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Distribution of leptospirosis among stray dogs in the Okinawa Islands, Japan: comparison of the microcapsule and microscopic agglutination tests.

Three hundred and thirty-eight sera collected from stray dogs in the Okinawa islands were examined for antibodies against Leptospira interrogans using the microscopic agglutination test (MAT) and the one-point microcapsule agglutination test (MCAT). Seventy-eight sera (23%) showed a positive reaction to at least one of the six serovar antigens, and 69 of these reacted with serovar canicola by microcapsule agglutination test. The mixed microcapsule agglutination test detected 68 of the microscopic agglutination test-positive sera, and the 10 remaining were negative by microcapsule agglutination test. On the other hand, a single microcapsule agglutination test which was sensitized with serovar canicola detected 77 of the microscopic agglutination test-positive sera and the remaining one was microcapsule agglutination test-negative.

Agglutination Tests↗

[16S rRNA gene PCR-SSCP analysis of the reference strains from 15 serovars (14 serogroups) of pathogenic leptospires in China].

The DNAs of reference strains from 15 serovars (14 serogroups) of pathogenic leptospires in China were amplified with 16S rRNA gene primers, and then single-strand conformation polymorphism (SSCP) of the products were analyzed in 12.5% nondenaturing mini polyacrylamide gel (containing 5% glycerol) combining with silver-staining. All products showed two bands on electrophoresis at different parameters of voltage or current and concentration of gel. It proved that serovar lai, serovar canicola, serovar pyrogenes, serovar autumnalis, serovar australis, serovar pomona, serovar linhai, serovar hebdomadis, serovar haemolytica, serovar wolffi and serovar paidjan have the identical pattern (Leptospira interrogans), while serovar javanica, serovar ballum, serovar tarassovi and serovar manhao I belong to another pattern (L. borgpetersenii). The result was consistent with the classification of genetic species by Yasuda et al (1987) and Ramadass et al (1992).

China↗

A serological investigation of leptospirosis in sows from central Thailand.

Leptospirosis is a major pig-associated zoonosis worldwide. There are also human occupational disease exposure risks. For better public health and sanitation, the prevalence of this disease was investigated. Four hundred sera were obtained from sows in central Thailand during August 2004 to January 2005 and examined with 20 reference leptospira serovars by a microscopic agglutination test (MAT). The results showed that 10% of samples gave a positive reaction. The most prevalent serovar was grippotyphosa which accounted for 55% of all positive cases. Other minor serovars were canicola and a non-pathogenic strain (patoc). It follows that, the sow may be one of the disease transmitting sources for humans in this area. The disease correlation between pigs, humans and the environments should be evaluated.

Animals↗

[Occurrence of Leptospira spp. soropositive stray dogs in Itapema, Santa Catarina, Brazil].

This study aimed to verify Leptospira spp. serovar infections in stray dogs in Itapema, Santa Catarina, Brazil. Serum samples were collected from 590 stray dogs and tested against 25 Leptospira spp. serovars using the microscopic agglutination test. Prevalence of anti-leptospiral antibodies against one or more serovars was 10.5%. The most frequent serovar was pyrogenes, positive in 26 (18.0%) samples, followed by canicola with 20 (13.8%) and icterohaemorragiae and copenhageni with 18 (12.5%, with antibody titers from 1:100 to 1:3,200). Significant prevalence (10.4 to 11.1%) was also detected against serovars castellonis, butembo, and grippothyphosa.

Agglutination Tests↗

Sero surveillance of leptospirosis among sewer workers in Pune.

Leptospirosis is an important occupational disease affecting people coming in contact with animals and their discharges. The occurrence of infection in ones workplaces is linked to the environment to which the worker is exposed and the adaptability of the organism in that working environment. Rodents usually abound in underground sewers and are carriers of leptospira. The urine of rodents and other animals present in that area is likely to contaminate these sewers. Leptospira are excreted in the urine of infected animals. Thus sewer workers are at a potential risk of leptospirosis. The prevalence of leptospirosis in these workers could thus indirectly predict the presence of the disease in animals in a particular geographical niche. Total seventy-eight sewer workers from 5 different municipal wards in Pune were examined to find out the evidence of past infection with leptospira using microagglutination test (MAT). The prevalence rate was found to be 16.6%. The serovars to which antibodies were detected include autumnalis (38.4%), pyrogenes (23.0%), canicola (15.3%) and pomona (15.3%). Evidence of leptospiral infection was found to be maximum in sewer workers in the areas of the city that were infested with rodents and stray animals.

Animals↗

[The serovars of Leptospira interrogans isolated from cases of human leptospirosis in São Paulo, Brazil].

