heat or chemically treated Leptospira interrogans serovar icterohaemorrhagiae CF-1, L. interrogans serovar canicola Moulton, L. interrogans serovar grippotyphosa SC4397, L. interrogans serovar hardjo Hardjoprajitno, and L. interrogans serovar pomona MLS. Temperatures used to determine alterations in antigens were 50, 56, 80, 100, and 121 degrees C. The chemicals used were formaldehyde, phenol, and thimerosal. Analysis was done using Laemmli gels and Western blots. We found that heating at 50 or 56 degrees C had the smallest effect on antigens, whereas heating at 80 or 100 degrees C caused the appearance of high-molecular-weight bands on Western blots. Heating at 121 degrees C caused almost all the bands to disappear, in both the stained gel and the Western blot. Both phenol and Formalin treatment altered proteins, whereas thimerosal treatment appeared to have little effect compared with heating at 50 degrees C.

Antigens, Bacterial↗

Evaluation of the hemagglutination test for epidemiologic studies of leptospiral antibodies in wild mammals.

Sera from 153 wild animals of 18 species were tested for antibodies against 12 serovars of Leptospira by the microscopic agglutination (MA) test. Seventy-five percent of the animals tested were seropositive against one or more of the 12 serovars used. The most commonly found serovars were pomona, autumnalis, pyrogenes, icterohaemorrhagiae, australis, and canicola. Of 62 carnivores representing 7 species, 55 (89%) were seropositive, as were 46 (60%) of 77 rodents from 9 species. Leptospira of the serovar copenhageni serogroup icterohaemorrhagiae were recovered from kidney tissues of a Norway rat (Rattus norvegicus). Of 443 wildlife sera tested by the indirect hemagglutination (IHA) test using cells sensitized with L. illini antigen, 47 (11%), mainly carnivores and deer, gave a heterophile reaction. Of the remaining 396 sera, 164 (41%) were seropositive for leptospirosis by the IHA test. To compare the IHA test with the MA test, 143 serum samples were tested by both methods. There was 84% concordance between the two tests.

Agglutination Tests↗

[Gene diagnostics of acute persistent Leptospira infection].

The results of clinical trials carried out in different foci have demonstrated high diagnostic value of analysis made with the use of the polymerase chain reaction (PCR) at early stages of Leptospira infection caused by infective agents of the serogroups Grippotyphosa, Canicola and Icterohaemorrhagiae. The possibility of leptospiremia lasting considerably longer than heretofore believed to be possible, as well as the persistence of leptospires in the liquor of patients after the acute phase of the disease is over, i.e. during the early and late convalescence periods, has been shown. This is indicative of good prospects of using the PCR analysis not only for early rapid diagnostics of Leptospira infections, but also for controlling the course of the infection, for prognosticating early and late complications of the disease, as well the mechanisms of pathogenesis.

Acute Disease↗

Laboratory and clinical features of experimental feline leptospirosis.

In order to assess the clinical, laboratorial and epidemiological aspects of feline leptospirosis, ten female and male adult cats were experimentally inoculated with pathogenic and autochthonous field isolate of Leptospira interrogans. Five of them were inoculated subcutaneously with serovar icterohaemorrhagiae (R-192) and the others five with serovar canicola. No clinical and laboratorial alterations were found in these animals. Antileptospiral agglutinins were detected in 90% of the infected cats, shortly after the 1st week after inoculation. The leptospiral agglutinins were detected for 8 to 12 weeks and the elimination of leptospires through urine was observed only in animals infected with serovar canicola, beginning 2 to 4 weeks after inoculation and lasting for 2 to 8 weeks. Isolations of leptospires from blood and kidneys were unsuccessful.

Animals↗

The bacteriological prevalence of leptospiral infection in cattle and buffaloes in West Malaysia.