Eighteen strains of L. interrogans isolated from human cases were serotyped by the agglutinin-absorption test at Instituto Adolfo Lutz in São Paulo, Brazil. Fourteen were identified as serovar copenhageni (icterohaemorrhagiae serogroup), 2 as canicola (canicola serogroup), 1 as castellonis (Ballum serogroup) and 1 as pomona serogroup (serovar not yet defined). The frequency of serovar copenhageni in 100% of the isolates in icterohaemorrhagiae serogroup is emphasized and more studies to verify the real serovars prevalence as subsidy to the epidemiology of this infection are suggested by the authors.

Agglutination Tests↗

Leptospira interrogans serovar bratislava infection in two dogs.

Two dogs with clinical histories suggestive of leptospirosis were examined serologically and culturally for evidence of leptospiral infection. Antibodies to Leptospira interrogans serovar bratislava were detected in serum from one dog, and the organism was isolated from urine of that dog. In a serologic survey of dogs in the state of Illinois, reactor rates to bratislava were higher than those to canicola or icterohaemorrhagiae. In cases of suspect canine leptospirosis, serovars such as bratislava, not contained in canine vaccines, should be considered in a differential diagnosis.

Animals↗

[Latex agglutination system for the rapid diagnosis of leptospirosis in Cuba].

OBJECTIVES: To assess the sensitivity, specificity, reproducibility, and stability of five latex agglutination systems for detecting antibodies against leptospira in human and animal sera, by using the Leptospira serotypes that are most widely prevalent in Cuba. METHODS: We performed an analytic and descriptive study with 706 human sera (65 tested positive for antibodies against leptospira with microagglutination (MAT) and hemagglutination (HA) techniques; 156 sera that tested negative with MAT and HA); 485 sera from 424 patients who had clinical or epidemiologic signs of leptospirosis; and 29 animal sera (16 from equines, 6 from bovines, 5 from porcines, 1 from a canine, and 1 from an ovine). All of the samples were tested with five latex conjugates made from whole cells of Leptospira interrogans, specifically the four serogroups that circulated most widely in Cuba from 2002 to 2004. The cells obtained from cultured cell lines yielded four specific conjugates (latex-canicola, latex-icterohemorrhagiae, latexpomona, and latex-sejroe), as well as one latex conjugate made from a combination of all four serogroups in equal quantities (latex-pool). In addition, samples were tested with the commercial latex agglutination Lepto Tek Tri Dot (bioMeriuex, France) kit. The stability and reproducibility of the latex conjugates were assessed through monthly controls over a period of 6 months with positive and negative sera. RESULTS: Of the systems that were assessed, the best combination of sensitivity and specificity was obtained with the latex-Pool conjugate (93,8% and 90,4%, respectively). The best combination of positive and negative predictive values was seen with the latex-Sejroe conjugate (90,9% and 95,8%), respectively), followed by the latex-Pool conjugate (94.2% and 96.6%, respectively). The positive and negative predictive values of the Lepto Tek Dri Dot commercial system were 78.5% and 88.4%, respectively. Among the 137 patient samples that tested positive for one of the serotypes when MAT was used, latex conjugates succeeded in correctly identifying 107 (78.1%), whereas the latex-Pool conjugate detected 116 (84.7%) positive sera. When animal sera were tested, the latex-Pool conjugate detected the greatest number of positive serum samples and showed the greatest concordance with MAT (93.1%). The conjugates studied showed good stability and reproducibility. CONCLUSIONS: Latex conjugates made from whole cells of the most widely circulating leptospira in Cuba showed a degree of concordance with MAT that was similar to or better than that seen with the Lepto Tek Dri Dot commercial system, both in human and animal sera. We recommend more widespread use of the latex-Pool conjugate in Cuba in the initial screening for antibodies against leptospira.

Animals↗

Leptospira serology in small ruminants on St. Croix, U.S. Virgin Islands.

A serological survey of 16 serovars of Leptospira interrogans, previously reported in tropical small ruminants, was undertaken to determine the serovars involved and the prevalence of these antibodies in sheep and goats on St. Croix, U. S. Virgin Islands (USVI). Seven of eight goat herds (108 animals) had at least two seropositive animals in each herd with an individual animal seroprevalence of 26%. The 53 sheep tested (one flock only) showed a 32% seroprevalence. Antibodies against seven serovars were detected in goats (autumnalis, ballum, bataviae, bratislava, canicola, icterohemorrhagiae, and pyrogenes). In addition, hardjo antibodies were detected in sheep. Serovar autumnalis accounted for about 30% of seropositive animals in each species. Many animals showed titers against more than one serovar. The number of seropositive animals suggests Leptospira may be a factor in the health of small ruminants on St. Croix.

Agglutination Tests↗

[Leptospira antibody detection in dog serum in the years 1985 to 1988].