A cross-sectional bacteriological survey of cattle in West Malaysia revealed 14.4% (32/222) had leptospiral infection. Isolates were obtained from all except one herd with prevalence of infection in herds ranging from 0-44.8%. A small number of buffalo urine samples were examined and all of them were found to be negative. A leptospiral isolate obtained from a bovine kidney proved to be a new serovar of Leptospira interrogans and the name unipertama was assigned to it. Six other leptospiral serovars were isolated, namely canicola, australis, javanica, ballum, pomona and hardjo. All six serovars were isolated for the first time in cattle in Malaysia. Cattle in Malaysia appear to be the maintenance host for serovar hardjo. The presence of the other serovars in cattle was probably due to contact with the maintenance hosts, pigs for serovar pomona and rodents for the other three serovars. It appears that the epidemiology of leptospiral infection in cattle in Malaysia is similar to that reported overseas.

Animals↗

Serological and microbiological findings on 3 farms with equine leptospiral abortions.

Blood and urine samples from horses on 3 central Kentucky horse farms with prior histories of leptospiral abortions were analysed. Blood samples were obtained from all available horses on each farm and tested for antibodies to 6 leptospira serovars. Urine samples were collected from non-gravid mares with serum antibody titres > or = 1:800 and examined for leptospires by dark-field microscopy, fluorescent antibody testing and culture. Adult horses had the greatest serological evidence of exposure to leptospira, followed by yearlings, then foals. Of horses with anti-leptospiral antibodies, 76.3% reacted against serovar bratislava, 49.1% against icterohaemorrhagiae, 20.1% against grippotyphosa, 16.3% against pomona genotype kennewicki, 15.9% against hardjo and 1.0% against canicola. These findings confirmed that horses in central Kentucky are exposed to multiple leptospira serovars. The seroprevalence of antibodies to the 6 selected serovars was consistent among the 3 selected farms. There was no direct correlation between the serovar that was the cause of abortion in the previous season and prevalence of positive titres to that serovar in horses tested on the farms in the following year. Furthermore, there was no evidence of long-term urinary shedding of leptospira by horses with high leptospira antibody titres.

Abortion, Veterinary↗

[An investigation on the distribution of leptospirae interrogans in water and soil in southwest of Yunnan Province].

The conservation of pathogenic leptospirae in water and soil is one of the important problems of geography-epidemiological survey of leptospirosis. We have conducted this investigation by using of filtration-culture isolation method in five counties in the west and southwest of Yunnan during 1988-1992. The results showed that the isolates from samples of water and soil collected between July and September in Menglian and Huaping county were leptospirae biflexa while in the samples of water and soil collected between November and December in three counties, 8 strains of L. interrogans were isolated from the samples of Gengma and Baoshan county. The total positive rate was 3.31% (8/242). The positive rate of water and soil was 2.14% (3/140) and 4.9% (5/102) respectively. Strains obtained in this investigation belong to six serogroups: Icterohaemorrhagiae and Pyrogenes in 2 respectively. Canicola, Grippotyphosa, Sejroe and Australis in 1 respectively. These serogroups of leptospirae obtained from water and soil were corresponded with the serogroups obtained from the patients and animals with leptospirosis in local area. This survey provided new data for expounding the structure of the focus of leptospirosis.

China↗

Protection against Leptospira interrogans sensu lato challenge by DNA immunization with the gene encoding hemolysin-associated protein 1.

The use of DNA constructs encoding leptospiral proteins is a promising new approach for vaccination against leptospirosis. In previous work we determined that immunization with hemolysis-associated protein 1 (Hap1) (LipL32) expressed by adenovirus induced significant protection against a virulent Leptospira challenge in gerbils. To avoid the use of the adenovirus vector, we checked for clinical protection against lethal challenge by DNA vaccination. A DNA vaccine expressing Hap1 was designed to enhance the direct gene transfer of this protein into gerbils. A challenge was performed 3 weeks after the last immunization with a virulent strain of serovar canicola. Our results show that the cross-protective effect with pathogenic strains of Leptospira, shared by Hap1, could be mediated by the DNA plasmid vector. This finding should facilitate the design and development of a new generation of vaccines against bacteria, particularly Leptospira interrogans sensu lato.