In 1985-1988, 993 serum samples of dogs from Southern Bavaria and 408 samples from Northern Bavaria and from several Lands of the Federal Republic of Germany were tested for antibodies against the serovars canicola, icterohaemorrhagiae, grippotyphosa, bratislava, pomona, saxkoebing, sejroe and hardjo by using the microscopic agglutination test (MAT). 683 seras (48.75%) out of altogether 1401 samples showed a reaction against one up to seven serovars. The mostly low canicola- and icterohaemorrhagiae titers, having been proved in over 30% of the samples, can be put down to the fact, that usually the dogs had been vaccinated. Most frequently titers were found with the serovars grippotyphosa and bratislava--in Southern Bavaria 28.3%, in Northern Bavaria and other Lands of the Federal Republic of Germany 18.6%. The prevalence of titers against serovar saxkoebing, with or without a reaction against other serovars out of homologous and heterologous serogroups, reach up to 3.2% in sendings coming from Southern Bavaria and in other sendings up to 6.1%.

Animals↗

Seroprevalence of bovine leptospirosis in Garanhuns Municipal District, Pernambuco State, Brazil.

The prevalence of Leptospira interrogans serovars in dairy cattle was determined by analyzing 464 serum samples from cows on 15 properties in Garanhuns municipal district, Pernambuco State, Brazil. A microscopic seroagglutination test including 12 serovars of Leptospira interrogans as antigens was used. Samples with titres 100 were considered positive. Two hundred and twenty-one (47.63%) of the samples were positive to one or more serovars. The prevalence of the serovars was hardjo (21.98%), bratislava (15.73%), castellonis (11.64%), tarassovi (10.56%), pyrogenes (1.72%), icterohaemorrhagiae (1.08%), pomona (0.86%), wolffi (0.86%), grippotyphosa (0.86%), djasiman (0.43%), canicola (0.21 %), and copenhageni (0.21%).

Agglutination Tests↗

Isolation of Leptospira from wild forest animals in Amazonian Brazil.

The role of wild, forest animals as reservoirs of Leptospira was investigated in Pará State, north Brazil. 696 animals were examined by culture of kidney tissue; isolates of serovar ballum were made from the rodent Proechimys sp. and the opossum Didelphis marsupialis; leptospires of the serogroups hebdomadis, grippotyphosa and cynopteri were isolated from the armadillo Dasypus novemcinctus, and as yet untyped leptospires were isolated from Proechimys and the procyonid carnivore Nasua nasua. Antibodies to serovars bataviae, butembo, canicola, castellonis, celledoni, grippotyphosa, panama, icterohaemorrhagiae and wolffi were demonstrated among 222 other animals examined by serological methods.

Animal Population Groups↗

Sero-epidemiology of canine leptospirosis in Trinidad: serovars, implications for vaccination and public health.

A sero-epidemiological study on canine leptospirosis was conducted in house, stray, farm and hunting dogs, as well as in suspect cases of clinical canine leptospirosis. Serum samples were collected from apparently healthy (vaccinated and non-vaccinated), house dogs. A questionnaire was administered to the owners to elicit information on risk factors for leptospirosis. The microscopic agglutination test was used to screen for leptospirosis using 17 international serovars. Reciprocal titres of between 100 and <800 were considered as evidence of past exposure while reciprocal titres of 800 or greater were classified as suggestive of acute/current infection. Of a total of 419 serum samples tested, 61 (14.6%) were seropositive for Leptospira agglutinins, 23 (5.5%) had mixed infections and 16 (3.8%) had current infection. Amongst 50 suspected cases of clinical leptospirosis, 24 (48.0%) were seropositive and only 13 (26.0%) had current infection compared with 10 (6.3%) and three (1.9%) of 160 apparently healthy house dogs respectively. The difference was statistically significant (P < 0.05; chi2). Twelve (25.5%) of 47 hunting dogs, 10 (20.4%) of 49 farm dogs and five (4.4%) of 113 stray dogs were seropositive (P < 0.05; chi2). Overall, a total of nine serovars were detected with serovars mankarso, icterohaemorrhagiae RGA, autumnalis and copenhageni being involved in 29 (47.5%), 20 (32.8%), 25 (41.0%) and 10 (16.4%) respectively in 61 seropositive dogs (P < 0.05; chi2). Serovar mankarso was most predominant in seropositive apparently healthy dogs, 37.8% (14/37), suspected clinical cases of leptospirosis, 62.5% (15/24) compared with serovar icterohaemorrhagiae with a frequency of 21.6% (8/37) and 50.0% (12/24), the difference being statistically significant (P < 0.05; chi2). Although all vaccines used for prevention of canine leptospirosis in the country contain serovars canicola and icterohaemorrhagiae, serovar mankarso was mostly associated with infection and disease and may be a good candidate for inclusion in the vaccine used locally. The public health risk posed to owners of dogs infected with Leptospira cannot be over-emphasized considering the zoonotic nature of the disease.