Animals↗

The use of methanol extract of Leptospira interrogans in complement fixation tests for leptospirosis.

Methanol extracts were obtained from L. interrogans serovars icterohaemorrhagiae and canicola and L. biflexa serovar patoc. Human sera from 167 normal individuals and 40 patients with different infectious diseases tested by complement fixation tests showed negative reactions. Sera from 100 patients with a suspicion of leptospirosis were tested by complement fixation tests and microscopic agglutination reactions. Agreement of 84% was found for those two reactions. Positive microscopic agglutination tests at a dilution 1:20-1:400 with negative complement fixation tests were observed in 5% of patients and negative microscopic agglutination with complement fixation tests in the range of 1:20-1:1280 were observed in 11% of the cases.

Agglutination Tests↗

[Leptospira interrogans in a canine population of Greater Buenos Aires: variables associated with seropositivity].

We determined the seroprevalence of leptospirosis in a suburban canine population for the purpose of analyzing the association between different individual and environmental variables and seropositivity for leptospirosis. The study, which was cross-sectional, was performed in July 1992 in a neighborhood of Greater Buenos Aires with approximately 9,500 inhabitants and a canine population of around 2,000 animals. We studied a random sample of 223 dogs and obtained a blood sample from each. Each animal's epidemiologic history was obtained by interviewing the housewife. Serologic measurements were performed by the microagglutination technique with the use of 10 different serotypes of Leptospira interrogans. Of the 223 dogs that were tested, 57% showed seropositivity; 82% of the positive sera coagglutinated with two or more serotypes. The most frequently detected serotypes were canicola and pyrogenes. Seroprevalence in females was less common than in males (P < 0.05) and in puppies less than 1 year old it was less common than in older animals (P < 0.01). Street behavior in the dog and the presence of stagnant water in front of the owner's dwelling were the most important of the risk factors examined. The associations between seropositivity on the one hand and contact with trash deposits, hunting behavior and the presence of rodents inside the dwelling on the other were not statistically significant. Different control measures are discussed.

Age Factors↗

[Significance of human leptospirosis in Mexico. Detection of Leptospira antibodies in a blood donor population].

The presence of specific serum antibodies has been used as a diagnostic test for human leptospirosis. The presence of these antibodies in humans is indicative of an active natural infection. Its detection after exposure denotes the presence of immunity. Serum samples from 206 adult blood donors were analyzed with a microscopic agglutination assay against 7 serovars of Leptospira interrogans. A total of 7% were positive with the following serovar distribution; shermani 53%, canicola 33%, pyrogens 20%, pomona 13% and icterohaemorrhagiae 6%. The highest frequency of seropositivity was found in the 20 year to 39 age group. These results in asymptomatic individuals show that leptospirosis is a frequent zoonosis in Mexico.

Adolescent↗

Protein and antigen profiles of prevalent serovars of Leptospira interrogans.

Whole-cell and detergent-soluble proteins, enriched for outer membrane antigens, of the Leptospira interrogans serovars present in commercially available pentavalent vaccines (hardjo, pomona, icterohaemorrhagiae, grippotyphosa, and canicola) were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting (immunoblotting). Protein and antigenic profiles of these serovars, representing several serogroups, were compared with similar profiles of the most common North American bovine pathogen, serovar hardjo type hardjo bovis. The reference strain of serovar balcanica and a hardjoprajitno bovine field isolate (serovar hardjo) were also assayed. Coomassie blue-stained gels revealed extensive protein similarities, and Western blots demonstrated antigenic relatedness throughout the low- and high-molecular-weight regions. Possible quantitative differences in protein expression among the strains were noted, as were similarities in the protein profiles of type hardjo bovis and the balcanica reference strain. A cocktail composed of these homologous antigens may serve as an efficacious subunit vaccine for leptospirosis.

Animals↗