Animals↗

Genetic differences among the LPS biosynthetic loci of serovars of Leptospira interrogans and Leptospira borgpetersenii.

The gene organization in the lipopolysaccharide biosynthetic (rfb) locus was analyzed in seven Leptospira interrogans serovars within serogroup Icterohemorrhagiae, seven non-Icterohemorrhagiae serovars and one Leptospira borgpetersenii serovar. Two groups of loci were delineated based on DNA hybridization and sequence analysis. Group 1 contained the two Hardjo subtypes, Hardjoprajitno and Hardjobovis. Group 2 (containing Copenhageni, Pomona, Naam, Mwogolo, Smithi, Lai, Canicola, Autumnalis, Pyrogenes, Australis and Icterohemorrhagiae) differed from Group 1 in its organization upstream of orf11, where five ORFs (32, 33, 34, 35, 37) were identified that were not contained in the Group 1 loci. These ORFs encoded a putative epimerase (orf32), a glycosyltransferase (orf33), two integral membrane proteins (orfs 34 and 35), and a galactosyltransferase (orf37). Serovars Australis, Pomona and Autumnalis did not contain orf37. Serovar Bataviae was excluded from the grouping because of its unique genetic organization upstream of orf13. In the Group 2 loci, comparison of the genetic layout at the 5' end revealed differences which included mutations disrupting reading frames in either or both orf34 and orf35 and apparent allelic differences between orf33 homologs that may be sufficient to account for the genetic basis of serovar identity.

Alleles↗

Broadly reacting precipitating and agglutinating antigen of leptospirae.

A saprophytic Leptospira biflexa strain of equine origin was found which cross-reacts with immune rabbit antisera to 14 pathogenic Leptospira interrogans serotypes. Sera from goats experimentally inoculated with the saprophyte showed multiple low-level cross-agglutination reactions against a battery of live L. interrogans serotypes. Sonically treated and saline-extracted suspensions of the L. biflexa strain and serotypes canicola, icterohaemorrhagiae, and pomona yielded a common precipitating protein antigen that was detected by immunodiffusion and immunoelectrophoresis with all of the antileptospiral sera examined. In cross-absorption and gel diffusion tests, the precipitinogen from each of the strains was shown to be identical. Formaldehyde treatments and heating at 100 degree C suggest that the cellular location of the common antigen is either somatic or subsurface, and Sephadex G-200 gel filtration enabled the isolation of the active fraction of the L. biflexa antigen. Monoprecipitin sera against the common antigen of L. biflexa were produced by immunizing rabbits with specific precipitates in agar. In gel diffusion and immunoelectrophoresis tests the antisera with each of the soluble antigens developed a single precipitin formation, and the antisera agglutinated formolized and heated whole-cell suspensions of serotypes canicola, icterohaemorrhagiae, and pomona at low dilutions. The soluble L. biflexa antigen was evaluated as an immunogen and in passive immunity tests for protection against death and kidney infection in hamsters. No cross-protection occurred when the hamsters were challenged with virulent leptospires. In contrast, the animals vaccinated or administered hamster immune serum before challenge died earlier than the control animals.

Animals↗

Serovar distribution of a DNA sequence involved in the antigenic relationship between Leptospira and equine cornea.

BACKGROUND: Horses infected with Leptospira present several clinical disorders, one of them being recurrent uveitis. A common endpoint of equine recurrent uveitis is blindness. Serovar pomona has often been incriminated, although others have also been reported. An antigenic relationship between this bacterium and equine cornea has been described in previous studies. A leptospiral DNA fragment that encodes cross-reacting epitopes was previously cloned and expressed in Escherichia coli. RESULTS: A region of that DNA fragment was subcloned and sequenced. Samples of leptospiral DNA from several sources were analysed by PCR with two primer pairs designed to amplify that region. Reference strains from serovars canicola, icterohaemorrhagiae, pomona, pyrogenes, wolffi, bataviae, sentot, hebdomadis and hardjo rendered products of the expected sizes with both pairs of primers. The specific DNA region was also amplified from isolates from Argentina belonging to serogroups Canicola and Pomona. Both L. biflexa serovar patoc and L. borgpetersenii serovar tarassovi rendered a negative result. CONCLUSIONS: The DNA sequence related to the antigen mimicry with equine cornea was not exclusively found in serovar pomona as it was also detected in several strains of Leptospira belonging to different serovars. The results obtained with L. biflexa serovar patoc strain Patoc I and L. borgpetersenii serovar tarassovi strain Perepelicin suggest that this sequence is not present in these strains, which belong to different genomospecies than those which gave positive results. This is an interesting finding since L. biflexa comprises nonpathogenic strains and serovar tarassovi has not been associated clinically with equine uveitis.

Animals↗